Two Doses of Multimeric-001 (M-001) Followed by Influenza Vaccine

• Sponsor: National Institute of Allergy and Infectious Diseases (NIAID) (NIH)
• Overall Status: Completed
• CT.gov ID: NCT03058692
• Collaborator: (none)
• Enrollment: 120
• Locations: 3
• Arms: 2
• Actual Duration (Months): 9.2
• Patients Per Site: 40
• Patients Per Site Per Month: 4.3

Study Details

Study Description

Brief Summary

This is a Phase II randomized, double-blind, placebo-controlled trial in 120 males and non-pregnant females, 18 to 49 years old, inclusive, who are in good health and meet all eligibility criteria. This clinical trial will be conducted at 3 United States sites and is designed to assess the safety, reactogenicity, and immunogenicity of two priming doses of M-001 followed by a seasonal quadrivalent inactivated influenza vaccine (IIV4). The duration of this trial for each subject will be approximately 7 months. The entire study duration will be approximately 24 months. The primary objectives are: 1) To assess the safety as measured by vaccine related adverse events, reactogenicity, and laboratory adverse events of two doses of M-001 vaccine, each dose administered approximately 21 days apart; and 2) To assess the T cell responses to M-001 component peptides following two doses of M-001.

Condition or Disease	Intervention/Treatment	Phase
Influenza; Influenza Immunisation	Biological: Influenza Multimeric-001 Vaccine; Other: Placebo; Biological: Quadrivalent Recombinant Seasonal Influenza Vaccine	Phase 2

Detailed Description

This is a Phase II randomized, double-blind, placebo-controlled trial in 120 males and non-pregnant females, 18 to 49 years old, inclusive, who are in good health and meet all eligibility criteria. This clinical trial will be conducted at 3 United States sites and is designed to assess the safety, reactogenicity, and immunogenicity of two priming doses of M-001 followed by a seasonal quadrivalent inactivated influenza vaccine (IIV4). The duration of this trial for each subject will be approximately 7 months. The entire study duration will be approximately 24 months. The primary objectives are: 1) To assess the safety as measured by vaccine related adverse events, reactogenicity, and laboratory adverse events of two doses of M-001 vaccine, each dose administered approximately 21 days apart; and 2) To assess the T cell responses to M-001 component peptides following two doses of M-001. The secondary objectives are: 1) To assess all serious adverse events (SAEs) following receipt of each dose of M-001 vaccine or placebo, each dose separated by approximately 21 days, through the end of the study; 2) To assess all unsolicited non-serious AEs following receipt of each dose of M-001 or placebo, each dose separated by approximately 21 days, through 21 days after each dose of M-001 or placebo; and 3) To assess the serum HAI and Neut antibody responses to the 2018-2019 IIV4 vaccine viruses.

Study Design

• Study Type: Interventional
• Actual Enrollment: 120 participants
• Allocation: Randomized
• Intervention Model: Parallel Assignment
• Masking: Double (Participant, Investigator)
• Primary Purpose: Prevention
• Official Title: A Phase II, Double-Blind, Multicenter, Randomized, Placebo-Controlled Trial to Assess the Safety, Reactogenicity and Immunogenicity of Two Doses of Multimeric-001 (M-001) Followed by Seasonal Quadrivalent Influenza Vaccine
• Actual Study Start Date: Apr 9, 2018
• Actual Primary Completion Date: Jan 14, 2019
• Actual Study Completion Date: Jan 14, 2019

Arms and Interventions

Arm	Intervention/Treatment
Experimental: M-001 + IIV4 0.4 ml injection of M-001 (1 mg dose) intramuscularly on Day 1 and Day 22, followed by 0.5 ml injection of IIV4 (60 mcg HA) intramuscularly on Day 172 (n=60)	Biological: Influenza Multimeric-001 Vaccine; The M-001 vaccine consists of 3 repetitions of 9 conserved linear epitopes that are prepared as a single recombinant protein. The M-001 vaccine is expected to protect against existing as well as future seasonal and pandemic virus strains.; Biological: Quadrivalent Recombinant Seasonal Influenza Vaccine; Quadrivalent Inactivated Influenza Vaccine (IIV4) for intramuscular injection is indicated for active immunization against influenza disease caused by influenza virus subtypes A and type B present in the vaccine.
Placebo Comparator: Placebo+ IIV4 0.4 ml injection of placebo intramuscularly on Day 1 and Day 22, followed by 0.5 ml injection of IIV4 (60 mcg HA) intramuscularly on Day 172 (n=60)	Other: Placebo; Placebo is saline injection; Biological: Quadrivalent Recombinant Seasonal Influenza Vaccine; Quadrivalent Inactivated Influenza Vaccine (IIV4) for intramuscular injection is indicated for active immunization against influenza disease caused by influenza virus subtypes A and type B present in the vaccine.

Outcome Measures

Primary Outcome Measures

1. Mean Percentage of Perforin+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

2. Mean Percentage of Cluster of Differentiation 107a Positive (CD107a+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

3. Mean Percentage of Interleukin-2 Positive (IL-2+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

4. Mean Percentage of Tumor Necrosis Factor Positive (TNF+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

5. Mean Percentage of Interferon Gamma Positive (IFNg+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

6. Mean Percentage of Perforin+CD107a+IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

7. Mean Percentage of Perforin+CD107a+IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

8. Mean Percentage of Perforin+CD107a+IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

9. Mean Percentage of Perforin+CD107a+IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

   The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

10. Mean Percentage of Perforin+CD107a+IL-2-TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

11. Mean Percentage of Perforin+CD107a+IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

12. Mean Percentage of Perforin+CD107a+IL-2-TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

13. Mean Percentage of Perforin+CD107a+IL-2-TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

14. Mean Percentage of Perforin+CD107a- IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

15. Mean Percentage of Perforin+CD107a- IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

16. Mean Percentage of Perforin+CD107a- IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

17. Mean Percentage of Perforin+CD107a- IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

18. Mean Percentage of Perforin+CD107a- IL-2- TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

19. Mean Percentage of Perforin+CD107a- IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

20. Mean Percentage of Perforin+CD107a- IL-2- TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

21. Mean Percentage of Perforin+CD107a- IL-2- TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

22. Mean Percentage of Perforin- CD107a+IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

23. Mean Percentage of Perforin- CD107a+IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

24. Mean Percentage of Perforin- CD107a+IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

25. Mean Percentage of Perforin- CD107a+IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

26. Mean Percentage of Perforin- CD107a+IL-2- TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

27. Mean Percentage of Perforin- CD107a+IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

28. Mean Percentage of Perforin- CD107a+IL-2- TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

29. Mean Percentage of Perforin- CD107a+IL-2- TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

30. Mean Percentage of Perforin- CD107a- IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

31. Mean Percentage of Perforin- CD107a- IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

32. Mean Percentage of Perforin- CD107a- IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

33. Mean Percentage of Perforin- CD107a- IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

34. Mean Percentage of Perforin- CD107a- IL-2- TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

35. Mean Percentage of Perforin- CD107a- IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

36. Mean Percentage of Perforin- CD107a- IL-2- TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

37. Mean Percentage of Perforin- CD107a- IL-2- TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides [Day 1 through Day 36]

    The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.

