A suction blister protocol to study human Tk-cell recall responses in vivo

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Documents

  • Line L. Holm
  • Milica Vukmanovic-Stejic
  • Thomas Blauenfeldt
  • Benfield, Thomas
  • Peter Andersen
  • Arne N. Akbar
  • Morten Ruhwald

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.

Original languageEnglish
Article numbere57554
JournalJournal of Visualized Experiments
Volume2018
Issue number138
Number of pages8
ISSN1940-087X
DOIs
Publication statusPublished - 2018

    Research areas

  • Antigen, Immunology, Immunology and infection, In vivo response, Issue 138, Memory T-cell, PPD, Recall, Skin challenge, Suction blister, T-cell, Tuberculin skin test, Tuberculosis, Vaccines

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