Influenza Immunity in Children

• Sponsor: University of Rochester (Other)
• Overall Status: Completed
• CT.gov ID: NCT02559505
• Collaborator: (none)
• Enrollment: 134
• Location: 1
• Arms: 8
• Actual Duration (Months): 57.1
• Patients Per Site Per Month: 2.3

Study Details

Study Description

Brief Summary

This study evaluates how different methods of early exposure to influenza (natural infection, live attenuated influenza vaccination, inactivated influenza vaccination) initially stimulate immunity and poise the immune system to respond to a future challenge with the inactivated influenza vaccine.

Condition or Disease	Intervention/Treatment	Phase
Influenza	Biological: Seasonal IIV 0.25 mL dose; Other: Natural influenza infection; Biological: Seasonal IIV 0.5 mL dose	N/A

Detailed Description

The proposed research addresses the fact that, despite high childhood morbidity from influenza and broad recommendations for vaccination, very little is known about how anti-influenza immunity is shaped by the method of initial exposure. The objective of this research is to understand how CD4 T cell and B cell responses are altered by the method of initial influenza priming, with the long-term goal of determining how a child's initial influenza encounter poises the immune system to respond to subsequent influenza challenges. The investigators central hypothesis is that differences in the mode of influenza antigen exposure in early childhood will generate long lasting, detectable changes in memory CD4 T cell and B cell specificity and function that influence the response to future influenza vaccinations and infections. This hypothesis will be tested by comparing 1) CD4 T cell and 2) antibody responses in cohorts of children initially exposed to influenza through either natural infection or inactivated or live attenuated vaccination. A combination of multiparameter assays will be used to determine the phenotype and functional potential of hemagglutinin (HA)- and nucleoprotein (NP)-specific CD4 T cells. The breadth and avidity of the neutralizing and non-neutralizing antibody responses and its distribution against head and stalk epitopes will also be evaluated. By determining how initial priming shapes the specificity and functional potential of the anti-influenza CD4 T cell and antibody responses, the investigators will gain the knowledge necessary to optimize current influenza vaccination strategies and develop novel influenza vaccines able to provide highly efficacious universal protection against both seasonal and potentially pandemic viral strains.

Study Design

• Study Type: Interventional
• Actual Enrollment: 134 participants
• Allocation: Non-Randomized
• Intervention Model: Parallel Assignment
• Masking: None (Open Label)
• Primary Purpose: Basic Science
• Official Title: Understanding How the Initial Encounter With Influenza Virus Poises Children for Protective Immunity
• Actual Study Start Date: Oct 1, 2015
• Actual Primary Completion Date: Jul 3, 2020
• Actual Study Completion Date: Jul 3, 2020

Arms and Interventions

Arm	Intervention/Treatment
Experimental: 6-12 months Seasonal IIV Children 6 - 12 months of age vaccinated with seasonal IIV	Biological: Seasonal IIV 0.25 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: Inactivated influenza vaccine
Experimental: 3-12 months natural infection Children 3-12 months of age presenting with natural influenza infection	Other: Natural influenza infection; Children enrolled on presentation to their primary care provider with a natural influenza infection; Biological: Seasonal IIV 0.5 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: inactivated influenza vaccine
Experimental: 13-35 months Seasonal IIV Children 13-35 months of age vaccinated with seasonal IIV	Biological: Seasonal IIV 0.25 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: Inactivated influenza vaccine
Experimental: 13-35 months natural infection Children 13-35 months of age presenting with natural influenza infection	Other: Natural influenza infection; Children enrolled on presentation to their primary care provider with a natural influenza infection; Biological: Seasonal IIV 0.5 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: inactivated influenza vaccine
Experimental: 3-5 years Seasonal IIV Children 3-5 years of age vaccinated with seasonal IIV	Biological: Seasonal IIV 0.5 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: inactivated influenza vaccine
Experimental: 3-5 years natural infection Children 3-5 years of age presenting with natural influenza infection	Other: Natural influenza infection; Children enrolled on presentation to their primary care provider with a natural influenza infection; Biological: Seasonal IIV 0.5 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: inactivated influenza vaccine
Experimental: 6-8 years Seasonal IIV Children 6-8 years of age vaccinated with seasonal IIV	Biological: Seasonal IIV 0.5 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: inactivated influenza vaccine
Experimental: 6-8 years natural infection Children 6-8 years of age presenting with natural influenza infection	Other: Natural influenza infection; Children enrolled on presentation to their primary care provider with a natural influenza infection; Biological: Seasonal IIV 0.5 mL dose; Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age; Other Names: inactivated influenza vaccine

