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1. mRNA-1273 vaccination induces polyfunctional memory CD4 and CD8 T cell responses in patients with solid cancers undergoing immunotherapy or/and chemotherapy

Author: Anastasia Gangaev, Yannick van Sleen, Nicole Brandhorst, Kelly Hoefakker, Bimal Prajapati, Amrita Singh, Annemarie Boerma, Marieke van der Heiden, Sjoukje F. Oosting, Astrid A. M. van der Veldt, T. Jeroen N. Hiltermann, Corine H. GeurtsvanKessel, Anne-Marie C. Dingemans, Egbert F. Smit, Elisabeth G. E. de Vries, John B. A. G. Haanen, Pia Kvistborg, and Debbie van Baarle
Subjects: SARS-CoV-2-specific T cells, COVID-19, cancer, immunotherapy, chemotherapy, COVID-19 vaccination, Immunologic diseases. Allergy, RC581-607
Abstract: IntroductionResearch has confirmed the safety and comparable seroconversion rates following SARS-CoV-2 vaccination in patients with solid cancers. However, the impact of cancer treatment on vaccine-induced T cell responses remains poorly understood.MethodsIn this study, we expand on previous findings within the VOICE trial by evaluating the functional and phenotypic composition of mRNA-1273-induced T cell responses in patients with solid tumors undergoing immunotherapy, chemotherapy, or both, compared to individuals without cancer. We conducted an ELISpot analysis on 386 participants to assess spike-specific T cell responses 28 days after full vaccination. Further in-depth characterization of using flow cytometry was performed on a subset of 63 participants to analyze the functional phenotype and differentiation state of spike-specific T cell responses.ResultsELISpot analysis showed robust induction of spike-specific T cell responses across all treatment groups, with response rates ranging from 75% to 80%. Flow cytometry analysis revealed a distinctive cytokine production pattern across cohorts, with CD4 T cells producing IFNγ, TNF, and IL-2, and CD8 T cells producing IFNγ, TNF, and CCL4. Variations were observed in the proportion of monofunctional CD4 T cells producing TNF, particularly higher in individuals without cancer and patients treated with chemotherapy alone, while those treated with immunotherapy or chemoimmunotherapy predominantly produced IFNγ. Despite these differences, polyfunctional spike-specific memory CD4 and CD8 T cell responses were comparable across cohorts. Notably, immunotherapy-treated patients exhibited an expansion of spike-specific CD4 T cells with a terminally differentiated effector memory phenotype.DiscussionThese findings demonstrate that systemic treatment in patients with solid tumors does not compromise the quality of polyfunctional mRNA-1273-induced T cell responses. This underscores the importance of COVID-19 vaccination in patients with solid cancers undergoing systemic treatment.
Published: 2024
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2. Differential effects of PD-1 and CTLA-4 blockade on the melanoma-reactive CD8 T cell response

Author: Gangaev, Anastasia, Rozeman, Elisa A, Rohaan, Maartje W, Isaeva, Olga I, Philips, Daisy, Patiwael, Sanne, van den Berg, Joost H, Ribas, Antoni, Schadendorf, Dirk, Schilling, Bastian, Schumacher, Ton N, Blank, Christian U, Haanen, John BAG, and Kvistborg, Pia
Subjects: Biomedical and Clinical Sciences, Oncology and Carcinogenesis, Immunology, Cancer, Clinical Research, 5.1 Pharmaceuticals, Development of treatments and therapeutic interventions, Adult, Aged, Aged, 80 and over, CD8-Positive T-Lymphocytes, CTLA-4 Antigen, Cohort Studies, Epitopes, T-Lymphocyte, Female, Hepatitis A Virus Cellular Receptor 2, Humans, Immune Checkpoint Inhibitors, In Vitro Techniques, Kinetics, Lymphocyte Activation, Male, Melanoma, Melanoma-Specific Antigens, Middle Aged, Programmed Cell Death 1 Receptor, Receptors, Antigen, T-Cell, alpha-beta, Receptors, CXCR5, PD-1, CTLA-4, melanoma-reactive CD8 T cells
Abstract: Immune checkpoint inhibitors targeting programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have revolutionized the treatment of melanoma patients. Based on early studies addressing the mechanism of action, it was assumed that PD-1 blockade mostly influences T cell responses at the tumor site. However, recent work has demonstrated that PD-1 blockade can influence the T cell compartment in peripheral blood. If the activation of circulating, tumor-reactive T cells would form an important mechanism of action of PD-1 blockade, it may be predicted that such blockade would alter either the frequency and/or the breadth of the tumor-reactive CD8 T cell response. To address this question, we analyzed CD8 T cell responses toward 71 melanoma-associated epitopes in peripheral blood of 24 melanoma patients. We show that both the frequency and the breadth of the circulating melanoma-reactive CD8 T cell response was unaltered upon PD-1 blockade. In contrast, a broadening of the circulating melanoma-reactive CD8 T cell response was observed upon CTLA-4 blockade, in concordance with our prior data. Based on these results, we conclude that PD-1 and CTLA-4 blockade have distinct mechanisms of action. In addition, the data provide an argument in favor of the hypothesis that anti-PD-1 therapy may primarily act at the tumor site.
Published: 2021

3. mRNA-1273 vaccination induces polyfunctional memory CD4 and CD8 T cell responses in patients with solid cancers undergoing immunotherapy or/and chemotherapy.

Author: Gangaev, Anastasia, van Sleen, Yannick, Brandhorst, Nicole, Hoefakker, Kelly, Prajapati, Bimal, Singh, Amrita, Boerma, Annemarie, van der Heiden, Marieke, Oosting, Sjoukje F., van der Veldt, Astrid A. M., Hiltermann, T. Jeroen N., GeurtsvanKessel, Corine H., Dingemans, Anne-Marie C., Smit, Egbert F., de Vries, Elisabeth G. E., Haanen, John B. A. G., Kvistborg, Pia, and van Baarle, Debbie
Subjects: T cells, CANCER chemotherapy, COVID-19 vaccines, CANCER cells, FLOW cytometry
Abstract: Introduction: Research has confirmed the safety and comparable seroconversion rates following SARS-CoV-2 vaccination in patients with solid cancers. However, the impact of cancer treatment on vaccine-induced T cell responses remains poorly understood. Methods: In this study, we expand on previous findings within the VOICE trial by evaluating the functional and phenotypic composition of mRNA-1273-induced T cell responses in patients with solid tumors undergoing immunotherapy, chemotherapy, or both, compared to individuals without cancer. We conducted an ELISpot analysis on 386 participants to assess spike-specific T cell responses 28 days after full vaccination. Further in-depth characterization of using flow cytometry was performed on a subset of 63 participants to analyze the functional phenotype and differentiation state of spike-specific T cell responses. Results: ELISpot analysis showed robust induction of spike-specific T cell responses across all treatment groups, with response rates ranging from 75% to 80%. Flow cytometry analysis revealed a distinctive cytokine production pattern across cohorts, with CD4 T cells producing IFNγ, TNF, and IL-2, and CD8 T cells producing IFNγ, TNF, and CCL4. Variations were observed in the proportion of monofunctional CD4 T cells producing TNF, particularly higher in individuals without cancer and patients treated with chemotherapy alone, while those treated with immunotherapy or chemoimmunotherapy predominantly produced IFNγ. Despite these differences, polyfunctional spike-specific memory CD4 and CD8 T cell responses were comparable across cohorts. Notably, immunotherapy-treated patients exhibited an expansion of spike-specific CD4 T cells with a terminally differentiated effector memory phenotype. Discussion: These findings demonstrate that systemic treatment in patients with solid tumors does not compromise the quality of polyfunctional mRNA-1273-induced T cell responses. This underscores the importance of COVID-19 vaccination in patients with solid cancers undergoing systemic treatment. [ABSTRACT FROM AUTHOR]
Published: 2024
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4. Identification and characterization of a SARS-CoV-2 specific CD8+ T cell response with immunodominant features

