Establishment of a new screening method for anti-HIV-1 drugs / 中华临床感染病杂志

ABSTRACT

Objective To establish and assess a new screening method for anti-HIV-1 drugs.Methods JLTRG cells were co-cultured with different proportions of H9/HTLV-Ⅲ B cells for 24,48,72 and 96 h.Intensity and density of green fluorescent protein were observed under fluorescence microscope,and were tested using flow cytometry.The optimal proportion of cells in co-culture system and the culture time were determined.The effectiveness of Enfuvirtide (T20) and Efavirenz (EFV),and their half maximal inhibitory concentrations (IC50) were determined by using cell co-culture system and half life of drugs.HIV load was detected using RT-PCR for HIV-1 p24 antigen,and its correlations with drug concentration and mean fluorescent intensity were analyzed by Spearman rank correlation analysis.Results Experiments demonstrated that JLTRG cells co-cultured with H9/HTLV-ⅢB cells at the proportion of 10 ∶ 1 for 72 hours was the best.Along with the concentrations of T20 and EFV changed,JLTRG cells were infected with HIV-1 in different degrees,and the IC50s of T20 and EFV were 10 nmol/L and 5 nmol/L,respectively.The concentrations of T20 and EFV were negatively correlated with mean fluorescent intensity and viral load (r =-1,-0.986 and-1,-1,P ＜ 0.01); and mean fluorescent intensity was positively correlated with viral load (r =0.986 and 1,P ＜ 0.01).Conclusion The drug screening method established in this study is efficient and easy to operate,which provides a new option for anti-HIV-1 drug screening.