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

The purpose of this study is to investigate the response of T lymphocytes from Japanese patients with pemphigus vulgaris(PV)to Desmoglein3，the autoantigen of PV，and to characterize the properties of Dsg3-specific T cells.1。We performed high resolution HLA class II typing in14 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 DRB1*14(14/14：100%)、DQB1*0503(12/14：85%)、or DQB1*0301(9/12：#china_person0#)、when compared with healthy individuals.2.The 11recombinant Dsg3fusion proteins(Dsg3.1-11)with Maltose Binding Protein(MBP)were prepared to encompass each40 amino acids staggered by 15 residues，starting from the EC1 of Dsg33。Autoreactive T cell responses to Dsg3were investigated in14PV patients and 20healthy controls by coculture of PBMC with the11recombinant Dsg3fusion proteins and the incorporation of[Ii D13文件D1H]-thymidine。Primary体外T cell responses(SI=1.9-2.7)to Dsg3.9(residues201-204)或Dsg3.10(residues226-265)were observed in11/14PV patients and 7/14 healthy individuals expressing the PV-associated HLA-DRB1*14 and DQB1*05。In contrast，PBMC from6normal controls carrying HLA class II alleles other than DRB1*14and DQB1*05were not stimulated by recombinant Dsg3fusion proteins.These observations demonstrate that T cell response to Dsg3can 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 Dsg3 molecule may contain epitopes that are recognized by Dsg3-specific T cell.This study will allow us to further develop and characterize the Dsg3-specific T cell clone in Japanese PV patients.

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.