Author: Anastasia Gangaev, Steven L. C. Ketelaars, Olga I. Isaeva, Sanne Patiwael, Anna Dopler, Kelly Hoefakker, Sara De Biasi, Lara Gibellini, Cristina Mussini, Giovanni Guaraldi, Massimo Girardis, Cami M. P. Talavera Ormeno, Paul J. M. Hekking, Neubury M. Lardy, Mireille Toebes, Robert Balderas, Ton N. Schumacher, Huib Ovaa, Andrea Cossarizza, and Pia Kvistborg
Subjects: Science
Abstract: Many viral antigens have been identified in patients with COVID-19 patients, but which of these result in meaningful immune responses is unclear. Here the authors identify a range of SARS-CoV-2 CD8+ T cell responses across patients including a response targeting an epitope of ORF1ab with immunodominant properties.
Published: 2021
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5. Identification and characterization of a SARS-CoV-2 specific CD8+ T cell response with immunodominant features

Author: Gangaev, Anastasia, Ketelaars, Steven L. C., Isaeva, Olga I., Patiwael, Sanne, Dopler, Anna, Hoefakker, Kelly, De Biasi, Sara, Gibellini, Lara, Mussini, Cristina, Guaraldi, Giovanni, Girardis, Massimo, Ormeno, Cami M. P. Talavera, Hekking, Paul J. M., Lardy, Neubury M., Toebes, Mireille, Balderas, Robert, Schumacher, Ton N., Ovaa, Huib, Cossarizza, Andrea, and Kvistborg, Pia
Published: 2021
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6. Alternative trafficking of Weibel‐Palade body proteins in CRISPR/Cas9‐engineered von Willebrand factor–deficient blood outgrowth endothelial cells

Author: Schillemans, Maaike, Kat, Marije, Westeneng, Jurjen, Gangaev, Anastasia, Hofman, Menno, Nota, Benjamin, van Alphen, Floris P.J., de Boer, Martin, van den Biggelaar, Maartje, Margadant, Coert, Voorberg, Jan, and Bierings, Ruben
Published: 2019
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7. Identification of presented SARS-CoV-2 HLA class I and HLA class II peptides using HLA peptidomics

Author: Adi Nagler, Shelly Kalaora, Chaya Barbolin, Anastasia Gangaev, Steven L.C. Ketelaars, Michal Alon, Joy Pai, Gil Benedek, Yfat Yahalom-Ronen, Noam Erez, Polina Greenberg, Gal Yagel, Aviyah Peri, Yishai Levin, Ansuman T. Satpathy, Erez Bar-Haim, Nir Paran, Pia Kvistborg, and Yardena Samuels
Subjects: SARS-CoV-2, HLA, Peptides, Peptidomics, out-of-frame-ORFs, immuno-reactive, Biology (General), QH301-705.5
Abstract: Summary: The human leukocyte antigen (HLA)-bound viral antigens serve as an immunological signature that can be selectively recognized by T cells. As viruses evolve by acquiring mutations, it is essential to identify a range of presented viral antigens. Using HLA peptidomics, we are able to identify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-derived peptides presented by highly prevalent HLA class I (HLA-I) molecules by using infected cells as well as overexpression of SARS-CoV-2 genes. We find 26 HLA-I peptides and 36 HLA class II (HLA-II) peptides. Among the identified peptides, some are shared between different cells and some are derived from out-of-frame open reading frames (ORFs). Seven of these peptides were previously shown to be immunogenic, and we identify two additional immunoreactive peptides by using HLA multimer staining. These results may aid the development of the next generation of SARS-CoV-2 vaccines based on presented viral-specific antigens that span several of the viral genes.
Published: 2021
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8. An “alert state” ribosome population acts as a master regulator of cytokine-mediated processes

Author: Dopler, Anna, primary, Alkan, Ferhat, additional, Malka, Yuval, additional, Hoefakker, Kelly, additional, Isaeva, Olga I., additional, Kerkhoff, Mandy, additional, Gangaev, Anastasia, additional, Silva, Joana, additional, Ramalho, Sofia, additional, Hoekman, Liesbeth, additional, Altelaar, Maarten, additional, van der Kammen, Rob, additional, Yewdell, Jonathan Wilson, additional, Kvistborg, Pia, additional, and Faller, William James, additional
Published: 2023
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9. Alternative trafficking of Weibel‐Palade body proteins in CRISPR/Cas9‐engineered von Willebrand factor–deficient blood outgrowth endothelial cells

Author: Maaike Schillemans, Marije Kat, Jurjen Westeneng, Anastasia Gangaev, Menno Hofman, Benjamin Nota, Floris P. J. van Alphen, Martin de Boer, Maartje van den Biggelaar, Coert Margadant, Jan Voorberg, and Ruben Bierings
Subjects: endothelial cells, gene knockout techniques, protein transport, secretory vesicles, von Willebrand factor, Diseases of the blood and blood-forming organs, RC633-647.5
Abstract: Abstract Background Synthesis of the hemostatic protein von Willebrand factor (VWF) drives formation of endothelial storage organelles called Weibel‐Palade bodies (WPBs). In the absence of VWF, angiogenic and inflammatory mediators that are costored in WPBs are subject to alternative trafficking routes. In patients with von Willebrand disease (VWD), partial or complete absence of VWF/WPBs may lead to additional bleeding complications, such as angiodysplasia. Studies addressing the role of VWF using VWD patient–derived blood outgrowth endothelial cells (BOECs) have reported conflicting results due to the intrinsic heterogeneity of patient‐derived BOECs. Objective To generate a VWF‐deficient endothelial cell model using clustered regularly interspaced short palindromic repeats (CRISPR) genome engineering of blood outgrowth endothelial cells. Methods We used CRISPR/CRISPR‐associated protein 9 editing in single‐donor cord blood–derived BOECs (cbBOECs) to generate clonal VWF−/− cbBOECs. Clones were selected using high‐throughput screening, VWF mutations were validated by sequencing, and cells were phenotypically characterized. Results Two VWF−/− BOEC clones were obtained and were entirely devoid of WPBs, while their overall cell morphology was unaltered. Several WPB proteins, including CD63, syntaxin‐3 and the cargo proteins angiopoietin (Ang)‐2, interleukin (IL)‐6, and IL‐8 showed alternative trafficking and secretion in the absence of VWF. Interestingly, Ang‐2 was relocated to the cell periphery and colocalized with Tie‐2. Conclusions CRISPR editing of VWF provides a robust method to create VWF‐ deficient BOECs that can be directly compared to their wild‐type counterparts. Results obtained with our model system confirmed alternative trafficking of several WPB proteins in the absence of VWF and support the theory that increased Ang‐2/Tie‐2 interaction contributes to angiogenic abnormalities in VWD patients.
Published: 2019
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10. 863 Identification and characterization of an immunodominant SARS-CoV-2-specific CD8 T cell response

