Heterogeneous loss of HIV transcription and proviral DNA from 8E5/LAV lymphoblastic leukemia cells revealed by RNA FISH:FLOW analyses

Retrovirology. 2016 Aug 11;13(1):55. doi: 10.1186/s12977-016-0289-2.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line, Tumor
  • DNA, Viral / analysis
  • DNA, Viral / genetics*
  • Flow Cytometry
  • Genome, Viral
  • HIV-1 / genetics*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma
  • Proviruses / genetics*
  • RNA, Viral / analysis*
  • Real-Time Polymerase Chain Reaction
  • Transcription Factors / metabolism*
  • Transcription, Genetic*


  • DNA, Viral
  • RNA, Viral
  • Transcription Factors