Abstract
HIV-1 subtype C viruses are responsible for 50% of global HIV burden. However, nearly all currently available reporter viruses widely used in HIV research are based on subtype B. We constructed and characterized a replication competent HIV-1 subtype C reporter virus expressing mGreenLantern. mGreenLantern sequences were inserted in-frame with nef ATG in HIV-1 (IndieC1) . As controls, we employed HIV-1 (IndieC1) , HIV-1 (ADA,) and HIV-1 (NLAD8-GFP-Nef) viruses. HIV-1 (IndieC1-mGreenLantern) (HIV-1 (IndieC1-mGL) ) exhibited characteristics of the parental HIV-1 (IndieC1) virus, including its infectivity in TZMbl reporter cells and replication competence in macrophages. To further characterize HIV-1 (IndieC1-mGL) virus, we tested its responsiveness to CCL2 levels, a characteristic feature of subtype B HIV-1 that is missing in subtype C. CCL2 immunodepletion inhibited the production of HIV-1 (ADA) and HIV-1 (NLAD8-GFP-Nef) as expected, but not that of HIV-1 (IndieC1-mGL) as previously reported. We also tested the effect of Methamphetamine, as its effect is mediated by NF-κB and since subtype C viruses carry an additional copy of NFκB. We found that methamphetamine increased the replication of all viruses tested in macrophages, however, its effect was much more robust for HIV-1 (IndieC1) and HIV-1 (IndieC1-mGL) . Our studies established that HIV-1 (IndieC1-mGL) retains all the characteristics of the parental HIV-1 (IndieC1) and can be a useful tool for HIV-1 subtype C investigations.