Ex vivo analysis of desmoglein 1-responsive T-helper (Th) 1 and Th2 cells in patients with pemphigus foliaceus and healthy individuals.
Pemphigus foliaceus (PF) is a severe autoimmune bullous disorder, characterized by autoantibodies (autoAb) against desmoglein 1 (Dsg1). As T cells may be critical in the pathology of PF, the aim of the present study was to identify and characterize autoaggressive T-helper cells reactive to Dsg1 in PF patients and healthy individuals. Eight patients with the clinical diagnosis of PF and six HLA class II-matched healthy individuals were examined. By magnetic cell-sorting (MACS) cytokine-secretion assay, Dsg1-responsive T-helper (Th) 1 and Th2 cells were isolated and cloned by limiting dilution. The generated T-cell clones (TCC) were characterized regarding proliferative response, TCR Vbeta-chain usage, and cytokine profile upon in vitro stimulation with Dsg1. Both Dsg1-reactive Th1 and Th2 cells were detected in PF patients and controls at similar frequencies. A total of 15 Th1 and Th2 clones were isolated from patients and 27 TCC from healthy controls. Analysis of TCR Vbeta-chain usage of autoreactive T cells from both groups revealed no predominance of a specific Vbeta chain. Noteworthy, the isolated TCC showed a polarized Th1- or Th2-like phenotype upon in vitro culture and stable expression of Th1 or Th2 cytokines during long-term in vitro culture. In summary, our data demonstrate that T-cell autoreactivity against Dsg1 is not restricted to patients with PF. Moreover, both Th1 and Th2 cells were present in patients and healthy donors, suggesting that the loss of B-cell tolerance against Dsg1 in PF is not exclusively determined by the presence of autoaggressive T cells.
Gebhard
Kerstin L
KL
Department of Dermatology, University of Erlangen, Erlangen, Germany.
Veldman
Christian M
CM
Wassmuth
Ralf
R
Schultz
Erwin
E
Schuler
Gerold
G
Hertl
Michael
M
D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D015820:Cadherins / Q000378:metabolism*; D016022:Case-Control Studies; D049109:Cell Proliferation; D002469:Cell Separation; D016207:Cytokines / Q000378:metabolism; D051183:Desmoglein 1; D000939:Epitopes / Q000737:chemistry; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007371:Interferon-gamma / Q000378:metabolism; D008280:Magnetics; D010392:Pemphigus / Q000378:metabolism*; D010641:Phenotype; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012871:Skin Diseases / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D018417:Th1 Cells / Q000166:cytology*; D018418:Th2 Cells / Q000166:cytology*
D015820:Cadherins; D016207:Cytokines; D051183:Desmoglein 1; D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007074:Immunoglobulin G; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; D007371:Interferon-gamma