Nishanian P G, Ojo-Amaize E, Uittenbogaart C H, Giorgi J V
Department of Microbiology and Immunology, UCLA School of Medicine 90024-1747.
J Immunol Methods. 1988 Mar 16;107(2):261-71. doi: 10.1016/0022-1759(88)90227-x.
Standardized samples of target cells expressing viral antigen are necessary to perform reproducible and comparable functional assays for immune responses to human immunodeficiency virus (HIV-1). HIV-infected cultured cell lines show wide variability in the number of HIV-expressing cells and in the amount of viral antigen per cell. They, as well as cells coated with the infectious virus, are a biohazard. 0.015% v/v beta-propiolactone (BPL) in buffered solution at pH 7.5 inactivates the infectivity of HIV but retains its antigenicity. BPL-inactivated HIV easily binds to T4 receptor bearing T-lymphocytes in a 30 min incubation at 22 degrees C. The process of adsorption is dose-dependent and results in a standardized and reproducible sample of target cells. Applications for the method are demonstrated and include: (1) preparation of target cells for assay of antibody-dependent cellular cytotoxicity (ADCC), and (2) characterization of antisera to HIV for their capacity to block HIV binding to cells.
对于人类免疫缺陷病毒（HIV-1）免疫反应而言，要进行可重复且具有可比性的功能测定，表达病毒抗原的靶细胞标准化样本必不可少。感染HIV的培养细胞系在表达HIV的细胞数量以及每个细胞的病毒抗原量方面存在很大差异。它们以及涂有感染性病毒的细胞都具有生物危害性。在pH 7.5的缓冲溶液中0.015% v/v的β-丙内酯（BPL）可使HIV的感染性失活，但保留其抗原性。BPL灭活的HIV在22℃孵育30分钟时很容易与带有T4受体的T淋巴细胞结合。吸附过程是剂量依赖性的，可产生标准化且可重复的靶细胞样本。文中展示了该方法的应用，包括：（1）制备用于抗体依赖性细胞毒性（ADCC）测定的靶细胞，以及（2）鉴定针对HIV的抗血清阻止HIV与细胞结合的能力。
