Udono H, Kuribayashi K, Fujita T, Mieno M, Shiku H, Nakayama E
Department of Oncology, Nagasaki University School of Medicine, Japan.
Immunobiology. 1989 Feb;178(4-5):390-400. doi: 10.1016/S0171-2985(89)80061-0.
Proliferation of the cloned cytotoxic T lymphocyte (CTL) line 10B-5 induced by anticlonotypic antibody and its blocking by anti-Lyt-2 mAb were studied. Clone 10B-5 was derived from spleen cells of a (BALB/c x C57BL/6)F1, (CB6F1)-nu/+ mouse immunized with UV female 1 sarcoma. 10B-5 cells lysed UV female 1 specifically and proliferated upon stimulation with UV female 1 and feeder cells in the presence of IL 2. Anti-clonotypic monoclonal antibody (mAb) N1-56 was produced by a hybridoma established by fusion of spleen cells from a CB6F1-nu/+ mouse that had been immunized by five injections of 10B-5 cells prefixed with 0.1% formaldehyde. N1-56 mAb immunoprecipitated 90 kd molecules cleavable to 45 kd molecules under reducing conditions, indicating its reaction with T cell antigen receptor (TCR). N1-56 mAb in its soluble form induced a proliferative response of 10B-5 cells. Thus, the antigen binding site of N1-56 mAb appeared to substitute for the specific antigen determinant on UV female 1 sarcoma. The F(ab')2, but not the Fab fragment of purified N1-56 mAb, stimulated proliferation, indicating that cross-linking of TCR molecules was necessary for stimulation. The proliferative response of 10B-5 cells induced by soluble N1-56 mAb was blocked by addition of anti-Lyt-2.2 mAb to the cultures. The specificity of Lyt-2 blocking was confirmed by an absorption test. The proliferative response of 10B-5 cells induced by Con A, but not that induced by IL 2, was blocked by anti-Lyt-2.2 mAb. These results indicated that blocking by anti-Lyt-2.2 mAb was at an early stage and that it could be bypassed by stimulation with IL2.
研究了抗独特型抗体诱导克隆细胞毒性T淋巴细胞（CTL）系10B - 5的增殖及其被抗Lyt - 2单克隆抗体阻断的情况。克隆10B - 5源自用紫外线照射的雌性1号肉瘤免疫的（BALB/c×C57BL/6）F1、（CB6F1）-nu/+小鼠的脾细胞。10B - 5细胞能特异性裂解紫外线照射的雌性1号肉瘤细胞，并在白细胞介素2存在的情况下，经紫外线照射的雌性1号肉瘤细胞和饲养细胞刺激后增殖。抗独特型单克隆抗体（mAb）N1 - 56由一个杂交瘤产生，该杂交瘤是通过将经五次注射用0.1%甲醛预处理的10B - 5细胞免疫的CB6F1 - nu/+小鼠的脾细胞融合而建立的。N1 - 56 mAb在还原条件下免疫沉淀出可裂解为45 kd分子的90 kd分子，表明其与T细胞抗原受体（TCR）发生反应。可溶性形式的N1 - 56 mAb诱导了10B - 5细胞的增殖反应。因此，N1 - 56 mAb的抗原结合位点似乎替代了紫外线照射的雌性1号肉瘤上的特异性抗原决定簇。纯化的N1 - 56 mAb的F(ab')2片段而非Fab片段刺激了增殖，表明TCR分子的交联对于刺激是必要的。向培养物中添加抗Lyt - 2.2 mAb可阻断可溶性N1 - 56 mAb诱导的10B - 5细胞的增殖反应。通过吸收试验证实了Lyt - 2阻断的特异性。抗Lyt - 2.2 mAb可阻断由刀豆蛋白A诱导的10B - 5细胞的增殖反应，但不能阻断由白细胞介素2诱导的增殖反应。这些结果表明，抗Lyt - 2.2 mAb的阻断作用发生在早期阶段，并且可以通过白细胞介素2刺激来绕过。