38. Number of Participants With Clinical Safety Laboratory Adverse Events After the First M-001 Vaccination [Day 9]

    Blood was collected after first vaccination for assessment by a central clinical laboratory. Clinical safety laboratory adverse events included white blood cells (WBC) </=3900/uL or >/=10,600/uL; platelets </=139,000/uL or >/=416,000/uL; hemoglobin </=11.4 g/dL (female) or </=12.4 g/dL (male); alanine aminotransferase (ALT) >/=44 IU/L (female) or >/=61 IU/L (male); creatinine >/=1.1 mg/dL (female) or >/=1.4 (male); and total bilirubin >/=1.30 mg/dL.

39. Number of Participants With Clinical Safety Laboratory Adverse Events After Second M-001 Vaccination [Day 22 through Day 29]

    Clinical safety laboratory adverse events included WBC less than or equal to 3900/uL or greater than or equal to 10,600/uL; platelets less than or equal to 139,000/uL or greater than or equal to 416,000/uL; hemoglobin less than or equal to 11.4 g/dL (female) or less than or equal to 12.4 g/dL (male); alanine aminotransferase (ALT) greater than or equal to 44 IU/L (female) or greater than or equal to 61 IU/L (male); creatinine greater than or equal to 1.1 mg/dL (female) or greater than or equal to 1.4 (male); and total bilirubin greater than or equal to 1.30 mg/dL.

40. Number of Participants Reporting Solicited Injection Site and Systemic Reactogenicity Events After the First M-001 Vaccination [Day 1 through Day 8]

    Participants maintained a memory aid and thermometer to record daily oral temperatures and the occurrence of systemic reactions of feverishness, fatigue, malaise, myalgia, arthralgia, headache, and nausea, as well as local injection site reactions of pain, tenderness, redness, and swelling for 8 days after vaccination (Day 0-7) based on their interference with daily activities, with a severity grade of mild meaning no interference, moderate as some interference and severe as significant interference/prevented daily activity. Fever was defined as an oral temperature of 38 degree Celsius or higher. Participants are counted if they reported experiencing the symptom at any severity on any of the 8 days post vaccination.

41. Number of Participants Reporting Solicited Injection Site and Systemic Reactogenicity Events After the Second M-001 Vaccination [Day 22 through Day 29]

    Participants maintained a memory aid and thermometer to record daily oral temperatures and the occurrence of systemic reactions of feverishness, fatigue, malaise, myalgia, arthralgia, headache, and nausea, as well as local injection site reactions of pain, tenderness, redness, and swelling for 8 days after vaccination (Day 0-7) based on their interference with daily activities, with a severity grade of mild meaning no interference, moderate as some interference and severe as significant interference/prevented daily activity. Fever was defined as an oral temperature of 38 degree Celsius or higher. Participants are counted if they reported experiencing the symptom at any severity on any of the 8 days post vaccination.

42. Number of Participants Reporting Vaccine-related Serious Adverse Events (SAEs) After M-001 Vaccination [Day 1 through Day 200]

    SAEs included any untoward medical occurrence that resulted in death; was life threatening; was a persistent/significant disability/incapacity; required inpatient hospitalization or prolongation or a congenital anomaly/birth defect. Relationship (related or unrelated to the study product) was determined by a site principal investigator blinded to the study product received by the participant.

Secondary Outcome Measures

1. Number of Participants Reporting Serious Adverse Events (SAEs) After M-001 Vaccination [Day 1 through Day 200]

   SAEs included any untoward medical occurrence that resulted in death; was life threatening; was a persistent/significant disability/incapacity; required inpatient hospitalization or prolongation or a congenital anomaly/birth defect. All events are included regardless of relationship to the study product.

2. Incidence of Unsolicited Non-serious Adverse Events (AEs) After M-001 Vaccination [Day 1 through Day 43]

   Unsolicited adverse events were collected from the time of first vaccination and at each follow-up visit through Day 43. Adverse events were defined as any untoward medical occurrence regardless of its causal relationship to the study treatment. Events were coded with MedDRA and are reported by System Organ Class.

3. The Percentage of Subjects Achieving HAI Seroconversion to IIV4 Vaccine Virus From Day 172 to Day 200 [Day 172 to Day 200]

   Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Seroconversion was defined as a Day 172 titer less than 10 and Day 200 titer greater than or equal to 40, or for those with a Day 172 titer of 10 or greater, a minimum 4-fold rise in Day 200 antibody titer.

4. The Percentage of Subjects Achieving Neutralizing Antibody Seroconversion to IIV4 Vaccine Virus From Day 172 to Day 200 [Day 172 to Day 200]

   Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Seroconversion was defined as a Day 172 titer less than 10 and Day 200 titer greater than or equal to 40, or for those with a Day 172 titer of 10 or greater, a minimum 4-fold rise in Day 200 antibody titer.

5. The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 1 [Day 1]

   Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

6. The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 43 [Day 43]

   Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

7. The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 172 [Day 172]

   Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

8. The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 200 [Day 200]

   Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

9. The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 1 [Day 1]

   Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

10. The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 43 [Day 43]

    Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

11. The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 172 [Day 172]

    Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

12. The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 200 [Day 200]

    Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 18 Years to 49 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

1. Provide written informed consent prior to initiation of any study procedures.

2. Are able to understand and comply with planned study procedures and be available for all study visits.

3. Are males or non-pregnant females, 18 to 49 years old, inclusive.

4. Are in good health*.

*As determined by medical history and targeted physical examination to evaluate acute or currently ongoing chronic medical diagnoses or conditions, defined as those that have been present for at least 90 days, which would affect the assessment of the safety of subjects or the immunogenicity of study vaccinations. Chronic medical diagnoses or conditions should be stable for the last 60 days prior to investigational vaccine study product administration. This includes no change in chronic prescription medication, dose, or frequency as a result of deterioration of the chronic medical diagnosis or condition in the 60 days prior to enrollment and investigational vaccine study product administration. Any prescription change that is due to change of health care provider, insurance company, etc., or that is done for financial reasons, as long as in the same class of medication, will not be considered a deviation of this inclusion criterion. Any change in prescription medication due to improvement of a disease outcome, as determined by the site principal investigator or appropriate sub-investigator, will not be considered a deviation of this inclusion criterion. Subjects may be on chronic or as needed (prn) medications if, in the opinion of the site principal investigator or appropriate sub-investigator, they pose no additional risk to subject safety or assessment of reactogenicity and immunogenicity and do not indicate a worsening of medical diagnosis or condition. Similarly, medication changes subsequent to enrollment and investigational study product vaccination are acceptable provided there was no deterioration in the subject's chronic medical condition that necessitated a medication change. All chronic medical condtions should pose no additional risk to the subject or interference with the evaluation of responses to study vaccination. Note: Topical, nasal, and inhaled medications (with the exception of inhaled corticosteroids as outlined in the Subject Exclusion Criteria), herbals, vitamins, and supplements are permitted.

1. Oral temperature is less than 100.0 degrees F.

2. Pulse** is 50 to 100 bpm, inclusive.

**Acceptable pulse range prior to IIV4 dose is 45 to 115 bpm, inclusive and no symptoms.

1. Systolic blood pressure*** is 85 to 150 mmHg, inclusive. ***Acceptable systolic blood pressure range prior to IIV4 dose is 80 to 155 mm Hg, inclusive and no symptoms.

