title: T cell line passage can select for pre-existing neutralization-sensitive variants from the quasispecies of primary human immunodeficiency virus type-1 isolates.
abstract: Primary human immunodeficiency type 1 viruses (HIV-1) resist antibody neutralization but become sensitive after passage through T cell lines. We and others previously reported an increased neutralization sensitivity of HIV-1 after prolonged culture on primary peripheral blood mononuclear cells (PBMC). Hence we hypothesized that adaptation to growth in T cell lines is in fact selection of a pre-existing neutralization-sensitive HIV-1 variant from the quasispecies in the PBMC culture. Indeed, increased neutralization sensitivity was associated with largely identical synonymous and non-synonymous mutations between progeny of parallel H9 passages from the same split inoculum from 2 of 3 viruses. H9 T cell line adaptation of molecular cloned HIV-1 was less successful and associated with only a few de novo mutations that varied between parallel H9-adapted progeny from the same split inoculum. We conclude that T cell line adaptation of HIV-1 can indeed select for a pre-existing variant but that this most likely depends on the viral diversity in the inoculum.
authors.affiliation: Department of Clinical Viro-Immunology, Sanquin Research and Landsteiner Laboratory of the Academic Medical Center, Center for Infection and Immunity Amsterdam, University of Amsterdam, Amsterdam, The Netherlands. e.quakkelaar@sanquin.nl
authors.forename: Esther D
authors.initials: ED
authors.lastname: Quakkelaar
authors.forename: Tim
authors.initials: T
authors.lastname: Beaumont
authors.forename: Ad C
authors.initials: AC
authors.lastname: van Nuenen
authors.forename: Floris P J
authors.initials: FP
authors.lastname: van Alphen
authors.forename: Brigitte D M
authors.initials: BD
authors.lastname: Boeser-Nunnink
authors.forename: Angélique B
authors.initials: AB
authors.lastname: van 't Wout
authors.forename: Hanneke
authors.initials: H
authors.lastname: Schuitemaker
mesh_terms: D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D014644:Genetic Variation; D015483:HIV Antibodies / Q000276:immunology*; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009500:Neutralization Tests; D010802:Phylogeny; D011533:Proviruses / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012641:Selection, Genetic; D016415:Sequence Alignment; D013601:T-Lymphocytes / Q000821:virology*
chemical_list: D015483:HIV Antibodies; D012367:RNA, Viral