Testing antiretroviral drug efficacy in conventional mice infected with chimeric HIV-1.
OBJECTIVE
We previously described chimeric HIV-1, EcoHIV, which can infect mouse cells in culture and cause spreading infection in conventional immunocompetant mice. We have now applied this system as a model for preclinical evaluation of anti-retroviral drugs.


METHODS
We used chimeric virus EcoHIV/NDK constructed on the backbone of subtype D NDK. EcoHIV/NDK expression in mice was characterized 5-10 days after infection by testing viral DNA, RNA, and protein burdens in spleen and macrophages by real-time PCR (QPCR), RT-PCR, and p24 ELISA. For antiviral evaluation, groups of 5-7 mice were pretreated with 2',3'-dideoxycytidine (ddC), abacavir, or vehicle; mice were then infected with EcoHIV/NDK, treatment maintained for additional 48 h, and tested for viral DNA and RNA burdens in spleens and macrophages by QPCR.


RESULTS
EcoHIV/NDK infected mice reproducibly showed viral burdens of up to 1.4 x 10 viral DNA copies and 200 pg p24 per 10 spleen cells and expressed spliced Vif RNA and mature p24 in macrophages 5-10 days after infection. Treatment of mice with 60 or 300 mg ddC/kg/day blocked EcoHIV/NDK infection in a dose-dependent manner with significantly lower viral DNA and RNA burdens at both drug doses (P < 0.001) in the spleens of infected mice. Abacavir tested at 100 mg/kg/day caused 96% inhibition of viral DNA synthesis in spleen and it almost completely abolished viral spliced RNA synthesis in spleens and macrophages.


CONCLUSIONS
The system of chimeric HIV-1 infection of mice permits rapid, statistically powerful, and inexpensive evaluation of antiretroviral drugs in vivo.
Hadas
Eran
E
Molecular Virology Division, St. Luke's-Roosevelt Hospital Center, Columbia University Medical Center, New York 10019, USA.
Borjabad
Alejandra
A
Chao
Wei
W
Saini
Manisha
M
Ichiyama
Kozi
K
Potash
Mary Jane
MJ
Volsky
David J
DJ
D000818:Animals; D019380:Anti-HIV Agents / Q000627:therapeutic use*; D002678:Chimera; D004279:DNA, Viral / Q000032:analysis; D015224:Dideoxynucleosides / Q000627:therapeutic use; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015658:HIV Infections / Q000188:drug therapy* / Q000517:prevention & control / Q000821:virology; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D017737:Macrophages, Peritoneal / Q000821:virology; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D013154:Spleen / Q000821:virology; D019562:Viral Load; D016047:Zalcitabine / Q000627:therapeutic use; D000099227:Abacavir
D019380:Anti-HIV Agents; D004279:DNA, Viral; D015224:Dideoxynucleosides; D016047:Zalcitabine; D000099227:Abacavir