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5. High-resolution mapping of cell-cycle dynamics during steady-state T-cell development and regeneration in vivo

Author

Kunze-Schumacher, Heike, Verheyden, Nikita A., Grewers, Zoe, Meyer-Hermann, Michael, Greiff, Victor, Robert, Philippe A., Krueger, Andreas, Kunze-Schumacher, Heike, Verheyden, Nikita A., Grewers, Zoe, Meyer-Hermann, Michael, Greiff, Victor, Robert, Philippe A., and Krueger, Andreas

Abstract

Control of cell proliferation is critical for the lymphocyte life cycle. However, little is known on how stage-specific alterations in cell-cycle behavior drive proliferation dynamics during T-cell development. Here, we employed in vivo dual-nucleoside pulse labeling combined with determination of DNA replication over time as well as fluorescent ubiquitination-based cell-cycle indicator mice to establish a quantitative high-resolution map of cell-cycle kinetics of thymocytes. We developed an agent-based mathematical model of T-cell developmental dynamics. To generate the capacity for proliferative bursts, cell-cycle acceleration followed a 'stretch model', characterized by simultaneous and proportional contraction of both G1 and S phase. Analysis of cell-cycle phase dynamics during regeneration showed tailored adjustments of cell-cycle phase dynamics. Taken together, our results highlight intrathymic cell-cycle regulation as an adjustable system to maintain physiologic tissue homeostasis and foster our understanding of dysregulation of the T-cell developmental program.

Published
2023

8. Molecular and cellular control of intrathymic T-cell development

Author

Kunze-Schumacher, Heike

Subjects
ddc:570
Abstract

T-cell development is a highly dynamic and stepwise process comprimising T lineage commitment, T-cell receptor (TCR) gene rearrangements and subsequent selection. From a quantitative point of view, only a few hundred progenitor cells migrate from the bone marrow into the thymus. Developing thymocytes (termed double negative (DN), CD4-CD8-) can be further divided into DN1-4 cells based on the expression of CD25 and CD44. These developmental events are interspersed by proliferative bursts which ultimately lead to the generation of millions of double positive (DP, CD4+CD8+) thymocytes that then undergo selection. As a consequence, a proportion of naïve T-cells evolves to ensure adaptive, but not autoreactive immunity. Previous studies of our lab focused on the quantification of thymus colonization and identified thymus entry to be dependent on expression of the chemokine receptors CCR7 and CCR9 (Krueger et al., 2010; Ziętara et al., 2015). CCR7/9 double knockout (DKO) mice are almost completely devoid of the most immature thymocyte populations (DN1 and DN2), but show near normal DN3 cellularity. Interestingly, a similar defect during early development but a virtually complete recovery of later stages and total thymocyte numbers was also observed in thymi of miR-17~92 deficient mice. Here, a failure of prethymic IL-7 signaling dampens early T-cell development (Regelin et al., 2015). For this reason, we hypothesized a tight regulation of thymocyte population size through alterations in the underlying cell cycle kinetics. In this thesis, we employed in vivo single- and dual-nucleoside pulse labeling combined with determination of DNA replication over time in different WT thymocyte subsets at steady-state. Based on this, we assessed alterations in cell cycle kinetics of CCR7/9 and miR-17~92 defcicient mice and identified compensatory mechanisms of thymocytes on the level of cell cycle phase distribution and cell cycle speed. In addition, single-cell RNA sequencing helped to obtain information on cell cycle dynamics of early thymocyte subsets, exemplarily shown for WT and CCR7/9 DKO mice. Lastly, we performed cell cycle analyses in a model of endogenous thymic repair upon sublethal total body irradiation which provided insight into intrathymic cell cycle regulation as an adjustable system to re-establish normal thymus cellularity. In the second part of the thesis, we addressed the role of miR-21 in the thymus. In various studies, we and others identified miRNAs as key posttranscriptional regulators of the immune system and especially for T-cell development (Regelin et al. 2015; Mildner et al. 2017; Li et al. 2007; Ebert et al. 2009; Ziętara et al. 2013; Schaffert et al. 2015). The dynamic expression of miR-21 during T-cell development (Neilson et al. 2007; Kirigin et al. 2012; Kuchen et al. 2010) prompted us to hypothesize that miR-21 has a regulatory function in the thymus. A miR 21-knockout mouse model allowed us to study the role of this miRNA for the development of T-cells in the thymus and the maintenance of T-cells in the periphery. In addition, we performed competitive bone marrow chimera experiments in the context of miR-21 deficiency and overexpression. Further insights were provided by exploring the function of miR-21 in negative selection in vivo as well as in T-cell differentiation in coculture experiments in vitro. To unravel implications of miR-21 to regulate cellular stress responses, we assessed the contribution of miR-21 in a model of endogenous regeneration of the thymus after sublethal irradiation. We could not provide evidence for a prominent role for miR-21 during T-cell development. Together, our experiments revealed that miR-21 is largely dispensable for physiologic T-cell development despite high and dynamic expression in the thymus (Kunze Schumacher et al., 2018). The apparent discrepancy between dynamic expression but lack of a regulatory function in the thymus led us to conclude that miR-21 is rather fine tuning T-cell responses than controlling a developmental event.

Published
2022

9. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Author

Cossarizza, Andrea, primary, Chang, Hyun‐Dong, additional, Radbruch, Andreas, additional, Abrignani, Sergio, additional, Addo, Richard, additional, Akdis, Mübeccel, additional, Andrä, Immanuel, additional, Andreata, Francesco, additional, Annunziato, Francesco, additional, Arranz, Eduardo, additional, Bacher, Petra, additional, Bari, Sudipto, additional, Barnaba, Vincenzo, additional, Barros‐Martins, Joana, additional, Baumjohann, Dirk, additional, Beccaria, Cristian G., additional, Bernardo, David, additional, Boardman, Dominic A., additional, Borger, Jessica, additional, Böttcher, Chotima, additional, Brockmann, Leonie, additional, Burns, Marie, additional, Busch, Dirk H., additional, Cameron, Garth, additional, Cammarata, Ilenia, additional, Cassotta, Antonino, additional, Chang, Yinshui, additional, Chirdo, Fernando Gabriel, additional, Christakou, Eleni, additional, Čičin‐Šain, Luka, additional, Cook, Laura, additional, Corbett, Alexandra J., additional, Cornelis, Rebecca, additional, Cosmi, Lorenzo, additional, Davey, Martin S., additional, De Biasi, Sara, additional, De Simone, Gabriele, additional, del Zotto, Genny, additional, Delacher, Michael, additional, Di Rosa, Francesca, additional, Di Santo, James, additional, Diefenbach, Andreas, additional, Dong, Jun, additional, Dörner, Thomas, additional, Dress, Regine J., additional, Dutertre, Charles‐Antoine, additional, Eckle, Sidonia B. G., additional, Eede, Pascale, additional, Evrard, Maximilien, additional, Falk, Christine S., additional, Feuerer, Markus, additional, Fillatreau, Simon, additional, Fiz‐Lopez, Aida, additional, Follo, Marie, additional, Foulds, Gemma A., additional, Fröbel, Julia, additional, Gagliani, Nicola, additional, Galletti, Giovanni, additional, Gangaev, Anastasia, additional, Garbi, Natalio, additional, Garrote, José Antonio, additional, Geginat, Jens, additional, Gherardin, Nicholas A., additional, Gibellini, Lara, additional, Ginhoux, Florent, additional, Godfrey, Dale I., additional, Gruarin, Paola, additional, Haftmann, Claudia, additional, Hansmann, Leo, additional, Harpur, Christopher M., additional, Hayday, Adrian C., additional, Heine, Guido, additional, Hernández, Daniela Carolina, additional, Herrmann, Martin, additional, Hoelsken, Oliver, additional, Huang, Qing, additional, Huber, Samuel, additional, Huber, Johanna E., additional, Huehn, Jochen, additional, Hundemer, Michael, additional, Hwang, William Y. K., additional, Iannacone, Matteo, additional, Ivison, Sabine M., additional, Jäck, Hans‐Martin, additional, Jani, Peter K., additional, Keller, Baerbel, additional, Kessler, Nina, additional, Ketelaars, Steven, additional, Knop, Laura, additional, Knopf, Jasmin, additional, Koay, Hui‐Fern, additional, Kobow, Katja, additional, Kriegsmann, Katharina, additional, Kristyanto, H., additional, Krueger, Andreas, additional, Kuehne, Jenny F., additional, Kunze‐Schumacher, Heike, additional, Kvistborg, Pia, additional, Kwok, Immanuel, additional, Latorre, Daniela, additional, Lenz, Daniel, additional, Levings, Megan K., additional, Lino, Andreia C., additional, Liotta, Francesco, additional, Long, Heather M., additional, Lugli, Enrico, additional, MacDonald, Katherine N., additional, Maggi, Laura, additional, Maini, Mala K., additional, Mair, Florian, additional, Manta, Calin, additional, Manz, Rudolf Armin, additional, Mashreghi, Mir‐Farzin, additional, Mazzoni, Alessio, additional, McCluskey, James, additional, Mei, Henrik E., additional, Melchers, Fritz, additional, Melzer, Susanne, additional, Mielenz, Dirk, additional, Monin, Leticia, additional, Moretta, Lorenzo, additional, Multhoff, Gabriele, additional, Muñoz, Luis Enrique, additional, Muñoz‐Ruiz, Miguel, additional, Muscate, Franziska, additional, Natalini, Ambra, additional, Neumann, Katrin, additional, Ng, Lai Guan, additional, Niedobitek, Antonia, additional, Niemz, Jana, additional, Almeida, Larissa Nogueira, additional, Notarbartolo, Samuele, additional, Ostendorf, Lennard, additional, Pallett, Laura J., additional, Patel, Amit A., additional, Percin, Gulce Itir, additional, Peruzzi, Giovanna, additional, Pinti, Marcello, additional, Pockley, A. Graham, additional, Pracht, Katharina, additional, Prinz, Immo, additional, Pujol‐Autonell, Irma, additional, Pulvirenti, Nadia, additional, Quatrini, Linda, additional, Quinn, Kylie M., additional, Radbruch, Helena, additional, Rhys, Hefin, additional, Rodrigo, Maria B., additional, Romagnani, Chiara, additional, Saggau, Carina, additional, Sakaguchi, Shimon, additional, Sallusto, Federica, additional, Sanderink, Lieke, additional, Sandrock, Inga, additional, Schauer, Christine, additional, Scheffold, Alexander, additional, Scherer, Hans U., additional, Schiemann, Matthias, additional, Schildberg, Frank A., additional, Schober, Kilian, additional, Schoen, Janina, additional, Schuh, Wolfgang, additional, Schüler, Thomas, additional, Schulz, Axel R., additional, Schulz, Sebastian, additional, Schulze, Julia, additional, Simonetti, Sonia, additional, Singh, Jeeshan, additional, Sitnik, Katarzyna M., additional, Stark, Regina, additional, Starossom, Sarah, additional, Stehle, Christina, additional, Szelinski, Franziska, additional, Tan, Leonard, additional, Tarnok, Attila, additional, Tornack, Julia, additional, Tree, Timothy I. M., additional, van Beek, Jasper J. P., additional, van de Veen, Willem, additional, van Gisbergen, Klaas, additional, Vasco, Chiara, additional, Verheyden, Nikita A., additional, von Borstel, Anouk, additional, Ward‐Hartstonge, Kirsten A., additional, Warnatz, Klaus, additional, Waskow, Claudia, additional, Wiedemann, Annika, additional, Wilharm, Anneke, additional, Wing, James, additional, Wirz, Oliver, additional, Wittner, Jens, additional, Yang, Jennie H. M., additional, and Yang, Juhao, additional

