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

日本寻常型天疱疮患者 Desmoglein-3 特异性 T 细胞的发育和表征

基本信息

批准号：
10670804
负责人：
SAKURAI Toshiharu
金额：
$ 2.43万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670804/
关键词：
Pemphigus vulgaris (PV)T lymphocyte Desmoglein 3 (Dsg 3)Human Leukocyte Antigen (HLA)

项目摘要

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.

本研究的目的是调查日本寻常型天疱疮 (PV) 患者的 T 淋巴细胞对 PV 自身抗原 Desmoglein 3 的反应，并表征 Dsg 3 特异性 T 细胞的特性。1．我们使用 PCR-RFLP（聚合酶链反应-限制性片段长度多态性）方法对 Amagai 博士介绍的 14 名日本 PV 患者进行了高分辨率 HLA II 类分型。与健康个体相比，DRB 1*14 (14/14:100%)、DQB1*0503 (12/14:85%) 或 DQB1*0301(9/12:64%) 存在显着相关性。 2. 制备 11 个带有麦芽糖结合蛋白 (MBP) 的重组 Dsg 3 融合蛋白 (Dsg 3. 1-11)，以涵盖每 40 个氨基酸，从 Dsg 33 的 EC 1 开始，每 40 个氨基酸错开 15 个残基。通过将 PBMC 与 11 个重组蛋白共培养，在 14 名 PV 患者和 20 名健康对照中研究了对 Dsg 3 的自身反应性 T 细胞反应。 Dsg 3 融合蛋白和[ィイD13ィエD1H]-胸苷的掺入。在表达 PV 相关 HLA-DRB1*14 和 DQB1*05 的 11/14 PV 患者和 7/14 健康个体中观察到对 Dsg 3.9（残基 201-204）或 Dsg 3.10（残基 226-265）的初级体外 T 细胞反应（SI=1.9-2.7）。相比之下，来自携带除DRB1*14和DQB1*05之外的HLA II类等位基因的6个正常对照的PBMC未受到重组Dsg 3融合蛋白的刺激。这些观察结果表明，在携带与PV中高度流行的主要组织相容性复合体II类等位基因相同或相似的PV患者和健康供体中可以检测到对Dsg 3的T细胞反应。这表明 Dsg 3 分子的 EC2-3 区域可能包含被 Dsg 3 特异性 T 细胞识别的表位。这项研究将使我们能够进一步开发和表征日本真性红细胞增多症患者中的 Dsg 3 特异性 T 细胞克隆。