TZM-gfp cells: a tractable fluorescent tool for analysis of rare and early HIV-1 infection.
David W GludishSaikat BoliarShannon CaldwellDumizulu L TemboElizabeth T ChimbayoKondwani C JamboHenry C MwandumbaDavid G RussellPublished in: Scientific reports (2020)
Here we describe TZM-gfp, a novel HIV-1 reporter cell derived from the same parental clone JC.53, used previously to generate the widely-utilized indicator cell line TZM-bl. We re-engineered JC.53 cells to express GFP under regulation of HIV Tat and Rev. We characterize the new reporter cell line to show that TZM-gfp cells are equally susceptible to HIV infection, exhibit minimal background signal, and can report HIV infection in rare cells from a bulk population of experimentally-infected human monocyte-derived macrophages. We demonstrate the utility and sensitivity of the cells in detection of even a single HIV-positive macrophage by fluorescence-assisted correlative electron microscopy, using the GFP signal to guide imaging of HIV virions in primary co-culture. Finally, we used TZM-gfp cells for viral capture during co-culture with human peripheral blood mononuclear cells, showing that TZM-gfp can support outgrowth and analyses of patient-derived primary HIV-1 isolates.
Keyphrases
- antiretroviral therapy
- hiv positive
- induced apoptosis
- hiv infected
- human immunodeficiency virus
- men who have sex with men
- cell cycle arrest
- hiv testing
- hiv aids
- endothelial cells
- endoplasmic reticulum stress
- south africa
- stem cells
- mesenchymal stem cells
- signaling pathway
- oxidative stress
- bone marrow
- adipose tissue
- living cells
- single molecule
- peripheral blood
- fluorescent probe
- loop mediated isothermal amplification
- induced pluripotent stem cells