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A whole blood intracellular cytokine assay optimised for field site studies demonstrates polyfunctionality of CD4+ T cells in acute scrub typhus

Inthawong, M. and Pinthong, N. and Thaiprakhong, A. and Wangrangsimakul, T. and Sunyakumthorn, P. and Hill, J. and Sonthayanon, P. and Paris, D. H. and Dunachie, S. J. and Kronsteiner, B.. (2023) A whole blood intracellular cytokine assay optimised for field site studies demonstrates polyfunctionality of CD4+ T cells in acute scrub typhus. PLoS Negl Trop Dis, 17 (3). e0010905.

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Abstract

BACKGROUND: Assessment of cellular immune responses by combining intracellular cytokine staining and immunophenotyping using flow cytometry enables the simultaneous measurement of T cell phenotype and effector function in response to pathogens and vaccines. The use of whole blood samples rather than peripheral blood mononuclear cells avoids both the need for immediate processing and loss of functional antigen presenting cells due to processing and cryopreservation. Using whole blood provides the possibility to stimulate peripheral T cells in situ, and is more suitable for studies where sample volume is limited, such as those involving children, the elderly and critically ill patients. The aim of this study was to provide a robust tool for the assessment of antigen-specific T cell responses in a field site setting with limited resources. METHODOLOGY/PRINCIPLE FINDINGS: We optimised a flow cytometry-based whole blood intracellular cytokine assay (WBA) with respect to duration of antigen stimulation and intracellular protein retention time. We demonstrate the ability of the WBA to capture polyfunctional T cell responses in the context of acute scrub typhus infection, by measuring IFN-gamma, TNF and IL-2 in CD4+ and CD8+ T cells in response to the causative agent O. tsutsugamushi (OT). Using an optimised OT antigen preparation, we demonstrate the presence of polyfunctional antigen-specific memory CD4+ T cells in the blood of scrub typhus patients. CONCLUSIONS/SIGNIFICANCE: In conclusion, this flow cytometry-based WBA is well-suited for use at field study sites, and enables the assessment of polyfunctional T cell responses to infectious agents and vaccines through delineation of antigen-specific cytokine secretion at the single cell level.
Faculties and Departments:09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH)
03 Faculty of Medicine > Departement Public Health > Sozial- und Präventivmedizin > Medicines Development (Paris)
09 Associated Institutions > Swiss Tropical and Public Health Institute (Swiss TPH) > Department of Medicine (MED) > Medicines Development (Paris)
UniBasel Contributors:Paris, Daniel Henry
Item Type:Article, refereed
Article Subtype:Research Article
ISSN:1935-2735 (Electronic)1935-2727 (Linking)
Note:Publication type according to Uni Basel Research Database: Journal article
Language:English
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Last Modified:09 May 2023 06:32
Deposited On:09 May 2023 06:32

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