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白细胞粘附抑制试验(LAI)是具有抗原特异性、方法简便而快速的一种免疫反应,并在肿瘤免疫学领域内得到广泛应用。抗原的制备将手术切除的新鲜肺癌组织以及同一患者的正常肺组织切细后,用0.01M pH7.2的磷酸盐缓冲盐水洗净,分别制成匀浆。注入5倍量的磷酸盐缓冲盐水,在4℃用磁力搅拌器搅拌24小时后,以40,0000×g离心60分钟,取上清液再用10倍量的磷酸盐缓冲液透析两次,再用40,000×g离心15分钟,其上清液即为肺癌相关抗原(LCAA)。以同法制备的正常肺组织的上清液作为对照。抗原可保存于-90℃备用。以Lowry法测定LCAA中的蛋白浓度。使用时将浓度调为300μg/ml。
Leukocyte adhesion inhibition test (LAI) is an antigen-specific, simple and rapid method of immune response, and is widely used in the field of tumor immunology. Preparation of Antigen Fresh surgically resected lung cancer tissue and normal lung tissue from the same patient were shredded and then washed with 0.01 M phosphate buffered saline, pH 7.2, and homogenized separately. Five times the amount of phosphate buffered saline was injected, stirred at 4 ° C for 24 hours with a magnetic stirrer, and centrifuged at 40,000 × g for 60 minutes. The supernatant was further dialyzed twice with 10-fold amount of phosphate buffer, After centrifugation at 40,000 × g for 15 minutes, the supernatant is lung cancer associated antigen (LCAA). The normal lung tissue prepared with the supernatant as a control. Antigens can be stored at -90 ℃ for later use. The protein concentration in LCAA was determined by the Lowry method. Use to adjust the concentration to 300μg / ml.