Published
2021
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10. Modeling the dynamics of T-cell development in the thymus

Author

Robert, Philippe A., Kunze-Schumacher, Heike, Greiff, Victor, and Krüger, Andreas

Subjects
ordinary differential equations (ODE), T-cell receptor (TCR), mathematical modeling, lcsh:Astrophysics, Review, T-cell development, agent-based models, lcsh:QC1-999, thymic selection, lcsh:QB460-466, multi-scale models, lcsh:Q, ddc:610, complex systems, lcsh:Science, lcsh:Physics
Abstract

The thymus hosts the development of a specific type of adaptive immune cells called T cells. T cells orchestrate the adaptive immune response through recognition of antigen by the highly variable T-cell receptor (TCR). T-cell development is a tightly coordinated process comprising lineage commitment, somatic recombination of Tcr gene loci and selection for functional, but non-self-reactive TCRs, all interspersed with massive proliferation and cell death. Thus, the thymus produces a pool of T cells throughout life capable of responding to virtually any exogenous attack while preserving the body through self-tolerance. The thymus has been of considerable interest to both immunologists and theoretical biologists due to its multi-scale quantitative properties, bridging molecular binding, population dynamics and polyclonal repertoire specificity. Here, we review experimental strategies aimed at revealing quantitative and dynamic properties of T-cell development and how they have been implemented in mathematical modeling strategies that were reported to help understand the flexible dynamics of the highly dividing and dying thymic cell populations. Furthermore, we summarize the current challenges to estimating in vivo cellular dynamics and to reaching a next-generation multi-scale picture of T-cell development.

Published
2021

11. High-resolution mapping of cell cycle dynamics during steady-state T cell development and regeneration in vivo

Author

Kunze-Schumacher, Heike, Verheyden, Nikita A., Grewers, Zoe, Meyer-Hermann, Michael, Greiff, Victor, Robert, Philippe A., and Krueger, Andreas

Abstract

Control of cell proliferation is critical for the lymphocyte life cycle. However, little is known about how stage-specific alterations in cell cycle behavior drive proliferation dynamics during T cell development. Here, we employed in vivodual-nucleoside pulse labeling combined with the determination of DNA replication over time as well as fluorescent ubiquitination-based cell cycle indicator mice to establish a quantitative high-resolution map of cell cycle kinetics of thymocytes. We developed an agent-based mathematical model of T cell developmental dynamics. To generate the capacity for proliferative bursts, cell cycle acceleration followed a “stretch model” characterized by the simultaneous and proportional contraction of both G1 and S phases. Analysis of cell cycle phase dynamics during regeneration showed tailored adjustments of cell cycle phase dynamics. Taken together, our results highlight intrathymic cell cycle regulation as an adjustable system to maintain physiologic tissue homeostasis and foster our understanding of dysregulation of the T cell developmental program.

Published
2025
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12. The Role of MicroRNAs in Development and Function of Regulatory T Cells – Lessons for a Better Understanding of MicroRNA Biology

Author

Kunze-Schumacher, Heike and Krueger, Andreas

Subjects
lcsh:Immunologic diseases. Allergy, regulatory T cell, Cell Differentiation, Lymphocyte Activation, T-Lymphocytes, Regulatory, Treg cell, MicroRNAs, Gene Expression Regulation, thymus, Animals, Humans, Gene Regulatory Networks, T-cell receptor, ddc:610, gene regulation, lcsh:RC581-607, miRNA
Abstract

MicroRNAs (miRNAs) have emerged as critical posttranscriptional regulators of the immune system, including function and development of regulatory T (Treg) cells. Although this critical role has been firmly demonstrated through genetic models, key mechanisms of miRNA function in vivo remain elusive. Here, we review the role of miRNAs in Treg cell development and function. In particular, we focus on the question what the study of miRNAs in this context reveals about miRNA biology in general, including context-dependent function and the role of individual targets vs. complex co-targeting networks. In addition, we highlight potential technical pitfalls and state-of-the-art approaches to improve the mechanistic understanding of miRNA biology in a physiological context.

Published
2020
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13. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Author