Author: Pia Kvistborg and Anastasia Gangaev
Subjects: Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Published: 2020
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11. Imatinib Regulates miR-483-3p and Mitochondrial Respiratory Complexes in Gastrointestinal Stromal Tumors

Author: Wen-Kuan Huang, Hao Shi, Pinar Akçakaya, Katarina Zeljic, Anastasia Gangaev, Stefano Caramuta, Chun-Nan Yeh, Robert Bränström, Catharina Larsson, and Weng-Onn Lui
Subjects: gastrointestinal stromal tumor (GIST), imatinib, microRNA, oxidative phosphorylation, miR-483, succinate dehydrogenase B, Biology (General), QH301-705.5, Chemistry, QD1-999
Abstract: Metabolic adaptation to increased oxidative phosphorylation (OXPHOS) has been found in gastrointestinal stromal tumor (GIST) upon imatinib treatment. However, the underlying mechanism of imatinib-induced OXPHOS is unknown. Discovering molecules that mediate imatinib-induced OXPHOS may lead to the development of therapeutic strategies synergizing the efficacy of imatinib. In this study, we explored the role of microRNAs in regulating OXPHOS in GIST upon imatinib treatment. Using a microarray approach, we found that miR-483-3p was one of the most downregulated miRNAs in imatinib-treated tumors compared to untreated tumors. Using an extended series of GIST samples, we further validated the downregulation of miR-483-3p in imatinib-treated GIST samples by RT-qPCR. Using both gain- and loss-of-function experiments, we showed that miR-483-3p could regulate mitochondrial respiratory Complex II expression, suggesting its role in OXPHOS regulation. Functionally, miR-483-3p overexpression could rescue imatinib-induced cell death. These findings provide the molecular link for imatinib-induced OXPHOS expression and the biological role of miR-483-3p in regulating cell viability upon imatinib treatment.
Published: 2021
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12. The Weibel-Palade Body Localized SNARE (Soluble NSF Attachment Protein Receptor) Syntaxin-3 Modulates Von Willebrand Factor Secretion From Endothelial Cells

