Maeba J, Lee S T, Paraskevas F
Department of Internal Medicine, University of Manitoba, Winnipeg, Canada.
J Immunol Methods. 1988 Jan 21;106(1):7-17. doi: 10.1016/0022-1759(88)90265-7.
A simple in vitro method is described for the induction of a potent mediator that interferes with suppressor cell function. The mediator consists of three easily identifiable components, Ig, class II determinants and antigen, that form a unique complex similar to, or identical with, the complexes detected in vivo within 3-6 h after immunization. The formation of the antisuppressor mediator in vitro takes place in two steps: the first involves a macrophage-T cell interaction which generates an 'intermediate complex' containing antigen and class II determinants. In the second step the addition of immunochemically purified IgG from normal mouse serum to the macrophage-T cell supernate generates potent antisuppressor activity, which is assayed by the conversion of suppression to immunity. It is suggested that the IgG interacts with the 'intermediate complex' giving rise to the final complex identical to that found in vivo 6 h after immunization. No activity is detected when IgG is added to a supernate of antigen-fed macrophages in the absence of T cells. Furthermore, the T cell plays an additional important role in the formation of the final complexes since it restricts the source of the IgG that will generate the antisuppressor activity. In other words the antisuppressor function is detected only if the IgG matches the donor of the T cell in the Igh locus. The T cell involved in the formation of the complex is the Ly1+ subpopulation. This method should allow elucidation of the genetic, cellular and molecular mechanisms in the activation of this important T cell pathway.
本文描述了一种简单的体外方法，用于诱导一种能干扰抑制性细胞功能的强效介质。该介质由三种易于识别的成分组成，即免疫球蛋白（Ig）、II类决定簇和抗原，它们形成一种独特的复合物，类似于或等同于免疫后3 - 6小时在体内检测到的复合物。体外抗抑制介质的形成分两步进行：第一步涉及巨噬细胞与T细胞的相互作用，产生一种包含抗原和II类决定簇的“中间复合物”。第二步，将来自正常小鼠血清的免疫化学纯化的IgG添加到巨噬细胞 - T细胞的上清液中，产生强效的抗抑制活性，通过将抑制转化为免疫来测定。有人认为，IgG与“中间复合物”相互作用，产生与免疫后6小时在体内发现的最终复合物相同的复合物。当IgG添加到无T细胞的抗原喂养巨噬细胞的上清液中时，未检测到活性。此外，T细胞在最终复合物的形成中还起着另外一个重要作用，因为它限制了将产生抗抑制活性的IgG的来源。换句话说，只有当IgG在免疫球蛋白重链（Igh）基因座上与T细胞的供体匹配时，才能检测到抗抑制功能。参与复合物形成的T细胞是Ly1 +亚群。这种方法应该能够阐明激活这一重要T细胞途径的遗传、细胞和分子机制。
