Construction of an infectious HIV type 1 molecular clone from an African patient with a subtype D/C Recombinant Virus.
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.
Shi
Binshan
B
Wadsworth Center, New York State Department of Health, Albany, New York 12208, USA.
Philpott
Sean M
SM
Weiser
Barbara
B
Kuiken
Carla
C
Brunner
Cheryl
C
Fang
Guowei
G
Fowke
Keith R
KR
Plummer
Francis A
FA
Rowland-Jones
Sarah
S
Bwayo
Job
J
Anzala
Aggrey O
AO
Kimani
Joshua
J
Kaul
Rupert
R
Burger
Harold
H
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
D004279:DNA, Viral