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J Virol Methods. 2014 Apr;199:17-24. doi: 10.1016/j.jviromet.2014.01.003. Epub 2014 Jan 10.

Enhanced and efficient detection of virus-driven cytokine expression by human NK and T cells.

Author information

1
Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
2
Department of Global Health, University of Washington, Seattle, WA 98195, USA; Department of Epidemiology, University of Washington, Seattle, WA 98195, USA.
3
Infectious Disease Institute, Makerere University, Kampala, Uganda.
4
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.
5
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.

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.

KEYWORDS:

Cellular immunity; Flow cytometry; Immune monitoring; Intracellular cytokine staining; Viral infection

PMID:
24418500
PMCID:
PMC3951839
DOI:
10.1016/j.jviromet.2014.01.003
[Indexed for MEDLINE]
Free PMC Article

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