人血单核细胞IL-1的制备与活性单位测定

Production Quantitation of Human Blood Monocytes IL-1

本文报告应用人血白细胞血小板悬液和静脉血(3—5ml)54份,分离单核细胞用于产生IL-1。用小鼠胸腺细胞增殖法测定IL-1活性及滴定其活性单位。结果用人血白细胞血小板悬液制备的IL-1测定其单位平均为1265U/ml,此含IL-1的细胞培养上清液用于进一步提纯IL-1。54份血标本测定IL-1活性单位平均为64.12±113.5U/ml,计算其正常范围是0-291U/ml。应用免疫点迹法测定培养上清液中IL-1的特异性,结果测定样品与标准IL-1在硝酸纤维膜上均呈现同样的斑点,说明它们有共同的特异性。

In present work monocytes obtained from human blood (200ml x 10, Beijing central blood bank) were used for the production of 1L-1 and the activities of IL-1 releaseed from monocytes of 54 volunteers blood samples (3-5ml) were determined by thymocyte proliferation of C57BL strain mouse. The production of IL-1 was performed by culture of the monocytes with RPMI-1640 ( 1 %NBS, 1 ug/ml LPS) for 40 hr. The supernatant was collected and the unit of IL-1 was measured (1265U/ml) . This supernatant will be used for futher purification. The results of 54 blood samples showed that the average of IL-1 was 64. 12 ±113.5U/ml (mean盡SE) and the normal range was between 0-29lU/ml. The specificity of human IL-l was tested by immune dot blot of anti-human IL-1 monoclonal antibody. The standard IL-1 and the IL-1 of supernatant bound membrane showed'dots'. It means that they have same specificity.