Development and Characterization of Desmoglein-3 Specific T cells from Japanese Patients with Pemphigus Vulgaris

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The purpose of this study is to investigate the response of T lymphocytes from Japanese patients with pemphigus vulgaris (PV) to Desmoglein 3, the autoantigen of PV, and to characterize the properties of Dsg 3-specific T cells.1. We performed high resolution HLA class II typing in 14 Japanese PV patients presented from Dr Amagai using the PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method. There was a significant association of either DRB 1*14 (14/14:100%), DQB1*0503 (12/14:85%), or DQB1*0301(9/12:64%), when compared with healthy individuals. 2. The 11 recombinant Dsg 3 fusion proteins (Dsg 3. 1-11) with Maltose Binding Protein (MBP) were prepared to encompass each 40 amino acids staggered by 15 residues, starting from the EC 1 of Dsg 33. Autoreactive T cell responses to Dsg 3 were investigated in 14 PV patients and 20 healthy controls by coculture of PBMC with the 11 recombinant Dsg 3 fusion proteins and the incorporation of [ィイD13ィエD1H]-thymidine. Primary in vitro T cell responses (SI=1.9-2.7) to Dsg 3.9 (residues 201-204) or Dsg 3.10 (residues 226-265) were observed in 11/14 PV patients and 7/14 healthy individuals expressing the PV-associated HLA-DRB1*14 and DQB1*05. In contrast, PBMC from 6 normal controls carrying HLA class II alleles other than DRB1*14 and DQB1*05 were not stimulated by recombinant Dsg 3 fusion proteins. These observations demonstrate that T cell response to Dsg 3 can be detected in PV patients and in healthy donors carrying major histocompatibility complex class II alleles identical or similar to those highly prevalent in PV. And that suggests EC2-3 region of the Dsg 3 molecule may contain epitopes that are recognized by Dsg 3-specific T cell. This study will allow us to further develop and characterize the Dsg 3-specific T cell clone in Japanese PV patients.