title: Construction and tropism characterisation of recombinant viruses exhibiting HIV-1 env gene from seminal strains.
abstract: Genetic differences between blood and mucosal-derived HIV-1 strains have been widely reported. As amplification of HIV-1 strains from mucosal samples including semen or saliva by co-culture has low sensitivity, we developed the construction of chimeric viruses expressing wild-type seminal HIV-1 envelope protein. Chimeric viruses were produced by co-transfection of a V1-V3 deleted pNL 43 vector and PCR fragments spanning the deleted region, amplified from HIV-1 RNA positive seminal plasma samples. After an initial testing of co-receptor usage by a tropism recombinant test, replication capacity and amplification of these recombinant viruses were assessed using PBMC. Four chimeric replicative strains, all using CXCR4 as coreceptor, were produced. The interaction between cell-free viral particles and reporter cell lines was assessed by confocal microscopy. These replicative chimeras exhibiting HIV-1 env from seminal strains represent useful tools for the in vitro study of the heterosexual transmission of HIV-1 and testing of microbicide activity.
authors.affiliation: Groupe Immunité des Muqueuses et Agents Pathogènes (GIMAP-EA 3064), Faculté de Médecine J. Lisfranc, Université Jean Monnet, IFRESIS et CHU de Saint-Etienne, 15 rue Ambroise Paré, 42023 Saint-Etienne cedex 02, France.
authors.forename: Philip
authors.initials: P
authors.lastname: Lawrence
authors.forename: Willy
authors.initials: W
authors.lastname: Berlier
authors.forename: Olivier
authors.initials: O
authors.lastname: Delezay
authors.forename: Sabine
authors.initials: S
authors.lastname: Palle
authors.forename: Thomas
authors.initials: T
authors.lastname: Olivier
authors.forename: Henia
authors.initials: H
authors.lastname: Saoudin
authors.forename: Stéphane
authors.initials: S
authors.lastname: Mottin
authors.forename: Frédéric
authors.initials: F
authors.lastname: Lucht
authors.forename: Bruno
authors.initials: B
authors.lastname: Pozzetto
authors.forename: Thomas
authors.initials: T
authors.lastname: Bourlet
mesh_terms: D002460:Cell Line; D015752:Genes, env; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D012367:RNA, Viral / Q000235:genetics; D016865:Reassortant Viruses / Q000235:genetics / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000378:metabolism; D012661:Semen / Q000821:virology; D016415:Sequence Alignment; D014162:Transfection; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*
chemical_list: D012367:RNA, Viral; D019718:Receptors, CXCR4; D054299:env Gene Products, Human Immunodeficiency Virus