Simon M M, Hochgeschwender U, Brugger U, Landolfo S
J Immunol. 1986 Apr 15;136(8):2755-62.
In this study we tested the effect of monoclonal antibodies (moAb) AN-18 to murine IFN-gamma on the generation of cytolytic T cells (CTL) from a homogeneous population of precursor cells (CTL-P). As responder cells, highly purified Lyt-2+ C57BL/6 lymph node T cells were used that had been positively selected by flow cytofluorometry on a cell sorter. Lyt-2+ cells were set up in bulk culture or in limiting dilution (LD) either with Con A or with P815 tumor cells as antigen and recombinant human interleukin 2 (rec.hIL 2) in the presence or absence of moAb AN-18 and tested for growth and development of CTL. The results show that moAb AN-18 but not the unrelated moAb AN-37 diminished or abrogated proliferative and cytolytic responses of Lyt-2+ lymphocytes to lectin and rec.hIL 2 in a dose-dependent manner. The inhibitory activity of the antibodies could be abolished by neutralizing moAb AN-18 with recombinant murine IFN-gamma (rec.mIFN-gamma) before their addition to culture. Kinetic analysis shows that the inhibitory effect of moAb AN-18 is only optimal when added at the beginning of culture or up to 48 hr after initiation. The frequencies of CTL-P responding either to Con A or to P815 tumor cells and rec.hIL 2 were reduced up to 10-fold in the presence of moAb AN-18. The inhibitory capacity of moAb AN-18 was also operative in cultures containing on the average one antigen-specific CTL-P. Together with the finding that activated CTL-P secrete IFN-gamma in response to rec.hIL 2 in a dose-dependent manner, the data suggest that endogenous IFN-gamma collaborates with exogenous IL 2 in the induction of CTL-P. The generation of CTL may therefore represent a case of autocrine growth regulation of normal lymphocytes, in which the same cell synthesizes and responds to its own factor.
在本研究中，我们测试了抗小鼠γ干扰素单克隆抗体（moAb）AN-18对来自前体细胞（CTL-P）同质群体的细胞毒性T细胞（CTL）生成的影响。作为反应细胞，使用了通过流式细胞荧光分选法在细胞分选仪上进行阳性选择的高度纯化的Lyt-2 + C57BL / 6淋巴结T细胞。将Lyt-2 +细胞以批量培养或有限稀释（LD）的方式，与刀豆蛋白A或P815肿瘤细胞作为抗原以及重组人白细胞介素2（rec.hIL 2）一起培养，在有或没有moAb AN-18的情况下，测试CTL的生长和发育。结果表明，moAb AN-18而非无关的moAb AN-37以剂量依赖的方式减弱或消除了Lyt-2 +淋巴细胞对凝集素和rec.hIL 2的增殖和细胞溶解反应。在将抗体添加到培养物之前，用重组小鼠γ干扰素（rec.mIFN-γ）中和moAb AN-18可以消除抗体的抑制活性。动力学分析表明，moAb AN-18的抑制作用仅在培养开始时或开始后48小时内添加时才是最佳的。在存在moAb AN-18的情况下，对刀豆蛋白A或P815肿瘤细胞以及rec.hIL 2作出反应的CTL-P频率降低了多达10倍。moAb AN-18的抑制能力在平均含有一个抗原特异性CTL-P的培养物中也起作用。连同活化的CTL-P以剂量依赖的方式响应rec.hIL 2分泌γ干扰素这一发现，数据表明内源性γ干扰素与外源性IL 2在CTL-P的诱导中协同作用。因此，CTL的生成可能代表正常淋巴细胞自分泌生长调节的一个例子，其中同一细胞合成并对自身因子作出反应。
