Replacement of the V3 region of gp120 with SDF-1 preserves the infectivity of T-cell line-tropic human immunodeficiency virus type 1.
Interaction between the human immunodeficiency virus type 1 (HIV-1) envelope and the relevant chemokine receptors is crucial for subsequent membrane fusion and viral entry. Although the V3 region of gp120 is known to determine the cell tropism as well as the coreceptor usage, the significance of the binding of the V3 region to the chemokine receptor has not been fully understood. To address this issue, we adopted the pseudotyped virus infection assay in which the V3 region of the T-cell line-tropic (T-tropic) NL4-3 envelope was replaced with a portion of stromal cell-derived factor 1 (SDF-1), the ligand of CXCR4. The V3 region of the NL4-3 envelope expression vector was replaced with three different stretches of SDF-1 cDNA. Expression of each chimeric envelope protein was confirmed by immunoprecipitation and Western blotting. Luciferase reporter viruses were prepared by cotransfection of the pNL4-3.Luc.E(-)R(-) vector and each chimeric envelope expression vector, and the infection assay was then carried out. We showed that pseudotyped viruses with one of the chimeric envelopes, NL4-3/SDF1-51, could infect U87.CD4.CXCR4 but not U87.CD4 or U87.CXCR4 cells and that this infection was inhibited by the ligand of CXCR4, SDF-1beta, by anti-human SDF-1 antibody, or by an anti-CD4 antibody, Leu3a, in a dose-dependent manner. Furthermore, chimeric NL4-3/SDF1-51 gp120 significantly inhibited binding of labeled SDF-1 to CXCR4. It was suggested that replacement of the V3 region of the NL4-3 envelope with SDF-1 preserved the CD4-dependent infectivity of T-tropic HIV-1. These results indicate that binding between the V3 region and the relevant coreceptor is important for viral entry, whether its amino acid sequence is indigenous to the virus or not.
Yonezawa
A
A
Department of Hematology/Oncology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Hori
T
T
Takaori-Kondo
A
A
Morita
R
R
Uchiyama
T
T
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015704:CD4 Antigens / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D002461:Cell Line, Transformed; D054407:Chemokine CCL4; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016207:Cytokines / Q000378:metabolism; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000502:physiology*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D019402:Macrophage Inflammatory Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000502:physiology*; D019718:Receptors, CXCR4 / Q000378:metabolism; D012995:Solubility; D013601:T-Lymphocytes / Q000821:virology*; D014771:Virion
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; C515255:CXCL12 protein, human; D054407:Chemokine CCL4; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D016207:Cytokines; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D008164:Luminescent Proteins; D019402:Macrophage Inflammatory Proteins; D010446:Peptide Fragments; D019718:Receptors, CXCR4; D049452:Green Fluorescent Proteins