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Inhibition of HIV-1 replication by anti-integrase single chain Fv antibody generated in situ

ZHANG Hui zhong1, ZHU Ming hua2, DUAN Ling xun3 1Department of Orthopeadics, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038,Shaanxi Province, China 2Department of Pathology, Second Military Medical University, Shanghai,China 3Thomas  
Aim To evaluate the therapeutic effects of anti HIV 1 integrase sFv(IN sFv) on the inhibition of HIV 1 replication. Methods Challenge experiments, utilizing the highly cytopathic viral strain NL4 3, were conducted with SupT1 cells and peripheral blood mononuclear cells(PBMCs) stably transduced with anti HIV 1 IN sFv gene. The spread of HIV 1 in the cultures was determined by quantitating the levels of HIV 1 p24 antigen released into the culture medium. To further confirm the mechanism of HIV 1 replication inhibition, semiquantitative 2 LTR PCR with nested primers was performed to amplify HIV 1 circled 2 LTR DNA at different time points after the anti HIV 1 IN sFv transduced and non transduced SupT1 cells were infected with NL4 3 virus at a high MOIs of 2.0. Results The Challenge results showed approximately 95%to 98%inhibition of HIV 1 p24 antigen generation in the cells transduced with IN sFv and IN sFv nls compared with the control cells. The 2 LTR PCR results demonstrated that the detectable levels of HIV 1 2 LTR DNA molecules occurred significantly earlier in cells expressing anti HIV 1 IN sFv than in the control cells. Conclusion The present data suggest that the IN sFv can significantly inhibit HIV 1 replication via the blocking of HIV 1 integration and is therefore a promising therapeutic agent for gene therapy of the HIV 1 infection.
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