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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • BCG Vaccine / pharmacology
  • BCG Vaccine / therapeutic use*
  • Blister / etiology*
  • Humans
  • Immunity, Cellular / immunology*
  • Mycobacterium tuberculosis / pathogenicity*
  • Skin / immunology*
  • T-Lymphocytes / immunology*
  • Tuberculosis / diagnosis*
  • Tuberculosis / immunology

Substances

  • BCG Vaccine