Comparative Analysis of the Th17 Cellular Response in Active and Inactive Pemphigus Vulgaris Patients

Sponsor

Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran (Other)

Overall Status

Recruiting

CT.gov ID

NCT04096222

Collaborator

Hospital General Dr. Manuel Gea González (Other)

Enrollment

Locations

20.9

Anticipated Duration (Months)

Patients Per Site

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

This study will compare the pattern of Th17 immune response in active and inactive pemphigus subjects. Skin and serum samples will be taken at the moment of enrollment.

Condition or Disease	Intervention/Treatment	Phase
Pemphigus Vulgaris Bullous Dermatoses Autoimmune Diseases

Detailed Description

Pemphigus is an autoimmune disease characterized by production of autoantibodies against desmogleins 1 and 3, which are part of the epidermis desmosomes. The first line of treatment are corticosteroids with or without the use of adjuvants (e.g. azathioprine, mycophenolate or rituximab). T lymphocytes are responsible for the initiation and maturation of the humoral response and the B cell activation required for the production of autoantibodies. In the last decade, the Th17 immune response has been implicated in the pathogenesis of pemphigus. Recently, the existence of tertiary lymphoid organ-like structures within the skin lesions was suggested. This structures contain T lymphocytes, B lymphocytes and plasma cells; these cells interact and create a local microenvironment for the production of autoantibodies. Most of the T cells in this structures are T helper CD4+ and express IL-21, and half of them produce IL-17.

In this study the investigators aim to evaluate comparatively the Th17 and T regulatory immune response in the lesional skin and serum of active and inactive pemphigus subjects that are treated with corticosteroids with or without adjuvants and a third group of healthy subjects. The investigators will study skin and serum due to the difference of lymphocytes and cytokines in both tissues. The primary hypothesis is: active pemphigus vulgaris subjects will have different levels of TH17 response in comparison to inactive patients.

The investigators will use descriptive statistics, association and correlation test of hypothesis.

Study Design

Study Type:

Observational

Anticipated Enrollment :

42 participants

Observational Model:

Case-Control

Time Perspective:

Other

Official Title:

Comparative Analysis of the Th17 Cellular Response in Active and Inactive Pemphigus Vulgaris Patients

Actual Study Start Date :

Jun 29, 2021

Anticipated Primary Completion Date :

Mar 28, 2023

Anticipated Study Completion Date :

Mar 28, 2023

Arms and Interventions

Arm	Intervention/Treatment
Active pemphigus vulgaris subjects Subjects with the diagnosis of pemphigus and with active or inactive disease in the skin will be invited to participate. After informed consent, at the enrollment visit the following interventions will be done: 1) two 4 mm punch biopsies in a target lesion, 2) A 34 ml blood sample will be taken, 3) photographs of the subject, 4) demographic, disease, comorbidity and treatment data will be documented, 5) PDAI and ABSIS, 6) Biometric data
Inactive pemphigus vulgaris subjects Subjects with the diagnosis of pemphigus and with active or inactive disease in the skin will be invited to participate. After informed consent, at the enrollment visit the following interventions will be done: 1) two 4 mm punch biopsies in a target lesion, 2) A 34 ml blood sample will be taken, 3) photographs of the subject, 4) demographic, disease, comorbidity and treatment data will be documented, 5) PDAI and ABSIS, 6) Biometric data
Healthy subjects Healthy subjects will be invited to participate. After informed consent, at the enrollment visit the following interventions will be done: 1) Demographic data will be documented, 2) biometric data, 3) A 34 ml blood sample will be taken

Outcome Measures

Primary Outcome Measures

Level of Th17 cytokines in skin of pemphigus vulgaris subjects [Enrollment]
The level of IL-17a, IL-21, IL-22, and IL-23 mRNA from skin biopsies at the time of enrollment and when the subject reaches 75% of PDAI improvement or after a year of follow-up (termination visit). The percentage of change in these two determinations will be calculated.

Other Outcome Measures

Pemphigus Disease Area Index (PDAI) [Enrollment]
PDAI has a total of 0-263 points, 250 are related to activity and 13 to damage. Disease severity is considered as follows: 1) moderate <= 14 points; b) significative 15-44 points; and c) extensive >=45
Disease activity [Enrollment]
Measured with Pemphigus Disease Area Index (PDAI) and Autoimmune Bullous Skin Disorder Intensity Score (ABSIS). PDAI has a total of 0-263 points, 250 are related to activity and 13 to damage. ABSIS has a total score of 0-206.
Autoimmune Bullous Skin Disorder Intensity Score (ABSIS) [Enrollment]
ABSIS has a total score of 0-206. Disease severity is considered as follows: 1) moderate <= 16 points; b) significative 17-52 points; and c) extensive >=53
Treatment [Enrollment]
The medications and doses used since the diagnosis and during the study
Proportion of Th17 and Treg populations on skin biopsies [Enrollment]
iopsies will be processed for immunohistochemistry for TH17 subsets (CD+IL17a+) and Treg subset (CD25+Foxp3+). The proportion of both subsets will be quantified with specialized software.
Level of Th17 cytokines in serum of pemphigus vulgaris subjects [Enrollment]
The level of IL-17a, IL-21, IL-22 and IL-23 determined by luminometry
Level of Th17 chemokines levels [Enrollment]
The level of CCL20 and CXCL8 mRNA from skin biopsies and in serum by luminometry
Level of Treg cytokines [Enrollment]
Determination in skin (RT-PCR) and serum (luminometry) of CCL20 and CXCL8
Level of Treg chemokines [Enrollment]
Determinations at the moment of enrollment and at the termination visit in skin (RT-PCR) and serum (luminometry)
P-glycoprotein transporter activity in mononuclear peripheral cells [Enrollment]
The activity will be a measure of the percentage of efflux of daunorubicin ar 37°C.
Percentage of Th17 peripheral cells with expression of P-glycoprotein on the surface [Enrollment]
Cells with the phenotype IL-17a+CCR6+CXR3hiCCR4loCCR10-CD161+PGP+ will be measured with flow cytometry
Levels of anti-desmogleins 1 and 3 [Enrollment]
Levels will be measures with an ELISA assay
Proportion of peripheral cellular subpopulations [Enrollment]
Determinations with flow cytometry
Level of cytokines in supernatant of cellular culture [Enrollment]
Determinations at the moment of enrollment with ELISA

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years and Older

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

Inclusion Criteria:

Current cutaneous activity of pemphigus
Subjects will be treated with corticosteroids with or without adjuvants
Accept and sign the informed consent

Exclusion Criteria:

Pregnancy
Concurrent autoimmune diseases with skin lesions
Concurrent diagnosis of cancer
Concurrent active infectious disease

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Hospital General Dr. Manuel Gea González	Mexico City		Mexico	14080
2	Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran	Mexico City		Mexico	14080