title: Direct measurement of CD8+ T cell responses in macaques infected with simian immunodeficiency virus.
abstract: The simian immunodeficiency virus (SIV) macaque model system has been used extensively to study AIDS pathogenesis and to test candidate vaccines for their ability to protect against homologous or heterologous challenge with pathogenic SIV or SHIV. Recent studies suggest that stimulation of HIV-1-specific CTL responses is important for effective vaccination against HIV-1. While quantitative measurements of SIV-specific cytotoxic T lymphocyte (CTL) responses have been facilitated by the use of tetrameric peptide complexes, this technique is currently limited to the study of Mamu-A*01-positive rhesus macaques. Furthermore, very few SIV-specific CTL epitopes have been identified, and there is limited identification of other MHC alleles in macaques. In this study, cytokine flow cytometry (CFC) was used to quantify SIV-specific CD8+ antigen-reactive T cells in macaques infected with SIV. We found a strong correlation (r = 0.96, P < 0.001) between CD8+ antigen-reactive T cells stained with the Mamu-A*01 p11C, C-M tetramer and production of intracellular TNF-alpha in the CFC assay. Furthermore, the CFC assay was used to identify a novel SIV-specific CTL epitope in Envelope (SIV Env, a.a. 486-494, sequence AEVAELYRL). The use of the CFC assay facilitates the study of antigen-reactive T cell responses in SIV infection and vaccination.
authors.affiliation: Aaron Diamond AIDS Research Center, The Rockefeller University, 455 First Avenue, New York, New York, 10016, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Donahoe
authors.forename: W J
authors.initials: WJ
authors.lastname: Moretto
authors.forename: R V
authors.initials: RV
authors.lastname: Samuel
authors.forename: K J
authors.initials: KJ
authors.lastname: Metzner
authors.forename: P A
authors.initials: PA
authors.lastname: Marx
authors.forename: T
authors.initials: T
authors.lastname: Hanke
authors.forename: R I
authors.initials: RI
authors.lastname: Connor
authors.forename: D F
authors.initials: DF
authors.lastname: Nixon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology*; D002460:Cell Line; D002478:Cells, Cultured; D003601:Cytotoxicity Tests, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D007424:Intracellular Fluid / Q000276:immunology / Q000821:virology; D008253:Macaca mulatta; D008969:Molecular Sequence Data; D016097:Simian Acquired Immunodeficiency Syndrome / Q000276:immunology* / Q000821:virology*; D015302:Simian Immunodeficiency Virus / Q000276:immunology*; D013194:Staining and Labeling
chemical_list: D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; C110764:Mamu-A 01 antigen