Kane K P, Mescher M F
Division of Membrane Biology, Medical Biology Institute, La Jolla, CA 92037.
J Immunol. 1990 Feb 1;144(3):824-9.
Although engagement of the TCR via antibody can be sufficient to trigger T cells, responses to Ag-bearing cells require additional "accessory" interactions in many cases. A method has been developed which allows preparation of surfaces bearing both purified class I alloantigen and coimmobilized antibodies. With this approach, it is possible to mimic such "accessory" interactions and to examine their quantitative effects on triggering via TCR-Ag interaction. Experiments are described which use this approach to examine triggering of degranulation by cloned, allogeneic CTL lines. Coimmobilization of antibodies specific for any of a variety of CTL surface proteins, including CD8, class I MHC proteins, CD45 (T200) and Thy-1, had the effect of decreasing the critical threshold density of Ag necessary to trigger responses, and decreasing by an order of magnitude the density required to stimulate a half-maximal response. Furthermore, in comparison with the 30-min lag seen with Ag alone, response was initiated immediately when an antibody specific for a CTL surface component was present. These results are consistent with the hypothesis that any CTL surface molecule having sufficient affinity for a component of the target surface can contribute to activation via the Ag-specific TCR; and at low Ag density could determine whether any response occurs.
尽管通过抗体与TCR结合足以触发T细胞,但在许多情况下,对携带抗原的细胞的反应还需要额外的“辅助”相互作用。已经开发出一种方法,可制备同时带有纯化的I类同种异体抗原和共固定抗体的表面。通过这种方法,可以模拟这种“辅助”相互作用,并研究它们对通过TCR-抗原相互作用触发的定量影响。本文描述了使用这种方法来检测克隆的同种异体CTL系脱颗粒触发情况的实验。共固定针对多种CTL表面蛋白(包括CD8、I类MHC蛋白、CD45(T200)和Thy-1)中任何一种的抗体,具有降低触发反应所需的抗原临界阈值密度的作用,并使刺激半最大反应所需的密度降低一个数量级。此外,与单独使用抗原时观察到的30分钟延迟相比,当存在针对CTL表面成分的抗体时,反应立即启动。这些结果与以下假设一致:任何对靶表面成分具有足够亲和力的CTL表面分子都可以通过抗原特异性TCR促进激活;并且在低抗原密度下可以决定是否发生任何反应。