乳酸脱氢酶释放法检测鸡自然杀伤细胞活性条件的优化

Test improvement of lactic dehydrogenase realease on natural killer cytotoxic indexes in broiler chickens

以肉仔鸡外周血自然杀伤(NK)细胞作为研究对象,探讨乳酸脱氢酶(LDH)释放法检测NK细胞活性中5个最为关键的影响因素,选用Yac-1淋巴瘤细胞作靶细胞.结果表明:效靶细胞比为15∶1,在含2%犊牛血清的RPMI 1640维持液中,效靶细胞于37℃、5%CO2条件下作用2 h,于37℃恒温条件下酶促反应10 min,检测波长为492 nm可获得满意结果;验证试验结果表明,优化的LDH释放法检测鸡外周血NK细胞活性的方法是可行的.

Five factors related to measurement of natural killer（NK） cell activities with lactic dehydrogenase（LDH） realease in peripheral blood of broilers were examined.Yac-1 cells were selected as target cells.The result shows that when ratio of effective cell to target cell was 15∶1,effective cell and target cell were suspended with 2% fetal calf serum-RPMI 1640 in 37 ℃ and were acted on 5% CO2 for 2 h,enzymatic catalyzed reaction was persisted in 10 min,wavelength was confirmed in 492 nm,NK cell activities in peripheral blood of broilers were well determined,indicating that the improved method can be used for the test of NK cytotoxicity of broilers.