Rolf M. Schwiete Foundation, Associazione Italiana per la Ricerca sul Cancro, German Research Foundation, National Institutes of Health (US), European Commission, Cossarizza, Andrea, Chang, Hyun‐Dong, Radbruch, Andreas, Abrignani, Sergio, Addo, Richard, Akdis, Mübeccel, Andrä, Immanuel, Andreata, Francesco, Annunziato, Francesco, Arranz, Eduardo, Bacher, Petra, Knop, Laura, Knopf, Jasmin, Koay, Hui-Fern, Kobow, Katja, Kriegsmann, Katharina, Kristyanto, H., Krueger, Andreas, Kuehne, Jenny F., Kunze-Schumacher, Heike, Maini, Mala K., Verheyden, Nikita A., Kvistborg, Pia, Kwok, Immanuel, Latorre, Daniela, Mair, Florian, Bari, Sudipto, Manta, Calin, Armin Manz, Rudolf, Mashreghi, Mir-Farzin, Mazzoni, Alessio, McCluskey, James, Borstel, Anouk von, Mei, Henrik E., Melchers, Fritz, Melzer, Susanne, Mielenz, Dirk, Monin, Leticia, Barnaba, Vincenzo, Moretta, Lorenzo, Multhoff, Gabriele, Muñoz, Luis Enrique, Muñoz-Ruiz, Miguel, Ward-Hartstonge, Kirsten A., Muscate, Franziska, Natalini, Ambra, Neumann, Katrin, Guan N., Lai, Niedobitek, Antonia, Niemz, Jana, Barros-Martins, Joana, Nogueira Almeida, Larissa, Notarbartolo, Samuele, Ostendorf, Lennard, Warnatz, Klaus, Pallett, Laura J., Patel, Amit A., Itir Percin, Gulce, Peruzzi, Giovanna, Pinti, Marcello, Pockley, A. Graham, Pracht, Katharina, Baumjohann, Dirk, Prinz, Immo, Pujol-Autonell, Irma, Waskow, Claudia, Pulvirenti, Nadia, Quatrini, Linda, Quinn, Kylie M., Radbruch, Helena, Rhys, Hefin, Rodrigo, Maria B., Romagnani, Chiara, Saggau, Carina, Beccaria, Cristian G., Sakaguchi, Shimon, Wiedemann, Annika, Sallusto, Federica, Sanderink, Lieke, Sandrock, Inga, Schauer, Christine, Scheffold, Alexander, Scherer, Hans U., Schiemann, Matthias, Schildberg, Frank A., Schober, Kilian, Bernardo, David, Wilharm, Anneke, Schoen, Janina, Schuh, Wolfgang, Schüler, Thomas, Schulz, Axel R., Schulz, Sebastian, Schulze, Julia, Simonetti, Sonia, Singh, Jeeshan, Sitnik, Katarzyna M., Stark, Regina, Wing, James, Boardman, Dominic A., Starossom, Sarah, Stehle, Christina, Szelinski, Franziska, Tan, Leonard, Tarnok, Attila, Tornack, Julia, Tree, Timothy I. M., Van Beek, Jasper J. P., Veen, Willem van de, Wirz, Oliver, Gisbergen, Klaas van, Borger, Jessica, Vasco, Chiara, Böttcher, Chotima, Lenz, Daniel, Wittner, Jens, Yang, Jennie H. M., Yang, Juhao, Brockmann, Leonie, Burns, Marie, Busch, Dirk H., Cameron, Garth, Cammarata, Ilenia, Cassotta, Antonino, Chang, Yinshui, Levings, Megan K., Chirdo, Fernando G., Christakou, Eleni, Cicin-Sain, Luka, Cook, Laura, Corbett, Alexandra J., Cornelis, Rebecca, Cosmi, Lorenzo, Davey, Martin S., Biasi, Sara De, Simone, Gabriele De, Lino, Andreia C., Zotto, Genny del, Delacher, Michael, Rosa, Francesca Di, Santo, James Di, Diefenbach, Andreas, Dong, Jun, Dörner, Thomas, Dress, Regine J., Dutertre, Charles-Antoine, Eckle, Sidonia B. G., Liotta, Francesco, Eede, Pascale, Evrard, Maximilien, Falk, Christine S., Feuerer, Markus, Fillatreau, Simon, Fiz-López, Aida, Follo, Marie, Foulds, Gemma A., Fröbel, Julia, Gagliani, Nicola, Long, Heather M., Galletti, Giovanni, Gangaev, Anastasia, Garbi, Natalio, Garrote, José Antonio, Geginat, Jens, Gherardin, Nicholas A., Gibellini, Lara, Ginhoux, Florent, Godfrey, Dale I., Gruarin, Paola, Lugli, Enrico, Haftmann, Claudia, Hansmann, Leo, Harpur, Christopher M., Hayday, Adrian C., Heine, Guido, Hernández, Daniela Carolina, Herrmann, Martin, Hoelsken, Oliver, Huang, Qing, Huber, Samuel, MacDonald, Katherine N., Huber, Johanna E., Huehn, Jochen, Hundemer, Michael, Hwang, William Y. K., Iannacone, Matteo, Ivison, Sabine M., Jäck, Hans-Martin, Jani, Peter K., Keller, Baerbel, Kessler, Nina, Maggi, Laura, Ketelaars, Steven, Rolf M. Schwiete Foundation, Associazione Italiana per la Ricerca sul Cancro, German Research Foundation, National Institutes of Health (US), European Commission, Cossarizza, Andrea, Chang, Hyun‐Dong, Radbruch, Andreas, Abrignani, Sergio, Addo, Richard, Akdis, Mübeccel, Andrä, Immanuel, Andreata, Francesco, Annunziato, Francesco, Arranz, Eduardo, Bacher, Petra, Knop, Laura, Knopf, Jasmin, Koay, Hui-Fern, Kobow, Katja, Kriegsmann, Katharina, Kristyanto, H., Krueger, Andreas, Kuehne, Jenny F., Kunze-Schumacher, Heike, Maini, Mala K., Verheyden, Nikita A., Kvistborg, Pia, Kwok, Immanuel, Latorre, Daniela, Mair, Florian, Bari, Sudipto, Manta, Calin, Armin Manz, Rudolf, Mashreghi, Mir-Farzin, Mazzoni, Alessio, McCluskey, James, Borstel, Anouk von, Mei, Henrik E., Melchers, Fritz, Melzer, Susanne, Mielenz, Dirk, Monin, Leticia, Barnaba, Vincenzo, Moretta, Lorenzo, Multhoff, Gabriele, Muñoz, Luis Enrique, Muñoz-Ruiz, Miguel, Ward-Hartstonge, Kirsten A., Muscate, Franziska, Natalini, Ambra, Neumann, Katrin, Guan N., Lai, Niedobitek, Antonia, Niemz, Jana, Barros-Martins, Joana, Nogueira Almeida, Larissa, Notarbartolo, Samuele, Ostendorf, Lennard, Warnatz, Klaus, Pallett, Laura J., Patel, Amit A., Itir Percin, Gulce, Peruzzi, Giovanna, Pinti, Marcello, Pockley, A. Graham, Pracht, Katharina, Baumjohann, Dirk, Prinz, Immo, Pujol-Autonell, Irma, Waskow, Claudia, Pulvirenti, Nadia, Quatrini, Linda, Quinn, Kylie M., Radbruch, Helena, Rhys, Hefin, Rodrigo, Maria B., Romagnani, Chiara, Saggau, Carina, Beccaria, Cristian G., Sakaguchi, Shimon, Wiedemann, Annika, Sallusto, Federica, Sanderink, Lieke, Sandrock, Inga, Schauer, Christine, Scheffold, Alexander, Scherer, Hans U., Schiemann, Matthias, Schildberg, Frank A., Schober, Kilian, Bernardo, David, Wilharm, Anneke, Schoen, Janina, Schuh, Wolfgang, Schüler, Thomas, Schulz, Axel R., Schulz, Sebastian, Schulze, Julia, Simonetti, Sonia, Singh, Jeeshan, Sitnik, Katarzyna M., Stark, Regina, Wing, James, Boardman, Dominic A., Starossom, Sarah, Stehle, Christina, Szelinski, Franziska, Tan, Leonard, Tarnok, Attila, Tornack, Julia, Tree, Timothy I. M., Van Beek, Jasper J. P., Veen, Willem van de, Wirz, Oliver, Gisbergen, Klaas van, Borger, Jessica, Vasco, Chiara, Böttcher, Chotima, Lenz, Daniel, Wittner, Jens, Yang, Jennie H. M., Yang, Juhao, Brockmann, Leonie, Burns, Marie, Busch, Dirk H., Cameron, Garth, Cammarata, Ilenia, Cassotta, Antonino, Chang, Yinshui, Levings, Megan K., Chirdo, Fernando G., Christakou, Eleni, Cicin-Sain, Luka, Cook, Laura, Corbett, Alexandra J., Cornelis, Rebecca, Cosmi, Lorenzo, Davey, Martin S., Biasi, Sara De, Simone, Gabriele De, Lino, Andreia C., Zotto, Genny del, Delacher, Michael, Rosa, Francesca Di, Santo, James Di, Diefenbach, Andreas, Dong, Jun, Dörner, Thomas, Dress, Regine J., Dutertre, Charles-Antoine, Eckle, Sidonia B. G., Liotta, Francesco, Eede, Pascale, Evrard, Maximilien, Falk, Christine S., Feuerer, Markus, Fillatreau, Simon, Fiz-López, Aida, Follo, Marie, Foulds, Gemma A., Fröbel, Julia, Gagliani, Nicola, Long, Heather M., Galletti, Giovanni, Gangaev, Anastasia, Garbi, Natalio, Garrote, José Antonio, Geginat, Jens, Gherardin, Nicholas A., Gibellini, Lara, Ginhoux, Florent, Godfrey, Dale I., Gruarin, Paola, Lugli, Enrico, Haftmann, Claudia, Hansmann, Leo, Harpur, Christopher M., Hayday, Adrian C., Heine, Guido, Hernández, Daniela Carolina, Herrmann, Martin, Hoelsken, Oliver, Huang, Qing, Huber, Samuel, MacDonald, Katherine N., Huber, Johanna E., Huehn, Jochen, Hundemer, Michael, Hwang, William Y. K., Iannacone, Matteo, Ivison, Sabine M., Jäck, Hans-Martin, Jani, Peter K., Keller, Baerbel, Kessler, Nina, Maggi, Laura, and Ketelaars, Steven

Abstract

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.

