Activated integrins identify functional antigen-specific CD8+ T cells within minutes after antigen stimulation

Stoyan Dimitrov; Cécile Gouttefangeas; Luciana Besedovsky; Anja T.R. Jensen; P. Anoop Chandran; Elisa Rusch; Ramona Businger; Michael Schindler; Tanja Lange; Jan Born; Hans Georg Rammensee

doi:10.1073/pnas.1720714115

Activated integrins identify functional antigen-specific CD8⁺ T cells within minutes after antigen stimulation

Stoyan Dimitrov^*, Cécile Gouttefangeas, Luciana Besedovsky, Anja T.R. Jensen, P. Anoop Chandran, Elisa Rusch, Ramona Businger, Michael Schindler, Tanja Lange, Jan Born, Hans Georg Rammensee

^*Corresponding author for this work

22 Citations (Scopus)

Abstract

Immediate β₂-integrin activation upon T cell receptor stimulation is critical for effective interaction between T cells and their targets and may therefore be used for the rapid identification and isolation of functional T cells. We present a simple and sensitive flow cytometry-based assay to assess antigen-specific T cells using fluorescent intercellular adhesion molecule (ICAM)-1 multimers that specifically bind to activated β₂-integrins. The method is compatible with surface and intracellular staining; it is applicable for monitoring of a broad range of virus-, tumor-, and vaccine-specific CD8⁺ T cells, and for isolating viable antigen-reacting cells. ICAM-1 binding correlateswith peptide-MHCmultimer binding but, notably, it identifies the fraction of antigen-specific CD8⁺ T cells with immediate and high functional capability (i.e., expressing high levels of cytotoxic markers and cytokines). Compared with the currently available methods, staining of activated β₂-integrins presents the unique advantage of requiring activation times of only several minutes, therefore delivering functional information nearly reflecting the in vivo situation. Hence, the ICAM-1 assay is most suitable for rapid and precise monitoring of functional antigen-specific T cell responses, including for patient samples in a variety of clinical settings, as well as for the isolation of functional T cells for adoptive cell-transfer immunotherapies.

Original language	English
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	115
Issue number	24
Pages (from-to)	E5536-E5545
ISSN	0027-8424
DOIs	https://doi.org/10.1073/pnas.1720714115
Publication status	Published - 12.06.2018

Funding

ACKNOWLEDGMENTS. We thank S. Stevanovićfor providing synthetic peptides; S. Heidu, J. Lehnholz, K. Witte, and E.-M. Schmidt for excellent technical assistance and advice; M. Szczepanski, J. C. P. Santiago, M. Esen, M. Buhl, and G. Marasca for sampling blood from the subjects and patients; all subjects and patients who participated in this study; and M. Hallschmid and R. Littwin for critical reading and proofreading of the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft Grants SFB 654 (to T.L. and H.-G.R.) and SFB 685 (to C.G. and H.-G.R.); grants from the German Federal Ministry of Education and Research (BMBF) to the German Center for Diabetes Research (DZD; 01GI0925) (to S.D. and J.B.); and European Research Council Grant AdG 339842, MUTAEDITING (to H.-G.R.).

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

UN SDGs

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

Access to Document

10.1073/pnas.1720714115

Cite this

Dimitrov, S., Gouttefangeas, C., Besedovsky, L., Jensen, A. T. R., Chandran, P. A., Rusch, E., Businger, R., Schindler, M., Lange, T., Born, J., & Rammensee, H. G. (2018). Activated integrins identify functional antigen-specific CD8⁺ T cells within minutes after antigen stimulation. Proceedings of the National Academy of Sciences of the United States of America, 115(24), E5536-E5545. https://doi.org/10.1073/pnas.1720714115

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title = "Activated integrins identify functional antigen-specific CD8+ T cells within minutes after antigen stimulation",

abstract = "Immediate β2-integrin activation upon T cell receptor stimulation is critical for effective interaction between T cells and their targets and may therefore be used for the rapid identification and isolation of functional T cells. We present a simple and sensitive flow cytometry-based assay to assess antigen-specific T cells using fluorescent intercellular adhesion molecule (ICAM)-1 multimers that specifically bind to activated β2-integrins. The method is compatible with surface and intracellular staining; it is applicable for monitoring of a broad range of virus-, tumor-, and vaccine-specific CD8+ T cells, and for isolating viable antigen-reacting cells. ICAM-1 binding correlateswith peptide-MHCmultimer binding but, notably, it identifies the fraction of antigen-specific CD8+ T cells with immediate and high functional capability (i.e., expressing high levels of cytotoxic markers and cytokines). Compared with the currently available methods, staining of activated β2-integrins presents the unique advantage of requiring activation times of only several minutes, therefore delivering functional information nearly reflecting the in vivo situation. Hence, the ICAM-1 assay is most suitable for rapid and precise monitoring of functional antigen-specific T cell responses, including for patient samples in a variety of clinical settings, as well as for the isolation of functional T cells for adoptive cell-transfer immunotherapies.",

author = "Stoyan Dimitrov and C{\'e}cile Gouttefangeas and Luciana Besedovsky and Jensen, \{Anja T.R.\} and Chandran, \{P. Anoop\} and Elisa Rusch and Ramona Businger and Michael Schindler and Tanja Lange and Jan Born and Rammensee, \{Hans Georg\}",

note = "Funding Information: ACKNOWLEDGMENTS. We thank S. Stevanovi{\'c}for providing synthetic peptides; S. Heidu, J. Lehnholz, K. Witte, and E.-M. Schmidt for excellent technical assistance and advice; M. Szczepanski, J. C. P. Santiago, M. Esen, M. Buhl, and G. Marasca for sampling blood from the subjects and patients; all subjects and patients who participated in this study; and M. Hallschmid and R. Littwin for critical reading and proofreading of the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft Grants SFB 654 (to T.L. and H.-G.R.) and SFB 685 (to C.G. and H.-G.R.); grants from the German Federal Ministry of Education and Research (BMBF) to the German Center for Diabetes Research (DZD; 01GI0925) (to S.D. and J.B.); and European Research Council Grant AdG 339842, MUTAEDITING (to H.-G.R.). Publisher Copyright: {\textcopyright} 2018 National Academy of Sciences.All Rights Reserved. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.",

