Isolation and characterization of two divergent infectious molecular clones of HIV type 1 longitudinally obtained from a seropositive patient by a progressive amplification procedure
Isolation and characterization of two divergent infectious molecular clones of HIV type 1 longitudinally obtained from a seropositive patient by a progressive amplification procedure
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DOI：
10.1089/aid.1997.13.743
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发表时间：
1997-06-10
影响因子：
1.5
通讯作者：
Gupta, P
中科院分区：
文献类型：
--
作者：
Chen, MS;Singh, MK;Gupta, P
感染性分子克隆的分离对于我们理解HIV诱导的发病机制是有价值的。使用标准噬菌体λ载体和新的渐进扩增程序，从处于疾病不同阶段的血清阳性受试者中纵向分离了两个HIV-1感染性分子克隆。结果表明，渐进扩增法比常规噬斑杂交法更简便、特异，两种HIV-1感染性克隆具有明显的细胞嗜性和致细胞病变特性，无症状期获得的HIV-1克隆具有嗜巨噬细胞性和非合胞体诱导特性。相反，在AIDS发展阶段获得的HIV-1克隆对T细胞和巨噬细胞具有双重嗜性，并诱导合胞体。对两个克隆的21个酶切位点进行了详细的分析，发现其中9个是独特的，这些独特的酶切位点主要位于包膜区。此外，对整个gp 120区的核苷酸序列分析支持了酶切分析的结果，并表明这两个克隆密切相关，差异仅限于可变区。V-3区氨基酸序列的差异可以解释T细胞嗜性和合胞体诱导特性的差异。来自同一患者在疾病不同阶段的两种不同感染性分子克隆的可用性可能有助于研究HIV-1发病机制。
Isolation of infectious molecular clones has been valuable to our understanding of HIV-induced pathogenesis, Two infectious molecular clones of HIV-1 were isolated longitudinally from a seropositive subject at different stages of the disease, using a standard bacteriophage lambda,vector and a novel progressive amplification procedure. We found the progressive amplification procedure was simpler and more specific than the conventional plaque hybridization assay, The two infectious HIV-1 clones had distinct cell tropism and cytopathic properties, The HIV-1 clone obtained at the asymptomatic stage of the disease was macrophage tropic and had a non-syncytium-inducing property. In contrast, the HIV-1 clone obtained at the stage of AIDS development was dual tropic for T cells and macrophages and induced syncytia. A detailed analysis of the restriction sites of the two clones showed 9 of 21 sites to be unique, These unique restriction sites were predominantly localized in the envelope region, Furthermore, the nucleotide sequence analysis of the entire gp120 region supported the results from the restriction analysis and showed that these two clones are closely related, and the differences are restricted to the variable domains, The difference in amino acid sequences in the V-3 region may explain the observed differences in T cell tropism and syncytium-inducing properties, Availability of two distinct infectious molecular clones from the same patient at different stages of the disease may be useful in studies on the mechanism of HIV-1 pathogenesis.