Role of self-Ia antigen in the recognition of self and not self by T cells: Study by use of cloned T cell lines.

自身 Ia 抗原在 T 细胞识别自身和非自身中的作用：使用克隆 T 细胞系进行研究。

基本信息

批准号：
60480179
负责人：
SAITO Kazuhisa
金额：
$ 2.82万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

项目摘要

1. Even by use of I-A^KK gene-transfected L cells as antigen presenting cells (APC), azobenzenearso- nate-tyrosine (ABA-tyr) specific T cell clones or T hybridomas (I-A^kk restricted) showed increase in DNA synthesis or IL 2 production in respones to ABA-tyr, and these responses were blocked by anti-I-A^KK antibody. 2. To study directly antigen recognition of T cells, we examined changes in Ca^<2+>2 influx and membrane fluidity of T cells after antigen recognition by stopped flow method using fluorescent probes. In the presence of I-A^KK gene-transfected L cells or liposomes conjugated with Ia^KK molecules, ABA-tyr specific T cells showed marked increase in Ca^<2+>2 influx and membrane fluidity within 2 seconds after stimulation with ABA-tyr, and these responses were blocked by anti-I-A^KK antibody. 3. To clarify whether ABA-tyr should bind to Ia molecules before being recognized by T cells, we compared the rate constants of Ca^<2+>2 influx when the order of mixing of T cells, ABA-tyr and APC was changed. The rate constant was almost the same even by change in the order of mixing. These results support the trimolecular complex model of antigen recognition by T cells. 4. Among auto-Ia-reactive T cell clones, noe clone elicited lichen planus-like lesion in the skin of syngeneic mice. Another clone injected i.v. into syngeneic mice caused production of anti-DNA antibody and auto-antibody to erythrocytes. We ate exploring the causes of autoimmune diseases by use of these auto-Ia-reactive T cell clones.

1. 即使使用 I-A^KK 基因转染的 L 细胞作为抗原呈递细胞 (APC)，偶氮苯胂-酪氨酸 (ABA-tyr) 特异性 T 细胞克隆或 T 杂交瘤（I-A^kk 限制性）也显示出 DNA 合成的增加或 IL 2 的产生响应 ABA-tyr，并且这些响应被抗 I-A^KK 抗体阻断。 2.为了直接研究T细胞的抗原识别，我们使用荧光探针通过停流法检查了抗原识别后T细胞的Ca^2+2流入和膜流动性的变化。在存在I-A^KK基因转染的L细胞或与Ia^KK分子缀合的脂质体的情况下，ABA-tyr特异性T细胞在用ABA-刺激后2秒内表现出Ca^2+2流入和膜流动性的显着增加。 tyr，并且这些反应被抗 I-A^KK 抗体阻断。 3.为了阐明ABA-tyr在被T细胞识别之前是否应该与Ia分子结合，我们比较了当T细胞、ABA-tyr和APC的混合顺序改变时Ca^<2+>2流入的速率常数。即使改变混合顺序，速率常数也几乎相同。这些结果支持 T 细胞识别抗原的三分子复合物模型。 4.在自身Ia反应性T细胞克隆中，noe克隆在同基因小鼠的皮肤中引起扁平苔藓样病变。另一个克隆体静脉注射进入同系小鼠体内引起抗DNA抗体和红细胞自身抗体的产生。我们利用这些自身 Ia 反应性 T 细胞克隆来探索自身免疫性疾病的原因。