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以氚标记胸腺嘧啶核苷(~3H-TdR)掺入法检测每分钟脉冲数(CPM),比较母牛分枝杆菌活菌悬液、辐射杀死菌悬液及高温杀死菌悬液对健康人外周血T淋巴细胞增殖反应的影响.以此作为评价治疗以细胞介导免疫为主的结核病免疫功能障碍免疫制剂效力的一个重要参数.在加入单一刺激因子实验中,对照组CPM为448±131;加入BCG、母牛分枝杆菌活菌悬液、辐射杀灭菌悬液以及高温杀死菌悬液的CPM分别为1037±194,2299±140,1819±528,994±186.这4种制剂的刺激指数均在2.0以上,尤其是母牛分枝杆菌活菌悬液、辐射杀死菌悬液分别为5.13,4.06.结果表明,母牛分枝杆菌3种制剂与BCG基本相似,在体外对T淋巴细胞增殖反应有明显的促进作用,其中以活菌悬液和辐射杀死菌悬液更为显著.另一试验中,以重组白细胞介素-2(rIL-2)作对照,BCG、母牛分枝杆菌悬液、辐射杀死菌悬液及高温杀死菌悬液分别加入rIL-2,目的在于观察菌体抗原加淋巴因子对T淋巴细胞增殖反应有无协同刺激作用.对照组的CPM为3721±1336,BCG加rIL-2组CPM为6904±1218;母牛分枝杆菌3种制剂的CPM分别为9544±1727;8530±744;8230±1035.结果说明,除BCG加rIL-2组外,母牛分枝杆菌3种制剂加rIL-2对T淋巴细胞增殖反应均有明显的协同促进作用.
The number of pulses per minute (CPM) was measured by incorporation of tritiated thymidine (~ 3H-TdR), the viable suspension of Mycobacterium vaccae was irradiated, the suspension was irradiated by radiation and the suspension of bacteria was killed by high temperature Healthy human peripheral blood T lymphocyte proliferation response.To evaluate the treatment of cell-mediated immune-based immunodominant immune function of tuberculosis is an important parameter.In a single stimulus test, the control group CPM 448 ± 131; CPM added BCG, viable Mycobacterium vaccae, radiation killed suspension and high temperature killed bacterial suspension were 1037 ± 194, 2299 ± 140, 1819 ± 528 and 994 ± 186 respectively The stimulation indexes of the preparations were both above 2.0, especially the viable suspension of Mycobacterium vaccae and the suspension of irradiation-killed bacteria were 5.13 and 4.06 respectively.The results showed that the three preparations of Mycobacterium vaccae were almost similar to BCG, In vitro, the proliferation of T lymphocytes was obviously promoted, especially the suspension of bacteria was killed by the live bacterial suspension and radiation.Another experiment, recombinant interleukin-2 (rIL-2) BCG, mycobacterium vaccae suspension, radiation to kill the bacterial suspension and high temperature killed bacteria were added rIL-2, the purpose is to observe the cell antigen plus lymphokines on T lymphocyte proliferation reaction with or without synergistic stimulation.The control group CPM was 3721 ± 1336, BCG plus rIL-2 group CPM was 6904 ± 1218; cow bifurcation The CPM of the three kinds of preparations of Bacillus were 9544 ± 1727, 8530 ± 744 and 8230 ± 1035 respectively.The results showed that the addition of rIL-2 to Mycobacterium vaccae to BCG plus rIL-2 had no significant effect on T lymphocyte proliferation Have obvious synergistic effect.