Substitution of HIV Type-1 Non-B env Genes in C2,a Subtype B Cassette System,Results in Functional Chimeric Viruses

Env gene glycoprotein products are essential to viral infectivity and important targets for a host's humoral and cellular immune responses. We have reported the construction of C2, an effective env gene cassetting system for assessing biological properties of HIV-1 subtype B env gene glycoprotein products within a constant genetic background (Zheng NN and Daniels RS: AIDS Res Hum Retroviruses 2001;17:1501–7506). Here we report the ability of C2 to produce chimeric subtype A, C, D, A/E, F, and J HIV-1 and studies of the viruses' biological properties. Virus RNAs were extracted and full-length env genes rescued by RT-PCR. Expressioncompetent env genes were cloned into the C2 cassette and chimeric recombinant viruses produced by transfecting 293T cells. For each subtype, X4 viruses yielded higher TCID₅₀ than R5 viruses and the TCID₅₀ of chimeric viruses were either the same as or lower than their parental viruses. The limited coreceptor usage of R5-tropic parent viruses was retained in the chimeric viruses. Generally, with the exception of the subtype C virus (SE12808), the X4-tropic parental viruses utilized CXCR4 and a wide range of additional coreceptors, while their respective chimeric viruses retained CXCR4 usage but showed a more limited range in respect of other coreceptors. The replication rates of non-B subtype chimeric viruses were generally lower (1.5- to 13.6-fold) than their respective parental viruses with the exception of C2-92UG029, an X4-tropic subtype A chimeric virus. This study demonstrates that C2 is a functional cassette capable of producing infectious chimeric viruses to allow study of the biological phenotypes and functions of HIV-1 subtype B and non-B subtype glycoproteins.