Kuribayashi K, Tanaka C, Matsubayashi Y, Masuda T, Udono H, Abe M, Nakayama E, Shiku H
Institute for Immunology, Faculty of Medicine, Kyoto University, Japan.
J Immunol. 1988 Dec 1;141(11):4074-80.
In this study, we first established several CTL clones of (BALB/c x C57BL/6)F1 origin that were specific for either syngeneic UV female 1 or UV male 1 fibrosarcoma cell lines. All the CTL clones had Thy-1+ Lyt-2+ L3T4- phenotypes and showed Kd restriction when lysing the corresponding target cells. Sera obtained from syngeneic animals immunized with three CTL clones, 10B-5 for UV female 1, and CTL9 and CTL10 for UV male 1, showed specific inhibition of target cell lysis with the corresponding CTL clones. The inhibitory activities were found in sera of the majority of immunized animals. Because the inhibitory activity resides in protein A-binding fraction, mAb were produced by hybridizing spleen cells of hyperimmune animals. N1-56 was thus obtained from a mouse immunized with 10B-5 CTL clone reactive with UV female 1. N1-56 was clonotype specific, reacting with 10B-5 but not with other CTL lines or leukemia cell lines. No N1-56+ cells were detectable in thymocytes, lymph node cells, or spleen cells of either naive or UV female 1-immune CB6F1 mice. Immunoprecipitation showed that N1-56 reacts with 90,000 Mr molecules on 10B-5 CTL clone under nonreducing conditions and 45,000 Mr molecules under reducing conditions, indicating its reactivities with idiotypic determinants of TCR on the CTL clone. N1-56 inhibited lytic activity of 10B-5, but neither N1-56 nor alpha-10B-5 hyperimmune serum inhibited that of alpha-UV female 1 mixed lymphocyte tumor cell culture cells. N1-56 induced proliferation of 10B-5 without addition of Ag.
在本研究中,我们首先建立了几个源自(BALB/c×C57BL/6)F1的CTL克隆,它们对同基因的紫外线照射的雌性1号或紫外线照射的雄性1号纤维肉瘤细胞系具有特异性。所有的CTL克隆都具有Thy-1⁺Lyt-2⁺L3T4⁻表型,并且在裂解相应靶细胞时表现出Kd限制。用三个CTL克隆免疫同基因动物获得的血清,即针对紫外线照射的雌性1号的10B-5以及针对紫外线照射的雄性1号的CTL9和CTL10,显示出对相应CTL克隆介导的靶细胞裂解的特异性抑制。在大多数免疫动物的血清中都发现了这种抑制活性。由于抑制活性存在于与蛋白A结合的部分,因此通过将超免疫动物的脾细胞杂交来制备单克隆抗体。用与紫外线照射的雌性1号反应的10B-5 CTL克隆免疫小鼠从而获得了N1-56。N1-56具有克隆型特异性,与10B-5反应,但不与其他CTL系或白血病细胞系反应。在未免疫的或紫外线照射的雌性1号免疫的CB6F1小鼠的胸腺细胞、淋巴结细胞或脾细胞中均未检测到N1-56⁺细胞。免疫沉淀显示,N1-56在非还原条件下与10B-5 CTL克隆上的90,000 Mr分子反应,在还原条件下与45,000 Mr分子反应,表明它与CTL克隆上TCR的独特型决定簇反应。N1-56抑制10B-5的裂解活性,但N1-56和α-10B-5超免疫血清均不抑制α-紫外线照射的雌性1号混合淋巴细胞肿瘤细胞培养细胞的裂解活性。在不添加抗原的情况下,N1-56诱导10B-5增殖。