2. Diastolic blood pressure**** is 55 to 95 mmHg, inclusive.

****Acceptable diastolic blood pressure range prior to IIV4 dose is 50 to 100 mm Hg, inclusive and no symptoms.

1. Women of childbearing potential* must use an acceptable method of contraception** from 30 days prior to vaccination until 60 days after the second of dose of M-001 or placebo.

2. Women of childbearing potential***** must use an acceptable method of contraception****** from 30 days prior to receipt of IIV vaccination, and must plan to use until 28 days after the IIV.

*****Not sterilized via tubal ligation, bilateral oophorectomy, hysterectomy, or successful Essure(R) placement (permanent, non-surgical, non-hormonal sterilization with history of documented radiological confirmation test achieved or with use of another approved birth control method if confirmation test not confirmed) and still menstruating or < 1 year of the last menses if menopausal).

******Includes, but is not limited to, non-male sexual relationships, abstinence from sexual intercourse with a male partner, monogamous relationship with a vasectomized partner, male condoms with the use of applied spermicide, intrauterine devices, NuvaRing(R), and licensed hormonal methods such as implants, injectables, contraceptive patches or oral contraceptives ("the pill"). Method of contraception will be captured on the appropriate data collection form.

1. Female subjects of childbearing potential must have a negative urine or serum pregnancy test within 24 hours prior to study vaccination.

Exclusion Criteria:

1. Have an acute illness*, as determined by the site PI or appropriate sub-investigator, within 72 hours prior to study vaccination.

• An acute illness which is nearly resolved with only minor residual symptoms remaining is allowable if, in the opinion of the site PI or appropriate sub-investigator, the residual symptoms will not interfere with the ability to assess safety parameters as required by the protocol.

1. Have any medical disease or condition that, in the opinion of the site PI or appropriate sub-investigator, is a contraindication to study participation**.

**Including acute or chronic medical disease or condition, defined as persisting for at least 90 days, that would place the subject at an unacceptable risk of injury, render the subject unable to meet the requirements of the protocol, or may interfere with the evaluation of responses or the subject's successful completion of this study.

1. Have immunosuppression as a result of an underlying illness or treatment, or use of anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study vaccination.

2. Have known active neoplastic disease or a history of any hematologic malignancy. Non-melanoma skin cancers that are not active are permitted.

3. Have known HIV, hepatitis B, or hepatitis C infection.

4. Have known hypersensitivity or allergy to eggs, egg or chicken protein, or other components of the study vaccine.

5. Have a history of severe reactions following previous immunization with licensed or unlicensed influenza vaccines.

6. Have a history of Guillain-Barré Syndrome.

7. Have a history of alcohol or drug abuse within 5 years prior to study vaccination.

8. Have any diagnosis, current or past, of schizophrenia, bipolar disease, or other psychiatric diagnosis that may interfere with subject compliance or safety evaluations.

9. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement for danger to self or others within 10 years prior to study vaccination.

10. Have taken oral or parenteral (including intraarticular) corticosteroids of any dose within 30 days prior to study vaccination.

11. Have taken high-dose*** dose inhaled corticosteroids within 30 days prior to study vaccination****.

***High-dose defined as per age as using inhaled high dose per reference chart https://www.nhlbi.nih.gov/health-pro/guidelines/current/asthma-guidelines/quick-refere nce-html#estimated-comparative-daily-doses.

****Topical and nasal steroids are permissible.

1. Received any licensed live vaccine or plan to receive a licensed live vaccine within 30 days prior or 21 days after each M-001 study vaccination.

2. Received a licensed inactivated vaccine within 14 days prior to or 21 days after each M-001 study vaccination.

3. Plans to or received the current 2018-2019 influenza vaccine (inactivated or live prior to ordering the study (the 2018-2019 influenza vaccine will be given during the trial.))

4. Received immunoglobulin or other blood products (with exception of Rho D immunoglobulin) within 90 days prior to study vaccination.

5. Received an experimental agent***** within 30 days prior to the first study vaccination, or expects to receive an experimental agent****** during the 7-month trial-reporting period.

*****Including vaccine, drug, biologic, device, blood product, or medication.

******Other than from participation in this study.

1. Are participating or plan to participate in another clinical trial with an interventional agent******* that will be received during the 7-month trial-reporting period.

*******Including agent (licensed or unlicensed vaccine, drug, biologic, device, blood product, or medication) during the 9-month study period.

1. Plan to travel outside the U.S. (continental U.S., Hawaii and Alaska) in the time between the first study vaccination and 21 days after the last study vaccination********.

********Study vaccination refers to investigational study product vaccination.

1. Female subjects who are breastfeeding or plan to breastfeed at any given time from the first study vaccination until 30 days after the last study vaccination.

2. Blood donation or planned blood donation within 30 days prior to the study vaccination through 30 days after the last blood drawn for this study.

3. Have signs or symptoms that could confound or confuse assessment of study vaccine reactogenicity*********.

*********The study vaccination should be postponed/deferred until signs or symptoms have resolved and if within the acceptable protocol-specified window for that visit.

1. Have a history of convulsions or encephalomyelitis within 90 days prior to study vaccination.

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	University of Iowa - Vaccine Research and Education Unit	Iowa City	Iowa	United States	52242-2600
2	Cincinnati Children's Hospital Medical Center - Infectious Diseases	Cincinnati	Ohio	United States	45229-3039
3	Baylor College of Medicine - Molecular Virology and Microbiology	Houston	Texas	United States	77030-3411

Sponsors and Collaborators

• National Institute of Allergy and Infectious Diseases (NIAID)

Investigators

None specified.

Study Documents (Full-Text)

• Statistical Analysis Plan - Sep 9, 2019
• Study Protocol - Dec 6, 2018

More Information

Publications

None provided.

• Responsible Party: National Institute of Allergy and Infectious Diseases (NIAID)
• ClinicalTrials.gov Identifier: NCT03058692
• Other Study ID Numbers: 14-0112; HHSN272201300015I
• First Posted: Feb 23, 2017
• Last Update Posted: Jun 18, 2020
• Last Verified: Nov 27, 2019

• Studies a U.S. FDA-regulated Drug Product: Yes
• Studies a U.S. FDA-regulated Device Product: No
• Product Manufactured in and Exported from the U.S.: No
• Keywords provided by National Institute of Allergy and Infectious Diseases (NIAID): Immunogenicity; Influenza Vaccine; M-001 vaccine; Reactogenicity; Safety
• Additional relevant MeSH terms: Influenza, Human; Vaccines

Study Results

Participant Flow

Recruitment Details	Participants were healthy adult males and non-pregnant females recruited from existing volunteer populations and from the communities at large around the clinical sites. Participants were enrolled between 09APR2018 and 28JUN2018.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml injection of M-001 (1 mg dose) intramuscularly on Day 1 and Day 22, followed by 0.5 ml injection of IIV4 (60 mcg HA) intramuscularly on Day 172 Influenza Multimeric-001 Vaccine: The M-001 vaccine consists of 3 repetitions of 9 conserved linear epitopes that are prepared as a single recombinant protein. The M-001 vaccine is expected to protect against existing as well as future seasonal and pandemic virus strains. Quadrivalent Recombinant Seasonal Influenza Vaccine: Quadrivalent Inactivated Influenza Vaccine (IIV4) for intramuscular injection is indicated for active immunization against influenza disease caused by influenza virus subtypes A and type B present in the vaccine.	0.4 ml injection of placebo intramuscularly on Day 1 and Day 22, followed by 0.5 ml injection of IIV4 (60 mcg HA) intramuscularly on Day 172 Placebo: Placebo is saline injection Quadrivalent Recombinant Seasonal Influenza Vaccine: Quadrivalent Inactivated Influenza Vaccine (IIV4) for intramuscular injection is indicated for active immunization against influenza disease caused by influenza virus subtypes A and type B present in the vaccine.
Period Title: Overall Study
STARTED	61	59
COMPLETED	58	57
NOT COMPLETED	3	2

