摘要
目的：探讨一种测定人与小鼠自然杀伤细胞（ＮＫ）杀伤活性的方法。方法：采用ＮＡＧ微量酶比色法，对此法应用的最适条件进行了系统的研究，对正常人不同年龄组进行数次实验，对Ｂａｌｂ／ｃ小鼠也进行了不同浓度的测定实验。结果：正常人的效应细胞与靶细胞（Ｋ５６２）比例为１０∶１时，效应细胞浓度为２×１０６，孵育时间为２０ｈ，以测定效应细胞和靶细胞培养上清中Ｎ乙酰βＤ氨基己糖酶活性，作为ＮＫ杀伤活性的指标，是测定系统的最适条件。小鼠（Ｂａｌｂ／ｃ）效靶比例与人的相似，最适比例为１０∶１或２０∶１。结论：本法有一定的实用价值，无同位素危害，具有较好的敏感性、稳定性、可重复性。
Abstract Objective:To study the activation of killing activity of natural killer(NK) of human and mouse.Methods:Cell functional state was estimated by using a chromogenic substrate(PnitrophenolNacetylβDglucosaminide,NAG) for an ubiquitous lysosomal enzyme,NacetylβDhexosaminidase.The method has been used in which microtiter reaction wells were directly scanned in the Microplate Reader.The systematic study has been performed for determining the appropriate condition of the method.The method was performed in different age groups of normal persons and mice(Balb/c) with different concentration measurement.Results:After target cell(K 562) and effect cell of normal persons were incubated for 20 hours under the 1/10 ratios of target cell to effect cell(2×106),activation of NacetylβDhexosaminidase of culture supernatant of target cell and effect cell were measured.The activation was the index of kill activation of NK cell.The condition was the appropriate condition of measurement system.The ratio of target cell to effect cell in mouse was similar to that in human.The appropriate ratio was 1/20 or 1/10.Conclusion:The main advantages of this assay are that its practical,sensitive,stable,reproducible and with no need of radioisotope.Subject headings:mice;killer cells,natural;colorimetry;lysosomes
出处
《中日友好医院学报》
1998年第1期21-24,共4页
Journal of China-Japan Friendship Hospital
