| Research Abstract |
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.
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