Author: Schillemans, Maaike, Karampini, Ellie, van den Eshof, Bart, Gangaev, Anastasia, Hofman, Menno, van Breevoort, Dorothee, Meems, Henriët, Janssen, Hans, Mulder, Aat A., Jost, Carolina R., Escher, Johanna C., Adam, Rüdiger, Carter, Tom, Koster, Abraham J., van den Biggelaar, Maartje, Voorberg, Jan, and Bierings, Ruben
Published: 2018
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13. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Author: Cossarizza, A, Chang, H, Radbruch, A, Abrignani, S, Addo, R, Akdis, M, Andra, I, Andreata, F, Annunziato, F, Arranz, E, Bacher, P, Bari, S, Barnaba, V, Barros-Martins, J, Baumjohann, D, Beccaria, C, Bernardo, D, Boardman, D, Borger, J, Bottcher, C, Brockmann, L, Burns, M, Busch, D, Cameron, G, Cammarata, I, Cassotta, A, Chang, Y, Chirdo, F, Christakou, E, Cicin-Sain, L, Cook, L, Corbett, A, Cornelis, R, Cosmi, L, Davey, M, De Biasi, S, De Simone, G, del Zotto, G, Delacher, M, Di Rosa, F, Santo, J, Diefenbach, A, Dong, J, Dorner, T, Dress, R, Dutertre, C, Eckle, S, Eede, P, Evrard, M, Falk, C, Feuerer, M, Fillatreau, S, Fiz-Lopez, A, Follo, M, Foulds, G, Frobel, J, Gagliani, N, Galletti, G, Gangaev, A, Garbi, N, Garrote, J, Geginat, J, Gherardin, N, Gibellini, L, Ginhoux, F, Godfrey, D, Gruarin, P, Haftmann, C, Hansmann, L, Harpur, C, Hayday, A, Heine, G, Hernandez, D, Herrmann, M, Hoelsken, O, Huang, Q, Huber, S, Huber, J, Huehn, J, Hundemer, M, Hwang, W, Iannacone, M, Ivison, S, Jack, H, Jani, P, Keller, B, Kessler, N, Ketelaars, S, Knop, L, Knopf, J, Koay, H, Kobow, K, Kriegsmann, K, Kristyanto, H, Krueger, A, Kuehne, J, Kunze-Schumacher, H, Kvistborg, P, Kwok, I, Latorre, D, Lenz, D, Levings, M, Lino, A, Liotta, F, Long, H, Lugli, E, Macdonald, K, Maggi, L, Maini, M, Mair, F, Manta, C, Manz, R, Mashreghi, M, Mazzoni, A, Mccluskey, J, Mei, H, Melchers, F, Melzer, S, Mielenz, D, Monin, L, Moretta, L, Multhoff, G, Munoz, L, Munoz-Ruiz, M, Muscate, F, Natalini, A, Neumann, K, Ng, L, Niedobitek, A, Niemz, J, Almeida, L, Notarbartolo, S, Ostendorf, L, Pallett, L, Patel, A, Percin, G, Peruzzi, G, Pinti, M, Pockley, A, Pracht, K, Prinz, I, Pujol-Autonell, I, Pulvirenti, N, Quatrini, L, Quinn, K, Radbruch, H, Rhys, H, Rodrigo, M, Romagnani, C, Saggau, C, Sakaguchi, S, Sallusto, F, Sanderink, L, Sandrock, I, Schauer, C, Scheffold, A, Scherer, H, Schiemann, M, Schildberg, F, Schober, K, Schoen, J, Schuh, W, Schuler, T, Schulz, A, Schulz, S, Schulze, J, Simonetti, S, Singh, J, Sitnik, K, Stark, R, Starossom, S, Stehle, C, Szelinski, F, Tan, L, Tarnok, A, Tornack, J, Tree, T, van Beek, J, van de Veen, W, van Gisbergen, K, Vasco, C, Verheyden, N, von Borstel, A, Ward-Hartstonge, K, Warnatz, K, Waskow, C, Wiedemann, A, Wilharm, A, Wing, J, Wirz, O, Wittner, J, Yang, J, Cossarizza A., Chang H. -D., Radbruch A., Abrignani S., Addo R., Akdis M., Andra I., Andreata F., Annunziato F., Arranz E., Bacher P., Bari S., Barnaba V., Barros-Martins J., Baumjohann D., Beccaria C. G., Bernardo D., Boardman D. A., Borger J., Bottcher C., Brockmann L., Burns M., Busch D. H., Cameron G., Cammarata I., Cassotta A., Chang Y., Chirdo F. G., Christakou E., Cicin-Sain L., Cook L., Corbett A. J., Cornelis R., Cosmi L., Davey M. S., De Biasi S., De Simone G., del Zotto G., Delacher M., Di Rosa F., Santo J. D., Diefenbach A., Dong J., Dorner T., Dress R. J., Dutertre C. -A., Eckle S. B. G., Eede P., Evrard M., Falk C. S., Feuerer M., Fillatreau S., Fiz-Lopez A., Follo M., Foulds G. A., Frobel J., Gagliani N., Galletti G., Gangaev A., Garbi N., Garrote J. A., Geginat J., Gherardin N. A., Gibellini L., Ginhoux F., Godfrey D. I., Gruarin P., Haftmann C., Hansmann L., Harpur C. M., Hayday A. C., Heine G., Hernandez D. C., Herrmann M., Hoelsken O., Huang Q., Huber S., Huber J. E., Huehn J., Hundemer M., Hwang W. Y. K., Iannacone M., Ivison S. M., Jack H. -M., Jani P. K., Keller B., Kessler N., Ketelaars S., Knop L., Knopf J., Koay H. -F., Kobow K., Kriegsmann K., Kristyanto H., Krueger A., Kuehne J. F., Kunze-Schumacher H., Kvistborg P., Kwok I., Latorre D., Lenz D., Levings M. K., Lino A. C., Liotta F., Long H. M., Lugli E., MacDonald K. N., Maggi L., Maini M. K., Mair F., Manta C., Manz R. A., Mashreghi M. -F., Mazzoni A., McCluskey J., Mei H. E., Melchers F., Melzer S., Mielenz D., Monin L., Moretta L., Multhoff G., Munoz L. E., Munoz-Ruiz M., Muscate F., Natalini A., Neumann K., Ng L. G., Niedobitek A., Niemz J., Almeida L. N., Notarbartolo S., Ostendorf L., Pallett L. J., Patel A. A., Percin G. I., Peruzzi G., Pinti M., Pockley A. G., Pracht K., Prinz I., Pujol-Autonell I., Pulvirenti N., Quatrini L., Quinn K. M., Radbruch H., Rhys H., Rodrigo M. B., Romagnani C., Saggau C., Sakaguchi S., Sallusto F., Sanderink L., Sandrock I., Schauer C., Scheffold A., Scherer H. U., Schiemann M., Schildberg F. A., Schober K., Schoen J., Schuh W., Schuler T., Schulz A. R., Schulz S., Schulze J., Simonetti S., Singh J., Sitnik K. M., Stark R., Starossom S., Stehle C., Szelinski F., Tan L., Tarnok A., Tornack J., Tree T. I. M., van Beek J. J. P., van