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

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

doi:10.3791/57554

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

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

^*Corresponding author for this work

Department of Clinical Medicine

2 Citations (Scopus)

29 Downloads (Pure)

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.

Original language	English
Article number	e57554
Journal	Journal of Visualized Experiments
Volume	2018
Issue number	138
Number of pages	8
ISSN	1940-087X
DOIs	https://doi.org/10.3791/57554
Publication status	Published - 2018

Keywords

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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.3791/57554Licence: CC BY-ND

A Suction Blister Protocol to Study Human T-cell Recall Responses In VivoFinal published version, 406 KBLicence: CC BY-ND

Cite this

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title = "A suction blister protocol to study human Tk-cell recall responses in vivo",

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-Gu{\'e}rin 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.",

keywords = "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",

author = "Holm, {Line L.} and Milica Vukmanovic-Stejic and Thomas Blauenfeldt and Thomas Benfield and Peter Andersen and Akbar, {Arne N.} and Morten Ruhwald",

year = "2018",

doi = "10.3791/57554",

language = "English",

volume = "2018",

journal = "Journal of Visualized Experiments",

issn = "1940-087X",

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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 -

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

Abstract

Keywords

UN SDGs

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