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doi = "10.1073/pnas.1720714115",

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Dimitrov, S, Gouttefangeas, C, Besedovsky, L, Jensen, ATR, Chandran, PA, Rusch, E, Businger, R, Schindler, M, Lange, T, Born, J & Rammensee, HG 2018, 'Activated integrins identify functional antigen-specific CD8⁺ T cells within minutes after antigen stimulation', Proceedings of the National Academy of Sciences of the United States of America, vol. 115, no. 24, pp. E5536-E5545. https://doi.org/10.1073/pnas.1720714115

Activated integrins identify functional antigen-specific CD8⁺ T cells within minutes after antigen stimulation. / Dimitrov, Stoyan; Gouttefangeas, Cécile; Besedovsky, Luciana et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, No. 24, 12.06.2018, p. E5536-E5545.

Research output: Journal Articles › Journal articles › Research › peer-review

TY - JOUR

T1 - Activated integrins identify functional antigen-specific CD8+ T cells within minutes after antigen stimulation

AU - Dimitrov, Stoyan

AU - Gouttefangeas, Cécile

AU - Besedovsky, Luciana

AU - Jensen, Anja T.R.

AU - Chandran, P. Anoop

AU - Rusch, Elisa

AU - Businger, Ramona

AU - Schindler, Michael

AU - Lange, Tanja

AU - Born, Jan

AU - Rammensee, Hans Georg

N1 - Funding Information: ACKNOWLEDGMENTS. We thank S. Stevanovićfor providing synthetic peptides; S. Heidu, J. Lehnholz, K. Witte, and E.-M. Schmidt for excellent technical assistance and advice; M. Szczepanski, J. C. P. Santiago, M. Esen, M. Buhl, and G. Marasca for sampling blood from the subjects and patients; all subjects and patients who participated in this study; and M. Hallschmid and R. Littwin for critical reading and proofreading of the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft Grants SFB 654 (to T.L. and H.-G.R.) and SFB 685 (to C.G. and H.-G.R.); grants from the German Federal Ministry of Education and Research (BMBF) to the German Center for Diabetes Research (DZD; 01GI0925) (to S.D. and J.B.); and European Research Council Grant AdG 339842, MUTAEDITING (to H.-G.R.). Publisher Copyright: © 2018 National Academy of Sciences.All Rights Reserved. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.

PY - 2018/6/12

Y1 - 2018/6/12

N2 - Immediate β2-integrin activation upon T cell receptor stimulation is critical for effective interaction between T cells and their targets and may therefore be used for the rapid identification and isolation of functional T cells. We present a simple and sensitive flow cytometry-based assay to assess antigen-specific T cells using fluorescent intercellular adhesion molecule (ICAM)-1 multimers that specifically bind to activated β2-integrins. The method is compatible with surface and intracellular staining; it is applicable for monitoring of a broad range of virus-, tumor-, and vaccine-specific CD8+ T cells, and for isolating viable antigen-reacting cells. ICAM-1 binding correlateswith peptide-MHCmultimer binding but, notably, it identifies the fraction of antigen-specific CD8+ T cells with immediate and high functional capability (i.e., expressing high levels of cytotoxic markers and cytokines). Compared with the currently available methods, staining of activated β2-integrins presents the unique advantage of requiring activation times of only several minutes, therefore delivering functional information nearly reflecting the in vivo situation. Hence, the ICAM-1 assay is most suitable for rapid and precise monitoring of functional antigen-specific T cell responses, including for patient samples in a variety of clinical settings, as well as for the isolation of functional T cells for adoptive cell-transfer immunotherapies.

AB - Immediate β2-integrin activation upon T cell receptor stimulation is critical for effective interaction between T cells and their targets and may therefore be used for the rapid identification and isolation of functional T cells. We present a simple and sensitive flow cytometry-based assay to assess antigen-specific T cells using fluorescent intercellular adhesion molecule (ICAM)-1 multimers that specifically bind to activated β2-integrins. The method is compatible with surface and intracellular staining; it is applicable for monitoring of a broad range of virus-, tumor-, and vaccine-specific CD8+ T cells, and for isolating viable antigen-reacting cells. ICAM-1 binding correlateswith peptide-MHCmultimer binding but, notably, it identifies the fraction of antigen-specific CD8+ T cells with immediate and high functional capability (i.e., expressing high levels of cytotoxic markers and cytokines). Compared with the currently available methods, staining of activated β2-integrins presents the unique advantage of requiring activation times of only several minutes, therefore delivering functional information nearly reflecting the in vivo situation. Hence, the ICAM-1 assay is most suitable for rapid and precise monitoring of functional antigen-specific T cell responses, including for patient samples in a variety of clinical settings, as well as for the isolation of functional T cells for adoptive cell-transfer immunotherapies.

UR - https://www.scopus.com/pages/publications/85049196486

U2 - 10.1073/pnas.1720714115

DO - 10.1073/pnas.1720714115

M3 - Journal articles

C2 - 29844168

AN - SCOPUS:85049196486

SN - 0027-8424

VL - 115

SP - E5536-E5545

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 24

ER -