title: Construction of an infectious HIV type 1 molecular clone from an African patient with a subtype D/C Recombinant Virus.
abstract: The majority of HIV-1 infections worldwide occur in Africa, where subtype B viruses are rare and intersubtype recombinants are common. Pathogenesis and vaccine studies need to focus on viruses derived from African patients, and infectious HIV-1 molecular clones can be useful tools. To clone non-B subtypes and recombinant viruses from patients, we cultivated HIV-1 from the plasma of a Kenyan long-term survivor. Viral DNA was cloned into a plasmid, which was transfected into COS cells; progeny virus was propagated in PBMCs. Sequence analyses revealed that both the patient's plasma HIV-1 RNA and the cloned DNA genomes were recombinants between subtypes D and C; subtype C sequences comprised the nef and LTR regions. The cloned virus used the CCR5 coreceptor and did not form syncytia in vitro. This infectious HIV-1 subtype D/C recombinant molecular clone obtained from a Kenyan long-term survivor promises to be useful to study pathogenesis and vaccine design.
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, New York 12208, USA.
authors.forename: Binshan
authors.initials: B
authors.lastname: Shi
authors.forename: Sean M
authors.initials: SM
authors.lastname: Philpott
authors.forename: Barbara
authors.initials: B
authors.lastname: Weiser
authors.forename: Carla
authors.initials: C
authors.lastname: Kuiken
authors.forename: Cheryl
authors.initials: C
authors.lastname: Brunner
authors.forename: Guowei
authors.initials: G
authors.lastname: Fang
authors.forename: Keith R
authors.initials: KR
authors.lastname: Fowke
authors.forename: Francis A
authors.initials: FA
authors.lastname: Plummer
authors.forename: Sarah
authors.initials: S
authors.lastname: Rowland-Jones
authors.forename: Job
authors.initials: J
authors.lastname: Bwayo
authors.forename: Aggrey O
authors.initials: AO
authors.lastname: Anzala
authors.forename: Joshua
authors.initials: J
authors.lastname: Kimani
authors.forename: Rupert
authors.initials: R
authors.lastname: Kaul
authors.forename: Harold
authors.initials: H
authors.lastname: Burger
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005260:Female; D020276:HIV Long-Term Survivors; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity*; D006801:Humans; D007630:Kenya; D008969:Molecular Sequence Data; D010957:Plasmids; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D014162:Transfection
chemical_list: D004279:DNA, Viral