title: Improved flow-based method for HIV/SIV envelope-specific memory B-cell evaluation in rhesus macaques.
abstract: The ability to elicit potent and long-lasting broadly neutralizing HIV envelope (Env)-specific antibodies has become a key goal for HIV vaccine development. Consequently, the ability to rapidly and efficiently monitor development of memory B cells in pre-clinical and clinical vaccine trails is critical for continued progress in vaccine design. We have developed an improved flow cytometry-based method for the rapid and efficient identification of gp120-specific memory B cells in peripheral blood, bone marrow, and mucosal tissues which allows their direct staining without the need for prior cell sorting or enrichment. We demonstrate staining of both HIV and SIV Env-specific memory B cells in PBMC, bone marrow, and rectal tissue of vaccinated and infected rhesus macaques. Validation of the method is illustrated by statistically significant correlations with memory B cell levels quantified by ELISPOT assay and with serum binding antibody titers determined by ELISA. In addition to quantification, this method will bring the power of flow cytometry to the study of homing and trafficking of Env-specific memory B cells.
authors: Section on Immune Biology of Retroviral Infection, Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, United States.
authors: Venkatramanan
authors: V
authors: Mohanram
authors: Section on Immune Biology of Retroviral Infection, Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, United States.
authors: Thorsten
authors: T
authors: Demberg
authors: Section on Immune Biology of Retroviral Infection, Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, United States.
authors: Iskra
authors: I
authors: Tuero
authors: Section on Immune Biology of Retroviral Infection, Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, United States.
authors: Diego
authors: D
authors: Vargas-Inchaustegui
authors: Human Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD 21702, United States.
authors: George N
authors: GN
authors: Pavlakis
authors: Human Retrovirus Pathogenesis Section, Vaccine Branch, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD 21702, United States.
authors: Barbara K
authors: BK
authors: Felber
authors: Section on Immune Biology of Retroviral Infection, Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, United States. Electronic address: guroffm@mail.nih.gov.
authors: Marjorie
authors: M
authors: Robert-Guroff
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay; D005434:Flow Cytometry / Q000379:methods*; D006678:HIV / Q000276:immunology*; D006801:Humans; D007156:Immunologic Memory; D007413:Intestinal Mucosa / Q000276:immunology; D008253:Macaca mulatta; D016097:Simian Acquired Immunodeficiency Syndrome / Q000175:diagnosis* / Q000276:immunology; D015302:Simian Immunodeficiency Virus / Q000276:immunology*; D014611:Vaccination; D054299:env Gene Products, Human Immunodeficiency Virus / Q000276:immunology*
chemical_list: D016915:AIDS Vaccines; D000914:Antibodies, Viral; D054299:env Gene Products, Human Immunodeficiency Virus
keywords: Envelope-specific memory B cells; HIV; SIV