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

我们克隆了人II型胶原蛋白（CII）特异性T细胞克隆（K-102）的T细胞受体（TCR）基因，因为T细胞接种的活性依赖于其TCR。结果表明，K-102细胞携带由vpha8.2 jalpha37和vbeta12dbeta1.1 jbeta1.1 1cbeta1组成的α β型TCR。我们试图制备针对Vbeta12的单克隆抗体（MoA），以检测Vbeta12在转染到合适载体上的受体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 MoAb，结果失败。然而，在与美国华盛顿大学Kanagawa博士的合作研究中，我们获得了抗vbeta12 MoAb （MR11-1）。MR11-1不仅与K-102细胞发生反应，而且对K-102细胞的体外抗原包括增殖反应有抑制作用。此外，体内给药MR11-1通过K-102细胞的转移和活性胶原诱导的关节炎（CA）抑制被动包括关节炎的发展。这些结果表明，参与CA的致病性T细胞主要使用Vbeta12作为其TCR。另一方面，为了研究转染Vbeta12基因是否能重建保留T细胞接种活性的功能性TCR，我们在具有与ca易感性DBA/1小鼠相同的H-2^q单倍型的SWR小鼠中制备了Vbeta12转基因小鼠，但由于包括Vbeta12在内的许多Vbeta基因的遗传缺陷而产生了ca抗性。利用该转基因小鼠的研究表明，Vbeta12在CA中起关键作用，但需要其他因素的共存，可能包括Valpha基因产物。由于这项研究的基础已经准备好，开发T细胞疫苗蛋白的工作目前正在进行中。

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.