Published
2021

16. Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

Author

Cossarizza, Andrea, primary, Chang, Hyun‐Dong, additional, Radbruch, Andreas, additional, Acs, Andreas, additional, Adam, Dieter, additional, Adam‐Klages, Sabine, additional, Agace, William W., additional, Aghaeepour, Nima, additional, Akdis, Mübeccel, additional, Allez, Matthieu, additional, Almeida, Larissa Nogueira, additional, Alvisi, Giorgia, additional, Anderson, Graham, additional, Andrä, Immanuel, additional, Annunziato, Francesco, additional, Anselmo, Achille, additional, Bacher, Petra, additional, Baldari, Cosima T., additional, Bari, Sudipto, additional, Barnaba, Vincenzo, additional, Barros‐Martins, Joana, additional, Battistini, Luca, additional, Bauer, Wolfgang, additional, Baumgart, Sabine, additional, Baumgarth, Nicole, additional, Baumjohann, Dirk, additional, Baying, Bianka, additional, Bebawy, Mary, additional, Becher, Burkhard, additional, Beisker, Wolfgang, additional, Benes, Vladimir, additional, Beyaert, Rudi, additional, Blanco, Alfonso, additional, Boardman, Dominic A., additional, Bogdan, Christian, additional, Borger, Jessica G., additional, Borsellino, Giovanna, additional, Boulais, Philip E., additional, Bradford, Jolene A., additional, Brenner, Dirk, additional, Brinkman, Ryan R., additional, Brooks, Anna E. S., additional, Busch, Dirk H., additional, Büscher, Martin, additional, Bushnell, Timothy P., additional, Calzetti, Federica, additional, Cameron, Garth, additional, Cammarata, Ilenia, additional, Cao, Xuetao, additional, Cardell, Susanna L., additional, Casola, Stefano, additional, Cassatella, Marco A., additional, Cavani, Andrea, additional, Celada, Antonio, additional, Chatenoud, Lucienne, additional, Chattopadhyay, Pratip K., additional, Chow, Sue, additional, Christakou, Eleni, additional, Čičin‐Šain, Luka, additional, Clerici, Mario, additional, Colombo, Federico S., additional, Cook, Laura, additional, Cooke, Anne, additional, Cooper, Andrea M., additional, Corbett, Alexandra J., additional, Cosma, Antonio, additional, Cosmi, Lorenzo, additional, Coulie, Pierre G., additional, Cumano, Ana, additional, Cvetkovic, Ljiljana, additional, Dang, Van Duc, additional, Dang‐Heine, Chantip, additional, Davey, Martin S., additional, Davies, Derek, additional, De Biasi, Sara, additional, Del Zotto, Genny, additional, Dela Cruz, Gelo Victoriano, additional, Delacher, Michael, additional, Della Bella, Silvia, additional, Dellabona, Paolo, additional, Deniz, Günnur, additional, Dessing, Mark, additional, Di Santo, James P., additional, Diefenbach, Andreas, additional, Dieli, Francesco, additional, Dolf, Andreas, additional, Dörner, Thomas, additional, Dress, Regine J., additional, Dudziak, Diana, additional, Dustin, Michael, additional, Dutertre, Charles‐Antoine, additional, Ebner, Friederike, additional, Eckle, Sidonia B. G., additional, Edinger, Matthias, additional, Eede, Pascale, additional, Ehrhardt, Götz R.A., additional, Eich, Marcus, additional, Engel, Pablo, additional, Engelhardt, Britta, additional, Erdei, Anna, additional, Esser, Charlotte, additional, Everts, Bart, additional, Evrard, Maximilien, additional, Falk, Christine S., additional, Fehniger, Todd A., additional, Felipo‐Benavent, Mar, additional, Ferry, Helen, additional, Feuerer, Markus, additional, Filby, Andrew, additional, Filkor, Kata, additional, Fillatreau, Simon, additional, Follo, Marie, additional, Förster, Irmgard, additional, Foster, John, additional, Foulds, Gemma A., additional, Frehse, Britta, additional, Frenette, Paul S., additional, Frischbutter, Stefan, additional, Fritzsche, Wolfgang, additional, Galbraith, David W., additional, Gangaev, Anastasia, additional, Garbi, Natalio, additional, Gaudilliere, Brice, additional, Gazzinelli, Ricardo T., additional, Geginat, Jens, additional, Gerner, Wilhelm, additional, Gherardin, Nicholas A., additional, Ghoreschi, Kamran, additional, Gibellini, Lara, additional, Ginhoux, Florent, additional, Goda, Keisuke, additional, Godfrey, Dale I., additional, Goettlinger, Christoph, additional, González‐Navajas, Jose M., additional, Goodyear, Carl S., additional, Gori, Andrea, additional, Grogan, Jane L., additional, Grummitt, Daryl, additional, Grützkau, Andreas, additional, Haftmann, Claudia, additional, Hahn, Jonas, additional, Hammad, Hamida, additional, Hämmerling, Günter, additional, Hansmann, Leo, additional, Hansson, Goran, additional, Harpur, Christopher M., additional, Hartmann, Susanne, additional, Hauser, Andrea, additional, Hauser, Anja E., additional, Haviland, David L., additional, Hedley, David, additional, Hernández, Daniela C., additional, Herrera, Guadalupe, additional, Herrmann, Martin, additional, Hess, Christoph, additional, Höfer, Thomas, additional, Hoffmann, Petra, additional, Hogquist, Kristin, additional, Holland, Tristan, additional, Höllt, Thomas, additional, Holmdahl, Rikard, additional, Hombrink, Pleun, additional, Houston, Jessica P., additional, Hoyer, Bimba F., additional, Huang, Bo, additional, Huang, Fang‐Ping, additional, Huber, Johanna E., additional, Huehn, Jochen, additional, Hundemer, Michael, additional, Hunter, Christopher A., additional, Hwang, William Y. K., additional, Iannone, Anna, additional, Ingelfinger, Florian, additional, Ivison, Sabine M, additional, Jäck, Hans‐Martin, additional, Jani, Peter K., additional, Jávega, Beatriz, additional, Jonjic, Stipan, additional, Kaiser, Toralf, additional, Kalina, Tomas, additional, Kamradt, Thomas, additional, Kaufmann, Stefan H. E., additional, Keller, Baerbel, additional, Ketelaars, Steven L. C., additional, Khalilnezhad, Ahad, additional, Khan, Srijit, additional, Kisielow, Jan, additional, Klenerman, Paul, additional, Knopf, Jasmin, additional, Koay, Hui‐Fern, additional, Kobow, Katja, additional, Kolls, Jay K., additional, Kong, Wan Ting, additional, Kopf, Manfred, additional, Korn, Thomas, additional, Kriegsmann, Katharina, additional, Kristyanto, Hendy, additional, Kroneis, Thomas, additional, Krueger, Andreas, additional, Kühne, Jenny, additional, Kukat, Christian, additional, Kunkel, Désirée, additional, Kunze‐Schumacher, Heike, additional, Kurosaki, Tomohiro, additional, Kurts, Christian, additional, Kvistborg, Pia, additional, Kwok, Immanuel, additional, Landry, Jonathan, additional, Lantz, Olivier, additional, Lanuti, Paola, additional, LaRosa, Francesca, additional, Lehuen, Agnès, additional, LeibundGut‐Landmann, Salomé, additional, Leipold, Michael D., additional, Leung, Leslie Y.T., additional, Levings, Megan K., additional, Lino, Andreia C., additional, Liotta, Francesco, additional, Litwin, Virginia, additional, Liu, Yanling, additional, Ljunggren, Hans‐Gustaf, additional, Lohoff, Michael, additional, Lombardi, Giovanna, additional, Lopez, Lilly, additional, López‐Botet, Miguel, additional, Lovett‐Racke, Amy E., additional, Lubberts, Erik, additional, Luche, Herve, additional, Ludewig, Burkhard, additional, Lugli, Enrico, additional, Lunemann, Sebastian, additional, Maecker, Holden T., additional, Maggi, Laura, additional, Maguire, Orla, additional, Mair, Florian, additional, Mair, Kerstin H., additional, Mantovani, Alberto, additional, Manz, Rudolf A., additional, Marshall, Aaron J., additional, Martínez‐Romero, Alicia, additional, Martrus, Glòria, additional, Marventano, Ivana, additional, Maslinski, Wlodzimierz, additional, Matarese, Giuseppe, additional, Mattioli, Anna Vittoria, additional, Maueröder, Christian, additional, Mazzoni, Alessio, additional, McCluskey, James, additional, McGrath, Mairi, additional, McGuire, Helen M., additional, McInnes, Iain B., additional, Mei, Henrik E., additional, Melchers, Fritz, additional, Melzer, Susanne, additional, Mielenz, Dirk, additional, Miller, Stephen D., additional, Mills, Kingston H.G., additional, Minderman, Hans, additional, Mjösberg, Jenny, additional, Moore, Jonni, additional, Moran, Barry, additional, Moretta, Lorenzo, additional, Mosmann, Tim R., additional, Müller, Susann, additional, Multhoff, Gabriele, additional, Muñoz, Luis Enrique, additional, Münz, Christian, additional, Nakayama, Toshinori, additional, Nasi, Milena, additional, Neumann, Katrin, additional, Ng, Lai Guan, additional, Niedobitek, Antonia, additional, Nourshargh, Sussan, additional, Núñez, Gabriel, additional, O'Connor, José‐Enrique, additional, Ochel, Aaron, additional, Oja, Anna, additional, Ordonez, Diana, additional, Orfao, Alberto, additional, Orlowski‐Oliver, Eva, additional, Ouyang, Wenjun, additional, Oxenius, Annette, additional, Palankar, Raghavendra, additional, Panse, Isabel, additional, Pattanapanyasat, Kovit, additional, Paulsen, Malte, additional, Pavlinic, Dinko, additional, Penter, Livius, additional, Peterson, Pärt, additional, Peth, Christian, additional, Petriz, Jordi, additional, Piancone, Federica, additional, Pickl, Winfried F., additional, Piconese, Silvia, additional, Pinti, Marcello, additional, Pockley, A. Graham, additional, Podolska, Malgorzata Justyna, additional, Poon, Zhiyong, additional, Pracht, Katharina, additional, Prinz, Immo, additional, Pucillo, Carlo E. M., additional, Quataert, Sally A., additional, Quatrini, Linda, additional, Quinn, Kylie M., additional, Radbruch, Helena, additional, Radstake, Tim R. D. J., additional, Rahmig, Susann, additional, Rahn, Hans‐Peter, additional, Rajwa, Bartek, additional, Ravichandran, Gevitha, additional, Raz, Yotam, additional, Rebhahn, Jonathan A., additional, Recktenwald, Diether, additional, Reimer, Dorothea, additional, Reis e Sousa, Caetano, additional, Remmerswaal, Ester B.M., additional, Richter, Lisa, additional, Rico, Laura G., additional, Riddell, Andy, additional, Rieger, Aja M., additional, Robinson, J. Paul, additional, Romagnani, Chiara, additional, Rubartelli, Anna, additional, Ruland, Jürgen, additional, Saalmüller, Armin, additional, Saeys, Yvan, additional, Saito, Takashi, additional, Sakaguchi, Shimon, additional, Sala‐de‐Oyanguren, Francisco, additional, Samstag, Yvonne, additional, Sanderson, Sharon, additional, Sandrock, Inga, additional, Santoni, Angela, additional, Sanz, Ramon Bellmàs, additional, Saresella, Marina, additional, Sautes‐Fridman, Catherine, additional, Sawitzki, Birgit, additional, Schadt, Linda, additional, Scheffold, Alexander, additional, Scherer, Hans U., additional, Schiemann, Matthias, additional, Schildberg, Frank A., additional, Schimisky, Esther, additional, Schlitzer, Andreas, additional, Schlosser, Josephine, additional, Schmid, Stephan, additional, Schmitt, Steffen, additional, Schober, Kilian, additional, Schraivogel, Daniel, additional, Schuh, Wolfgang, additional, Schüler, Thomas, additional, Schulte, Reiner, additional, Schulz, Axel Ronald, additional, Schulz, Sebastian R., additional, Scottá, Cristiano, additional, Scott‐Algara, Daniel, additional, Sester, David P., additional, Shankey, T. Vincent, additional, Silva‐Santos, Bruno, additional, Simon, Anna Katharina, additional, Sitnik, Katarzyna M., additional, Sozzani, Silvano, additional, Speiser, Daniel E., additional, Spidlen, Josef, additional, Stahlberg, Anders, additional, Stall, Alan M., additional, Stanley, Natalie, additional, Stark, Regina, additional, Stehle, Christina, additional, Steinmetz, Tobit, additional, Stockinger, Hannes, additional, Takahama, Yousuke, additional, Takeda, Kiyoshi, additional, Tan, Leonard, additional, Tárnok, Attila, additional, Tiegs, Gisa, additional, Toldi, Gergely, additional, Tornack, Julia, additional, Traggiai, Elisabetta, additional, Trebak, Mohamed, additional, Tree, Timothy I.M., additional, Trotter, Joe, additional, Trowsdale, John, additional, Tsoumakidou, Maria, additional, Ulrich, Henning, additional, Urbanczyk, Sophia, additional, van de Veen, Willem, additional, van den Broek, Maries, additional, van der Pol, Edwin, additional, Van Gassen, Sofie, additional, Van Isterdael, Gert, additional, van Lier, René A.W., additional, Veldhoen, Marc, additional, Vento‐Asturias, Salvador, additional, Vieira, Paulo, additional, Voehringer, David, additional, Volk, Hans‐Dieter, additional, von Borstel, Anouk, additional, von Volkmann, Konrad, additional, Waisman, Ari, additional, Walker, Rachael V., additional, Wallace, Paul K., additional, Wang, Sa A., additional, Wang, Xin M., additional, Ward, Michael D., additional, Ward‐Hartstonge, Kirsten A, additional, Warnatz, Klaus, additional, Warnes, Gary, additional, Warth, Sarah, additional, Waskow, Claudia, additional, Watson, James V., additional, Watzl, Carsten, additional, Wegener, Leonie, additional, Weisenburger, Thomas, additional, Wiedemann, Annika, additional, Wienands, Jürgen, additional, Wilharm, Anneke, additional, Wilkinson, Robert John, additional, Willimsky, Gerald, additional, Wing, James B., additional, Winkelmann, Rieke, additional, Winkler, Thomas H., additional, Wirz, Oliver F., additional, Wong, Alicia, additional, Wurst, Peter, additional, Yang, Jennie H. M., additional, Yang, Juhao, additional, Yazdanbakhsh, Maria, additional, Yu, Liping, additional, Yue, Alice, additional, Zhang, Hanlin, additional, Zhao, Yi, additional, Ziegler, Susanne Maria, additional, Zielinski, Christina, additional, Zimmermann, Jakob, additional, and Zychlinsky, Arturo, additional