de Veen W., van Gisbergen K., Vasco C., Verheyden N. A., von Borstel A., Ward-Hartstonge K. A., Warnatz K., Waskow C., Wiedemann A., Wilharm A., Wing J., Wirz O., Wittner J., Yang J. H. M., Yang J., Cossarizza, A, Chang, H, Radbruch, A, Abrignani, S, Addo, R, Akdis, M, Andra, I, Andreata, F, Annunziato, F, Arranz, E, Bacher, P, Bari, S, Barnaba, V, Barros-Martins, J, Baumjohann, D, Beccaria, C, Bernardo, D, Boardman, D, Borger, J, Bottcher, C, Brockmann, L, Burns, M, Busch, D, Cameron, G, Cammarata, I, Cassotta, A, Chang, Y, Chirdo, F, Christakou, E, Cicin-Sain, L, Cook, L, Corbett, A, Cornelis, R, Cosmi, L, Davey, M, De Biasi, S, De Simone, G, del Zotto, G, Delacher, M, Di Rosa, F, Santo, J, Diefenbach, A, Dong, J, Dorner, T, Dress, R, Dutertre, C, Eckle, S, Eede, P, Evrard, M, Falk, C, Feuerer, M, Fillatreau, S, Fiz-Lopez, A, Follo, M, Foulds, G, Frobel, J, Gagliani, N, Galletti, G, Gangaev, A, Garbi, N, Garrote, J, Geginat, J, Gherardin, N, Gibellini, L, Ginhoux, F, Godfrey, D, Gruarin, P, Haftmann, C, Hansmann, L, Harpur, C, Hayday, A, Heine, G, Hernandez, D, Herrmann, M, Hoelsken, O, Huang, Q, Huber, S, Huber, J, Huehn, J, Hundemer, M, Hwang, W, Iannacone, M, Ivison, S, Jack, H, Jani, P, Keller, B, Kessler, N, Ketelaars, S, Knop, L, Knopf, J, Koay, H, Kobow, K, Kriegsmann, K, Kristyanto, H, Krueger, A, Kuehne, J, Kunze-Schumacher, H, Kvistborg, P, Kwok, I, Latorre, D, Lenz, D, Levings, M, Lino, A, Liotta, F, Long, H, Lugli, E, Macdonald, K, Maggi, L, Maini, M, Mair, F, Manta, C, Manz, R, Mashreghi, M, Mazzoni, A, Mccluskey, J, Mei, H, Melchers, F, Melzer, S, Mielenz, D, Monin, L, Moretta, L, Multhoff, G, Munoz, L, Munoz-Ruiz, M, Muscate, F, Natalini, A, Neumann, K, Ng, L, Niedobitek, A, Niemz, J, Almeida, L, Notarbartolo, S, Ostendorf, L, Pallett, L, Patel, A, Percin, G, Peruzzi, G, Pinti, M, Pockley, A, Pracht, K, Prinz, I, Pujol-Autonell, I, Pulvirenti, N, Quatrini, L, Quinn, K, Radbruch, H, Rhys, H, Rodrigo, M, Romagnani, C, Saggau, C, Sakaguchi, S, Sallusto, F, Sanderink, L, Sandrock, I, Schauer, C, Scheffold, A, Scherer, H, Schiemann, M, Schildberg, F, Schober, K, Schoen, J, Schuh, W, Schuler, T, Schulz, A, Schulz, S, Schulze, J, Simonetti, S, Singh, J, Sitnik, K, Stark, R, Starossom, S, Stehle, C, Szelinski, F, Tan, L, Tarnok, A, Tornack, J, Tree, T, van Beek, J, van de Veen, W, van Gisbergen, K, Vasco, C, Verheyden, N, von Borstel, A, Ward-Hartstonge, K, Warnatz, K, Waskow, C, Wiedemann, A, Wilharm, A, Wing, J, Wirz, O, Wittner, J, Yang, J, Cossarizza A., Chang H. -D., Radbruch A., Abrignani S., Addo R., Akdis M., Andra I., Andreata F., Annunziato F., Arranz E., Bacher P., Bari S., Barnaba V., Barros-Martins J., Baumjohann D., Beccaria C. G., Bernardo D., Boardman D. A., Borger J., Bottcher C., Brockmann L., Burns M., Busch D. H., Cameron G., Cammarata I., Cassotta A., Chang Y., Chirdo F. G., Christakou E., Cicin-Sain L., Cook L., Corbett A. J., Cornelis R., Cosmi L., Davey M. S., De Biasi S., De Simone G., del Zotto G., Delacher M., Di Rosa F., Santo J. D., Diefenbach A., Dong J., Dorner T., Dress R. J., Dutertre C. -A., Eckle S. B. G., Eede P., Evrard M., Falk C. S., Feuerer M., Fillatreau S., Fiz-Lopez A., Follo M., Foulds G. A., Frobel J., Gagliani N., Galletti G., Gangaev A., Garbi N., Garrote J. A., Geginat J., Gherardin N. A., Gibellini L., Ginhoux F., Godfrey D. I., Gruarin P., Haftmann C., Hansmann L., Harpur C. M., Hayday A. C., Heine G., Hernandez D. C., Herrmann M., Hoelsken O., Huang Q., Huber S., Huber J. E., Huehn J., Hundemer M., Hwang W. Y. K., Iannacone M., Ivison S. M., Jack H. -M., Jani P. K., Keller B., Kessler N., Ketelaars S., Knop L., Knopf J., Koay H. -F., Kobow K., Kriegsmann K., Kristyanto H., Krueger A., Kuehne J. F., Kunze-Schumacher H., Kvistborg P., Kwok I., Latorre D., Lenz D., Levings M. K., Lino A. C., Liotta F., Long H. M., Lugli E., MacDonald K. N., Maggi L., Maini M. K., Mair F., Manta C., Manz R. A., Mashreghi M. -F., Mazzoni A., McCluskey J., Mei H. E., Melchers F., Melzer S., Mielenz D., Monin L., Moretta L., Multhoff G., Munoz L. E., Munoz-Ruiz M., Muscate F., Natalini A., Neumann K., Ng L. G., Niedobitek A., Niemz J., Almeida L. N., Notarbartolo S., Ostendorf L., Pallett L. J., Patel A. A., Percin G. I., Peruzzi G., Pinti M., Pockley A. G., Pracht K., Prinz I., Pujol-Autonell I., Pulvirenti N., Quatrini L., Quinn K. M., Radbruch H., Rhys H., Rodrigo M. B., Romagnani C., Saggau C., Sakaguchi S., Sallusto F., Sanderink L., Sandrock I., Schauer C., Scheffold A., Scherer H. U., Schiemann M., Schildberg F. A., Schober K., Schoen J., Schuh W., Schuler T., Schulz A. R., Schulz S., Schulze J., Simonetti S., Singh J., Sitnik K. M., Stark R., Starossom S., Stehle C., Szelinski F., Tan L., Tarnok A., Tornack J., Tree T. I. M., van Beek J. J. P., van de Veen W., van Gisbergen K., Vasco C., Verheyden N. A., von Borstel A., Ward-Hartstonge K. A., Warnatz K., Waskow C., Wiedemann A., Wilharm A., Wing J., Wirz O., Wittner J., Yang J. H. M., and Yang J.
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

