title: Chemokine receptor-usage of clinical HIV-1 isolates obtained from patients with HIV-1 infection in late clinical stages using PHA-blast.
abstract: In the present sudy, chemokine receptor-usage of primary HIV-1 isolates was examined using U87-CD4 cells expressing chemokine receptors CCR3, CCR5 and CXCR4. HIV-1 was isolated from the peripheral blood mononuclear cells (PBMC) and/or plasma of eight HIV-1-infected individuals in late CDC-II and CDC-IV clinical stages using PHA-blast prepared from the PBMC of healthy blood donors. The primary HIV-1 isolates from patients in late CDC-II stage rarely infected monocyte-derived macrophages in the present study, whereas most isolates from patients in the CDC-IV stage infected the macrophages. In the experiments using U87-CD4 cells expressing chemokine receptors, the isolates from patients in the late CDC-II stage infected U87-CD4 cells expressing CXCR4, but not U87-CD4 cells expressing CCR5. In contrast, most isolates from patients in the CDC-IV stage infected both U87-CD4 cells expressing CXCR4 or CCR5. The isolates which infected both U87-CD4 cells were supposed to contain dual tropic HIV-1 or a mixture of CXCR4-tropic and CCR5-tropic HIV-1s. Analysis of the deduced amino acid sequence of the V3 region in proviral env gene showed that the V3 region in U87-CD4 cells infected with CXCR4-tropic HIV-1 isolates was largely different from that in the cells infected with CCR5-tropic isolates, but were highly similar to that in cells infected with dual tropic isolates. These results suggest that PHA-blasts may preferentially support the replication of the CXCR4-tropic and dual tropic HIV-1s, and that CXCR4-tropic and dual tropic HIV-1s are also present in peripheral blood from patients in the late stage of the asymptomatic phase.
authors.affiliation: Institute of Clinical Research, Kure National Hospital, Hiroshima, Japan. icr1@kure-nh.go.jp
authors.forename: S
authors.initials: S
authors.lastname: Matsuda
authors.forename: M
authors.initials: M
authors.lastname: Miyata
mesh_terms: D000595:Amino Acid Sequence; D015704:CD4 Antigens / Q000235:genetics; D002460:Cell Line; D015686:Gene Products, env / Q000235:genetics; D015699:HIV Envelope Protein gp120 / Q000235:genetics; D015658:HIV Infections / Q000503:physiopathology / Q000821:virology*; D015497:HIV-1 / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008934:Mitogens / Q000494:pharmacology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D010835:Phytohemagglutinins / Q000494:pharmacology; D054397:Receptors, CCR3; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism*; D019707:Receptors, Chemokine / Q000235:genetics / Q000378:metabolism*; D015728:Receptors, HIV / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: C515350:CCR3 protein, human; D015704:CD4 Antigens; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D008934:Mitogens; D010446:Peptide Fragments; D010835:Phytohemagglutinins; D054397:Receptors, CCR3; D019713:Receptors, CCR5; D019718:Receptors, CXCR4; D019707:Receptors, Chemokine; D015728:Receptors, HIV