8E5/LAV cells harbor a single HIV provirus, and are used frequently to generate standards for HIV genome quantification. Using flow cytometry-based in situ mRNA hybridization validated by qPCR, we find that different batches of 8E5 cells contain varying numbers of cells lacking viral mRNA and/or viral genomes. These findings raise concerns for studies employing 8E5 cells for quantitation, and highlight the value of mRNA FISH and flow cytometry in the detection and enumeration of HIV-positive cells.
Keywords: 8E5; FISH:FACS; FISH:FLOW; HIV RNA in situ hybridization; HIV cell-associated RNA; HIV flow cytometry assay; HIV proviral loss; HIV proviral silencing; Reservoir quantitation.