title: Defining HIV and SIV Reservoirs in Lymphoid Tissues.
abstract: A primary obstacle to an HIV-1 cure is long-lived viral reservoirs, which must be eliminated or greatly reduced. Cure strategies have largely focused on monitoring changes in T cell reservoirs in peripheral blood (PB), even though the lymphoid tissues (LT) are primary sites for viral persistence. To track and discriminate viral reservoirs within tissue compartments we developed a specific and sensitive next-generationin situhybridization approach to detect vRNA, including vRNA+ cells and viral particles ("RNAscope"), vDNA+ cells ("DNAscope") and combined vRNA and vDNA with immunohistochemistry to detect and phenotype active and latently infected cells in the same tissue section. RNAscope is highly sensitive with greater speed of analysis compared to traditionalin situhybridization. The highly sensitive and specific DNAscope detected SIV/HIV vDNA+ cells, including duplexed detection of vDNA and vRNA or immunophenotypic markers in the same section. Analysis of LT samples from macaques prior to and during combination antiretroviral therapy demonstrated that B cell follicles are an important anatomical compartment for both latent and active viral persistence during treatment. These new tools should allow new insights into viral reservoir biology and evaluation of cure strategies.
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Claire
authors.initials: C
authors.lastname: Deleage
authors.affiliation: Department of Microbiology and Immunology, Medical School, University of Minnesota, Minneapolis, MN 55455.
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Wietgrefe
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Gregory
authors.initials: G
authors.lastname: Del Prete
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: David R
authors.initials: DR
authors.lastname: Morcock
authors.affiliation: Pathology and Histotechnology Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Xing Pei
authors.initials: XP
authors.lastname: Hao
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Michael
authors.initials: M
authors.lastname: Piatak
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Julian
authors.initials: J
authors.lastname: Bess
authors.affiliation: Department of Medicine. Medical School, University of Minnesota, Minneapolis, MN 55455.
authors.forename: Jodi L
authors.initials: JL
authors.lastname: Anderson
authors.affiliation: Department of Microbiology and Immunology, Medical School, University of Minnesota, Minneapolis, MN 55455.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Perkey
authors.affiliation: School of Public Health, Division of Biostatistics, University of Minnesota, Minneapolis, MN 55455.
authors.forename: Cavan
authors.initials: C
authors.lastname: Reilly
authors.affiliation: Division of Experimental Medicine, Department of Medicine, University of California, San Francisco, CA 94110.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: McCune
authors.affiliation: Department of Microbiology and Immunology, Medical School, University of Minnesota, Minneapolis, MN 55455.
authors.forename: Ashley T
authors.initials: AT
authors.lastname: Haase
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Lifson
authors.affiliation: Department of Medicine. Medical School, University of Minnesota, Minneapolis, MN 55455.
authors.forename: Timothy W
authors.initials: TW
authors.lastname: Schacker
authors.affiliation: AIDS and Cancer Virus Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702.
authors.forename: Jacob D
authors.initials: JD
authors.lastname: Estes
keywords: B cell follicles; DNAscope; HIV; RNAscope; SIV; follicular dendritic cells; in situ hybridization; reservoir