Published
2019
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17. Releasing the concept of HLA‐allele specific peptide anchors in viral infections: A non‐canonical naturally presented human cytomegalovirus‐derivedHLA‐A*24:02restricted peptide drives exquisite immunogenicity

Author

Pump, Wiebke C., primary, Schulz, Rebecca, additional, Huyton, Trevor, additional, Kunze‐Schumacher, Heike, additional, Martens, Jörg, additional, Hò, Gia‐Gia T., additional, Blasczyk, Rainer, additional, and Bade‐Doeding, Christina, additional

Published
2019
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18. miR-181a/b-1 controls thymic selection of Treg cells and tunes their suppressive capacity

Author

Łyszkiewicz, Marcin, primary, Winter, Samantha J., additional, Witzlau, Katrin, additional, Föhse, Lisa, additional, Brownlie, Rebecca, additional, Puchałka, Jacek, additional, Verheyden, Nikita A., additional, Kunze-Schumacher, Heike, additional, Imelmann, Esther, additional, Blume, Jonas, additional, Raha, Solaiman, additional, Sekiya, Takashi, additional, Yoshimura, Akihiko, additional, Frueh, Jochen T., additional, Ullrich, Evelyn, additional, Huehn, Jochen, additional, Weiss, Siegfried, additional, Gutierrez, Maximiliano G., additional, Prinz, Immo, additional, Zamoyska, Rose, additional, Ziętara, Natalia, additional, and Krueger, Andreas, additional

Published
2019
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19. Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

