title: Selection following isolation of human immunodeficiency virus type 1 in peripheral blood mononuclear cells and herpesvirus saimiri-transformed T cells is comparable.
abstract: In attempts to improve isolation rates and virus yields for human immunodeficiency virus (HIV), the use of herpesvirus saimiri-immortalized T cells (HVS T cells) has been investigated as an alternative to/improvement over peripheral blood mononuclear cells (PBMCs). Here we characterize isolates rescued, in the two cell types, from two asymptomatic, long-term non-progressing HIV-1-infected individuals. All rescued viruses replicated in PBMCs and HVS T cells only, displaying a non-syncytium inducing (NSI) phenotype, and using CCR5 as co-receptor. Furthermore, PBMC/HVS T cell virus pairs displayed similar neutralization profiles. Full-length, expression-competent env genes were rescued from all virus isolates and directly from the patient samples using proviral DNA and viral RNA as templates. Compared with the sequences retrieved directly from the patient samples, both cell types showed similar selection characteristics. Whilst the selections were distinct for individual patient samples, they shared a common characteristic in selecting for viruses with increased negative charge across the V2 domain of the viral glycoproteins. The latter was observed at the env gene sequencing level for three other patients whose HIV strains were isolated in PBMCs only. This further supports a common selection for viral sequences that display a macrophage-tropic/NSI phenotype and shows that HVS T cells are a viable alternative to PBMCs for HIV-1 isolation.
authors.affiliation: Virology Division, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK1.
authors.forename: Natalie N
authors.initials: NN
authors.lastname: Zheng
authors.affiliation: School of Biological and Applied Sciences, University of North London, 166-220 Holloway Road, London N7 8DB, UK2.
authors.forename: Cherelyn
authors.initials: C
authors.lastname: Vella
authors.affiliation: Department of HIV and Genitourinary Medicine, The Guy's, King's and St Thomas' School of Medicine, King's College Hospital, Denmark Hill, London SE5 9RS, UK3.
authors.forename: Philippa J
authors.initials: PJ
authors.lastname: Easterbrook
authors.affiliation: Virology Division, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK1.
authors.forename: Rod S
authors.initials: RS
authors.lastname: Daniels
mesh_terms: D000595:Amino Acid Sequence; D002461:Cell Line, Transformed; D018920:Coculture Techniques; D004279:DNA, Viral; D015686:Gene Products, env / Q000235:genetics; D015752:Genes, env; D015726:Giant Cells; D006023:Glycoproteins / Q000235:genetics; D015658:HIV Infections / Q000097:blood / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006567:Herpesvirus 2, Saimiriine; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000473:pathology / Q000821:virology*; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D011533:Proviruses / Q000235:genetics; D012367:RNA, Viral; D019713:Receptors, CCR5; D016415:Sequence Alignment; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology / Q000821:virology*
chemical_list: D004279:DNA, Viral; D015686:Gene Products, env; D006023:Glycoproteins; D012367:RNA, Viral; D019713:Receptors, CCR5