Establishment of pharmacological evaluation system for non-nucleoside reverse-transcriptase inhibitors resistant HIV-1

Consistent non-nucleoside reverse-transcriptase inhibitors (NNRTIs) resistant HIV-1 strains    occurred due to the clinical use for more than ten years of efavirenz (EFV), nevirapine (NVP), and delavirdine (DLV).  In this study, we established nine cell-based pharmacological models according to most NNRTIs-resistant clinical tested strains.  Resistant mutations were introduced into vector, pNL4-3.Luc.R^-E^-, by overlapping PCR.  Then, pseudovirions were produced by co-transfection of VSV-G plasmid and pNL4-3.Luc.R^-E^--mut.  All nine recombinant VSVG/HIV-mut pseudovirions (VSVG/HIV-wt, VSVG/HIV_-K103N, VSVG/HIV_-Y181C, VSVG/ HIV_-L100I,K103N, VSVG/HIV_-Y188L, VSVG/HIV_-K103N,Y181C, VSVG/HIV_-K103N,P225H, VSVG/HIV_-K103N,Y188L, VSVG/ HIV_-K103N,G109A and  
VSVG/HIV_-K103N,V108I) had high efficient infectivity.  Furthermore, they all showed resistant characteristics to EFV and NVP with IC₅₀ changes consisting with clinical reports, not to nucleoside  reverse-transcriptase inhibitors (AZT and d4T).  This series safe cell-based model, which could be carried out in BSL-2 laboratory, can be used for evaluating NNRTIs candidates.