Author

Cossarizza, Andrea, Chang, Hyun-Dong, Radbruch, Andreas, Acs, Andreas, Adam, Dieter, Adam-Klages, Sabine, Agace, William W., Aghaeepour, Nima, Akdis, Muebeccel, Allez, Matthieu, Almeida, Larissa Nogueira, Alvisi, Giorgia, Anderson, Graham, Andrae, Immanuel, Annunziato, Francesco, Anselmo, Achille, Bacher, Petra, Baldari, Cosima T., Bari, Sudipto, Barnaba, Vincenzo, Barros-Martins, Joana, Battistini, Luca, Bauer, Wolfgang, Baumgart, Sabine, Baumgarth, Nicole, Baumjohann, Dirk, Baying, Bianka, Bebawy, Mary, Becher, Burkhard, Beisker, Wolfgang, Benes, Vladimir, Beyaert, Rudi, Blanco, Alfonso, Boardman, Dominic A., Bogdan, Christian, Borger, Jessica G., Borsellino, Giovanna, Boulais, Philip E., Bradford, Jolene A., Brenner, Dirk, Brinkman, Ryan R., Brooks, Anna E. S., Busch, Dirk H., Buescher, Martin, Bushnell, Timothy P., Calzetti, Federica, Cameron, Garth, Cammarata, Ilenia, Cao, Xuetao, Cardell, Susanna L., Casola, Stefano, Cassatella, Marco A., Cavani, Andrea, Celada, Antonio, Chatenoud, Lucienne, Chattopadhyay, Pratip K., Chow, Sue, Christakou, Eleni, Cicin-Sain, Luka, Clerici, Mario, Colombo, Federico S., Cook, Laura, Cooke, Anne, Cooper, Andrea M., Corbett, Alexandra J., Cosma, Antonio, Cosmi, Lorenzo, Coulie, Pierre G., Cumano, Ana, Cvetkovic, Ljiljana, Dang, Van Duc, Dang-Heine, Chantip, Davey, Martin S., Davies, Derek, De Biasi, Sara, Del Zotto, Genny, Dela Cruz, Gelo Victoriano, Delacher, Michael, Della Bella, Silvia, Dellabona, Paolo, Deniz, Guennur, Dessing, Mark, Di Santo, James P., Diefenbach, Andreas, Dieli, Francesco, Dolf, Andreas, Doerner, Thomas, Dress, Regine J., Dudziak, Diana, Dustin, Michael, Dutertre, Charles-Antoine, Ebner, Friederike, Eckle, Sidonia B. G., Edinger, Matthias, Eede, Pascale, Ehrhardt, Goetz R. A., Eich, Marcus, Engel, Pablo, Engelhardt, Britta, Erdei, Anna, Esser, Charlotte, Everts, Bart, Evrard, Maximilien, Falk, Christine S., Fehniger, Todd A., Felipo-Benavent, Mar, Ferry, Helen, Feuerer, Markus, Filby, Andrew, Filkor, Kata, Fillatreau, Simon, Follo, Marie, Foerster, Irmgard, Foster, John, Foulds, Gemma A., Frehse, Britta, Frenette, Paul S., Frischbutter, Stefan, Fritzsche, Wolfgang, Galbraith, David W., Gangaev, Anastasia, Garbi, Natalio, Gaudilliere, Brice, Gazzinelli, Ricardo T., Geginat, Jens, Gerner, Wilhelm, Gherardin, Nicholas A., Ghoreschi, Kamran, Gibellini, Lara, Ginhoux, Florent, Goda, Keisuke, Godfrey, Dale I., Goettlinger, Christoph, Gonzalez-Navajas, Jose M., Goodyear, Carl S., Gori, Andrea, Grogan, Jane L., Grummitt, Daryl, Gruetzkau, Andreas, Haftmann, Claudia, Hahn, Jonas, Hammad, Hamida, Haemmerling, Guenter, Hansmann, Leo, Hansson, Goran, Harpur, Christopher M., Hartmann, Susanne, Hauser, Andrea, Hauser, Anja E., Haviland, David L., Hedley, David, Hernandez, Daniela C., Herrera, Guadalupe, Herrmann, Martin, Hess, Christoph, Hoefer, Thomas, Hoffmann, Petra, Hogquist, Kristin, Holland, Tristan, Hollt, Thomas, Holmdahl, Rikard, Hombrink, Pleun, Houston, Jessica P., Hoyer, Bimba F., Huang, Bo, Huang, Fang-Ping, Huber, Johanna E., Huehn, Jochen, Hundemer, Michael, Hunter, Christopher A., Hwang, William Y. K., Iannone, Anna, Ingelfinger, Florian, Ivison, Sabine M., Jaeck, Hans-Martin, Jani, Peter K., Javega, Beatriz, Jonjic, Stipan, Kaiser, Toralf, Kalina, Tomas, Kamradt, Thomas, Kaufmann, Stefan H. E., Keller, Baerbel, Ketelaars, Steven L. C., Khalilnezhad, Ahad, Khan, Srijit, Kisielow, Jan, Klenerman, Paul, Knopf, Jasmin, Koay, Hui-Fern, Kobow, Katja, Kolls, Jay K., Kong, Wan Ting, Kopf, Manfred, Korn, Thomas, Kriegsmann, Katharina, Kristyanto, Hendy, Kroneis, Thomas, Krueger, Andreas, Kuehne, Jenny, Kukat, Christian, Kunkel, Desiree, Kunze-Schumacher, Heike, Kurosaki, Tomohiro, Kurts, Christian, Kvistborg, Pia, Kwok, Immanuel, Landry, Jonathan, Lantz, Olivier, Lanuti, Paola, LaRosa, Francesca, Lehuen, Agnes, LeibundGut-Landmann, Salome, Leipold, Michael D., Leung, Leslie Y. T., Levings, Megan K., Lino, Andreia C., Liotta, Francesco, Litwin, Virginia, Liu, Yanling, Ljunggren, Hans-Gustaf, Lohoff, Michael, Lombardi, Giovanna, Lopez, Lilly, Lopez-Botet, Miguel, Lovett-Racke, Amy E., Lubberts, Erik, Luche, Herve, Ludewig, Burkhard, Lugli, Enrico, Lunemann, Sebastian, Maecker, Holden T., Maggi, Laura, Maguire, Orla, Mair, Florian, Mair, Kerstin H., Mantovani, Alberto, Manz, Rudolf A., Marshall, Aaron J., Martinez-Romero, Alicia, Martrus, Gloria, Marventano, Ivana, Maslinski, Wlodzimierz, Matarese, Giuseppe, Mattioli, Anna Vittoria, Maueroder, Christian, Mazzoni, Alessio, McCluskey, James, McGrath, Mairi, McGuire, Helen M., McInnes, Iain B., Mei, Henrik E., Melchers, Fritz, Melzer, Susanne, Mielenz, Dirk, Miller, Stephen D., Mills, Kingston H. G., Minderman, Hans, Mjosberg, Jenny, Moore, Jonni, Moran, Barry, Moretta, Lorenzo, Mosmann, Tim R., Mueller, Susann, Multhoff, Gabriele, Munoz, Luis Enrique, Munz, Christian, Nakayama, Toshinori, Nasi, Milena, Neumann, Katrin, Ng, Lai Guan, Niedobitek, Antonia, Nourshargh, Sussan, Nunez, Gabriel, O'Connor, Jose-Enrique, Ochel, Aaron, Oja, Anna, Ordonez, Diana, Orfao, Alberto, Orlowski-Oliver, Eva, Ouyang, Wenjun, Oxenius, Annette, Palankar, Raghavendra, Panse, Isabel, Pattanapanyasat, Kovit, Paulsen, Malte, Pavlinic, Dinko, Penter, Livius, Peterson, Paert, Peth, Christian, Petriz, Jordi, Piancone, Federica, Pickl, Winfried F., Piconese, Silvia, Pinti, Marcello, Pockley, A. Graham, Podolska, Malgorzata Justyna, Poon, Zhiyong, Pracht, Katharina, Prinz, Immo, Pucillo, Carlo E. M., Quataert, Sally A., Quatrini, Linda, Quinn, Kylie M., Radbruch, Helena, Radstake, Tim R. D. J., Rahmig, Susann, Rahn, Hans-Peter, Rajwa, Bartek, Ravichandran, Gevitha, Raz, Yotam, Rebhahn, Jonathan A., Recktenwald, Diether, Reimer, Dorothea, Reis e Sousa, Caetano, Remmerswaal, Ester B. M., Richter, Lisa, Rico, Laura G., Riddell, Andy, Rieger, Aja M., Robinson, J. Paul, Romagnani, Chiara, Rubartelli, Anna, Ruland, Juergen, Saalmueller, Armin, Saeys, Yvan, Saito, Takashi, Sakaguchi, Shimon, Sala-de-Oyanguren, Francisco, Samstag, Yvonne, Sanderson, Sharon, Sandrock, Inga, Santoni, Angela, Sanz, Ramon Bellmas, Saresella, Marina, Sautes-Fridman, Catherine, Sawitzki, Birgit, Schadt, Linda, Scheffold, Alexander, Scherer, Hans U., Schiemann, Matthias, Schildberg, Frank A., Schimisky, Esther, Schlitzer, Andreas, Schlosser, Josephine, Schmid, Stephan, Schmitt, Steffen, Schober, Kilian, Schraivogel, Daniel, Schuh, Wolfgang, Schueler, Thomas, Schulte, Reiner, Schulz, Axel Ronald, Schulz, Sebastian R., Scotta, Cristiano, Scott-Algara, Daniel, Sester, David P., Shankey, T. Vincent, Silva-Santos, Bruno, Simon, Anna Katharina, Sitnik, Katarzyna M., Sozzani, Silvano, Speiser, Daniel E., Spidlen, Josef, Stahlberg, Anders, Stall, Alan M., Stanley, Natalie, Stark, Regina, Stehle, Christina, Steinmetz, Tobit, Stockinger, Hannes, Takahama, Yousuke, Takeda, Kiyoshi, Tan, Leonard, Tarnok, Attila, Tiegs, Gisa, Toldi, Gergely, Tornack, Julia, Traggiai, Elisabetta, Trebak, Mohamed, Tree, Timothy I. M., Trotter, Joe, Trowsdale, John, Tsoumakidou, Maria, Ulrich, Henning, Urbanczyk, Sophia, van de Veen, Willem, van den Broek, Maries, van der Pol, Edwin, Van Gassen, Sofie, Van Isterdael, Gert, van Lier, Rene A. W., Veldhoen, Marc, Vento-Asturias, Salvador, Vieira, Paulo, Voehringer, David, Volk, Hans-Dieter, von Borstel, Anouk, von Volkmann, Konrad, Waisman, Ari, Walker, Rachael V., Wallace, Paul K., Wang, Sa A., Wang, Xin M., Ward, Michael D., Ward-Hartstonge, Kirsten A., Warnatz, Klaus, Warnes, Gary, Warth, Sarah, Waskow, Claudia, Watson, James V., Watzl, Carsten, Wegener, Leonie, Weisenburger, Thomas, Wiedemann, Annika, Wienands, Juergen, Wilharm, Anneke, Wilkinson, Robert John, Willimsky, Gerald, Wing, James B., Winkelmann, Rieke, Winkler, Thomas H., Wirz, Oliver F., Wong, Alicia, Wurst, Peter, Yang, Jennie H. M., Yang, Juhao, Yazdanbakhsh, Maria, Yu, Liping, Yue, Alice, Zhang, Hanlin, Zhao, Yi, Ziegler, Susanne Maria, Zielinski, Christina, Zimmermann, Jakob, Zychlinsky, Arturo, Cossarizza, Andrea, Chang, Hyun-Dong, Radbruch, Andreas, Acs, Andreas, Adam, Dieter, Adam-Klages, Sabine, Agace, William W., Aghaeepour, Nima, Akdis, Muebeccel, Allez, Matthieu, Almeida, Larissa Nogueira, Alvisi, Giorgia, Anderson, Graham, Andrae, Immanuel, Annunziato, Francesco, Anselmo, Achille, Bacher, Petra, Baldari, Cosima T., Bari, Sudipto, Barnaba, Vincenzo, Barros-Martins, Joana, Battistini, Luca, Bauer, Wolfgang, Baumgart, Sabine, Baumgarth, Nicole, Baumjohann, Dirk, Baying, Bianka, Bebawy, Mary, Becher, Burkhard, Beisker, Wolfgang, Benes, Vladimir, Beyaert, Rudi, Blanco, Alfonso, Boardman, Dominic A., Bogdan, Christian, Borger, Jessica G., Borsellino, Giovanna, Boulais, Philip E., Bradford, Jolene A., Brenner, Dirk, Brinkman, Ryan R., Brooks, Anna E. S., Busch, Dirk H., Buescher, Martin, Bushnell, Timothy P., Calzetti, Federica, Cameron, Garth, Cammarata, Ilenia, Cao, Xuetao, Cardell, Susanna L., Casola, Stefano, Cassatella, Marco A., Cavani, Andrea, Celada, Antonio, Chatenoud, Lucienne, Chattopadhyay, Pratip K., Chow, Sue, Christakou, Eleni, Cicin-Sain, Luka, Clerici, Mario, Colombo, Federico S., Cook, Laura, Cooke, Anne, Cooper, Andrea M., Corbett, Alexandra J., Cosma, Antonio, Cosmi, Lorenzo, Coulie, Pierre G., Cumano, Ana, Cvetkovic, Ljiljana, Dang, Van Duc, Dang-Heine, Chantip, Davey, Martin S., Davies, Derek, De Biasi, Sara, Del Zotto, Genny, Dela Cruz, Gelo Victoriano, Delacher, Michael, Della Bella, Silvia, Dellabona, Paolo, Deniz, Guennur, Dessing, Mark, Di Santo, James P., Diefenbach, Andreas, Dieli, Francesco, Dolf, Andreas, Doerner, Thomas, Dress, Regine J., Dudziak, Diana, Dustin, Michael, Dutertre, Charles-Antoine, Ebner, Friederike, Eckle, Sidonia B. G., Edinger, Matthias, Eede, Pascale, Ehrhardt, Goetz R. A., Eich, Marcus, Engel, Pablo, Engelhardt, Britta, Erdei, Anna, Esser, Charlotte, Everts, Bart, Evrard, Maximilien, Falk, Christine S., Fehniger, Todd A., Felipo-Benavent, Mar, Ferry, Helen, Feuerer, Markus, Filby, Andrew, Filkor, Kata, Fillatreau, Simon, Follo, Marie, Foerster, Irmgard, Foster, John, Foulds, Gemma A., Frehse, Britta, Frenette, Paul S., Frischbutter, Stefan, Fritzsche, Wolfgang, Galbraith, David W., Gangaev, Anastasia, Garbi, Natalio, Gaudilliere, Brice, Gazzinelli, Ricardo T., Geginat, Jens, Gerner, Wilhelm, Gherardin, Nicholas A., Ghoreschi, Kamran, Gibellini, Lara, Ginhoux, Florent, Goda, Keisuke, Godfrey, Dale I., Goettlinger, Christoph, Gonzalez-Navajas, Jose M., Goodyear, Carl S., Gori, Andrea, Grogan, Jane L., Grummitt, Daryl, Gruetzkau, Andreas, Haftmann, Claudia, Hahn, Jonas, Hammad, Hamida, Haemmerling, Guenter, Hansmann, Leo, Hansson, Goran, Harpur, Christopher M., Hartmann, Susanne, Hauser, Andrea, Hauser, Anja E., Haviland, David L., Hedley, David, Hernandez, Daniela C., Herrera, Guadalupe, Herrmann, Martin, Hess, Christoph, Hoefer, Thomas, Hoffmann, Petra, Hogquist, Kristin, Holland, Tristan, Hollt, Thomas, Holmdahl, Rikard, Hombrink, Pleun, Houston, Jessica P., Hoyer, Bimba F., Huang, Bo, Huang, Fang-Ping, Huber, Johanna E., Huehn, Jochen, Hundemer, Michael, Hunter, Christopher A., Hwang, William Y. K., Iannone, Anna, Ingelfinger, Florian, Ivison, Sabine M., Jaeck, Hans-Martin, Jani, Peter K., Javega, Beatriz, Jonjic, Stipan, Kaiser, Toralf, Kalina, Tomas, Kamradt, Thomas, Kaufmann, Stefan H. E., Keller, Baerbel, Ketelaars, Steven L. C., Khalilnezhad, Ahad, Khan, Srijit, Kisielow, Jan, Klenerman, Paul, Knopf, Jasmin, Koay, Hui-Fern, Kobow, Katja, Kolls, Jay K., Kong, Wan Ting, Kopf, Manfred, Korn, Thomas, Kriegsmann, Katharina, Kristyanto, Hendy, Kroneis, Thomas, Krueger, Andreas, Kuehne, Jenny, Kukat, Christian, Kunkel, Desiree, Kunze-Schumacher, Heike, Kurosaki, Tomohiro, Kurts, Christian, Kvistborg, Pia, Kwok, Immanuel, Landry, Jonathan, Lantz, Olivier, Lanuti, Paola, LaRosa, Francesca, Lehuen, Agnes, LeibundGut-Landmann, Salome, Leipold, Michael D., Leung, Leslie Y. T., Levings, Megan K., Lino, Andreia C., Liotta, Francesco, Litwin, Virginia, Liu, Yanling, Ljunggren, Hans-Gustaf, Lohoff, Michael, Lombardi, Giovanna, Lopez, Lilly, Lopez-Botet, Miguel, Lovett-Racke, Amy E., Lubberts, Erik, Luche, Herve, Ludewig, Burkhard, Lugli, Enrico, Lunemann, Sebastian, Maecker, Holden T., Maggi, Laura, Maguire, Orla, Mair, Florian, Mair, Kerstin H., Mantovani, Alberto, Manz, Rudolf A., Marshall, Aaron J., Martinez-Romero, Alicia, Martrus, Gloria, Marventano, Ivana, Maslinski, Wlodzimierz, Matarese, Giuseppe, Mattioli, Anna Vittoria, Maueroder, Christian, Mazzoni, Alessio, McCluskey, James, McGrath, Mairi, McGuire, Helen M., McInnes, Iain B., Mei, Henrik E., Melchers, Fritz, Melzer, Susanne, Mielenz, Dirk, Miller, Stephen D., Mills, Kingston H. G., Minderman, Hans, Mjosberg, Jenny, Moore, Jonni, Moran, Barry, Moretta, Lorenzo, Mosmann, Tim R., Mueller, Susann, Multhoff, Gabriele, Munoz, Luis Enrique, Munz, Christian, Nakayama, Toshinori, Nasi, Milena, Neumann, Katrin, Ng, Lai Guan, Niedobitek, Antonia, Nourshargh, Sussan, Nunez, Gabriel, O'Connor, Jose-Enrique, Ochel, Aaron, Oja, Anna, Ordonez, Diana, Orfao, Alberto, Orlowski-Oliver, Eva, Ouyang, Wenjun, Oxenius, Annette, Palankar, Raghavendra, Panse, Isabel, Pattanapanyasat, Kovit, Paulsen, Malte, Pavlinic, Dinko, Penter, Livius, Peterson, Paert, Peth, Christian, Petriz, Jordi, Piancone, Federica, Pickl, Winfried F., Piconese, Silvia, Pinti, Marcello, Pockley, A. Graham, Podolska, Malgorzata Justyna, Poon, Zhiyong, Pracht, Katharina, Prinz, Immo, Pucillo, Carlo E. M., Quataert, Sally A., Quatrini, Linda, Quinn, Kylie M., Radbruch, Helena, Radstake, Tim R. D. J., Rahmig, Susann, Rahn, Hans-Peter, Rajwa, Bartek, Ravichandran, Gevitha, Raz, Yotam, Rebhahn, Jonathan A., Recktenwald, Diether, Reimer, Dorothea, Reis e Sousa, Caetano, Remmerswaal, Ester B. M., Richter, Lisa, Rico, Laura G., Riddell, Andy, Rieger, Aja M., Robinson, J. Paul, Romagnani, Chiara, Rubartelli, Anna, Ruland, Juergen, Saalmueller, Armin, Saeys, Yvan, Saito, Takashi, Sakaguchi, Shimon, Sala-de-Oyanguren, Francisco, Samstag, Yvonne, Sanderson, Sharon, Sandrock, Inga, Santoni, Angela, Sanz, Ramon Bellmas, Saresella, Marina, Sautes-Fridman, Catherine, Sawitzki, Birgit, Schadt, Linda, Scheffold, Alexander, Scherer, Hans U., Schiemann, Matthias, Schildberg, Frank A., Schimisky, Esther, Schlitzer, Andreas, Schlosser, Josephine, Schmid, Stephan, Schmitt, Steffen, Schober, Kilian, Schraivogel, Daniel, Schuh, Wolfgang, Schueler, Thomas, Schulte, Reiner, Schulz, Axel Ronald, Schulz, Sebastian R., Scotta, Cristiano, Scott-Algara, Daniel, Sester, David P., Shankey, T. Vincent, Silva-Santos, Bruno, Simon, Anna Katharina, Sitnik, Katarzyna M., Sozzani, Silvano, Speiser, Daniel E., Spidlen, Josef, Stahlberg, Anders, Stall, Alan M., Stanley, Natalie, Stark, Regina, Stehle, Christina, Steinmetz, Tobit, Stockinger, Hannes, Takahama, Yousuke, Takeda, Kiyoshi, Tan, Leonard, Tarnok, Attila, Tiegs, Gisa, Toldi, Gergely, Tornack, Julia, Traggiai, Elisabetta, Trebak, Mohamed, Tree, Timothy I. M., Trotter, Joe, Trowsdale, John, Tsoumakidou, Maria, Ulrich, Henning, Urbanczyk, Sophia, van de Veen, Willem, van den Broek, Maries, van der Pol, Edwin, Van Gassen, Sofie, Van Isterdael, Gert, van Lier, Rene A. W., Veldhoen, Marc, Vento-Asturias, Salvador, Vieira, Paulo, Voehringer, David, Volk, Hans-Dieter, von Borstel, Anouk, von Volkmann, Konrad, Waisman, Ari, Walker, Rachael V., Wallace, Paul K., Wang, Sa A., Wang, Xin M., Ward, Michael D., Ward-Hartstonge, Kirsten A., Warnatz, Klaus, Warnes, Gary, Warth, Sarah, Waskow, Claudia, Watson, James V., Watzl, Carsten, Wegener, Leonie, Weisenburger, Thomas, Wiedemann, Annika, Wienands, Juergen, Wilharm, Anneke, Wilkinson, Robert John, Willimsky, Gerald, Wing, James B., Winkelmann, Rieke, Winkler, Thomas H., Wirz, Oliver F., Wong, Alicia, Wurst, Peter, Yang, Jennie H. M., Yang, Juhao, Yazdanbakhsh, Maria, Yu, Liping, Yue, Alice, Zhang, Hanlin, Zhao, Yi, Ziegler, Susanne Maria, Zielinski, Christina, Zimmermann, Jakob, and Zychlinsky, Arturo

