Kuchroo V K, Billings P R, Levine C, Martin C A, Kubo R T, Dorf M E
Department of Pathology, Harvard Medical School, Boston, MA 02115.
J Immunol. 1990 Aug 15;145(4):1059-65.
Eight different Ts cell hybridomas (including inducer (Ts1) and effector (Ts3) suppressor cells) specific for the 4-hydroxy-3-nitrophenyl acetyl (NP) hapten were tested for their ability to respond to Ag or anti-CD3 antibody in a growth-inhibition assay. Results suggest that the expression of the TCR-CD3 complex on Ts hybridomas is required for the Ag or anti-CD3-mediated growth inhibition. One of the CD3+, Ts hybridomas (CKB-Ts3-9.H3) was tested in detail; this CD4- effector suppressor cell hybridoma showed specific inhibition of growth in the presence of NP or NIP-coupled protein conjugates but not in the presence of other irrelevant hapten-protein conjugates. In addition, growth of this hybridoma was specifically inhibited by anti-CD3 and anti-TCR-alpha/beta antibodies but not by control hamster antibodies. In order to study the role of MHC molecules in Ag-mediated growth inhibition, Ts cell hybridomas were incubated with Ag (NP-keyhole limpet hemocyanin) in the presence of spleen cells from various H-2 congenic strains. The results suggest that the Ts hybridomas that express donor Ts-derived TCR beta-chain recognize Ag in an MHC-restricted manner, whereas the two Ts3 hybridomas that utilize BW5147-derived TCR-beta recognize Ag in H-2 unrestricted way. Co-incubation of anti-CD3 and anti-TCR-alpha/beta antibodies with specific Ag enhanced the Ag-mediated growth inhibition, whereas anti-LFA-1 antibody completely blocked the Ag-mediated effect. The combined data suggest that, like Th hybridomas, expression of CD3-associated-TCR complex is essential for the Ag responsiveness of Ts cell hybridomas.
针对4-羟基-3-硝基苯乙酰(NP)半抗原的8种不同的Ts细胞杂交瘤(包括诱导型(Ts1)和效应型(Ts3)抑制细胞)在生长抑制试验中检测了它们对抗原或抗CD3抗体的反应能力。结果表明,Ts杂交瘤上TCR-CD3复合物的表达是抗原或抗CD3介导的生长抑制所必需的。对其中一种CD3+ Ts杂交瘤(CKB-Ts3-9.H3)进行了详细检测;这种CD4-效应抑制细胞杂交瘤在存在NP或NIP偶联蛋白缀合物时显示出特异性的生长抑制,但在存在其他无关半抗原-蛋白缀合物时则没有。此外,这种杂交瘤的生长受到抗CD3和抗TCR-α/β抗体的特异性抑制,但不受对照仓鼠抗体的抑制。为了研究MHC分子在抗原介导的生长抑制中的作用,将Ts细胞杂交瘤与抗原(NP-钥孔戚血蓝蛋白)在来自各种H-2同基因品系的脾细胞存在下孵育。结果表明,表达供体Ts来源的TCR β链的Ts杂交瘤以MHC限制的方式识别抗原,而利用BW5147来源的TCR-β的两种Ts3杂交瘤以H-2非限制的方式识别抗原。抗CD3和抗TCR-α/β抗体与特异性抗原共同孵育增强了抗原介导的生长抑制,而抗LFA-1抗体完全阻断了抗原介导的效应。综合数据表明,与Th杂交瘤一样,CD3相关TCR复合物的表达对于Ts细胞杂交瘤的抗原反应性至关重要。