Mima T, Ohshima S, Sasai M, Nishioka K, Shimizu M, Murata N, Yasunami R, Matsuno H, Suemura M, Kishimoto T, Saeki Y
Molecular Medicine, Osaka University Medical School, Osaka, 565, Japan.
Biochem Biophys Res Commun. 1999 Sep 16;263(1):172-80. doi: 10.1006/bbrc.1999.1128.
Previously, we demonstrated the presence of at least two distinct subpopulations of patients with rheumatoid arthritis (RA) employing a cell-transfer experiment using severe combined immunodeficient (SCID) mice. One group of patients, whose T cells derived from the rheumatoid joints, induced synovial hyperplasia (SH) in the SCID mice (the positive group). The other group did not display the induction of SH (the negative group). TCR/Vbeta gene usage analysis indicated that some dominant T cell subpopulations were oligoclonally expanding only in the rheumatoid joints, and not in the periphery of the patients of the positive group. Moreover, these T cell subpopulations were not seen in the joints of patients in the negative group or in non-RA patients. In addition, the preferential uses of certain TCR/Vbetas (Vbeta8, Vbeta12, Vbeta13, and Vbeta14) genes were demonstrated in these T cells. In this study, to investigate whether these T cells are driven by a certain antigen(s), the third complementarity determining regions (CDR3s) of TCR/Vbeta, especially Vbeta8 and Vbeta14 PCR products, were cloned and sequenced. As a result, a dominant CDR3 sequence, CASS-PRERAT-YEQ, was found in Vbeta14+ T cells from the rheumatoid joint of a patient (Patient 1) of the positive group with a Vbeta14 skew. The identical CDR3 sequence also predominated in Vbeta14+ T cells from the rheumatoid joint of another patient (Patient 7) of the positive group with a Vbeta14 skew. In addition, in the patients (Patients 4, 7, 8) of the positive group with a Vbeta8 skew, other dominant CDR3 sequences, CASS-ENS-YEQ and CASS-LTEP-DTQ, were found as in the case of Vbeta14. However, no identical CDR3 sequences were detected dominantly in the joints of the patients in the negative group or in non-RA patients. A Vbeta14+ T cell clone (TCL), named G3, with the identical CDR3 sequence, CASS-PRERAT-YEQ, was isolated successfully from Patient 1, and cell transfer of G3 with autologous irradiated peripheral mononuclear cells induced SH in the SCID mice. Taken together, these results suggest that T cells inducing SH, thought to be pathogenic for RA, might be driven by a certain shared antigen(s).
此前，我们通过使用严重联合免疫缺陷（SCID）小鼠进行细胞转移实验，证明类风湿关节炎（RA）患者中至少存在两个不同的亚群。一组患者，其来自类风湿关节的T细胞在SCID小鼠中诱导了滑膜增生（SH）（阳性组）。另一组则未表现出SH的诱导（阴性组）。TCR/Vβ基因使用分析表明，一些优势T细胞亚群仅在类风湿关节中呈寡克隆扩增，而在阳性组患者的外周血中未出现。此外，在阴性组患者的关节或非RA患者的关节中未发现这些T细胞亚群。另外，在这些T细胞中发现了某些TCR/Vβ（Vβ8、Vβ12、Vβ13和Vβ14）基因的优先使用情况。在本研究中，为了调查这些T细胞是否由某种抗原驱动，对TCR/Vβ的第三个互补决定区（CDR3），特别是Vβ8和Vβ14的PCR产物进行了克隆和测序。结果，在阳性组一名患者（患者1）的类风湿关节的Vβ14+ T细胞中发现了一个优势CDR3序列CASS-PRERAT-YEQ，该患者存在Vβ14偏斜。在另一名存在Vβ14偏斜的阳性组患者（患者7）的类风湿关节的Vβ14+ T细胞中，相同的CDR3序列也占主导地位。此外，在存在Vβ8偏斜的阳性组患者（患者4、7、8）中，与Vβ14情况相同，还发现了其他优势CDR3序列CASS-ENS-YEQ和CASS-LTEP-DTQ。然而，在阴性组患者的关节或非RA患者的关节中未检测到占主导地位的相同CDR