Abstract

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.

Published
2019

20. miR-181a/b-1 controls thymic selection of Treg cells and tunes their suppressive capacity

Author

Bhandoola, Avinash, Łyszkiewicz, Marcin, Winter, Samantha J., Witzlau, Katrin, Föhse, Lisa, Brownlie, Rebecca, Puchałka, Jacek, Verheyden, Nikita A., Kunze-Schumacher, Heike, Imelmann, Esther, Blume, Jonas, Raha, Solaiman, Sekiya, Takashi, Yoshimura, Akihiko, Früh, Jochen, Ullrich, Evelyn, Hühn, Jochen, Weiss, Siegfried, Gutierrez, Maximiliano, Prinz, Immo, Zamoyska, Rose, Zie̜tara, Natalia Karolina, Krueger, Andreas, Bhandoola, Avinash, Łyszkiewicz, Marcin, Winter, Samantha J., Witzlau, Katrin, Föhse, Lisa, Brownlie, Rebecca, Puchałka, Jacek, Verheyden, Nikita A., Kunze-Schumacher, Heike, Imelmann, Esther, Blume, Jonas, Raha, Solaiman, Sekiya, Takashi, Yoshimura, Akihiko, Früh, Jochen, Ullrich, Evelyn, Hühn, Jochen, Weiss, Siegfried, Gutierrez, Maximiliano, Prinz, Immo, Zamoyska, Rose, Zie̜tara, Natalia Karolina, and Krueger, Andreas

Abstract

The interdependence of selective cues during development of regulatory T cells (Treg cells) in the thymus and their suppressive function remains incompletely understood. Here, we analyzed this interdependence by taking advantage of highly dynamic changes in expression of microRNA 181 family members miR-181a-1 and miR-181b-1 (miR-181a/b-1) during late T-cell development with very high levels of expression during thymocyte selection, followed by massive down-regulation in the periphery. Loss of miR-181a/b-1 resulted in inefficient de novo generation of Treg cells in the thymus but simultaneously permitted homeostatic expansion in the periphery in the absence of competition. Modulation of T-cell receptor (TCR) signal strength in vivo indicated that miR-181a/b-1 controlled Treg-cell formation via establishing adequate signaling thresholds. Unexpectedly, miR-181a/b-1–deficient Treg cells displayed elevated suppressive capacity in vivo, in line with elevated levels of cytotoxic T-lymphocyte–associated 4 (CTLA-4) protein, but not mRNA, in thymic and peripheral Treg cells. Therefore, we propose that intrathymic miR-181a/b-1 controls development of Treg cells and imposes a developmental legacy on their peripheral function.

