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J Vis Exp. 2018 Aug 11;(138). doi: 10.3791/57554.

A Suction Blister Protocol to Study Human T-cell Recall Responses In Vivo.

Author information

1
Department of Infectious Disease Immunology, Center for Vaccine Research, Statens Serum Institut; Department of Infectious Diseases, Hvidovre Hospital; Institute of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen; line.lindebo.holm@gmail.com.
2
Division of Infection and Immunity, University College London.
3
Department of Infectious Disease Immunology, Center for Vaccine Research, Statens Serum Institut.
4
Department of Infectious Diseases, Hvidovre Hospital; Institute of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen.

Abstract

Cutaneous antigen-recall models allow for studies of human memory responses in vivo. When combined with skin suction blister (SB) induction, this model offers accessibility to rare populations of antigen-specific T-cells representative of the cellular memory response as well as the cytokine microenvironment in situ. This report describes the practical procedure of a cutaneous recall, an SB induction, and a harvest of antigen-specific T-cells. To exemplify the method, the tuberculin skin test is used for antigenic recall in individuals who, prior to this study, underwent a Bacillus Calmette-Guérin vaccination against an infection with Mycobacterium tuberculosis. Finally, examples of multiplex and flow cytometric analyses of SB specimens are provided, illustrating high fractions of antigen-specific polyfunctional CD4+ T-cells available by this sampling method compared with cells isolated from the blood. The method described here is safe and minimally invasive, provides a unique opportunity to study both innate and adaptive immune responses in vivo, and may be beneficial to a broad community of researchers working with cell-mediated immunity and human memory responses, in the context of vaccine development.

PMID:
30148487
PMCID:
PMC6126709
DOI:
10.3791/57554
[Indexed for MEDLINE]
Free PMC Article

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