天然杀伤活性的测定——(1)体外8h(125)I-UdR释放试验

Detection of Natural Killing Activity 1. In Vitro 8h^(125)I-UdR-Release Test

用^(125)I—UdR标记靶YAC—1细胞建立8h^(125)I-UdR释放试验(8IRA),体外测定小鼠脾脏细胞NK活性。靶细胞标记的理想条件是:YAC—1细胞2×10~5/ml;加^(1250I-UdR74～111kBq/L;FUdR3×10^(-5)mol/L;在37℃、5%CO_2湿环境中培养5～7h,其^(125)I-UdR掺入量(标记率)为0.5cpm/细胞以上。影响^(125)I-UdR释放因素的研究表明,采用8IRA时,自发释放率在20%以下,而且NK活性的测定结果也稳定。

Under our laboratory conditions, 8h-^(125)I-UdR-Release-test was established with target YAC-1 cells labeled with ^(125)I-UdR for detecting Natural killing(N K)activity in vitro. The optimal conditions for target labelling were: YAC-1 cells 2 ×10~5/ml; ^(125)I-UdR 74-111 kBq/L; FUdR 3×10^(-5)mol/L & incubating for 5-7h at 37℃; 5% CO_2 & wet atmosphere, resulting in ^(125)I-UdR incorporation of more than 0.5 cpm/cell. The factors influencing the release of ^(125)I-UdR were also studied, and indicated that when 8h-^(125)I-UdR-Release-Test was used, the spontaneous release of ^(125)I-UdR was below 20% & measurement of N K activity was stable & believable as well.