A suction blister protocol to study human Tk-cell recall responses in vivo
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A suction blister protocol to study human Tk-cell recall responses in vivo. / Holm, Line L.; Vukmanovic-Stejic, Milica; Blauenfeldt, Thomas; Benfield, Thomas; Andersen, Peter; Akbar, Arne N.; Ruhwald, Morten.
I: Journal of Visualized Experiments, Bind 2018, Nr. 138, e57554, 2018.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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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
VL - 2018
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
SN - 1940-087X
IS - 138
M1 - e57554
ER -
ID: 214760368