Baseline Characteristics

Arm/Group Title	M-001 + IIV4	Placebo + IIV4	Total
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172	Total of all reporting groups
Overall Participants	61	59	120
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%
Between 18 and 65 years	61 100%	59 100%	120 100%
>=65 years	0 0%	0 0%	0 0%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	32.6 (7.9)	33.5 (7.8)	33.0 (7.8)
Sex: Female, Male (Count of Participants)
Female	42 68.9%	33 55.9%	75 62.5%
Male	19 31.1%	26 44.1%	45 37.5%
Ethnicity (NIH/OMB) (Count of Participants)
Hispanic or Latino	5 8.2%	6 10.2%	11 9.2%
Not Hispanic or Latino	56 91.8%	53 89.8%	109 90.8%
Unknown or Not Reported	0 0%	0 0%	0 0%
Race (NIH/OMB) (Count of Participants)
American Indian or Alaska Native	0 0%	0 0%	0 0%
Asian	8 13.1%	5 8.5%	13 10.8%
Native Hawaiian or Other Pacific Islander	0 0%	0 0%	0 0%
Black or African American	3 4.9%	3 5.1%	6 5%
White	49 80.3%	50 84.7%	99 82.5%
More than one race	1 1.6%	1 1.7%	2 1.7%
Unknown or Not Reported	0 0%	0 0%	0 0%
Region of Enrollment (participants) [Number]
United States	61 100%	59 100%	120 100%

Outcome Measures

1. Primary Outcome

Title	Mean Percentage of Perforin+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172
Measure Participants	58	59
Perforin+ CD4+ at Day 1	1.180	0.781
Perforin+ CD4+ Day 36	1.090	0.779
Perforin+ CD8+ Day 1	16.1	12.4
Perforin+ CD8+ Day 36	16.3	13.8

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.53
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.56
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

2. Primary Outcome

Title	Mean Percentage of Cluster of Differentiation 107a Positive (CD107a+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
CD107a+ CD4+ at Day 1	0.255	0.264
CD107a+ CD4+ Day 36	0.242	0.257
CD107a+ CD8+ Day 1	0.348	0.367
CD107a+ CD8+ Day 36	0.352	0.384

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.74
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.73
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

3. Primary Outcome

Title	Mean Percentage of Interleukin-2 Positive (IL-2+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
IL-2+ CD4+ at Day 1	0.0682	0.0634
IL-2+ CD4+ Day 36	0.0749	0.0692
IL-2+ CD8+ Day 1	0.1260	0.0939
IL-2+ CD8+ Day 36	0.1010	0.1170

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.66
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.48
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

4. Primary Outcome

Title	Mean Percentage of Tumor Necrosis Factor Positive (TNF+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
TNF+ CD4+ at Day 1	0.437	0.530
TNF+ CD4+ Day 36	0.450	0.395
TNF+ CD8+ Day 1	0.0750	0.0716
TNF+ CD8+ Day 36	0.0676	0.0596

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.32
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	>0.99
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

5. Primary Outcome

Title	Mean Percentage of Interferon Gamma Positive (IFNg+) CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
INFg+ CD4+ at Day 1	0.0418	0.0333
INFg+ CD4+ Day 36	0.0512	0.0436
INFg+ CD8+ Day 1	0.0558	0.0400
INFg+ CD8+ Day 36	0.0529	0.0476

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.0078
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.94
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

6. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172
Measure Participants	58	59
Perforin+CD107a+IL-2+TNF+IFNg+ CD4+ at Day 1	0	0.000067
Perforin+CD107a+IL-2+TNF+IFNg+ CD4+ Day 36	0	0
Perforin+CD107a+IL-2+TNF+IFNg+ CD8+ Day 1	0	0
Perforin+CD107a+IL-2+TNF+IFNg+ CD8+ Day 36	0.0000698	0

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Other Statistical Analysis		No non-zero results were reported for either group so statistical testing was not performed.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.5
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

7. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2+TNF+IFNg- CD4+ Day 1	0	0
Perforin+CD107a+IL-2+TNF+IFNg- CD4+ Day 36	0	0
Perforin+CD107a+IL-2+TNF+IFNg- CD8+ Day 1	0	0
Perforin+CD107a+IL-2+TNF+IFNg- CD8+ Day 36	0	0

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Other Statistical Analysis		No non-zero results were reported for either group so statistical testing was not performed.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Other Statistical Analysis		No non-zero results were reported for either group so statistical testing was not performed.

8. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2+TNF- IFNg+ CD4+ Day 1	0	0
Perforin+CD107a+IL-2+TNF- IFNg+ CD4+ Day 36	0	0
Perforin+CD107a+IL-2+TNF- IFNg+ CD8+ Day 1	0	0
Perforin+CD107a+IL-2+TNF- IFNg+ CD8+ Day 36	0	0

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Other Statistical Analysis		No non-zero results were reported for either group so statistical testing was not performed.

9. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2+TNF- IFNg- CD4+ Day 1	0	0
Perforin+CD107a+IL-2+TNF- IFNg- CD4+ Day 36	0	0
Perforin+CD107a+IL-2+TNF- IFNg- CD8+ Day 1	0.00033	0.0000461
Perforin+CD107a+IL-2+TNF- IFNg- CD8+ Day 36	0	0.0001540

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Other Statistical Analysis		No non-zero results were reported for either group so statistical testing was not performed.

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.5
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

10. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2-TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2-TNF+IFNg+ CD4+ at Day 1	0.0000741	0.000107
Perforin+CD107a+IL-2-TNF+IFNg+ CD4+ Day 36	0.0001600	0.000344
Perforin+CD107a+IL-2-TNF+IFNg+ CD8+ Day 1	0.00271	0.000269
Perforin+CD107a+IL-2-TNF+IFNg+ CD8+ Day 36	0.00187	0.00122

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	>0.99
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.74
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

11. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2-TNF+IFNg- CD4+ at Day 1	0.000036	0
Perforin+CD107a+IL-2-TNF+IFNg- CD4+ Day 36	0	0.000161
Perforin+CD107a+IL-2-TNF+IFNg- CD8+ Day 1	0.00288	0.00172
Perforin+CD107a+IL-2-TNF+IFNg- CD8+ Day 36	0.00326	0.00134

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.50
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.90
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

12. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2-TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2-TNF- IFNg+ CD4+ at Day 1	0	0
Perforin+CD107a+IL-2-TNF- IFNg+ CD4+ Day 36	0.0000566	0.000118
Perforin+CD107a+IL-2-TNF- IFNg+ CD8+ Day 1	0.000444	0.000409
Perforin+CD107a+IL-2-TNF- IFNg+ CD8+ Day 36	0.000323	0.000197

