Characterization of the single cycle replication of HIV-1 expressing Gaussia luciferase in human PBMCs, macrophages, and in CD4+ T cell-grafted nude mouse

Noncovalent SUMO-interaction motifs in HIV integrase play important roles in SUMOylation, cofactor binding, and virus replication

“…A HIV-1 single-cycle replicating virus was produced in 293 T cells as described previously [45]. Briefly, 293 T cells were co-transfected with an RT/IN /deleted HIV-1 provirus Bru∆RI/Gluc and each CMV-Vpr-RT-IN (wt or SIM M2 and M3 mutant) expression plasmid.…”

“…To test the effect of the IN mutants on viral infection, equal amounts of single cycle replicating viruses (adjusted by virion-associated p24 levels) were used to infect C8166 T cells 2 h, then washed and cells were cultured at 37 °C. At different time points after infection, the supernatant from each sample supernatants were collected and used for Gaussia luciferase Assay and HIV-1 p24 assay as described previously [45].…”

Synthesis and Antiretroviral Activity of 6-Acetyl-coumarin Derivatives against HIV-1 Infection

“…The in vitro toxicity study of synthesized compounds was performed using Trypan Blue exclusion method [33] while the antiretroviral activity of synthesized compounds against HIV-1 infection was determined by measuring the Gaussia luciferase (Gluc) activity [34] . The present study indicated that synthesized compounds have potential of inhibiting the HIV-1 infection in low micromolar concentration.…”

“…Antiretroviral drug, raltegravir (RAL) was obtained from the NIH AIDS Research and Reference Reagent Program. HIV-1 HxBru-Gluc virus was generated by transfecting 293T cells with corresponding HIV-1 proviral DNAs, as described previously [34,43] .…”

“…Three days post-infection, supernatant were collected and the levels of virus infection were monitored by measuring the Gluc activity in the supernatants from each infected cell culture using a POLARstar OPTIMA microplate luminometer (BMG Labtech, Germany). Gluc activity measurement was used as biomarker to detect the HIV infection [34,44] .…”