A Murine Viral Outgrowth Assay to Detect Residual HIV Type 1 in Patients With Undetectable Viral Loads

• Published in: The Journal of infectious diseases Vol. 212; no. 9; pp. 1387 - 1396
• Main Authors: Pate, Kelly A. Metcalf, Pohlmeyer, Christopher W., Walker-Sperling, Victoria E., Foote, Jeremy B., Najarro, Kevin M., Cryer, Catherine G., Salgado, Maria, Gama, Lucio, Engle, Elizabeth L., Shirk, Erin N., Queen, Suzanne E., Chioma, Stanley, Vermillion, Meghan S., Bullock, Brandon, Li, Ming, Lyons, Claire E., Adams, Robert J., Zink, M. Christine, Clements, Janice E., Mankowski, Joseph L., Blankson, Joel N.
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01.11.2015
• Subjects: Animals; Antiretroviral Therapy, Highly Active; CD4-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - virology; Disease Models, Animal; HIV Infections - diagnosis; HIV Infections - drug therapy; HIV-1 - growth & development; HIV-1 - isolation & purification; HIV/AIDS; Humans; Interleukin-2 - blood; Leukocytes, Mononuclear - virology; Macaca; Major and Brief Reports; Male; Mice; Simian immunodeficiency virus; Simian Immunodeficiency Virus - growth & development; Simian Immunodeficiency Virus - isolation & purification; Swine influenza virus; Viral Load; Viremia - veterinary; SIV; humanized mouse; cure; HIV; quantitative viral outgrowth assay (QVOA)
• ISSN: 0022-1899; 1537-6613
• DOI: 10.1093/infdis/jiv230

Background. Sensitive assays are needed for detection of residual human immunodeficiency virus (HIV) in patients with undetectable plasma viral loads to determine whether eradication strategies are effective. The gold standard quantitative viral outgrowth assay (QVOA) underestimates the magnitude of the viral reservoir. We sought to determine whether xenograft of leukocytes from HIV type 1 (HIV)-infected patients with undetectable plasma viral loads into immunocompromised mice would result in viral amplification. Methods. Peripheral blood mononuclear cells or purified CD4⁺ T cells from HIV or simian immunodeficiency virus (SIV)-infected subjects with undetectable plasma viral loads were adoptively transferred into NOD.Cg-PrkdcscidIl2rgtmlWjl/SzJ (NSG) mice. The mice were monitored for viremia following depletion of human CD8⁺ T cells to minimize antiviral activity. In some cases, humanized mice were also treated with activating anti-CD3 antibody. Results. With this murine viral outgrowth assay (MVOA), we successfully amplified replication-competent HIV or SIV from all subjects tested, including 5 HIV-positive patients receiving suppressive antiretroviral therapy (ART) and 6 elite controllers or suppressors who were maintaining undetectable viral loads without ART, including an elite suppressor from whom we were unable to recover virus by QVOA. Conclusions. Our results suggest that the MVOA has the potential to serve as a powerful tool to identify residual HIV in patients with undetectable viral loads.