Fikkert Valery, Cherepanov Peter, Van Laethem Kristel, Hantson Anke, Van Remoortel Barbara, Pannecouque Christophe, De Clercq Erik, Debyser Zeger, Vandamme Anne-Mieke, Witvrouw Myriam
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium.
Antimicrob Agents Chemother. 2002 Dec;46(12):3954-62. doi: 10.1128/AAC.46.12.3954-3962.2002.
We describe the development of chimeric virus technology (CVT) for human immunodeficiency virus (HIV) type 1 (HIV-1) env genes gp120, gp41, and gp160 for evaluation of the susceptibilities of HIV to entry inhibitors. This env CVT allows the recombination of env sequences derived from different strains into a proviral wild-type HIV-1 clone (clone NL4.3) from which the corresponding env gene has been deleted. An HIV-1 strain (strain NL4.3) resistant to the fusion inhibitor T20 (strain NL4.3/T20) was selected in vitro in the presence of T20. AMD3100-resistant strain NL3.4 (strain NL4.3/AMD3100) was previously selected by De Vreese et al. (K. De Vreese et al., J. Virol. 70:689-696, 1996). NL4.3/AMD3100 contains several mutations in its gp120 gene (De Vreese et al., J. Virol. 70:689-696, 1996), whereas NL4.3/T20 has mutations in both gp120 and gp41. Phenotypic analysis revealed that NL4.3/AMD3100 lost its susceptibility to dextran sulfate, AMD3100, AMD2763, T134, and T140 but not its susceptibility to T20, whereas NL4.3/T20 lost its susceptibility only to the inhibitory effect of T20. The recombination of gp120 of NL4.3/AMD3100 and gp41 of NL4.3/T20 or recombination of the gp160 genes of both strains into a wild-type background reproduced the phenotypic (cross-)resistance profiles of the corresponding strains selected in vitro. These data imply that mutations in gp120 alone are sufficient to reproduce the resistance profile of NL4.3/AMD3100. The same can be said for gp41 in relation to NL4.3/T20. In conclusion, we demonstrate the use of env CVT as a research tool in the delineation of the region important for the phenotypic (cross-)resistance of HIV strains to entry inhibitors. In addition, we obtained a proof of principle that env CVT can become a helpful diagnostic tool in assessments of the phenotypic resistance of clinical HIV isolates to HIV entry inhibitors.
我们描述了用于人类免疫缺陷病毒1型（HIV-1）包膜基因gp120、gp41和gp160的嵌合病毒技术（CVT）的开发，以评估HIV对进入抑制剂的敏感性。这种包膜CVT允许将来自不同毒株的包膜序列重组到一个已缺失相应包膜基因的前病毒野生型HIV-1克隆（克隆NL4.3）中。在T20存在的情况下，在体外选择了对融合抑制剂T20耐药的HIV-1毒株（毒株NL4.3/T20）。抗AMD3100的毒株NL3.4（毒株NL4.3/AMD3100）先前由De Vreese等人筛选得到（K. De Vreese等人，《病毒学杂志》70:689 - 696，1996年）。NL4.3/AMD3100在其gp120基因中含有多个突变（De Vreese等人，《病毒学杂志》70:689 - 696，1996年），而NL4.3/T20在gp120和gp41中均有突变。表型分析表明，NL4.3/AMD3100对硫酸葡聚糖、AMD3100、AMD2763、T134和T140失去了敏感性，但对T20仍保持敏感性，而NL4.3/T20仅对T20的抑制作用失去了敏感性。将NL4.3/AMD3100的gp120与NL4.3/T20的gp41重组，或将两种毒株的gp160基因重组到野生型背景中，重现了体外筛选的相应毒株的表型（交叉）耐药谱。这些数据表明，仅gp120中的突变就足以重现NL4.3/AMD3100的耐药谱。就NL4.3/T20而言，gp41也是如此。总之，我们证明了包膜CVT作为一种研究工具，可用于描绘HIV毒株对进入抑制剂的表型（交叉）耐药的重要区域。此外，我们还获得了一项原理证明，即包膜CVT可成为评估临床HIV分离株对HIV进入抑制剂的表型耐药性的有用诊断工具。
