The inhibitory effect of CCR5Delta32 protein on cell surface expression of the HIV-1 coreceptor CCR5 and CXCR4 / 中国免疫学杂志

ABSTRACT

Objective: To demonstrate that expression of the CCRSDelta32 protein in PBMCs able to down-regulate surface expression of the HIV-1 coreceptor CCR5 and CXCR4.Methods:CCR5Delta32 gene was amplified from human peripheral blood mononuclear cells(PBMCs)genomic DNA by using PCR, and then cloned into lentiviral vector pLenti6/V5-D-TOPO.Recombinant lentiviral particles were produced by packaging using 293T cells.Human PBMCs were transfected with the constructed recombinant lentiviral particles and the expression of CCR5Delta32 was detected by Western blot.The level of CCR5 and CXCR4 expression on transfected PBMCs was detected by FACS analysis.Results: The recombinant lentiviral vector pLenti-CCR5Delta32 was constructed successfully, and the target protein was expressed in PBMCs.FACS analysis showed that CCR5Delta32 protein expressed in PBMCs was able to down-regulate cell surface expression of CCR5 and CXCR4.Conclusion: This study is expected to be used for the gene therapy on AIDS, which deserves further study.