Infectious Pseudovirus Construction and Neutralization Susceptibility Analysis of Clinical Isolates of HIV-1 in China
We aimed to isolate and construct infectious human immunodeficiency virus(HIV)-1 Envelope(Env)pseudoviruses from HIV-1-infected Chinese donors and evaluate their neutralization susceptibilities.The HIV-1 env gene was amplified by reverse transcription and nested-polymerase chain reaction(PCR),and then cloned into the eukaryotic expression vector pVAX1 by homologous recombination.The neutralization susceptibility of HIV-1 pseudovirus was measured by the TZM-bl/pseudovirus assay.The key mutation site mediating virus escape was analyzed based on available structural information on broadly neutralizing antibodies.Eight and three HIV-1 pseudoviruses carrying the CRF07_BC env gene were obtained from two Chinese individuals(donorA and donorB).Sequencing analysis showed that the amino-acid homology between pseudoviruses from donorA and donorB was 83.2％to 85.2％.These 11 pseudoviruses could be neutralized by VRC01 and 10E8,but escaped neutralization by 35O22 and VRC34.01.N133D and R170E mutations might have impacted the neutralizing activity of PG16 against pseudoviruses derived from donor B.S291F and E336K mutations might have led to donorA pseudoviruses escaping from VRC-PG05.The G324E mutation might have further decreased the neutralizing activity of PGT121 against donorA pseudoviruses.In this study,we identified 11 infectious pseudoviruses carrying the HIV-1 CRF07_BC env gene from two Chinese HIV-1-infected donors and evaluated their neutralization susceptibilities for seven monoclonal broadly neutralizing antibodies.These findings revealed the key mutations mediating evasion of the HIV-1 antibody,provided a reference and guideline for Chinese HIV-1-infected donors receiving monoclonal antibodies,and enriched the research of HIV-1 pseudoviruses and neutralizing antibodies.
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