Construction of a macrophage-tropic subtype C HIV-1 mGreenLantern reporter virus for studies on HIV-1 replication and the impact of methamphetamine

bioRxiv [Preprint]. 2024 Oct 24:2024.10.24.619504. doi: 10.1101/2024.10.24.619504.

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.

Publication types

  • Preprint