Published
2019

24. miR‐191 modulates B‐cell development and targets transcription factors E2A, Foxp1, and Egr1

Author

Blume, Jonas, primary, Ziętara, Natalia, additional, Witzlau, Katrin, additional, Liu, Yanshan, additional, Sanchez, Oskar Ortiz, additional, Puchałka, Jacek, additional, Winter, Samantha J., additional, Kunze‐Schumacher, Heike, additional, Saran, Namita, additional, Düber, Sandra, additional, Roy, Bishnudeo, additional, Weiss, Siegfried, additional, Klein, Christoph, additional, Wurst, Wolfgang, additional, Łyszkiewicz, Marcin, additional, and Krueger, Andreas, additional

Published
2018
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26. Germline deletion reveals a non-essential role of the atypical MAPK6/ERK3

Author

Ronkina, Natalia, Schuster-Gossler, Karin, Hansmann, Florian Heinrich, Kunze-Schumacher, Heike, Sandrock, Inga, Yakovleva, Tatiana, Lafera, Juri, Baumgärtner, Wolfgang, Krueger, Andreas, Prinz, Immo, Gossler, Achim, Kotlyarov, Alexey, Gaestel, Matthias, Ronkina, Natalia, Schuster-Gossler, Karin, Hansmann, Florian Heinrich, Kunze-Schumacher, Heike, Sandrock, Inga, Yakovleva, Tatiana, Lafera, Juri, Baumgärtner, Wolfgang, Krueger, Andreas, Prinz, Immo, Gossler, Achim, Kotlyarov, Alexey, and Gaestel, Matthias

Abstract

MAPK6/ERK3 is an atypical member of the MAPKs. An essential role has been suggested by the perinatal lethal phenotype of ERK3 knockout mice carrying a lacZ insertion in exon 2 due to pulmonary disfunction and by defects in function, activation and positive selection of T cells. To study the role of ERK3 in vivo, we generated mice carrying a conditional Erk3 allele with exon3 flanked by LoxP sites. Loss of ERK3 protein was validated after deletion of Erk3 in the female germ line using zona pellucida 3 (Zp3)-cre and a clear reduction of the protein kinase MK5 is detected, providing first evidence for the existence of the ERK3/MK5 signaling complex in vivo. In contrast to the previously reported Erk3 knockout phenotype, these mice are viable and fertile, do not display pulmonary hypoplasia, acute respiratory failure, abnormal T cell development, reduction of thymocyte numbers or altered T cells selection. Hence, ERK3 is dispensable for pulmonary and T-cell functions. The perinatal lethality, lung and T-cell defects of the previous ERK3 knockout mice are likely due to ERK3-unrelated effects of the inserted lacZ-neomycin-resistance-cassette. The knockout mouse of the closely related atypical MAPK ERK4/MAPK4 is also normal suggesting redundant functions of both protein kinases.

Published
2018

27. miRNA miR-21 is largely dispensable for intrathymic T-cell development

Author

Kunze-Schumacher, Heike, Winter, Samantha J., Imelmann, Esther, Krueger, Andreas, Kunze-Schumacher, Heike, Winter, Samantha J., Imelmann, Esther, and Krueger, Andreas

Abstract

Development of T cells in the thymus is tightly controlled to continually produce functional, but not autoreactive, T cells. miRNAs provide a layer of post-transcriptional gene regulation to this process, but the role of many individual miRNAs in T-cell development remains unclear. miR-21 is prominently expressed in immature thymocytes followed by a steep decline in more mature cells. We hypothesized that such a dynamic expression was indicative of a regulatory function in intrathymic T-cell development. To test this hypothesis, we analyzed T-cell development in miR-21-deficient mice at steady state and under competitive conditions in mixed bone-marrow chimeras. We complemented analysis of knock-out animals by employing over-expression in vivo. Finally, we assessed miR-21 function in negative selection in vivo as well as differentiation in co-cultures. Together, these experiments revealed that miR-21 is largely dispensable for physiologic T-cell development. Given that miR-21 has been implicated in regulation of cellular stress responses, we assessed a potential role of miR-21 in endogenous regeneration of the thymus after sublethal irradiation. Again, miR-21 was completely dispensable in this process. We concluded that, despite prominent and highly dynamic expression in thymocytes, miR-21 expression was not required for physiologic T-cell development or endogenous regeneration.

Published
2018

28. Releasing the concept of HLA‐allele specific peptide anchors in viral infections: A non‐canonical naturally presented human cytomegalovirus‐derived HLA‐A*24:02 restricted peptide drives exquisite immunogenicity.

Author

Pump, Wiebke C., Schulz, Rebecca, Huyton, Trevor, Kunze‐Schumacher, Heike, Martens, Jörg, Hò, Gia‐Gia T., Blasczyk, Rainer, and Bade‐Doeding, Christina

Subjects
ALLELES, KILLER cell receptors, VIRUS diseases, HEMATOPOIETIC stem cell transplantation, LYTIC cycle, HUMAN cytomegalovirus
Abstract

T‐cell receptors possess the unique ability to survey and respond to their permanently modified ligands, self HLA‐I molecules bound to non‐self peptides of various origin. This highly specific immune function is impaired following hematopoietic stem cell transplantation (HSCT) for a timespan of several months needed for the maturation of T‐cells. Especially, the progression of HCMV disease in immunocompromised patients induces life‐threatening situations. Therefore, the need for a new immune system that delivers vital and potent CD8+ T‐cells carrying TCRs that recognize even one human cytomegalovirus (HCMV) peptide/HLA molecule and clear the viral infection long term becomes obvious. The transcription and translation of HCMV proteins in the lytic cycle is a precisely regulated cascade of processes, therefore, it is a highly sensitive challenge to adjust the exact time point of HCMV‐peptide recruitment over self‐peptides. We utilized soluble HLA technology in HCMV‐infected fibroblasts and sequenced naturally sHLA‐A*24:02 presented HCMV‐derived peptides. One peptide of 14 AAs length derived from the IE2 antigen induced the strongest T‐cell responses; this peptide can be detected with a low ranking score in general peptide prediction databanks. These results highlight the need for elaborate and HLA‐allele specific peptide selection. [ABSTRACT FROM AUTHOR]

Published
2019
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29. MicroRNA miR‐181a/b‐1 controls MAIT cell development.

Author

Winter, Samantha J, Kunze‐Schumacher, Heike, Imelmann, Esther, Grewers, Zoe, Osthues, Tabea, and Krueger, Andreas

Subjects
*MICRORNA, *T cells, *TRANSCRIPTION factors, *PATHOGENIC microorganisms, *DEVELOPMENTAL cytology, *LABORATORY mice
Abstract

Mucosal‐associated invariant T (MAIT) cells constitute a major fraction of innate‐like T cells in humans with critical roles in defense against microbial pathogens and in maintaining mucosal integrity. However, the molecular mechanisms underlying MAIT cell development remain largely elusive. Here we investigated the role of miR‐181a/b‐1, a pair of microRNAs that serve as rheostat of TCR signal strength, in this process. Loss of miR‐181a/b‐1 in mice resulted in a profound arrest in early MAIT cell development. As a consequence, in the absence of miR‐181a/b‐1, thymic MAIT cells failed to acquire functional maturity based on expression of transcription factors PLZF, T‐bet and RORγt. Temporal analysis of development using a molecular timer in combination with loss of miR‐181a/b‐1 revealed that MAIT cells complete functional maturation in the periphery and indicates that functionally mature MAIT cells in the thymus are long‐term resident cells. Thus, our study provides insight into the dynamics of MAIT cell development in vivo. Of note, deletion of miR‐181a/b‐1 alone completely mirrored loss of all miRNAs. Winter et al. demonstrate that intrathymic development of MAIT cells is controlled by miRNA miR‐181a/b‐1. See also: News & Commentary by Koay & Godfrey [ABSTRACT FROM AUTHOR]

Published
2019
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32. Soluble HLA Technology as a Strategy to Evaluate the Impact of HLA Mismatches

Author

Kunze-Schumacher, Heike, Blasczyk, Rainer, and Bade-Doeding, Christina

Subjects
Article Subject
Abstract

HLA class I incompatibilities still remain one of the main barriers for unrelated bone marrow transplantation (BMT); hence the molecular understanding of how to mismatch patients and donors and still have successful clinical outcomes will guide towards the future of unrelated BMT. One way to estimate the magnitude of polymorphisms within the PBR is to determine which peptides can be selected by individual HLA alleles and subsequently presented for recognition by T cells. The features (structure, length, and sequence) of different peptides each confer an individual pHLA landscape and thus directly shape the individual immune response. The elution and sequencing of peptides by mass spectrometric analysis enable determining the bona fide repertoire of presented peptides for a given allele. This is an effective and simple way to compare the functions of allelic variants and make a first assessment of their degree of permissivity. We describe the methodology used for peptide sequencing and the limitations of peptide prediction tools compared to experimental methods. We highlight the altered peptide features that are observed between allelic variants and the need to discover the altered peptide repertoire in situations of “artificial” graft versus host disease (GvHD) that occur in HLA-specific hypersensitive immune responses to drugs.

Published
2014
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33. miR‐191 modulates B‐cell development and targets transcription factors E2A, Foxp1, and Egr1.

Author

Blume, Jonas, Ziętara, Natalia, Witzlau, Katrin, Liu, Yanshan, Sanchez, Oskar Ortiz, Puchałka, Jacek, Winter, Samantha J., Kunze‐Schumacher, Heike, Saran, Namita, Düber, Sandra, Roy, Bishnudeo, Weiss, Siegfried, Klein, Christoph, Wurst, Wolfgang, Łyszkiewicz, Marcin, and Krueger, Andreas

Abstract

The interdependence of posttranscriptional gene regulation via miRNA and transcriptional regulatory networks in lymphocyte development is poorly understood. Here, we identified miR‐191 as direct upstream modulator of a transcriptional module comprising the transcription factors Foxp1, E2A, and Egr1. Deletion as well as ectopic expression of miR‐191 resulted in developmental arrest in B lineage cells, indicating that fine tuning of the combined expression levels of Foxp1, E2A, and Egr1, which in turn control somatic recombination and cytokine‐driven expansion, constitutes a prerequisite for efficient B‐cell development. In conclusion, we propose that miR‐191 acts as a rheostat in B‐cell development by fine tuning a key transcriptional program. Through CRISPR/Cas9‐mediated deletion and retrogenic overexpression of miR‐191, we demonstrate that a defined window of expression of this miRNA is required to allow for efficient B‐cell development beyond the proB/preBI stage. Furthermore, we suggest that control of B‐cell development by miR‐191 is mediated through its targets Foxp1, E2A, and/or Egr1. [ABSTRACT FROM AUTHOR]

Published
2019
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35. OR25

Author

Kraemer, Thomas, primary, Huyton, Trevor, additional, Kunze-Schumacher, Heike, additional, Abels, Wiebke, additional, Blasczyk, Rainer, additional, and Bade-Doeding, Christina, additional

Published
2014
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36. OR20

Author

Kunze-Schumacher, Heike, primary, Trevor, Huyton, additional, Blasczyk, Rainer, additional, and Bade-Doeding, Christina, additional

Published
2014
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