Li X, Amandoron E, Wainberg M A, Parniak M A
Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, Montreal, Quebec, Canada.
J Med Virol. 1993 Mar;39(3):251-9. doi: 10.1002/jmv.1890390314.
We produced a series of monoclonal antibodies against the human immunodeficiency virus (HIV-1) reverse transcriptase by immunizing mice with either purified recombinant HIV-1 p66 protein or with recombinant vaccinia virus which expresses HIV-1 pol sequences. The antibodies generated were specific for the reverse transcriptase protein, and recognized only the p51 and p66 subunits of the enzyme in each of the HIV-1 viral lysates and lysates of HIV-1 infected cells. The antibodies did not cross-react with HIV-2 reverse transcriptase. Most important, several of the antibodies are unique, in that they are the first that can bind to sites close to the N-terminal. This latter region has been suggested to form part of the polymerase domain of the reverse transcriptase. None of the antibodies could neutralize either the RNA-dependent DNA polymerase or RNase H activities of either p66 or p51/66 proteins. The binding patterns of these various antibodies to p66 and p51/66 were dependent on each of three independent variables: the source of antigen amployed, the individual specificity of the antibody, and the method employed to detect reactivity. These monoclonal antibodies provide useful reagents for the study of reverse transcriptase native structure-function relationships.
我们通过用纯化的重组HIV-1 p66蛋白或表达HIV-1 pol序列的重组痘苗病毒免疫小鼠，制备了一系列针对人类免疫缺陷病毒（HIV-1）逆转录酶的单克隆抗体。所产生的抗体对逆转录酶蛋白具有特异性，并且在每种HIV-1病毒裂解物和HIV-1感染细胞的裂解物中仅识别该酶的p51和p66亚基。这些抗体与HIV-2逆转录酶无交叉反应。最重要的是，其中几种抗体是独特的，因为它们是首批能够结合靠近N端位点的抗体。后一个区域被认为是逆转录酶聚合酶结构域的一部分。这些抗体均不能中和p66或p51/66蛋白的RNA依赖性DNA聚合酶或RNase H活性。这些不同抗体与p66和p51/66的结合模式取决于三个独立变量中的每一个：所用抗原的来源、抗体的个体特异性以及用于检测反应性的方法。这些单克隆抗体为研究逆转录酶天然结构-功能关系提供了有用的试剂。
