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

TY - JOUR

T1 - A suction blister protocol to study human Tk-cell recall responses in vivo

AU - Holm, Line L.

AU - Vukmanovic-Stejic, Milica

AU - Blauenfeldt, Thomas

AU - Benfield, Thomas

AU - Andersen, Peter

AU - Akbar, Arne N.

AU - Ruhwald, Morten

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

KW - Antigen

KW - Immunology

KW - Immunology and infection

KW - In vivo response

KW - Issue 138

KW - Memory T-cell

KW - PPD

KW - Recall

KW - Skin challenge

KW - Suction blister

KW - T-cell

KW - Tuberculin skin test

KW - Tuberculosis

KW - Vaccines

U2 - 10.3791/57554

DO - 10.3791/57554

M3 - Journal article

C2 - 30148487

AN - SCOPUS:85054552242

SN - 1940-087X

VL - 2018

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

IS - 138

M1 - e57554

ER -