14. Weibel-Palade Body Localized Syntaxin-3 Modulates Von Willebrand Factor Secretion From Endothelial Cells

Author: Schillemans, Maaike, Karampini, Ellie, van den Eshof, Bart L., Gangaev, Anastasia, Hofman, Menno, van Breevoort, Dorothee, Meems, Henriët, Janssen, Hans, Mulder, Aat A., Jost, Carolina R., Escher, Johanna C., Adam, Rüdiger, Carter, Tom, Koster, Abraham J., van den Biggelaar, Maartje, Voorberg, Jan, and Bierings, Ruben
Published: 2018
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15. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Author: Andrea Cossarizza, Hyun‐Dong Chang, Andreas Radbruch, Sergio Abrignani, Richard Addo, Mübeccel Akdis, Immanuel Andrä, Francesco Andreata, Francesco Annunziato, Eduardo Arranz, Petra Bacher, Sudipto Bari, Vincenzo Barnaba, Joana Barros‐Martins, Dirk Baumjohann, Cristian G. Beccaria, David Bernardo, Dominic A. Boardman, Jessica Borger, Chotima Böttcher, Leonie Brockmann, Marie Burns, Dirk H. Busch, Garth Cameron, Ilenia Cammarata, Antonino Cassotta, Yinshui Chang, Fernando Gabriel Chirdo, Eleni Christakou, Luka Čičin‐Šain, Laura Cook, Alexandra J. Corbett, Rebecca Cornelis, Lorenzo Cosmi, Martin S. Davey, Sara De Biasi, Gabriele De Simone, Genny del Zotto, Michael Delacher, Francesca Di Rosa, James Di Santo, Andreas Diefenbach, Jun Dong, Thomas Dörner, Regine J. Dress, Charles‐Antoine Dutertre, Sidonia B. G. Eckle, Pascale Eede, Maximilien Evrard, Christine S. Falk, Markus Feuerer, Simon Fillatreau, Aida Fiz‐Lopez, Marie Follo, Gemma A. Foulds, Julia Fröbel, Nicola Gagliani, Giovanni Galletti, Anastasia Gangaev, Natalio Garbi, José Antonio Garrote, Jens Geginat, Nicholas A. Gherardin, Lara Gibellini, Florent Ginhoux, Dale I. Godfrey, Paola Gruarin, Claudia Haftmann, Leo Hansmann, Christopher M. Harpur, Adrian C. Hayday, Guido Heine, Daniela Carolina Hernández, Martin Herrmann, Oliver Hoelsken, Qing Huang, Samuel Huber, Johanna E. Huber, Jochen Huehn, Michael Hundemer, William Y. K. Hwang, Matteo Iannacone, Sabine M. Ivison, Hans‐Martin Jäck, Peter K. Jani, Baerbel Keller, Nina Kessler, Steven Ketelaars, Laura Knop, Jasmin Knopf, Hui‐Fern Koay, Katja Kobow, Katharina Kriegsmann, H. Kristyanto, Andreas Krueger, Jenny F. Kuehne, Heike Kunze‐Schumacher, Pia Kvistborg, Immanuel Kwok, Daniela Latorre, Daniel Lenz, Megan K. Levings, Andreia C. Lino, Francesco Liotta, Heather M. Long, Enrico Lugli, Katherine N. MacDonald, Laura Maggi, Mala K. Maini, Florian Mair, Calin Manta, Rudolf Armin Manz, Mir‐Farzin Mashreghi, Alessio Mazzoni, James McCluskey, Henrik E. Mei, Fritz Melchers, Susanne Melzer, Dirk Mielenz, Leticia Monin, Lorenzo Moretta, Gabriele Multhoff, Luis Enrique Muñoz, Miguel Muñoz‐Ruiz, Franziska Muscate, Ambra Natalini, Katrin Neumann, Lai Guan Ng, Antonia Niedobitek, Jana Niemz, Larissa Nogueira Almeida, Samuele Notarbartolo, Lennard Ostendorf, Laura J. Pallett, Amit A. Patel, Gulce Itir Percin, Giovanna Peruzzi, Marcello Pinti, A. Graham Pockley, Katharina Pracht, Immo Prinz, Irma Pujol‐Autonell, Nadia Pulvirenti, Linda Quatrini, Kylie M. Quinn, Helena Radbruch, Hefin Rhys, Maria B. Rodrigo, Chiara Romagnani, Carina Saggau, Shimon Sakaguchi, Federica Sallusto, Lieke Sanderink, Inga Sandrock, Christine Schauer, Alexander Scheffold, Hans U. Scherer, Matthias Schiemann, Frank A. Schildberg, Kilian Schober, Janina Schoen, Wolfgang Schuh, Thomas Schüler, Axel R. Schulz, Sebastian Schulz, Julia Schulze, Sonia Simonetti, Jeeshan Singh, Katarzyna M. Sitnik, Regina Stark, Sarah Starossom, Christina Stehle, Franziska Szelinski, Leonard Tan, Attila Tarnok, Julia Tornack, Timothy I. M. Tree, Jasper J. P. van Beek, Willem van de Veen, Klaas van Gisbergen, Chiara Vasco, Nikita A. Verheyden, Anouk von Borstel, Kirsten A. Ward‐Hartstonge, Klaus Warnatz, Claudia Waskow, Annika Wiedemann, Anneke Wilharm, James Wing, Oliver Wirz, Jens Wittner, Jennie H. M. Yang, Juhao Yang, Rolf M. Schwiete Foundation, Associazione Italiana per la Ricerca sul Cancro, German Research Foundation, National Institutes of Health (US), European Commission, AII - Inflammatory diseases, Cossarizza, A, Chang, H, Radbruch, A, Abrignani, S, Addo, R, Akdis, M, Andra, I, Andreata, F, Annunziato, F, Arranz, E, Bacher, P, Bari, S, Barnaba, V, Barros-Martins, J, Baumjohann, D, Beccaria, C, Bernardo, D, Boardman, D, Borger, J, Bottcher, C, Brockmann, L, Burns, M, Busch, D, Cameron, G, Cammarata, I, Cassotta, A, Chang, Y, Chirdo, F, Christakou, E, Cicin-Sain, L, Cook, L, Corbett, A, Cornelis, R, Cosmi, L, Davey, M, De Biasi, S, De Simone, G, del Zotto, G, Delacher, M, Di Rosa, F, Santo, J, Diefenbach, A, Dong, J, Dorner, T, Dress, R, Dutertre, C, Eckle, S, Eede, P, Evrard, M, Falk, C, Feuerer, M, Fillatreau, S, Fiz-Lopez, A, Follo, M, Foulds, G, Frobel, J, Gagliani, N, Galletti, G, Gangaev, A, Garbi, N, Garrote, J, Geginat, J, Gherardin, N, Gibellini, L, Ginhoux, F, Godfrey, D, Gruarin, P, Haftmann, C, Hansmann, L, Harpur, C, Hayday, A, Heine, G, Hernandez, D, Herrmann, M, Hoelsken, O, Huang, Q, Huber, S, Huber, J, Huehn, J, Hundemer, M, Hwang, W, Iannacone, M, Ivison, S, Jack, H, Jani, P, Keller, B, Kessler, N, Ketelaars, S, Knop, L, Knopf, J, Koay, H, Kobow, K, Kriegsmann, K, Kristyanto, H, Krueger, A, Kuehne, J, Kunze-Schumacher, H, Kvistborg, P, Kwok, I, Latorre, D, Lenz, D, Levings, M, Lino, A, Liotta, F, Long, H, Lugli, E, Macdonald, K, Maggi, L, Maini, M, Mair, F, Manta, C, Manz, R, Mashreghi, M, Mazzoni, A, Mccluskey, J, Mei, H, Melchers, F, Melzer, S, Mielenz, D, Monin, L, Moretta, L, Multhoff, G, Munoz, L, Munoz-Ruiz, M, Muscate, F, Natalini, A, Neumann, K, Ng, L, Niedobitek, A, Niemz, J, Almeida, L, Notarbartolo, S, Ostendorf, L, Pallett, L, Patel, A, Percin, G, Peruzzi, G, Pinti, M, Pockley, A, Pracht, K, Prinz, I, Pujol-Autonell, I, Pulvirenti, N, Quatrini, L, Quinn, K, Radbruch, H, Rhys, H, Rodrigo, M, Romagnani, C, Saggau, C, Sakaguchi, S, Sallusto, F, Sanderink, L, Sandrock, I, Schauer, C, Scheffold, A, Scherer, H, Schiemann, M, Schildberg, F, Schober, K, Schoen, J, Schuh, W, Schuler, T, Schulz, A, Schulz, S, Schulze, J, Simonetti, S, Singh, J, Sitnik, K, Stark, R, Starossom, S, Stehle, C, Szelinski, F, Tan, L, Tarnok, A, Tornack, J, Tree, T, van