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.87
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.57
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

13. Primary Outcome

Title	Mean Percentage of Perforin+CD107a+IL-2-TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a+IL-2-TNF- IFNg- CD4+ at Day 1	0.00104	0.000728
Perforin+CD107a+IL-2-TNF- IFNg- CD4+ Day 36	0.00123	0.000928
Perforin+CD107a+IL-2-TNF- IFNg- CD8+ Day 1	0.0317	0.0237
Perforin+CD107a+IL-2-TNF- IFNg- CD8+ Day 36	0.0368	0.0263

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.82
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.076
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

14. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2+TNF+IFNg+ CD4+ at Day 1	0.0000621	0.0000457
Perforin+CD107a- IL-2+TNF+IFNg+ CD4+ Day 36	0.0000888	0.0001010
Perforin+CD107a- IL-2+TNF+IFNg+ CD8+ Day 1	0.000123	0
Perforin+CD107a- IL-2+TNF+IFNg+ CD8+ Day 36	0.000184	0.000128

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	>0.99
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.75
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

15. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2+TNF+IFNg- CD4+ at Day 1	0.0000375	0
Perforin+CD107a- IL-2+TNF+IFNg- CD4+ Day 36	0.0000327	0
Perforin+CD107a- IL-2+TNF+IFNg- CD8+ Day 1	0.0005150	0.000322
Perforin+CD107a- IL-2+TNF+IFNg- CD8+ Day 36	0.0000524	0

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.50
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.50
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

16. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2+TNF- IFNg+ CD4+ at Day 1	0	0.000101
Perforin+CD107a- IL-2+TNF- IFNg+ CD4+ Day 36	0	0.000061
Perforin+CD107a- IL-2+TNF- IFNg+ CD8+ Day 1	0	0.0003780
Perforin+CD107a- IL-2+TNF- IFNg+ CD8+ Day 36	0	0.0000966

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	>.99
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	>.99
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

17. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2+TNF- IFNg- CD4+ at Day 1	0.00399	0.00307
Perforin+CD107a- IL-2+TNF- IFNg- CD4+ Day 36	0.00458	0.00354
Perforin+CD107a- IL-2+TNF- IFNg- CD8+ Day 1	0.0742	0.0434
Perforin+CD107a- IL-2+TNF- IFNg- CD8+ Day 36	0.0546	0.0612

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.81
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.47
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

18. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2- TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2- TNF+IFNg+ CD4+ at Day 1	0.0000639	0.000372
Perforin+CD107a- IL-2- TNF+IFNg+ CD4+ Day 36	0	0.000485
Perforin+CD107a- IL-2- TNF+IFNg+ CD8+ Day 1	0.00122	0.00135
Perforin+CD107a- IL-2- TNF+IFNg+ CD8+ Day 36	0.00305	0.00168

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.0061
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.28
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

19. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2- TNF+IFNg- CD4+ at Day 1	0.001100	0.000199
Perforin+CD107a- IL-2- TNF+IFNg- CD4+ Day 36	0.000564	0.000454
Perforin+CD107a- IL-2- TNF+IFNg- CD8+ Day 1	0.00880	0.00426
Perforin+CD107a- IL-2- TNF+IFNg- CD8+ Day 36	0.00947	0.00595

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.36
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.32
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

20. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2- TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2- TNF- IFNg+ CD4+ at Day 1	0.000421	0.000436
Perforin+CD107a- IL-2- TNF- IFNg+ CD4+ Day 36	0.00027	0.000438
Perforin+CD107a- IL-2- TNF- IFNg+ CD8+ Day 1	0.00654	0.00177
Perforin+CD107a- IL-2- TNF- IFNg+ CD8+ Day 36	0.00376	0.00370

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.42
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.26
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

21. Primary Outcome

Title	Mean Percentage of Perforin+CD107a- IL-2- TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin+CD107a- IL-2- TNF- IFNg- CD4+ at Day 1	1.180	0.775
Perforin+CD107a- IL-2- TNF- IFNg- CD4+ Day 36	1.080	0.772
Perforin+CD107a- IL-2- TNF- IFNg- CD8+ Day 1	16.0	12.3
Perforin+CD107a- IL-2- TNF- IFNg- CD8+ Day 36	16.2	13.7

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.53
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.56
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

22. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2+TNF+IFNg+ CD4+ at Day 1	0.00135	0.00122
Perforin- CD107a+IL-2+TNF+IFNg+ CD4+ Day 36	0.00156	0.00138
Perforin- CD107a+IL-2+TNF+IFNg+ CD8+ Day 1	0.001170	0.001000
Perforin- CD107a+IL-2+TNF+IFNg+ CD8+ Day 36	0.000742	0.000465

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.97
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.62
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

23. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2+TNF+IFNg- CD4+ at Day 1	0.00230	0.00200
Perforin- CD107a+IL-2+TNF+IFNg- CD4+ Day 36	0.00212	0.00144
Perforin- CD107a+IL-2+TNF+IFNg- CD8+ Day 1	0.000197	0.0005900
Perforin- CD107a+IL-2+TNF+IFNg- CD8+ Day 36	0.000346	0.0000719

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.15
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.18
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

24. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2+TNF- IFNg+ CD4+ at Day 1	0	0.000156
Perforin- CD107a+IL-2+TNF- IFNg+ CD4+ Day 36	0.0000725	0.000078
Perforin- CD107a+IL-2+TNF- IFNg+ CD8+ Day 1	0	0
Perforin- CD107a+IL-2+TNF- IFNg+ CD8+ Day 36	0	0

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.94
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value
	Comments
	Method
	Comments
Other Statistical Analysis		No non-zero results were reported for either group so statistical testing was not performed.

25. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2+TNF- IFNg- CD4+ at Day 1	0.000809	0.000263
Perforin- CD107a+IL-2+TNF- IFNg- CD4+ Day 36	0.000653	0.000223
Perforin- CD107a+IL-2+TNF- IFNg- CD8+ Day 1	0.000377	0.000840
Perforin- CD107a+IL-2+TNF- IFNg- CD8+ Day 36	0.000912	0.000369

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.14
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.26
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

26. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2- TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2- TNF+IFNg+ CD4+ at Day 1	0.00269	0.00248
Perforin- CD107a+IL-2- TNF+IFNg+ CD4+ Day 36	0.00309	0.00262
Perforin- CD107a+IL-2- TNF+IFNg+ CD8+ Day 1	0.00513	0.00617
Perforin- CD107a+IL-2- TNF+IFNg+ CD8+ Day 36	0.00604	0.00374

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.72
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.31
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

27. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2- TNF+IFNg- CD4+ at Day 1	0.0147	0.0160
Perforin- CD107a+IL-2- TNF+IFNg- CD4+ Day 36	0.0123	0.0127
Perforin- CD107a+IL-2- TNF+IFNg- CD8+ Day 1	0.00678	0.00725
Perforin- CD107a+IL-2- TNF+IFNg- CD8+ Day 36	0.00427	0.00630

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.31
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.22
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

28. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2- TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2- TNF- IFNg+ CD4+ at Day 1	0.000511	0.000694
Perforin- CD107a+IL-2- TNF- IFNg+ CD4+ Day 36	0.000731	0.001910
Perforin- CD107a+IL-2- TNF- IFNg+ CD8+ Day 1	0.001580	0.002160
Perforin- CD107a+IL-2- TNF- IFNg+ CD8+ Day 36	0.001780	0.002240

