Lymphocyte-mediated cytotoxicity against human allogeneic and autologous lymphoid targets after concanavalin A-activation of cytotoxic effector cells.
Lymphocyte-mediated cytotoxicity against human allogeneic and autologous lymphoid targets after concanavalin A-activation of cytotoxic effector cells.
复制标题
刀豆球蛋白 A 激活细胞毒性效应细胞后，淋巴细胞介导的针对人同种异体和自体淋巴靶标的细胞毒性。
作者：
G. Hunninghake;A. Fauci
目前的研究表明，悬浮液的人外周血淋巴细胞可以被激活的刀豆蛋白A成为细胞毒性效应细胞显示针对广泛的同种异体抗原轴承淋巴靶点的特异性。反应的免疫学性质通过以下事实证明：除了在细胞毒性试验期间存在凝集素（PHA）外，自体靶标不裂解。某些培养条件对于最佳反应至关重要，包括细胞密度、培养基中2-巯基乙醇的存在以及某些批次的人AB血清，这些血清强烈支持细胞毒性淋巴细胞的产生。细胞毒性细胞仅在培养2至3天后产生，并且在培养24小时或4天后不存在。培养基中Con A的最佳浓度为10 - 20 µg/ml。所用的靶细胞是在无刺激的情况下培养2至3天的人淋巴细胞。细胞毒性试验是一项4小时试验，需要相对较高的效应细胞与靶细胞比率（250至500：1）才能获得对同种异体淋巴靶细胞的最大杀伤作用，而较低的比率足以对抗非淋巴靶细胞，如Chang肝细胞。Con A产生的淋巴细胞从特定的个人是一致的能力，安装一个高或低的细胞毒性反应对同种异体的目标。这与HL-A单倍型与靶细胞的相似性或不相似性程度无关。此外，在Con A产生的细胞毒性试验中为低应答者的个体的MLC胚细胞生成应答和MLC产生的同种异体抗原特异性细胞毒性应答与Con A产生的细胞毒性试验中为高应答者的个体一样高。此外，高应答者和低应答者的淋巴细胞对Con A表现出相似程度的胚细胞生成应答。本研究提供了一个简单的和可重复的系统，从人外周血细胞毒性效应细胞的产生和测定后，多克隆活化的刀豆蛋白A，并提供了一个模型，可用于研究的机制和调节因素参与激活的人细胞毒性淋巴细胞。
The present study has demonstrated that suspensions of human peripheral blood lymphocytes can be activated by Con A to become cytotoxic effector cells displaying specificity against a broad range of alloantigen-bearing lymphoid targets. The immunologic nature of the reaction was demonstrated by the fact that autologous targets were not lysed except when an agglutinin (PHA) was present during the cytotoxicity assay. Certain culture conditions were critical for optimal responses including the density of cells, the presence of 2-mercaptoethanol in the culture media, and certain batches of human AB serum which strongly supported the generation of cytotoxic lymphocytes. Cytotoxic cells were generated only after 2 to 3 days of culture and were not present at 24 hr or after 4 days of culture. The optimal concentration of Con A in the culture media was 10 to 20 µg/ml. The target cells employed were human lymphocytes maintained in culture without stimulation for 2 to 3 days. The cytotoxicity assay was a 4-hr assay and relatively high effector-to-target cell ratios (250 to 500:1) were necessary to obtain maximal killing against allogeneic lymphoid targets, whereas lower ratios were adequate against nonlymphoid targets such as Chang liver cells. Con A-generated lymphocytes from particular individuals were consistent in their capacity to mount a high or low cytotoxic response against allogeneic targets. This was independent of the degree of HL-A haplotype similarity or dissimilarity with the target cell. In addition, individuals who were low responders in the Con A-generated cytotoxicity mounted MLC blastogenic responses and MLC-generated alloantigen specific cytotoxic responses as high as those individuals who were high responders in the Con A-generated cytotoxicity assay. Also, lymphocytes from both high and low responders manifested similar degrees of blastogenic responses to Con A. The present study provides a simple and reproducible system for the generation and assay of cytotoxic effector cells from human peripheral blood after polyclonal activation by Con A and provides a model which can be used in studies of the mechanisms and regulatory factors involved in the activation of human cytotoxic lymphocytes.