Groenink M, Fouchier R A, de Goede R E, de Wolf F, Gruters R A, Cuypers H T, Huisman H G, Tersmette M
Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
J Virol. 1991 Apr;65(4):1968-75. doi: 10.1128/JVI.65.4.1968-1975.1991.
Previously, we and others have demonstrated a relation between the clinical course of human immunodeficiency virus type 1 (HIV-1) infection and biological properties of HIV-1 variants such as replication rate, syncytium-inducing (SI) capacity, and cytotropism. For the molecular analysis of the biological variability in these properties, we generated a panel of phenotypically distinct yet genetically highly homologous infectious molecular clones. These clones were derived from HIV-1 isolates, mostly recovered by direct clonal isolation, from a single individual in whom a transition from non-SI to SI isolates had been identified over time. Of 17 molecular clones tested, 8 were infectious. The clones exhibited differences in SI capacity and T-cell line tropism. Their phenotypes corresponded to those of their parental isolates, formally demonstrating that biological variability of HIV-1 isolates can be attributed to single molecular clones. With these clones we demonstrated that SI capacity and tropism for the H9 T-cell line, almost invariably coupled in primary HIV-1 isolates, are discernible properties. Also different requirements appeared to exist for H9 and Sup T1 cell line tropism. We obtained evidence that T-cell line tropism is not caused by differences in level of HIV-1 expression but most probably is restricted at the level of virus entry. Restriction mapping of four clones with divergent phenotypes revealed a high degree of nucleotide sequence homology (over 96.3%), indicating the usefulness of these clones for the tracking of genetic variability critical for differences in biological phenotype.
此前,我们和其他研究人员已经证明了人类免疫缺陷病毒1型(HIV-1)感染的临床病程与HIV-1变异株的生物学特性之间的关系,这些特性包括复制率、合胞体诱导(SI)能力和嗜亲性。为了对这些特性的生物学变异性进行分子分析,我们构建了一组表型不同但基因高度同源的感染性分子克隆。这些克隆源自HIV-1分离株,大多是通过直接克隆分离从一名个体中获得的,该个体中随着时间推移已鉴定出从非SI分离株向SI分离株的转变。在测试的17个分子克隆中,8个具有感染性。这些克隆在SI能力和T细胞系嗜亲性方面表现出差异。它们的表型与其亲本分离株的表型相对应,正式证明了HIV-1分离株的生物学变异性可归因于单个分子克隆。利用这些克隆,我们证明了在原发性HIV-1分离株中几乎总是相关联的SI能力和对H9 T细胞系的嗜亲性是可区分的特性。对于H9和Sup T1细胞系嗜亲性似乎也存在不同的要求。我们获得的证据表明,T细胞系嗜亲性不是由HIV-1表达水平的差异引起的,而很可能在病毒进入水平受到限制。对四个具有不同表型的克隆进行的限制性图谱分析显示出高度的核苷酸序列同源性(超过96.3%),表明这些克隆对于追踪对生物学表型差异至关重要的遗传变异性很有用。