title: Enhanced and efficient detection of virus-driven cytokine expression by human NK and T cells.
abstract: Cutting edge immune monitoring techniques increasingly measure multiple functional outputs for various cell types, such as intracellular cytokine staining (ICS) assays that measure cytokines expressed by T cells. To date, however, there is no precise method to measure virus-specific cytokine production by both T cells as well as NK cells in the same well, which is important to a greater extent given recent identification of NK cells expressing a memory phenotype. This study describes an adaptable and efficient ICS assay platform that can be used to detect antigen-driven cytokine production by human T cells and NK cells, termed "viral ICS". Importantly, this assay uses limited amount of cryopreserved PBMCs along with autologous heat-inactivated serum, thereby allowing for this assay to be performed when sample is scarce as well as geographically distant from the laboratory. Compared to a standard ICS assay that detects antigen-specific T cell cytokine expression alone, the viral ICS assay is comparable in terms of both HIV-specific CD4 and CD8T cell cytokine response rates and magnitude of response, with the added advantage of ability to detect virus-specific NK cell responses.
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Pattacini
authors.affiliation: Department of Global Health, University of Washington, Seattle, WA 98195, USA; Department of Epidemiology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Pamela M
authors.initials: PM
authors.lastname: Murnane
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
authors.forename: Tayler R
authors.initials: TR
authors.lastname: Fluharty
authors.affiliation: Infectious Disease Institute, Makerere University, Kampala, Uganda.
authors.forename: Elly
authors.initials: E
authors.lastname: Katabira
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
authors.forename: Stephen C
authors.initials: SC
authors.lastname: De Rosa
authors.affiliation: Department of Global Health, University of Washington, Seattle, WA 98195, USA; Department of Epidemiology, University of Washington, Seattle, WA 98195, USA; Department of Medicine, University of Washington, Seattle, WA 98195, USA.
authors.forename: Jared M
authors.initials: JM
authors.lastname: Baeten
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA; Department of Global Health, University of Washington, Seattle, WA 98195, USA. Electronic address: jlund@fhcrc.org.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Lund
mesh_terms: D016207:Cytokines / Q000096:biosynthesis*; D005434:Flow Cytometry / Q000379:methods*; D006678:HIV / Q000276:immunology*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology* / Q000821:virology; D013194:Staining and Labeling / Q000379:methods*; D013601:T-Lymphocytes / Q000276:immunology* / Q000821:virology; D014773:Virology / Q000379:methods*
chemical_list: D016207:Cytokines
keywords: Cellular immunity; Flow cytometry; Immune monitoring; Intracellular cytokine staining; Viral infection