Beek, J, van de Veen, W, van Gisbergen, K, Vasco, C, Verheyden, N, von Borstel, A, Ward-Hartstonge, K, Warnatz, K, Waskow, C, Wiedemann, A, Wilharm, A, Wing, J, Wirz, O, Wittner, J, Yang, J, Publica, Cossarizza, A., Chang, H. -D., Radbruch, A., Abrignani, S., Addo, R., Akdis, M., Andra, I., Andreata, F., Annunziato, F., Arranz, E., Bacher, P., Bari, S., Barnaba, V., Barros-Martins, J., Baumjohann, D., Beccaria, C. G., Bernardo, D., Boardman, D. A., Borger, J., Bottcher, C., Brockmann, L., Burns, M., Busch, D. H., Cameron, G., Cammarata, I., Cassotta, A., Chang, Y., Chirdo, F. G., Christakou, E., Cicin-Sain, L., Cook, L., Corbett, A. J., Cornelis, R., Cosmi, L., Davey, M. S., De Biasi, S., De Simone, G., del Zotto, G., Delacher, M., Di Rosa, F., Santo, J. D., Diefenbach, A., Dong, J., Dorner, T., Dress, R. J., Dutertre, C. -A., Eckle, S. B. G., Eede, P., Evrard, M., Falk, C. S., Feuerer, M., Fillatreau, S., Fiz-Lopez, A., Follo, M., Foulds, G. A., Frobel, J., Gagliani, N., Galletti, G., Gangaev, A., Garbi, N., Garrote, J. A., Geginat, J., Gherardin, N. A., Gibellini, L., Ginhoux, F., Godfrey, D. I., Gruarin, P., Haftmann, C., Hansmann, L., Harpur, C. M., Hayday, A. C., Heine, G., Hernandez, D. C., Herrmann, M., Hoelsken, O., Huang, Q., Huber, S., Huber, J. E., Huehn, J., Hundemer, M., Hwang, W. Y. K., Iannacone, M., Ivison, S. M., Jack, H. -M., Jani, P. K., Keller, B., Kessler, N., Ketelaars, S., Knop, L., Knopf, J., Koay, H. -F., Kobow, K., Kriegsmann, K., Kristyanto, H., Krueger, A., Kuehne, J. F., Kunze-Schumacher, H., Kvistborg, P., Kwok, I., Latorre, D., Lenz, D., Levings, M. K., Lino, A. C., Liotta, F., Long, H. M., Lugli, E., Macdonald, K. N., Maggi, L., Maini, M. K., Mair, F., Manta, C., Manz, R. A., Mashreghi, M. -F., Mazzoni, A., Mccluskey, J., Mei, H. E., Melchers, F., Melzer, S., Mielenz, D., Monin, L., Moretta, L., Multhoff, G., Munoz, L. E., Munoz-Ruiz, M., Muscate, F., Natalini, A., Neumann, K., Ng, L. G., Niedobitek, A., Niemz, J., Almeida, L. N., Notarbartolo, S., Ostendorf, L., Pallett, L. J., Patel, A. A., Percin, G. I., Peruzzi, G., Pinti, M., Pockley, A. G., Pracht, K., Prinz, I., Pujol-Autonell, I., Pulvirenti, N., Quatrini, L., Quinn, K. M., Radbruch, H., Rhys, H., Rodrigo, M. B., Romagnani, C., Saggau, C., Sakaguchi, S., Sallusto, F., Sanderink, L., Sandrock, I., Schauer, C., Scheffold, A., Scherer, H. U., Schiemann, M., Schildberg, F. A., Schober, K., Schoen, J., Schuh, W., Schuler, T., Schulz, A. R., Schulz, S., Schulze, J., Simonetti, S., Singh, J., Sitnik, K. M., Stark, R., Starossom, S., Stehle, C., Szelinski, F., Tan, L., Tarnok, A., Tornack, J., Tree, T. I. M., van Beek, J. J. P., van de Veen, W., van Gisbergen, K., Vasco, C., Verheyden, N. A., von Borstel, A., Ward-Hartstonge, K. A., Warnatz, K., Waskow, C., Wiedemann, A., Wilharm, A., Wing, J., Wirz, O., Wittner, J., Yang, J. H. M., and Yang, J.
Subjects: Immunology, citometry, Flow Cytometry, Infections, ddc, Autoimmune Diseases, Animals, Chronic Disease, Humans, Mice, Neoplasms, Practice Guidelines as Topic, Immunology and Allergy, ddc:610, Function and Dysfunction of the Nervous System, guideline
Abstract: © 2021 The Authors., 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., Hyun-Dong Chang is supported by the Dr. Rolf M. Schwiete Foundation. Susanne Melzer and Attila Tarnok thank De Novo Software for providing FCS Express. Enrico Lugli is supported by a grant from the Associazione Italiana per la Ricerca sul Cancro (AIRC IG20676). Gabriele De Simone and Giovanni Galletti were supported by Fellowships from the Fondazione Italiana per la Ricerca sul Cancro-Associazione Italiana per la Ricerca sul Cancro (FIRC-AIRC). Jun Dong is supported by Deutsche Forschungsgemeinschft (DFG, German Research Foundation) Projektnummer 389687267 and Chinesisch-Deutsches Zentrum für Wissenschaftsförderung [Sino-German Center for Research Promotion (SGC)] grant C-0072. Nicola Gagliani, Samuel Huber and Franziska Muscate are supported by DFG fundings: SFB841,GA 2441/3-1, HU 1714/10-1. The tetramer APC-conjugated H-2K (d) HIV-1 gag197-205 AMQMLKETI used in TDS assay for mouse blood T cells was obtained through the NIH Tetramer Facility. Larissa Nogueira Almeida was supported by DFG research grant MA 2273/14-1. Supported by the following grants: AIRC 5X1000 2018 id. 21147 (Lorenzo Moretta); AIRC IG 2017 id. 19920 (Lorenzo Moretta); RC-2020 OPBG (Lorenzo Moretta); AIRC and European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No 800924 (Linda Quatrini). Dirk Baumjohann was supported by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Emmy Noether Programme BA 5132/1-2 (252623821) and Germany's Excellence Strategy EXC2151 (390873048).
Published: 2021