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.16
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.44
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

29. Primary Outcome

Title	Mean Percentage of Perforin- CD107a+IL-2- TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a+IL-2- TNF- IFNg- CD4+ at Day 1	0.231	0.240
Perforin- CD107a+IL-2- TNF- IFNg- CD4+ Day 36	0.220	0.236
Perforin- CD107a+IL-2- TNF- IFNg- CD8+ Day 1	0.294	0.323
Perforin- CD107a+IL-2- TNF- IFNg- CD8+ Day 36	0.295	0.341

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.67
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.48
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

30. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2+TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2+TNF+IFNg+ CD4+ at Day 1	0.00328	0.00365
Perforin- CD107a- IL-2+TNF+IFNg+ CD4+ Day 36	0.00955	0.00289
Perforin- CD107a- IL-2+TNF+IFNg+ CD8+ Day 1	0.000238	0.000693
Perforin- CD107a- IL-2+TNF+IFNg+ IFNg- CD8+ Day 36	0.000288	0.000486

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	<0.001
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.62
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

31. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2+TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2+TNF+IFNg- CD4+ at Day 1	0.01170	0.01240
Perforin- CD107a- IL-2+TNF+IFNg- CD4+ Day 36	0.01640	0.01130
Perforin- CD107a- IL-2+TNF+IFNg- CD8+ Day 1	0.00161	0.00184
Perforin- CD107a- IL-2+TNF+IFNg- IFNg- CD8+ Day 36	0.00129	0.00102

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.0093
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.51
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

32. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2+TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2+TNF- IFNg+ CD4+ at Day 1	0.000169	0.000438
Perforin- CD107a- IL-2+TNF- IFNg+ CD4+ Day 36	0.000434	0.000372
Perforin- CD107a- IL-2+TNF- IFNg+ CD8+ Day 1	0.000397	0.000339
Perforin- CD107a- IL-2+TNF- IFNg+ CD8+ Day 36	0.000487	0.000378

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.54
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.43
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

33. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2+TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2+TNF- IFNg- CD4+ at Day 1	0.0445	0.0400
Perforin- CD107a- IL-2+TNF- IFNg- CD4+ Day 36	0.0395	0.0478
Perforin- CD107a- IL-2+TNF- IFNg- CD8+ Day 1	0.0468	0.0444
Perforin- CD107a- IL-2+TNF- IFNg- CD8+ Day 36	0.0417	0.0522

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.44
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.014
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

34. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2- TNF+IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2- TNF+IFNg+ CD4+ at Day 1	0.00935	0.00881
Perforin- CD107a- IL-2- TNF+IFNg+ CD4+ Day 36	0.01520	0.00731
Perforin- CD107a- IL-2- TNF+IFNg+ CD8+ Day 1	0.00488	0.00460
Perforin- CD107a- IL-2- TNF+IFNg+CD8+ Day 36	0.00257	0.00242

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	<0.001
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.96
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

35. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2- TNF+IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2- TNF+IFNg- CD4+ at Day 1	0.390	0.483
Perforin- CD107a- IL-2- TNF+IFNg- CD4+ Day 36	0.389	0.354
Perforin- CD107a- IL-2- TNF+IFNg- CD8+ Day 1	0.0388	0.0415
Perforin- CD107a- IL-2- TNF+IFNg- CD8+ Day 36	0.0341	0.0348

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.65
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.53
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

36. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2- TNF- IFNg+ CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2- TNF- IFNg+ CD4+ at Day 1	0.0238	0.0147
Perforin- CD107a- IL-2- TNF- IFNg+ CD4+ Day 36	0.0199	0.0255
Perforin- CD107a- IL-2- TNF- IFNg+ CD8+ Day 1	0.0314	0.0209
Perforin- CD107a- IL-2- TNF- IFNg+ CD8+ Day 36	0.0318	0.0308

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.35
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD8+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.66
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

37. Primary Outcome

Title	Mean Percentage of Perforin- CD107a- IL-2- TNF- IFNg- CD4+ and CD8+ T Cells After Stimulation With M-001 Component Peptides
Description	The percentages of CD4 and CD8 T cells expressing markers were determined using fluorescence-based flow cytometric assays from cryopreserved PBMCs collected and isolated from participants at Day 1 prior to initial vaccination and again at 14 days after the second vaccination. The percentages were calculated as the number of cells positive for these proteins divided by the total number of CD4+ or CD8+ T cells counted in each sample.
Time Frame	Day 1 through Day 36

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	58	59
Perforin- CD107a- IL-2- TNF- IFNg- CD4+ at Day 1	98.1	98.4
Perforin- CD107a- IL-2- TNF- IFNg- CD4+ Day 36	98.2	98.5
Perforin- CD107a- IL-2- TNF- IFNg- CD8+ Day 1	83.5	87.2
Perforin- CD107a- IL-2- TNF- IFNg- CD8+ Day 36	83.2	85.8

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.86
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	M-001 + IIV4, Placebo + IIV4
	Comments	This reports the analysis for response in CD4+ T cells at Day 36. The null hypothesis for the non-parametric test assumed both groups are from the same population, i.e., are homogeneous and have the same distribution. The study was not designed/powered to detect differences between groups in the frequencies of marker combinations, considering the issue of multiplicity.
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.53
	Comments	As this trial was not confirmatory, as common for early phase trials, p-values were not adjusted for multiple comparisons, rather included to aid interpretation of the estimates. A priori threshold for statistical significance was not defined.
	Method	Wilcoxon (Mann-Whitney)
	Comments

38. Primary Outcome

Title	Number of Participants With Clinical Safety Laboratory Adverse Events After the First M-001 Vaccination
Description	Blood was collected after first vaccination for assessment by a central clinical laboratory. Clinical safety laboratory adverse events included white blood cells (WBC) </=3900/uL or >/=10,600/uL; platelets </=139,000/uL or >/=416,000/uL; hemoglobin </=11.4 g/dL (female) or </=12.4 g/dL (male); alanine aminotransferase (ALT) >/=44 IU/L (female) or >/=61 IU/L (male); creatinine >/=1.1 mg/dL (female) or >/=1.4 (male); and total bilirubin >/=1.30 mg/dL.
Time Frame	Day 9

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received the first dose of study product and have a result reported for the parameter.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	61	59
WBC	5 8.2%	7 11.9%
platelets	1 1.6%	0 0%
hemoglobin	3 4.9%	1 1.7%
ALT	2 3.3%	0 0%
creatinine	0 0%	0 0%
total bilirubin	2 3.3%	2 3.4%

39. Primary Outcome

Title	Number of Participants With Clinical Safety Laboratory Adverse Events After Second M-001 Vaccination
Description	Clinical safety laboratory adverse events included WBC less than or equal to 3900/uL or greater than or equal to 10,600/uL; platelets less than or equal to 139,000/uL or greater than or equal to 416,000/uL; hemoglobin less than or equal to 11.4 g/dL (female) or less than or equal to 12.4 g/dL (male); alanine aminotransferase (ALT) greater than or equal to 44 IU/L (female) or greater than or equal to 61 IU/L (male); creatinine greater than or equal to 1.1 mg/dL (female) or greater than or equal to 1.4 (male); and total bilirubin greater than or equal to 1.30 mg/dL.
Time Frame	Day 22 through Day 29

