DeKruyff R, Clayberger C, Cantor H, Dorf M E
J Immunol. 1985 Oct;135(4):2243-8.
We have described a trinitrophenyl (TNP)-specific inducer clone, clone Ly-1-T1, which responds to a variety of different stimuli, including a) soluble TNP-protein conjugates plus syngeneic (H-2d) spleen cells, b) TNP directly coupled to syngeneic or allogeneic spleen cells, and c) activated I-A identical B cells in the absence of nominal antigen. In the present study we used a panel of antibodies to investigate the recognition structures involved in the activation of clone Ly-1-T1 by these different stimuli. We show that allogeneic spleen cells must be conjugated by using relatively high concentrations of TNBS to be efficient stimulators of the clone. In contrast, syngeneic spleen cells conjugated by using a much wider range of concentrations will activate the clone. The response of the clone to TNP-coupled allogeneic spleen cells is inhibited by anti-L3T4 and anti-Ia antibodies. In contrast, stimulation of the clone with syngeneic spleen cells coupled by using the same concentrations of TNBS is not inhibited with either anti-Ia or anti-L3T4 antibody. The inhibition pattern observed with anti-Ia and anti-L3T4 antibodies was also determined by the nature of the accessory population used to present soluble TNP-protein conjugates. Anti-I-Ad antibodies blocked the activation of clone Ly-1-T1 by TNP-protein plus splenic adherent cells, indicating the involvement of polymorphic I-A determinants in this response. Anti-L3T4 antibody had little or no effect on this response, suggesting that a significant L3T4-Ia interaction is not required. Finally, the response of the clone to activated B cells in the presence or absence of TNP-protein is exquisitely sensitive to inhibition by anti-L3T4 as well as anti-I-A antibodies. The data suggest that the requirement for an L3T4-I interaction depends on the combination of antigen and accessory cell type used to stimulate the clone.
我们已描述了一种三硝基苯基(TNP)特异性诱导克隆,即克隆Ly-1-T1,它能对多种不同刺激作出反应,包括:a)可溶性TNP-蛋白质偶联物加同基因(H-2d)脾细胞;b)直接偶联到同基因或异基因脾细胞上的TNP;以及c)在无名义抗原情况下被激活的I-A相同的B细胞。在本研究中,我们使用一组抗体来研究这些不同刺激激活克隆Ly-1-T1所涉及的识别结构。我们发现,异基因脾细胞必须通过使用相对高浓度的TNBS进行偶联,才能成为该克隆的有效刺激物。相比之下,使用更广泛浓度范围进行偶联的同基因脾细胞会激活该克隆。克隆对TNP偶联的异基因脾细胞的反应受到抗L3T4和抗Ia抗体的抑制。相比之下,用相同浓度的TNBS偶联的同基因脾细胞刺激该克隆时,抗Ia或抗L3T4抗体均不会产生抑制作用。抗Ia和抗L3T4抗体所观察到的抑制模式也取决于用于呈递可溶性TNP-蛋白质偶联物的辅助群体的性质。抗I-Ad抗体阻断了TNP-蛋白质加脾黏附细胞对克隆Ly-1-T1的激活,表明多态性I-A决定簇参与了这一反应。抗L3T4抗体对该反应几乎没有影响,这表明不需要显著的L3T4-Ia相互作用。最后,无论有无TNP-蛋白质,克隆对激活的B细胞的反应都对抗L3T4以及抗I-A抗体的抑制极为敏感。数据表明,对L3T4-I相互作用的需求取决于用于刺激克隆的抗原和辅助细胞类型的组合。