Analysis of simian immunodeficiency virus-specific CD8+ T-cells in rhesus macaques by peptide-MHC-I tetramer staining

Lucas Gonzalez-Nieto, Aline Domingues, Michael Ricciardi, Martin J. Gutman, Helen S. Maxwell, Nuria Pedreño-Lopez, Varian Bailey, Diogo M. Magnani, Mauricio Martins

Research output: Contribution to journalArticle

10 Scopus citations

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.

Original languageEnglish (US)
Article numbere54881
JournalJournal of Visualized Experiments
Volume2016
Issue number118
DOIs
StatePublished - Dec 23 2016

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Keywords

  • Immunology
  • Issue 118
  • Macaque
  • MHC
  • Rhesus
  • SIV
  • Tetramer
  • Vaccine

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Gonzalez-Nieto, L., Domingues, A., Ricciardi, M., Gutman, M. J., Maxwell, H. S., Pedreño-Lopez, N., Bailey, V., Magnani, D. M., & Martins, M. (2016). Analysis of simian immunodeficiency virus-specific CD8+ T-cells in rhesus macaques by peptide-MHC-I tetramer staining. Journal of Visualized Experiments, 2016(118), [e54881]. https://doi.org/10.3791/54881