T cell receptors recognizing type II collagen in HLA-DR-transgenic mice characterized by highly restricted Vβ usage
T cell receptors recognizing type II collagen in HLA-DR-transgenic mice characterized by highly restricted Vβ usage
复制标题
DOI：
10.1002/art.20289
复制
发表时间：
2004-06-01
影响因子：
--
通讯作者：
Stuart, JM
中科院分区：
文献类型：
--
作者：
He, XW;Rosloniec, EF;Stuart, JM
客观的。确定 HLA-DR 转基因小鼠中识别 II 型胶原 (CII) 的 T 细胞受体 (TCR) 结构，并检查具有某些 V-β 链的 T 细胞在胶原诱导的关节炎 (CIA) 中的作用。方法。 T 细胞杂交瘤是从 DR1 和 DR4 转基因小鼠中建立的，并根据它们对 CII 和含有 T 细胞决定簇的 CII 肽的反应进行选择。提取 RNA 并反转录成互补 DNA，然后使用适当的 V-β 和 V-α 亚家族特异性引物进行扩增。纯化聚合酶链式反应产物并直接测序。为了确定具有某些 V-β-链的 T 细胞在 CIA 中的作用，施用了 V-β-亚家族特异性抗体并确定了关节炎的发展和特征。结果。分析了 23 个克隆不同的 T 细胞杂交瘤的 TCR，这些杂交瘤源自 DR1 转基因小鼠，并且对含有免疫显性决定簇的 CII 肽有反应。这些杂交瘤主要使用 TCR V(β)14 和 V(β)8 基因片段（分别为 70% 和 30%）。在来自 DR4 转基因小鼠的 CII 反应性 T 细胞杂交瘤中也发现了 V-β 使用的相同限制。 V-α 基因的使用也受到限制，尽管这一点不如 V-β 基因明显。相反，杂交瘤表达了不同的第三互补决定区。删除携带 V(β)14 和携带 V(β)8 的 T 细胞可显着降低 CIA 的发生率和严重程度。结论。这些数据表明，DR1 和 DR4 不仅结合并呈递相同的 CII 免疫显性肽，而且还刺激高度受限的 T 细胞亚群。
Objective. To determine the T cell receptor (TCR) structure recognizing type II collagen (CII) in HLA-DR-transgenic mice, and to examine the role of T cells with certain V-beta-chains in collagen-induced arthritis (CIA).Methods. T cell hybridomas were established from DR1- and DR4-transgenic mice and selected for their responses to CII and CII peptide containing the T cell determinants. RNA was extracted and reverse transcribed into complementary DNA, which was then amplified using appropriate V-beta- and V-alpha-subfamily-specific primers. The polymerase chain reaction products were purified and directly sequenced. To determine the role of T cells with certain V-beta-chains in CIA, V-beta-subfamily-specific antibodies were administered and the development and characteristics of arthritis were determined.Results. TCRs of 23 clonally distinct T cell hybridomas that were derived from DR1-transgenic mice and that were reactive to the CII peptide containing the immunodominant determinant were analyzed. These hybridomas predominantly used the TCR V(beta)14 and V(beta)8 gene segments (70% and 30%, respectively). The same restriction in V-beta usage was also found in CII-reactive T cell hybridomas from DR4-transgenic mice. There was also restricted use of V-alpha genes, although this was less marked than that of V-beta. In contrast, the hybridomas expressed a diverse third complementarity-determining region. Deletion of both V(beta)14-bearing and V(beta)8-bearing T cells significantly reduced the incidence and severity of CIA.Conclusion. These data demonstrate that DR1 and DR4 not only bind and present the same CII immunodominant peptide, but also stimulate a highly restricted subset of T cells.