Construction and tropism characterisation of recombinant viruses exhibiting HIV-1 env gene from seminal strains.
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.
Lawrence
Philip
P
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.
Berlier
Willy
W
Delezay
Olivier
O
Palle
Sabine
S
Olivier
Thomas
T
Saoudin
Henia
H
Mottin
Stéphane
S
Lucht
Frédéric
F
Pozzetto
Bruno
B
Bourlet
Thomas
T
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*
D012367:RNA, Viral; D019718:Receptors, CXCR4; D054299:env Gene Products, Human Immunodeficiency Virus