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received the second dose of study product and have a result reported for the parameter.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	54	55
WBC	2 3.3%	3 5.1%
platelets	0 0%	0 0%
hemoglobin	2 3.3%	0 0%
ALT	2 3.3%	0 0%
creatinine	0 0%	0 0%
total bilirubin	2 3.3%	2 3.4%

40. Primary Outcome

Title	Number of Participants Reporting Solicited Injection Site and Systemic Reactogenicity Events After the First M-001 Vaccination
Description	Participants maintained a memory aid and thermometer to record daily oral temperatures and the occurrence of systemic reactions of feverishness, fatigue, malaise, myalgia, arthralgia, headache, and nausea, as well as local injection site reactions of pain, tenderness, redness, and swelling for 8 days after vaccination (Day 0-7) based on their interference with daily activities, with a severity grade of mild meaning no interference, moderate as some interference and severe as significant interference/prevented daily activity. Fever was defined as an oral temperature of 38 degree Celsius or higher. Participants are counted if they reported experiencing the symptom at any severity on any of the 8 days post vaccination.
Time Frame	Day 1 through Day 8

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received the first study product.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	61	59
Fever	0 0%	1 1.7%
Feverishness	1 1.6%	3 5.1%
Fatigue	14 23%	14 23.7%
Malaise	5 8.2%	5 8.5%
Myalgia	6 9.8%	6 10.2%
Arthralgia	1 1.6%	1 1.7%
Headache	13 21.3%	15 25.4%
Nausea	7 11.5%	1 1.7%
Injection site pain	12 19.7%	2 3.4%
Injection site tenderness	24 39.3%	9 15.3%
Injection site itchiness/pruritus	2 3.3%	1 1.7%
Injection site ecchymosis	2 3.3%	3 5.1%
Injection site erythema	14 23%	11 18.6%
Injection site induration/swelling	3 4.9%	4 6.8%

41. Primary Outcome

Title	Number of Participants Reporting Solicited Injection Site and Systemic Reactogenicity Events After the Second M-001 Vaccination
Description	Participants maintained a memory aid and thermometer to record daily oral temperatures and the occurrence of systemic reactions of feverishness, fatigue, malaise, myalgia, arthralgia, headache, and nausea, as well as local injection site reactions of pain, tenderness, redness, and swelling for 8 days after vaccination (Day 0-7) based on their interference with daily activities, with a severity grade of mild meaning no interference, moderate as some interference and severe as significant interference/prevented daily activity. Fever was defined as an oral temperature of 38 degree Celsius or higher. Participants are counted if they reported experiencing the symptom at any severity on any of the 8 days post vaccination.
Time Frame	Day 22 through Day 29

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received the second study product.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	55	55
Fever	1 1.6%	0 0%
Feverishness	1 1.6%	2 3.4%
Fatigue	8 13.1%	7 11.9%
Malaise	6 9.8%	4 6.8%
Myalgia	4 6.6%	2 3.4%
Arthralgia	0 0%	2 3.4%
Headache	8 13.1%	8 13.6%
Nausea	2 3.3%	1 1.7%
Injection site pain	13 21.3%	2 3.4%
Injection site tenderness	16 26.2%	11 18.6%
Injection site itchiness/pruritus	2 3.3%	1 1.7%
Injection site ecchymosis	0 0%	3 5.1%
Injection site erythema	15 24.6%	7 11.9%
Injection site induration/swelling	2 3.3%	2 3.4%

42. Primary Outcome

Title	Number of Participants Reporting Vaccine-related Serious Adverse Events (SAEs) After M-001 Vaccination
Description	SAEs included any untoward medical occurrence that resulted in death; was life threatening; was a persistent/significant disability/incapacity; required inpatient hospitalization or prolongation or a congenital anomaly/birth defect. Relationship (related or unrelated to the study product) was determined by a site principal investigator blinded to the study product received by the participant.
Time Frame	Day 1 through Day 200

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received at least one dose of study product.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	61	59
Count of Participants [Participants]	0 0%	0 0%

43. Secondary Outcome

Title	Number of Participants Reporting Serious Adverse Events (SAEs) After M-001 Vaccination
Description	SAEs included any untoward medical occurrence that resulted in death; was life threatening; was a persistent/significant disability/incapacity; required inpatient hospitalization or prolongation or a congenital anomaly/birth defect. All events are included regardless of relationship to the study product.
Time Frame	Day 1 through Day 200

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received at least one dose of study product.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	61	59
Count of Participants [Participants]	1 1.6%	0 0%

44. Secondary Outcome

Title	Incidence of Unsolicited Non-serious Adverse Events (AEs) After M-001 Vaccination
Description	Unsolicited adverse events were collected from the time of first vaccination and at each follow-up visit through Day 43. Adverse events were defined as any untoward medical occurrence regardless of its causal relationship to the study treatment. Events were coded with MedDRA and are reported by System Organ Class.
Time Frame	Day 1 through Day 43

Outcome Measure Data

Analysis Population Description

The Safety Analysis population includes all participants who received at least one dose of study product.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	61	59
Blood and lymphatic system disorders	0 0%	2 3.4%
Eye Disorders	1 1.6%	1 1.7%
Gastrointestinal disorders	2 3.3%	1 1.7%
Immune system disorders	0 0%	1 1.7%
Infections and Infestations	11 18%	12 20.3%
Injury, poisoning and procedural complications	2 3.3%	2 3.4%
Musculoskeletal and connective tissue disorders	4 6.6%	1 1.7%
Nervous system disorders	3 4.9%	2 3.4%
Reproductive system and breast disorders	1 1.6%	0 0%
Respiratory, thoracic and mediastinal disorders	3 4.9%	2 3.4%
Skin and subcutaneous tissue disorders	3 4.9%	3 5.1%
Vascular disorders	0 0%	1 1.7%

45. Secondary Outcome

Title	The Percentage of Subjects Achieving HAI Seroconversion to IIV4 Vaccine Virus From Day 172 to Day 200
Description	Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Seroconversion was defined as a Day 172 titer less than 10 and Day 200 titer greater than or equal to 40, or for those with a Day 172 titer of 10 or greater, a minimum 4-fold rise in Day 200 antibody titer.
Time Frame	Day 172 to Day 200

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all participants who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	52	50
B/Colorado/6/2017	13.5 22.1%	6.0 10.2%
B/Phuket/3073/2013	11.5 18.9%	8.0 13.6%
A/Michigan/45/2015 X-275	15.4 25.2%	16.0 27.1%
A/Singapore/INFIMH-16-0019/2016 NIB-104	25.0 41%	20.0 33.9%

46. Secondary Outcome

Title	The Percentage of Subjects Achieving Neutralizing Antibody Seroconversion to IIV4 Vaccine Virus From Day 172 to Day 200
Description	Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Seroconversion was defined as a Day 172 titer less than 10 and Day 200 titer greater than or equal to 40, or for those with a Day 172 titer of 10 or greater, a minimum 4-fold rise in Day 200 antibody titer.
Time Frame	Day 172 to Day 200

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all participants who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	52	50
B/Colorado/6/2017	17.3 28.4%	6.0 10.2%
B/Phuket/3073/2013	13.5 22.1%	8.0 13.6%
A/Michigan/45/2015 X-275	19.2 31.5%	16.0 27.1%
A/Singapore/INFIMH-16-0019/2016 NIB-104	46.2 75.7%	26.0 44.1%

