Ting C C, Hargrove M E
Division of Cancer Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Cell Immunol. 1992 Mar;140(1):130-44. doi: 10.1016/0008-8749(92)90182-o.
alpha CD3 induced the generation of activated killer cells from resting T cells. Pretreatment of the splenic responders with PMA, a phorbol ester, depleted protein kinase C and induced unresponsiveness to the generation of alpha CD3-induced activated killer (CD3-AK) cells. Addition of exogenous IL-4 (1 U/ml) restored the cytotoxic response, with the maximal effect achieved with 30 to 100 U/ml. The phenotypes of CD3-AK cells maintained in IL-2 or in IL-4, with or without PMA, were the same: Thy1+ and CD8+. These results were reproduced with purified T cells and purified CD8+ cells, indicating that both the effectors and precursors were CD8+ cells and IL-4 had a selective effect to upregulate the CD8+ cells. Similar results were obtained by using SSP (staurosporine), another PKC inhibitor. At 2 days prior to testing, switching the lymphokine added to 2-week PMA- and IL-2-maintained CD3-AK cells reversed their cytolytic activity: switching from IL-2 to IL-4 restored cytolytic activity, and switching from IL-4 to IL-2 reduced cytolytic activity. The cytolytic activity of these CD3-AK cells correlated with their ability to produce BLT-esterase. In the absence of PMA, CD3-AK cells cultured in either IL-2 or IL-4 were cytolytic and contained high levels of BLT-esterase. In contrast, in the presence of PMA, only the IL-4-maintained CD3-AK cells were cytolytic and produced significant amounts of BLT-esterase. The effect of IL-4 was abrogated by the alpha IL-4 antibody 11B11, which reduced the cytolytic activity of CD3-AK and the ability to produce BLT-esterase. The requirement of IL-2 was less stringent and its major role appeared to be maintaining the cell growth. These findings indicate that IL-4 may participate in the regulation of a PKC-independent pathway for the generation of CD3-AK cells by regulating the production of cytolytic granules.
α-CD3可诱导静息T细胞产生活化杀伤细胞。用佛波酯PMA预处理脾反应细胞，可耗尽蛋白激酶C并诱导其对α-CD3诱导的活化杀伤（CD3-AK）细胞的产生无反应。添加外源性白细胞介素-4（1 U/ml）可恢复细胞毒性反应，30至100 U/ml时达到最大效果。在有或无PMA的情况下，在白细胞介素-2或白细胞介素-4中维持的CD3-AK细胞的表型相同：Thy1+和CD8+。用纯化的T细胞和纯化的CD8+细胞重复了这些结果，表明效应细胞和前体细胞均为CD8+细胞，且白细胞介素-4对上调CD8+细胞有选择性作用。使用另一种蛋白激酶C抑制剂星孢菌素（SSP）也得到了类似结果。在测试前2天，将添加到用PMA和白细胞介素-2维持2周的CD3-AK细胞中的淋巴因子进行切换，可逆转其溶细胞活性：从白细胞介素-2切换到白细胞介素-4可恢复溶细胞活性，从白细胞介素-4切换到白细胞介素-2则降低溶细胞活性。这些CD3-AK细胞的溶细胞活性与其产生BLT酯酶的能力相关。在无PMA的情况下，在白细胞介素-2或白细胞介素-4中培养的CD3-AK细胞具有溶细胞性且含有高水平的BLT酯酶。相反，在有PMA的情况下，只有在白细胞介素-4中维持的CD3-AK细胞具有溶细胞性并产生大量的BLT酯酶。α-白细胞介素-4抗体11B11可消除白细胞介素-4的作用，该抗体可降低CD3-AK的溶细胞活性和产生BLT酯酶的能力。对白细胞介素-2的需求不那么严格，其主要作用似乎是维持细胞生长。这些发现表明，白细胞介素-4可能通过调节溶细胞颗粒的产生参与CD3-AK细胞产生的蛋白激酶C非依赖性途径的调节。
