Development by genetic engineering of T cell vaccination protein which suppresses autoimmune arthritis and the analysis of its mechanism

抑制自身免疫性关节炎的T细胞疫苗蛋白的基因工程开发及其机制分析

• 批准号: 03670253
• 负责人: KAKIMOTO Kiichi
• 金额: $ 0.38万
• 依托单位: KUMAMOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670253/
• 关键词: Collagen-induced arthritis; T cell clone; T cell vaccination; T cell receptor; Monoclonal antibody; Transgenic mice; コラ-ゲン誘導関節炎; T細胞クロ-ン; T細胞抗原受容体(TCR)V_β; 抗V_βモノクロ-ナル抗体; TCRトランスジェニックマウス

We carried out cloning of T cell receptor(TCR) gene of human type II collagen(CII)-specific T cell clone(K-102), since the activity of T cell vaccination was dependent on its TCR. The results showed that K-102 cells carried alphabeta type TCR which was composed of Valpha8.2Jalpha37 and Vbeta12Dbeta1.1Jbeta1.1Cbeta1. We tried to prepare monoclonal antibody(MoA) against Vbeta12 in order to check the expression of Vbeta12 on the recipient T cells transfected with Vbeta12 inserted into an appropriate vector. Mice were immunized with BL-17 cells, T cell hybridoma of K-102 cells in an attempt to prepare anti Vbeta12 MoAb resulting in failing in it. However, we could obtain anti-Vbeta12 MoAb (MR11-1) from Dr. Kanagawa of Washington University, USA in the cooperative study with him who succeeded in preparation of the MoAb. MR11-1 reacted not only with K-102 cells, but also showed suppressive effect on in vitro antigen-incluced proliferative response of K-102 cells. Besides, in vivo administration of MR11-1 suppressed the development of passively-incluced arthritis by the transfer of K-102 cells and active collagen-induced arthritis (CA). These results suggest that pathogenic T cells involved in CA predominantly use Vbeta12 as their TCR. On the other hand, in order to study whether the transfection of Vbeta12 gene can reconstruct functional TCR which retains T cell vaccination activity, we prepared Vbeta12 transgenic mice in SWR mice which has the same H-2^q haplotype as CA-prone DBA/1 mice but are CA-resistant due to genetic deficiency of many Vbeta genes including Vbeta12. The study by the use of this transgenic mice suggested the critical role of Vbeta12 in CA but showed requirement of the coexistence of other factors possibly including Valpha gene product. Since the basis for the study was now prepared, the work for the development of T cell vaccination protein is currently under way.

由于T细胞免疫的活性依赖于其T细胞受体(TCR)，因此我们克隆了人II型胶原(CII)特异性T细胞克隆(K-102)的T细胞受体(TCR)基因。结果表明，K-102细胞携带Alphabeta型TCR，由Valpha8.2Jalpha37和Vbeta12Dbeta1.1Jbeta1.1Cbeta1组成。我们试图制备抗Vbeta12的单抗，以检测Vbeta12在插入适当载体的受体T细胞上的表达。用BL-17细胞、K-102细胞的T细胞杂交瘤免疫小鼠，试图制备抗Vbeta12单抗，但失败。然而，我们可以从美国华盛顿大学的神奈川博士那里获得抗Vbeta12 Moab(MR11-1)，与成功制备Moab的博士合作研究。MR11-1不仅与K-102细胞发生反应，而且对K-102细胞的体外抗原诱导的增殖反应有抑制作用。此外，体内应用MR11-1通过转移K-102细胞和活动性胶原诱导性关节炎(CA)来抑制被动包涵性关节炎的发展。这些结果表明，CA的致病T细胞主要以Vbeta12为TCR。另一方面，为了研究转导Vbeta12基因能否重建具有T细胞免疫活性的功能性TCR，我们在SWR小鼠中制备了Vbeta12转基因小鼠，该小鼠与CA倾向的DBA/1小鼠具有相同的H-2^Q单倍型，但由于包括Vbeta12在内的许多Vbeta基因的遗传缺陷而对CA具有抗性。利用该转基因小鼠的研究表明，Vbeta12在CA中起着关键作用，但也需要其他因素的共存，可能包括Valpha基因产物。由于研究的基础现在已经准备好，开发T细胞疫苗蛋白的工作目前正在进行中。

项目成果

Mori.L.: "Expression of a transgenic T cell receptor β chain enhances collagen-induced arthritis." J.Exp.Med.176. 381-388 (1992)

Mori.L.：“转基因 T 细胞受体 β 链的表达可增强胶原诱导的关节炎。”J.Exp.Med.176（1992）。

Kakimoto,K.: "The effect of antiーaclkesion molecule antibody on the development of collagenーinduced arthritis" Cellular Immunology. (1922)

Kakimoto, K.：“抗关节分子抗体对胶原诱导的关节炎发展的影响”细胞免疫学（1922）。