Outcome Measures

Primary Outcome Measures

1. Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects [Visit 2 (day 8-14 post enrollment)]

   % H3- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining

2. Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects [Visit 3 (day 20-28 post enrollment)]

   % H3- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining

3. Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects [Visit 4 (day of vaccination year 2)]

   % H3- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining

4. Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects [Visit 5 (day 8-14 post-vaccination year 2)]

   % H3 protein- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining

5. Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects [Visit 6 (day 20-28 post-vaccination year 2)]

   % H3 Protein- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining

Secondary Outcome Measures

1. Mean Change in Percent of IFNg+ CD69+ CD4 T Cells Between Vaccinated Subjects in Different Age Subsets [Baseline to day 24 study year 1]

   CD4 T cell quantity and specificity will be measured using intracellular cytokine staining. We report here the mean change in percent of cells reactive to the influenza HA protein and H3 protein.

2. Mean Change in Percent of IFNg+ CD69+ CD4 T Cells Between Vaccinated Subjects in Different Age Subsets [Baseline to day 24 study year 2]

   CD4 T cell quantity and specificity will be measured using intracellular cytokine staining. We report here the mean change in percent of cells reactive to the influenza HA protein and H3 protein.

Other Outcome Measures

1. Change From Baseline to Day 10 and Day 24 in PBMC Gene Expression [Days 10 and 24 post vaccination]

   Changes in PBMC gene expression patterns due to prior influenza exposure will be assessed using RNA-seq analysis

Eligibility Criteria

Criteria

• Ages Eligible for Study: 3 Months to 8 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• Age

• Between 6 and 12 months to participate in the vaccination arm of cohort 1 (cohort 1A)

• Between 3 and 12 months to participate in the natural infection arm of cohort 1 (cohort 1B)

• Between 13 and 35 months of age to participate in either the vaccination or natural infection arm of cohort 2

• Between 36 months and 5 years of age to participate in either the vaccination or natural infection arm of cohort 3

• Between 6 years and 8 years of age to participate in either the vaccination or natural infection arm of cohort 4

• Gestational age of ≥37 weeks at birth

• Parent/guardian can provide informed consent

• Available for the duration of the study

• History of previous IIV administration ONLY for participation in the vaccination arm of cohorts 2, 3, or 4

• Acute illness documented to be due to influenza virus ONLY for participation in the natural infection arms of cohorts 1-4

Exclusion Criteria:

• Immunosuppression as a result of an underlying illness or condition (including HIV or a primary immunodeficiency syndrome)

• Active neoplastic disease

• Use of potentially immunosuppressive medications currently or within the past year (including chemotherapeutic agents) or chronic (>2 weeks) use of oral or inhaled steroid therapy

• A diagnosis of asthma requiring chronic controller medication

• Previous administration of influenza vaccine in the current influenza season ONLY for subjects receiving an influenza vaccination

• Receipt of immunoglobulin or another blood product within the year prior to study enrollment

• An acute illness within the previous 3 days or temperature >38o on screening EXCEPT for participation in the natural infection arms of cohorts 1-4

• A contraindication to influenza vaccination EXCEPT infants between 3 and 5 months presenting with natural influenza infection whose only contraindication is their current age

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	University of Rochester	Rochester	New York	United States	14642

Sponsors and Collaborators

• University of Rochester

Investigators

• Principal Investigator: Jennifer L Nayak, MD, University of Rochester

Study Documents (Full-Text)

• Study Protocol and Statistical Analysis Plan - Jan 21, 2016
• Informed Consent Form - Sep 4, 2015

More Information

Publications

None provided.

