title: HIV sequence variation associated with env antisense adoptive T-cell therapy in the hNSG mouse model.
abstract: The first use of lentiviral vectors in humans involved transduction of mature T-cells with an human immunodeficiency virus (HIV)-derived env antisense (envAS) vector to protect cells from HIV infection. In that study, only a minority of the patient T-cell population could be gene-modified, raising the question of whether the altered cells could affect replicating HIV populations. We investigated this using humanized NOD/SCID IL-2Rgamma(null) (hNSG) mice reconstituted with approximately 4-11% envAS-modified human T-cells. Mice were challenged with HIV-1(NL4-3), which has an env perfectly complementary to envAS, or with HIV-1(BaL), which has a divergent env. No differences were seen in viral titer between mice that received envAS-modified cells and control mice that did not. Using 454/Roche pyrosequencing, we analyzed the mutational spectrum in HIV populations in serum-from 33 mice we recovered 84,074 total reads comprising 31,290 unique sequence variants. We found enrichment of A-to-G transitions and deletions in envAS-treated mice, paralleling a previous tissue culture study where most target cells contained envAS, even though minority of cells were envAS-modified here. Unexpectedly, this enrichment was only detected after the challenge with HIV-1(BaL), where the viral genome would form an imperfect duplex with envAS, and not HIV-1(NL4-3), where a perfectly matched duplex would form.
authors.affiliation: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
authors.forename: Rithun
authors.initials: R
authors.lastname: Mukherjee
authors.forename: Gabriela
authors.initials: G
authors.lastname: Plesa
authors.forename: Scott
authors.initials: S
authors.lastname: Sherrill-Mix
authors.forename: Max W
authors.initials: MW
authors.lastname: Richardson
authors.forename: James L
authors.initials: JL
authors.lastname: Riley
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
mesh_terms: D000818:Animals; D001483:Base Sequence; D014644:Genetic Variation; D015658:HIV Infections / Q000628:therapy* / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016219:Immunotherapy, Adoptive; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016372:RNA, Antisense / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation*; D014161:Transduction, Genetic; D014779:Virus Replication / Q000235:genetics; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics*
chemical_list: D016372:RNA, Antisense; D012367:RNA, Viral; D054299:env Gene Products, Human Immunodeficiency Virus