Monoclonal antibodies and chemiluminescence immunoassay for detection of the surface protein of human T-cell lymphotropic virus.
Monoclonal antibodies and chemiluminescence immunoassay for detection of the surface protein of human T-cell lymphotropic virus.
复制标题
单克隆抗体和化学发光免疫分析法检测人 T 细胞嗜淋巴细胞病毒表面蛋白。
DOI：
10.1128/jcm.30.2.351-358.1992
复制
发表时间：
1992
影响因子：
9.4
通讯作者：
Poiesz,BJ
中科院分区：
文献类型：
--
作者：
Papsidero,LD;Dittmer,RP;Vaickus,L;Poiesz,BJ
抗人T细胞嗜淋巴细胞病毒I型（HTLV-I）的单克隆抗体（MAbs）识别病毒env基因编码蛋白的五个不同的抗原结构域。通过使用重组env蛋白和合成肽作为定位抗原，确定最具免疫原性的区域代表逆转录病毒表面蛋白的中心部分（结构域2;氨基酸165至191）。然而，只有一种单克隆抗体能够与天然病毒蛋白发生强烈反应。该抗体（克隆6C 2）针对逆转录病毒env基因的结构域4（氨基酸210至306）内的表位，并与HTLV-I和HTLV-II中的包膜蛋白反应，如通过免疫沉淀、固相结合和免疫印迹所确定的。不存在对其他人逆转录病毒（包括1型和2型人免疫缺陷病毒）包膜组分的反应性。流式细胞术数据表明，MAb 6C 2与慢性感染HTLV-I或HTLV-II的细胞系反应，也与新鲜成人T细胞白血病细胞上表达的表面抗原反应，随后用白细胞介素-2上调。通过化学发光免疫测定程序，皮克量的病毒表面蛋白可以检测到在未浓缩的上清液中的HTLV感染的细胞系和诊断培养物。细胞释放到培养上清液中的env和gag蛋白水平与细胞表面病毒外壳蛋白的表达百分比没有直接关系。此外，条件培养基中p19与gp 46的摩尔比因菌株而异，可能反映了病毒组装或包装机制的差异。单克隆抗体6C 2在研究逆转录病毒env基因蛋白的生物化学和免疫学行为以及研究HTLV-Ⅰ和HTLV-Ⅱ与受体的相互作用方面具有重要价值。
Monoclonal antibodies (MAbs) raised against human T-cell lymphotropic virus type I (HTLV-I) recognized five distinct antigenic domains of viral env gene-encoded proteins. By using recombinant env proteins and synthetic peptides as mapping antigens, it was determined that the most immunogenic region represented a central portion of the retroviral surface protein (domain 2; amino acids 165 to 191). However, only a single MAb was able to react strongly with native viral proteins. This antibody (clone 6C2) was directed to an epitope within domain 4 (amino acids 210 to 306) of the retroviral env gene and reacted with envelope proteins in both HTLV-I and HTLV-II, as determined by immunoprecipitation, solid-phase binding, and immunoblotting. No reactivity against envelope components of other human retroviruses, including human immunodeficiency virus types 1 and 2, was present. Flow cytometry data demonstrated that MAb 6C2 reacted with cell lines chronically infected with HTLV-I or HTLV-II and also with surface antigens expressed on fresh adult T-cell leukemia cells, following up-regulation with interleukin-2. By a chemiluminescence immunoassay procedure, picogram amounts of viral surface protein could be detected in the unconcentrated supernatants of HTLV-infected cell lines and in diagnostic cultures. Levels of env and gag proteins released by cells into culture supernatants were not directly related to percent expression of cell surface viral-coat proteins. Further, the molar ratio of p19 to gp46 in conditioned media varied from strain to strain, possibly reflecting differences in viral assembly or packaging mechanisms. MAb 6C2 will be of value in characterizing the biochemical and immunological behavior of retroviral env gene proteins and in studying the interaction of HTLV-I and HTLV-II with their receptors.