• Responsible Party: Jennifer Nayak, Associate Professor, University of Rochester
• ClinicalTrials.gov Identifier: NCT02559505
• Other Study ID Numbers: RSRB00058437
• First Posted: Sep 24, 2015
• Last Update Posted: Sep 2, 2021
• Last Verified: Aug 1, 2021

• Keywords provided by Jennifer Nayak, Associate Professor, University of Rochester: natural influenza infection; live attenuated influenza vaccination (LAIV); inactivated influenza vaccination (IIV); infants; children
• Additional relevant MeSH terms: Influenza, Human; Vaccines

Study Results

Participant Flow

Arm/Group Title	Acute	Vaccinated
Arm/Group Description	Children enrolled on presentation to their primary care provider with a natural influenza infection.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age
Period Title: Overall Study
STARTED	49	84
COMPLETED	22	62
NOT COMPLETED	27	22

Baseline Characteristics

Arm/Group Title	6-12 Months of Age: Vaccinated	3-12 Months of Age: Acute	13-35 Months of Age: Vaccinated	13-35 Months of Age: Acute	3-5 Years of Age: Vaccinated	3-5 Years of Age: Acute	6-8 Years of Age: Vaccinated	6-8 Years of Age: Acute	Total
Arm/Group Description	Children 6 - 12 months of age vaccinated with seasonal IIV Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 3-12 months of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 13-35 months of age vaccinated with seasonal IIV Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 13-35 months of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 3-5 years of age vaccinated with seasonal IIV Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 3-5 years of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 6-8 years of age vaccinated with seasonal IIV Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Children 6-8 years of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age	Total of all reporting groups
Overall Participants	13	8	30	19	18	12	23	9	132
Age, Customized (participants) [Number]
Number [participants]	13 100%	8 100%	30 100%	19 100%	18 100%	12 100%	23 100%	9 100%	132 100%
Sex: Female, Male (Count of Participants)
Female	9 69.2%	3 37.5%	14 46.7%	8 42.1%	8 44.4%	6 50%	10 43.5%	6 66.7%	64 48.5%
Male	4 30.8%	5 62.5%	16 53.3%	11 57.9%	10 55.6%	6 50%	13 56.5%	3 33.3%	68 51.5%
Ethnicity (NIH/OMB) (Count of Participants)
Hispanic or Latino	3 23.1%	0 0%	3 10%	4 21.1%	0 0%	2 16.7%	0 0%	3 33.3%	15 11.4%
Not Hispanic or Latino	10 76.9%	8 100%	27 90%	15 78.9%	18 100%	10 83.3%	23 100%	6 66.7%	117 88.6%
Unknown or Not Reported	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%
Race (NIH/OMB) (Count of Participants)
American Indian or Alaska Native	0 0%	0 0%	1 3.3%	0 0%	0 0%	0 0%	0 0%	0 0%	1 0.8%
Asian	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%
Native Hawaiian or Other Pacific Islander	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%	0 0%
Black or African American	4 30.8%	3 37.5%	12 40%	11 57.9%	0 0%	3 25%	1 4.3%	5 55.6%	39 29.5%
White	6 46.2%	4 50%	10 33.3%	3 15.8%	14 77.8%	8 66.7%	15 65.2%	2 22.2%	62 47%
More than one race	2 15.4%	1 12.5%	7 23.3%	3 15.8%	4 22.2%	0 0%	7 30.4%	1 11.1%	25 18.9%
Unknown or Not Reported	1 7.7%	0 0%	0 0%	2 10.5%	0 0%	1 8.3%	0 0%	1 11.1%	5 3.8%
Region of Enrollment (participants) [Number]
United States	13 100%	8 100%	30 100%	19 100%	18 100%	12 100%	23 100%	9 100%	132 100%

Outcome Measures

1. Primary Outcome

Title	Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects
Description	% H3- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining
Time Frame	Visit 2 (day 8-14 post enrollment)

Outcome Measure Data

Analysis Population Description

6 acute participants were infected with influenza strains that were not H3 and therefore were not included in the data analyzed for the primary outcome. Only vaccinated participants that age matched to the acute subjects were analyzed, resulting in 16 acutely infected and 28 vaccinated subjects included in the data analysis.

Arm/Group Title	Acute Infected	Vaccinated
Arm/Group Description	Children enrolled on presentation to their primary care provider with a natural influenza infection	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age
Measure Participants	16	28
Nucleoprotein	.010 (0.0023)	0.003 (0.0018)
H3 protein	0.009 (0.0019)	0.002 (0.0015)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.005
	Comments	p-value for H3 reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.024
	Comments	p-value for NP reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

2. Primary Outcome

Title	Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects
Description	% H3- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining
Time Frame	Visit 3 (day 20-28 post enrollment)

Outcome Measure Data

Analysis Population Description

6 acute participants were infected with influenza strains that were not H3 and therefore were not included in the data analyzed for the primary outcome. Only vaccinated participants that age matched to the acute subjects were analyzed, resulting in 16 acutely infected and 28 vaccinated subjects included in the data analysis.

