Yunoki M, Maotani-Imai K, Kusuda H, Motoyama M, Miyake S, Imai H, Shin Y S, Kato S, Sano K, Morita C
Section of Serology, Hokkaido University, Sapporo, Japan.
Arch Virol. 1991;116(1-4):143-58. doi: 10.1007/BF01319238.
A total of 81 cell clones persistently infected with the LAV-1 or HTLV-IIIB strain of human immunodeficiency virus type 1 (HIV-1) was isolated from cells which were obtained by serial passage of some proliferating MT-4 cells after a drastic cytolysis of most cells by HIV-1-infection. These cell clones were classified into 8 types (I to VIII) in terms of the expression of HIV-1 antigens, syncytium formation capacity, and reverse transcriptase activity and infectivity of virus particles in the culture fluid. Type I cell clones were producers of infectious HIV-1 particles, while types II to VIII cell clones did not produce infectious HIV-1 or were producers of uninfectious defective HIV-1 particles. Immunoprecipitation followed by SDS-polyacrylamide gel electrophoresis (PAGE) showed that the gag precursor protein in L-2 cell clone (type IV) was not cleaved to mature gag proteins, while the env precursor protein on L-3 cell clone (type III) was not cleaved to mature env protein. H-7 cell clone (type VIII) did not express any HIV-1 antigen. All these cell clones after the superinfection with infectious HIV-1 synthesized intact gag and env proteins, which were, at least in part, related to the HIV-1 genome persistently present in the cell clones before the superinfection, resulting in production of infectious HIV-1. For example, it was found that L-2 cell clone contained a single copy of the LAV-1 genome per haploid cell and produced doughnut-shaped particles. On the other hand, the cell clone isolated from the L-2 cell clone superinfected with infectious HTLV-IIIB contained the integrated HTLV-IIIB genome in addition to the LAV-1 genome present before the superinfection, and produced intact HIV-1 particles in addition to doughnut-shaped particles from a single cell. These results indicate that complementation and/or genetic recombination events in the superinfected cells may account for the production of infectious intact HIV-1 virions.
从一些增殖的MT - 4细胞经人免疫缺陷病毒1型（HIV - 1）感染导致大多数细胞剧烈溶解后经连续传代获得的细胞中，分离出总共81个持续感染HIV - 1的LAV - 1或HTLV - IIIB毒株的细胞克隆。根据HIV - 1抗原的表达、合胞体形成能力以及培养液中病毒颗粒的逆转录酶活性和感染性，这些细胞克隆被分为8种类型（I至VIII）。I型细胞克隆是有感染性的HIV - 1颗粒的生产者，而II至VIII型细胞克隆不产生有感染性的HIV - 1或产生无感染性的缺陷HIV - 1颗粒。免疫沉淀后进行SDS - 聚丙烯酰胺凝胶电泳（PAGE）显示，L - 2细胞克隆（IV型）中的gag前体蛋白未裂解为成熟的gag蛋白，而L - 3细胞克隆（III型）中的env前体蛋白未裂解为成熟的env蛋白。H - 7细胞克隆（VIII型）不表达任何HIV - 1抗原。所有这些细胞克隆在感染有感染性的HIV - 1后都合成了完整的gag和env蛋白，这些蛋白至少部分与超感染前细胞克隆中持续存在的HIV - 1基因组相关，从而导致产生有感染性的HIV - 1。例如，发现L - 2细胞克隆每个单倍体细胞含有一份LAV - 1基因组，并产生甜甜圈形状的颗粒。另一方面，从感染有感染性的HTLV - IIIB的L - 2细胞克隆中分离出的细胞克隆，除了超感染前存在的LAV - 1基因组外，还含有整合的HTLV - IIIB基因组，并且从单个细胞中除了产生甜甜圈形状的颗粒外还产生完整的HIV - 1颗粒。这些结果表明，超感染细胞中的互补和/或基因重组事件可能是产生有感染性的完整HIV - 1病毒体的原因。
