Molecular analysis of cytokine polymorphism in conjunction with the study of immune response to common recall antigens, superantigens and xenobiotic compounds in autoimmune skin diseases

Abstract

Autoimmune diseases in susceptible individuals depend largely on multiple factors the genetic makeup, environmental antigen challenge and the immunological status of the afflicted. There is a paucity of knowledge on the risk posed by various environmental toxins, microbial antigen or superantigen stimulants which may modify the clinical expression of the autoimmune skin diseases and the response of the patient to treatment strategies. The participation of cytokines in the induction and effector phases of the immune and inflammatory response shows that their polymorphism may be playing a critical role in the development of autoimmune diseases. Objectives: Present study is an attempt to elucidate the role of various microbial antigens or superantigens (sAgs) produced by certain pathogens, and organochlorine pesticides (OCPs) in altering effector T cells and cytokine expression in patients of three autoimmune skin diseases viz., pemphigus, systemic sclerosis (SSc) and systemic lupus erythematosus (SLE). Efforts were also made to investigate single nucleotide polymorphism of an array of cytokines at DNA/genomic level to enhance our understanding of their role in disease progression in these autoimmune skin diseases. Methods: Fifty three patients diagnosed to have autoimmune skin disease pemphigus (20), systemic sclerosis (20) and systemic lupus erythematosus (13) were enrolled in the study. Peripheral venous blood (~10 ml) was collected aseptically from each patient and used for isolation of peripheral blood mononuclear cells (PBMCs), pesticide residue quantification and DNA extraction. Six months after induction of clinical remission by immunosupressives/steroids, repeat blood samples were collected from each patient for PBMCs isolation. Healthy age-matched volunteers were also enrolled for comparisons. PBMCs were stimulated in-vitro with different concentrations of microbial antigens or sAgs which included Staphylococcal enterotoxin B (SEB), Streptococcal pyrogenic exotoxin A (SPEA) and Cytomegalovirus (CMV)