title: Analysis of Simian Immunodeficiency Virus-specific CD8+ T-cells in Rhesus Macaques by Peptide-MHC-I Tetramer Staining.
abstract: Peptide-major histocompatibility complex class I (pMHC-I) tetramers have been an invaluable tool to study CD8+T-cell responses. Because these reagents directly bind to T-cell receptors on the surface of CD8+T-lymphocytes, fluorochrome-labeled pMHC-I tetramers enable the accurate detection of antigen (Ag)-specific CD8+T-cells without the need for in vitro re-stimulation. Moreover, when combined with multi-color flow cytometry, pMHC-I tetramer staining can reveal key aspects of Ag-specific CD8+T-cells, including differentiation stage, memory phenotype, and activation status. These types of analyses have been especially useful in the field of HIV immunology where CD8+T-cells can affect progression to AIDS. Experimental infection of rhesus macaques with simian immunodeficiency virus (SIV) provides an invaluable tool to study cellular immunity against the AIDS virus. As a result, considerable progress has been made in defining and characterizing T-cell responses in this animal model. Here we present an optimized protocol for enumerating SIV-specific CD8+ T-cells in rhesus macaques by pMHC-I tetramer staining. Our assay permits the simultaneous quantification and memory phenotyping of two pMHC-I tetramer+ CD8+T-cell populations per test, which might be useful for tracking SIV-specific CD8+T-cell responses generated by vaccination or SIV infection. Considering the relevance of nonhuman primates in biomedical research, this methodology is applicable for studying CD8+T-cell responses in multiple disease settings.
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Lucas
authors.initials: L
authors.lastname: Gonzalez-Nieto
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Aline
authors.initials: A
authors.lastname: Domingues
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Michael
authors.initials: M
authors.lastname: Ricciardi
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Gutman
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Helen S
authors.initials: HS
authors.lastname: Maxwell
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Nuria
authors.initials: N
authors.lastname: Pedreño-Lopez
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Varian
authors.initials: V
authors.lastname: Bailey
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Diogo M
authors.initials: DM
authors.lastname: Magnani
authors.affiliation: Department of Pathology, University of Miami; mmartins@med.miami.edu.
authors.forename: Mauricio A
authors.initials: MA
authors.lastname: Martins
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000821:virology*; D005434:Flow Cytometry; D015395:Histocompatibility Antigens Class I / Q000737:chemistry*; D016130:Immunophenotyping; D008253:Macaca mulatta; D010455:Peptides; D015302:Simian Immunodeficiency Virus; D013194:Staining and Labeling
chemical_list: D015395:Histocompatibility Antigens Class I; D010455:Peptides