https://www.selleckchem.com/products/bgj398-nvp-bgj398.html We will also discuss how to reinforce the M1-polarization protocol to obtain a reliable model of reversible latency of infectious HIV-1 in primary M1-MDM.Monocytes/macrophages play critical roles in HIV transmission, viral spread (early in infection), and as a reservoir of virus throughout infection. In the current research area in HIV, there has been a recent resurgence of interest in the biology of monocyte subsets and macrophages and their role in HIV pathogenesis, and as long-lived HIV reservoir. Thus, sensitive and specific techniques are needed to measure the impact of these cells in the establishment of the "hard-core" reservoir, and in their capacity to cause a low-level virus production during cART. Here, a protocol is presented for cell culture and HIV-1 infection of granulocyte-macrophage colony-stimulating factor (GM-CSF) differentiated human monocyte-derived macrophages.Antiretroviral therapy (ART) has transformed the deadly human immunodeficiency virus type I (HIV-1) epidemic into a manageable chronic condition. Current ART is not curative and treatment interruption leads to viral rebound in people living with HIV-1 (PLWH). The main cause of viral rebound is the persistence of HIV reservoirs in long-lived memory CD4+ T cells. Accurate techniques to identify and quantify viral reservoirs are required to monitor therapeutic approaches designed to cure HIV infection. Th17-polarized CD4+ T cells are located at mucosal sites of HIV entry and are preferentially targeted for infection and viral reservoir persistence. They constitute an important reservoir in both blood and colon. In this chapter we describe a step-by-step flow cytometry-based protocol to isolate a fraction of Th17-enriched cells from PBMC based on their expression of the Th17 surface marker CCR6. The isolation of memory CCR6+CD4+ T cells allows subsequent PCR/RT-PCR-based HIV DNA/RNA quantifications, as well as their culture for quan