Swierkosz J E, Rock K, Marrack P, Kappler J W
J Exp Med. 1978 Feb 1;147(2):554-70. doi: 10.1084/jem.147.2.554.
A method was established for isolating antigen-specific murine helper T cells by selective binding to antigen-pulsed macrophage (Mphi) monolayers. Sheep erythrocyte (SRBC)-primed T cells, which remained strongly adherent to SRBC-pulsed syngeneic Mphi after 20 h in culture, were markedly enriched for helper activity when tested in the in vitro antitrinitrophenol (TNP) response to TNP-SRBC. Successful binding and enrichment occurred only if the Mphi were pulsed with the specific antigen to which the T-cell donors had been primed. The genetic control governing helper function in this system was then examined by using primed F1 T cells isolated on Mphi monolayers from congenic strains bearing parental H-2 haplotypes. SRBC-primed BDF1 (H-2b X H-2d) T cells, which bound to SRBC-pulsed H-2d Mphi, subsequently functioned as helper cells in cultures containing H-2d B cells and Mphi, but not in those containing H-2b B cells and Mphi. They remained unable to collaborate with B cells of the H-2B haplotype even in the presence of additional H-2d Mphi, indicating that H-2 restriction occurs at least at the level of the B cell. Similary, primed BDF1 T cells isolated on H-2b Mphi cooperated preferentially with H-2b B cells and Mphi. In both cases, the haplotype preference of the T cell was not due to alloreactive suppressor activity. These results suggest that primed F1 mice contain individual populations of helper T cells, each of which recognize antigen in association with a parental H-2 gene product(s) expressed during both Mphi-T cell and T cell-B cell interactions.
建立了一种通过与抗原脉冲巨噬细胞(Mphi)单层选择性结合来分离抗原特异性小鼠辅助性T细胞的方法。用绵羊红细胞(SRBC)致敏的T细胞,在培养20小时后仍强烈粘附于SRBC脉冲的同基因Mphi,当在体外对三硝基苯酚(TNP)-SRBC的抗TNP反应中进行测试时,其辅助活性显著富集。只有当Mphi用T细胞供体已致敏的特异性抗原进行脉冲时,才会成功发生结合和富集。然后,通过使用从携带亲本H-2单倍型的同类系菌株的Mphi单层上分离的致敏F1 T细胞,研究了该系统中控制辅助功能的遗传控制。用SRBC致敏的BDF1(H-2b×H-2d)T细胞,与SRBC脉冲的H-2d Mphi结合,随后在含有H-2d B细胞和Mphi的培养物中作为辅助细胞发挥作用,但在含有H-2b B细胞和Mphi的培养物中则不然。即使存在额外的H-2d Mphi,它们仍无法与H-2B单倍型的B细胞协作,这表明H-2限制至少发生在B细胞水平。同样,在H-2b Mphi上分离的致敏BDF1 T细胞优先与H-2b B细胞和Mphi协作。在这两种情况下,T细胞的单倍型偏好并非由于同种异体反应性抑制活性。这些结果表明,致敏的F1小鼠含有单个辅助性T细胞群体,每个群体在Mphi-T细胞和T细胞-B细胞相互作用过程中都能识别与亲本H-2基因产物相关的抗原。