Dolei A, Serra C, Arca M V, Tilocca F, Riva E, Antonelli G, Dianzani F, Toniolo A
Institute of Microbiology and Virology, University of Sassari, Italy.
AIDS Res Hum Retroviruses. 1994 Sep;10(9):1089-95. doi: 10.1089/aid.1994.10.1089.
We asked whether HIV-1 had the capacity to establish a persistent infection of cultured human diploid fibroblasts. Human strains of normal diploid embryo lung fibroblasts were infected with HIV-1 of the HTLV-IIIB and HIV-1P1 strains. Infection was followed over time, to analyze HIV expression. Virus production (intra- and extracellular virus) was evaluated as follows: ability to form syncytia in the C8166 T cell line, production of p24 and other viral antigens (ELISA and indirect immunofluorescence), search for a gag sequence in cell DNA by the polymerase chain reaction followed by hybridization to an HIV-1-specific probe (SK19). Cell-free culture supernatant was used as a virus source to infect de novo fibroblasts and C8166 T cells. Infection of cultured fibroblasts with either the HTLV-IIIB or HIV-1P1 strain led regularly to the establishment of persistently infected cultures. Fibroblast cells were capable of continuous virus production for at least 10 months. The released virus was capable of reinfecting cultured fibroblasts and of producing cytopathic effects in the C8166 T cell line. However, when compared to wild-type strains, the infectious virus derived from fibroblasts showed a prolonged replication cycle and a decreased ability to form syncytia in the T cell line. Therefore, HIV-1 can establish a persistent and productive infection in normal lung fibroblasts. The data are consistent with the hypothesis that in vivo, at least in the lung, fibroblasts may represent a virus reservoir and that infection of these cells may lead to the production of attenuated variants of HIV.
我们研究了HIV-1是否有能力在培养的人二倍体成纤维细胞中建立持续性感染。用HTLV-IIIB和HIV-1P1毒株的HIV-1感染正常人二倍体胚胎肺成纤维细胞系。随着时间推移跟踪感染情况，以分析HIV的表达。病毒产生（细胞内和细胞外病毒）评估如下：在C8166 T细胞系中形成多核巨细胞的能力、p24和其他病毒抗原的产生（酶联免疫吸附测定和间接免疫荧光法）、通过聚合酶链反应在细胞DNA中寻找gag序列，随后与HIV-1特异性探针（SK19）杂交。无细胞培养上清液用作病毒源，重新感染成纤维细胞和C8166 T细胞。用HTLV-IIIB或HIV-1P1毒株感染培养的成纤维细胞均能定期建立持续性感染培养物。成纤维细胞能够持续产生病毒至少10个月。释放的病毒能够重新感染培养的成纤维细胞，并在C8166 T细胞系中产生细胞病变效应。然而，与野生型毒株相比，源自成纤维细胞的感染性病毒显示出延长的复制周期，在T细胞系中形成多核巨细胞的能力降低。因此，HIV-1能够在正常肺成纤维细胞中建立持续性和生产性感染。这些数据与以下假设一致：在体内，至少在肺部，成纤维细胞可能是病毒储存库，这些细胞的感染可能导致HIV减毒株的产生。