Arm/Group Title	Acute Infected	Vaccinated
Arm/Group Description	Children enrolled on presentation to their primary care provider with a natural influenza infection.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age.
Measure Participants	16	28
Nucleoprotein	0.012 (0.0025)	0.003 (0.0017)
H3 Protein	0.006 (0.0021)	0.004 (0.0014)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.408
	Comments	p-value for H3 reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.004
	Comments	p-value for NP reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

3. Primary Outcome

Title	Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects
Description	% H3- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining
Time Frame	Visit 4 (day of vaccination year 2)

Outcome Measure Data

Analysis Population Description

6 acute participants were infected with influenza strains that were not H3 and therefore were not included in the data analyzed for the primary outcome. Only vaccinated participants that age matched to the acute subjects were analyzed, resulting in 16 acutely infected and 28 vaccinated subjects included in the data analysis.

Arm/Group Title	Acute Infected	Vaccinated
Arm/Group Description	Children enrolled on presentation to their primary care provider with a natural influenza infection	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age
Measure Participants	16	28
Nucleoprotein	0.003 (0.0030)	0.003 (0.0018)
H3 Protein	0.007 (0.0022)	0.003 (0.0015)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.991
	Comments	p-value for H3 reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.334
	Comments	p-value for NP reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

4. Primary Outcome

Title	Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects
Description	% H3 protein- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining
Time Frame	Visit 5 (day 8-14 post-vaccination year 2)

Outcome Measure Data

Analysis Population Description

6 acute participants were infected with influenza strains that were not H3 and therefore were not included in the data analyzed for the primary outcome. Only vaccinated participants that age matched to the acute subjects were analyzed, resulting in 16 acutely infected and 28 vaccinated subjects included in the data analysis.

Arm/Group Title	Acute Infected	Vaccinated
Arm/Group Description	Children enrolled on presentation to their primary care provider with a natural influenza infection	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age
Measure Participants	16	28
Nucleoprotein	0.010 (0.0031)	0.003 (0.0018)
H3 Protein	0.008 (0.0026)	0.004 (0.0015)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.226
	Comments	p-value for H3 reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.036
	Comments	p-value for NP reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

5. Primary Outcome

Title	Mean Percent of IFNg+ CD69+ CD4 T Cells Between Acute (H3N2) Infected and Vaccinated Subjects
Description	% H3 Protein- and nucleoprotein (NP)-specific CD4 T cells were measured using intracellular cytokine staining
Time Frame	Visit 6 (day 20-28 post-vaccination year 2)

Outcome Measure Data

Analysis Population Description

6 acute participants were infected with influenza strains that were not H3 and therefore were not included in the data analyzed for the primary outcome. Only vaccinated participants that age matched to the acute subjects were analyzed, resulting in 16 acutely infected and 28 vaccinated subjects included in the data analysis.

Arm/Group Title	Acute Infected	Vaccinated
Arm/Group Description	Children enrolled on presentation to their primary care provider with a natural influenza infection.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age
Measure Participants	16	28
Nucleoprotein	0.014 (0.0029)	0.003 (0.0019)
H3 Protein	0.008 (0.0024)	0.007 (0.0015)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.680
	Comments	p-value for H3 reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

Statistical Analysis 2

Statistical Analysis Overview	Comparison Group Selection	Acute Infected, Vaccinated
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.002
	Comments	p-value for H3 reactivity, difference of Acute Infected vs. Vaccinated.
	Method	ANCOVA
	Comments

6. Secondary Outcome

Title	Mean Change in Percent of IFNg+ CD69+ CD4 T Cells Between Vaccinated Subjects in Different Age Subsets
Description	CD4 T cell quantity and specificity will be measured using intracellular cytokine staining. We report here the mean change in percent of cells reactive to the influenza HA protein and H3 protein.
Time Frame	Baseline to day 24 study year 1

Outcome Measure Data

Analysis Population Description

Statistical analysis was completed on all patients who attended all six clinical visits in addition to those who had 3 or more visits that had withdrawn later.