47. Secondary Outcome

Title	The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 1
Description	Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 1

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	55	55
B/Colorado/6/2017	98.3 161.1%	93.2 158%
B/Phuket/3073/2013	93.3 153%	93.2 158%
A/Michigan/45/2015 X-275	80.0 131.1%	81.4 138%
A/Singapore/INFIMH-16-0019/2016 NIB-104	43.3 71%	33.9 57.5%

48. Secondary Outcome

Title	The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 43
Description	Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 43

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	53	53
B/Colorado/6/2017	96.2 157.7%	92.5 156.8%
B/Phuket/3073/2013	94.3 154.6%	92.5 156.8%
A/Michigan/45/2015 X-275	79.2 129.8%	79.2 134.2%
A/Singapore/INFIMH-16-0019/2016 NIB-104	39.6 64.9%	35.8 60.7%

49. Secondary Outcome

Title	The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 172
Description	Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 172

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	53	52
B/Colorado/6/2017	94.3 154.6%	88.5 150%
B/Phuket/3073/2013	94.3 154.6%	88.5 150%
A/Michigan/45/2015 X-275	81.1 133%	78.8 133.6%
A/Singapore/INFIMH-16-0019/2016 NIB-104	39.6 64.9%	30.8 52.2%

50. Secondary Outcome

Title	The Percentage of Participants With an HAI Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 200
Description	Blood was collected from participants for testing in the HAI assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 200

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	52	50
B/Colorado/6/2017	98.1 160.8%	96.0 162.7%
B/Phuket/3073/2013	98.1 160.8%	96.0 162.7%
A/Michigan/45/2015 X-275	94.2 154.4%	88.0 149.2%
A/Singapore/INFIMH-16-0019/2016 NIB-104	67.3 110.3%	60 101.7%

51. Secondary Outcome

Title	The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 1
Description	Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 1

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	60	59
B/Colorado/6/2017	30.0 49.2%	22.0 37.3%
B/Phuket/3073/2013	71.7 117.5%	74.6 126.4%
A/Michigan/45/2015 X-275	100 163.9%	96.6 163.7%
A/Singapore/INFIMH-16-0019/2016 NIB-104	91.7 150.3%	93.2 158%

52. Secondary Outcome

Title	The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 43
Description	Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 43

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	53	53
B/Colorado/6/2017	20.8 34.1%	28.3 48%
B/Phuket/3073/2013	77.4 126.9%	69.8 118.3%
A/Michigan/45/2015 X-275	98.1 160.8%	98.1 166.3%
A/Singapore/INFIMH-16-0019/2016 NIB-104	90.6 148.5%	92.5 156.8%

53. Secondary Outcome

Title	The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 172
Description	Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 172

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	53	52
B/Colorado/6/2017	24.5 40.2%	26.9 45.6%
B/Phuket/3073/2013	67.9 111.3%	69.2 117.3%
A/Michigan/45/2015 X-275	98.1 160.8%	96.2 163.1%
A/Singapore/INFIMH-16-0019/2016 NIB-104	86.8 142.3%	92.3 156.4%

54. Secondary Outcome

Title	The Percentage of Participants With a Neutralizing Antibody Titer of 40 or Greater and GMTs vs. IIV4 Vaccine Viruses at Day 200
Description	Blood was collected from participants for testing in the neutralizing antibody assay with the vaccine viruses as the assay antigens. Each sample was tested at least twice according to standard operating procedures and the result of each replicate reported. A participant is counted if the geometric mean of the replicate values was 40 or greater.
Time Frame	Day 200

Outcome Measure Data

Analysis Population Description

The modified intent-to-treat (mITT) population includes all subjects who received at least one dose of study vaccine and contributed at least one post-study vaccination venous blood sample for immunogenicity testing for which valid results were reported.

Arm/Group Title	M-001 + IIV4	Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.	0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172.
Measure Participants	52	50
B/Colorado/6/2017	69.2 113.4%	50.0 84.7%
B/Phuket/3073/2013	86.5 141.8%	82.0 139%
A/Michigan/45/2015 X-275	100 163.9%	100 169.5%
A/Singapore/INFIMH-16-0019/2016 NIB-104	98.1 160.8%	98.0 166.1%

Adverse Events

Time Frame	Solicited events were collected for 8 days after each vaccination, unsolicited non-serious AEs through 28 days after the second study vaccination, and SAEs through approximately Day 200.
Adverse Event Reporting Description	For events solicited on the Memory Aid in the 8 days after vaccination, a participant was considered to have one event if it was reported as experienced at any time in the 8-day period. Each 8-day period was considered a separate event.
Arm/Group Title	M-001 + IIV4		Placebo + IIV4
Arm/Group Description	0.4 ml (1 mg) M-001 IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172		0.4 ml Placebo IM on Day 1 and Day 22, followed by 0.5 ml IIV4 IM on Day 172
All Cause Mortality
	M-001 + IIV4		Placebo + IIV4
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/61 (0%)		0/59 (0%)
Serious Adverse Events
	M-001 + IIV4		Placebo + IIV4
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	1/61 (1.6%)		0/59 (0%)
Ear and labyrinth disorders
Deafness	1/61 (1.6%)	1	0/59 (0%)	0
Other (Not Including Serious) Adverse Events
	M-001 + IIV4		Placebo + IIV4
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	52/61 (85.2%)		44/59 (74.6%)
Gastrointestinal disorders
Nausea	9/61 (14.8%)	9	1/59 (1.7%)	2
General disorders
Fatigue	18/61 (29.5%)	22	16/59 (27.1%)	21
Feeling Hot	2/61 (3.3%)	2	4/59 (6.8%)	5
Injection Site Erythema	22/61 (36.1%)	29	14/59 (23.7%)	18
Injection Site Haemorrhage	2/61 (3.3%)	2	5/59 (8.5%)	6
Injection Site Induration	5/61 (8.2%)	5	5/59 (8.5%)	6
Injection Site Pain	33/61 (54.1%)	46	14/59 (23.7%)	20
Malaise	9/61 (14.8%)	11	7/59 (11.9%)	9
Infections and infestations
Respiratory Tract Infection	3/61 (4.9%)	3	5/59 (8.5%)	5
Investigations
Blood Bilirubin Increased	2/61 (3.3%)	4	3/59 (5.1%)	4
Haemoglobin Decreased	5/61 (8.2%)	6	1/59 (1.7%)	1
White Blood Cell Count Decreased	3/61 (4.9%)	4	5/59 (8.5%)	5
White Blood Cell Count Increased	4/61 (6.6%)	4	4/59 (6.8%)	5
Musculoskeletal and connective tissue disorders
Arthralgia	1/61 (1.6%)	1	3/59 (5.1%)	3
Myalgia	9/61 (14.8%)	10	7/59 (11.9%)	8
Nervous system disorders
Headache	17/61 (27.9%)	21	18/59 (30.5%)	23

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

Principal Investigators are NOT employed by the organization sponsoring the study.

The only disclosure restriction on the PI is that the sponsor can review results communications prior to public release and can embargo communications regarding trial results for a period that is less than or equal to 60 days. The sponsor cannot require changes to the communication and cannot extend the embargo.