Deep phenotyping characterization of human unconventional CD8+NKG2A/C+ T cells among T and NK cells by spectral flow cytometry

Aurelio Orta-Resendiz; Caroline Petitdemange; Sandrine Schmutz; Béatrice Jacquelin; Sophie Novault; Nicolas Huot; Michaela Müller-Trutwin

doi:10.1016/j.xpro.2023.102734

Deep phenotyping characterization of human unconventional CD8⁺NKG2A/C⁺ T cells among T and NK cells by spectral flow cytometry

STAR Protoc. 2023 Dec 15;4(4):102734. doi: 10.1016/j.xpro.2023.102734. Epub 2023 Nov 29.

Authors

Aurelio Orta-Resendiz¹, Caroline Petitdemange², Sandrine Schmutz³, Béatrice Jacquelin², Sophie Novault⁴, Nicolas Huot², Michaela Müller-Trutwin⁵

Affiliations

¹ Institut Pasteur, Université Paris Cité, HIV, Inflammation and Persistence Unit, 75015 Paris, France. Electronic address: [email protected].
² Institut Pasteur, Université Paris Cité, HIV, Inflammation and Persistence Unit, 75015 Paris, France.
³ Cytometry and Biomarkers, Center for Technological Resources and Research, Institut Pasteur, 75015 Paris, France. Electronic address: [email protected].
⁴ Cytometry and Biomarkers, Center for Technological Resources and Research, Institut Pasteur, 75015 Paris, France.
⁵ Institut Pasteur, Université Paris Cité, HIV, Inflammation and Persistence Unit, 75015 Paris, France. Electronic address: [email protected].

Abstract

Here, we present a protocol for setting three spectral flow cytometry panels for the characterization of human unconventional CD8⁺NKG2A/C⁺ T cells as well as other T and natural killer cell subsets. We describe steps for standardizing, preparing, and staining the cells, the experimental setup, and the final data analysis. This protocol should be advantageous in various settings including immunophenotyping of limited samples, immune function evaluation/monitoring, as well as research in oncology, autoimmune, and infectious diseases.

Keywords: Antibody; Flow Cytometry; Immunology.

MeSH terms

CD8-Positive T-Lymphocytes
Flow Cytometry / methods
Humans
Immunophenotyping
Killer Cells, Natural*
T-Lymphocytes*