Arm/Group Title	Vaccinated Age Subset (6-12 mo)	Vaccinated Age Subset (13-35 mo)	Vaccinated Age Subset (3-5 yr)	Vaccinated Age Subset (6-8 yr)
Arm/Group Description	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 - 12 months.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 13 - 35 months.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 3 - 5 years.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 - 8 years.
Measure Participants	8	12	14	22
HAB	0.00112 (0.00290)	0.00497 (0.00966)	0.00622 (0.01098)	0.01713 (0.02657)
H3	0.0006729 (0.0017802)	0.0002589 (0.005017)	0.0053456 (0.0101449)	0.001674 (0.008319)

7. Secondary Outcome

Title	Mean Change in Percent of IFNg+ CD69+ CD4 T Cells Between Vaccinated Subjects in Different Age Subsets
Description	CD4 T cell quantity and specificity will be measured using intracellular cytokine staining. We report here the mean change in percent of cells reactive to the influenza HA protein and H3 protein.
Time Frame	Baseline to day 24 study year 2

Outcome Measure Data

Analysis Population Description

Statistical analysis was completed on all patients who attended all six clinical visits in addition to those who had 3 or more visits that had withdrawn later.

Arm/Group Title	Vaccinated Age Subset (6-12 mo)	Vaccinated Age Subset (13-35 mo)	Vaccinated Age Subset (3-5 yr)	Vaccinated Age Subset (6-8 yr)
Arm/Group Description	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 - 12 months.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 13 - 35 months.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 3 - 5 years.	Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 - 8 years.
Measure Participants	8	12	14	22
HAB	0.00552 (0.01436)	0.00331 (0.01216)	0.00003 (0.01096)	0.005698 (0.01729)
H3	0.00350 (0.00372)	0.00177 (0.00371)	0.00188 (0.00650)	-0.0017093 (0.01015)

8. Other Pre-specified Outcome

Title	Change From Baseline to Day 10 and Day 24 in PBMC Gene Expression
Description	Changes in PBMC gene expression patterns due to prior influenza exposure will be assessed using RNA-seq analysis
Time Frame	Days 10 and 24 post vaccination

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title

Arm/Group Description

Adverse Events

Time Frame	up to 18 months
Arm/Group Title	6-12 Months: Vaccinated		3-12 Months: Acute		13-35 Months: Vaccinated		13-35 Months: Acute		3-5 Years: Vaccinated		3-5 Years: Acute		6-8 Years: Vaccinated		6-8 Years: Acute
Arm/Group Description	Children 6 - 12 months of age vaccinated with seasonal IIV Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 3-12 months of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 13-35 months of age vaccinated with seasonal IIV Seasonal IIV 0.25 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 13-35 months of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 3-5 years of age vaccinated with seasonal IIV Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 3-5 years of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 6-8 years of age vaccinated with seasonal IIV Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age		Children 6-8 years of age presenting with natural influenza infection Natural influenza infection: Children enrolled on presentation to their primary care provider with a natural influenza infection Seasonal IIV 0.5 mL dose: Fluzone (Sanofi Pasteur) 0.25 mL administered intramuscularly to children between 6 and 35 months of age
All Cause Mortality
	6-12 Months: Vaccinated		3-12 Months: Acute		13-35 Months: Vaccinated		13-35 Months: Acute		3-5 Years: Vaccinated		3-5 Years: Acute		6-8 Years: Vaccinated		6-8 Years: Acute
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/13 (0%)		0/8 (0%)		0/30 (0%)		0/19 (0%)		0/18 (0%)		0/12 (0%)		0/23 (0%)		0/9 (0%)
Serious Adverse Events
	6-12 Months: Vaccinated		3-12 Months: Acute		13-35 Months: Vaccinated		13-35 Months: Acute		3-5 Years: Vaccinated		3-5 Years: Acute		6-8 Years: Vaccinated		6-8 Years: Acute
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/13 (0%)		0/8 (0%)		0/30 (0%)		0/19 (0%)		0/18 (0%)		0/12 (0%)		0/23 (0%)		0/9 (0%)
Other (Not Including Serious) Adverse Events
	6-12 Months: Vaccinated		3-12 Months: Acute		13-35 Months: Vaccinated		13-35 Months: Acute		3-5 Years: Vaccinated		3-5 Years: Acute		6-8 Years: Vaccinated		6-8 Years: Acute
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/13 (0%)		0/8 (0%)		0/30 (0%)		0/19 (0%)		0/18 (0%)		0/12 (0%)		0/23 (0%)		0/9 (0%)

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

All Principal Investigators ARE employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.