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

Author: Lara Gibellini, Sussan Nourshargh, Susanna Cardell, Wlodzimierz Maslinski, Mar Felipo-Benavent, Florian Mair, Hans-Martin Jäck, Lilly Lopez, Klaus Warnatz, John Trowsdale, Diana Ordonez, Marcus Eich, William Hwang, Anne Cooke, Dirk Mielenz, Alberto Orfao, Winfried F. Pickl, Vladimir Benes, Alice Yue, T. Vincent Shankey, Maria Tsoumakidou, Virginia Litwin, Gelo Victoriano Dela Cruz, Andrea Cavani, Sara De Biasi, Larissa Nogueira Almeida, Jonathan J M Landry, Claudia Haftmann, Charlotte Esser, Ana Cumano, Anneke Wilharm, Francesco Dieli, Rudi Beyaert, Alessio Mazzoni, Burkhard Ludewig, Carlo Pucillo, Dirk H. Busch, Joe Trotter, Stipan Jonjić, Marc Veldhoen, Josef Spidlen, Aja M. Rieger, Dieter Adam, Srijit Khan, Todd A. Fehniger, Giuseppe Matarese, Maximilien Evrard, Christian Maueröder, Steffen Schmitt, Kristin A. Hogquist, Barry Moran, Raghavendra Palankar, Markus Feuerer, S Schmid, Susann Rahmig, Amy E. Lovett-Racke, James V. Watson, Megan K. Levings, Susanne Melzer, Dinko Pavlinic, Christopher M. Harpur, Christina Stehle, A. Graham Pockley, Toshinori Nakayama, Attila Tárnok, Juhao Yang, Michael Lohoff, Paulo Vieira, Francisco Sala-de-Oyanguren, Christian Kurts, Anastasia Gangaev, Alfonso Blanco, Hans Scherer, Regine J. Dress, Bruno Silva-Santos, Kiyoshi Takeda, Bimba F. Hoyer, Ilenia Cammarata, Daryl Grummitt, Isabel Panse, Günnur Deniz, Bianka Baying, Friederike Ebner, Esther Schimisky, Leo Hansmann, Thomas Kamradt, Edwin van der Pol, Daniel Scott-Algara, Anna Iannone, Giorgia Alvisi, Sebastian R. Schulz, Francesco Liotta, Irmgard Förster, Beatriz Jávega, Hans-Peter Rahn, Caetano Reis e Sousa, Livius Penter, Xuetao Cao, David P. Sester, Keisuke Goda, Peter Wurst, Iain B. McInnes, Ricardo T. Gazzinelli, Federica Piancone, Gerald Willimsky, Yotam Raz, Pärt Peterson, Wolfgang Fritzsche, Yvonne Samstag, Martin Büscher, Thomas Schüler, Susanne Hartmann, Robert J. Wilkinson, Anna E. S. Brooks, Steven L. C. Ketelaars, Catherine Sautès-Fridman, Anna Rubartelli, Petra Bacher, Katja Kobow, Marco A. Cassatella, Andrea Hauser, Henrik E. Mei, Kilian Schober, Silvia Della Bella, Graham Anderson, Michael D. Ward, Garth Cameron, Sebastian Lunemann, Katharina Kriegsmann, Katarzyna M. Sitnik, Brice Gaudilliere, Chantip Dang-Heine, Marcello Pinti, Paul Klenerman, Frank A. Schildberg, Joana Barros-Martins, Laura G. Rico, Hanlin Zhang, Christian Münz, Thomas Dörner, Jakob Zimmermann, Andrea M. Cooper, Jonni S. Moore, Andreas Diefenbach, Yanling Liu, Wolfgang Bauer, Tobit Steinmetz, Katharina Pracht, Leonard Tan, Peter K. Jani, Alan M. Stall, Petra Hoffmann, Christine S. Falk, Jasmin Knopf, Simon Fillatreau, Hans-Dieter Volk, Luis E. Muñoz, David L. Haviland, William W. Agace, Jonathan Rebhahn, Ljiljana Cvetkovic, Mohamed Trebak, Jordi Petriz, Mario Clerici, Diether J. Recktenwald, Anders Ståhlberg, Tristan Holland, Helen M. McGuire, Sa A. Wang, Christian Kukat, Thomas Kroneis, Laura Cook, Wan Ting Kong, Xin M. Wang, Britta Engelhardt, Pierre Coulie, Genny Del Zotto, Sally A. Quataert, Kata Filkor, Gabriele Multhoff, Bartek Rajwa, Federica Calzetti, Hans Minderman, Cosima T. Baldari, Jens Geginat, Hervé Luche, Gert Van Isterdael, Linda Schadt, Sophia Urbanczyk, Giovanna Borsellino, Liping Yu, Dale I. Godfrey, Achille Anselmo, Rachael C. Walker, Andreas Grützkau, David W. Hedley, Birgit Sawitzki, Silvia Piconese, Maria Yazdanbakhsh, Burkhard Becher, Ramon Bellmas Sanz, Michael Delacher, Hyun-Dong Chang, Immanuel Andrä, Hans-Gustaf Ljunggren, José-Enrique O'Connor, Ahad Khalilnezhad, Sharon Sanderson, Federico Colombo, Götz R. A. Ehrhardt, Inga Sandrock, Enrico Lugli, Christian Bogdan, James B. Wing, Susann Müller, Tomohiro Kurosaki, Derek Davies, Ester B. M. Remmerswaal, Kylie M. Quinn, Christopher A. Hunter, Andreas Radbruch, Timothy P. Bushnell, Anna Erdei, Sabine Adam-Klages, Pascale Eede, Van Duc Dang, Rieke Winkelmann, Thomas Korn, Gemma A. Foulds, Dirk Baumjohann, Matthias Schiemann, Manfred Kopf, Jan Kisielow, Lisa Richter, Jochen Huehn, Gloria Martrus, Alexander Scheffold, Jessica G. Borger, Sidonia B G Eckle, John Bellamy Foster, Anna Katharina Simon, Alicia Wong, Mübeccel Akdis, Gisa Tiegs, Toralf Kaiser, James McCluskey, Anna Vittoria Mattioli, Aaron J. Marshall, Hui-Fern Koay, Eva Orlowski-Oliver, Anja E. Hauser, J. Paul Robinson, Jay K. Kolls, Luca Battistini, Mairi McGrath, Jane L. Grogan, Natalio Garbi, Timothy Tree, Kingston H. G. Mills, Stefan H. E. Kaufmann, Wolfgang Schuh, Ryan R. Brinkman, Tim R. Mosmann, Vincenzo Barnaba, Andreas Dolf, Lorenzo Cosmi, Bo Huang, Andreia C. Lino, Baerbel Keller, René A. W. van Lier, Alexandra J. Corbett, Paul S. Frenette, Pleun Hombrink, Helena Radbruch, Sofie Van Gassen, Olivier Lantz, Lorenzo Moretta, Désirée Kunkel, Kirsten A. Ward-Hartstonge, Armin Saalmüller, Leslie Y. T. Leung, Salvador Vento-Asturias, Paola Lanuti, Alicia Martínez-Romero, Sarah Warth, Zhiyong Poon, Diana Dudziak, Andrea Cossarizza, Kovit Pattanapanyasat, Konrad von Volkmann, Jessica P. Houston, Agnès Lehuen, Andrew Filby, Pratip K. Chattopadhyay, Stefano Casola, Annika Wiedemann, Hannes Stockinger, Jürgen Ruland, Arturo Zychlinsky, Claudia Waskow, Katrin Neumann, Ari Waisman, Lucienne Chatenoud, Sudipto Bari, Kamran Ghoreschi, David W. Galbraith, Yvan Saeys, Hamida Hammad, Andrea Gori, Miguel López-Botet, Gabriel Núñez, Sabine Ivison, Michael Hundemer, Dorothea Reimer, Mark C. Dessing, Günter J. Hämmerling, Rudolf A. Manz, Tomas Kalina, Jonas Hahn, Holden T. Maecker, Hendy Kristyanto, Martin S. Davey, Henning Ulrich, Michael L. Dustin, Takashi Saito, Yousuke Takahama, Milena Nasi, Johanna Huber, Jürgen Wienands, Paolo Dellabona, Andreas Schlitzer, Michael D. Leipold, Kerstin H. Mair, Christian Peth, Immo Prinz, Chiara Romagnani, José M. González-Navajas, Josephine Schlosser, Marina Saresella, Matthias Edinger, Dirk Brenner, Nicole Baumgarth, Rikard Holmdahl, Fang-Ping Huang, Guadalupe Herrera, Malte Paulsen, Gergely Toldi, Luka Cicin-Sain, Reiner Schulte, Christina E. Zielinski, Thomas Winkler, Christoph Goettlinger, Philip E. Boulais, Jennie H M Yang, Antonio Celada, Heike Kunze-Schumacher, Julia Tornack, Florian Ingelfinger, Jenny Mjösberg, Andy Riddell, Leonie Wegener, Thomas Höfer, Christoph Hess, James P. Di Santo, Anna E. Oja, J. Kühne, Willem van de Veen, Mary Bebawy, Alberto Mantovani, Bart Everts, Giovanna Lombardi, Laura Maggi, Anouk von Borstel, Pia Kvistborg, Elisabetta Traggiai, A Ochel, Nima Aghaeepour, Charles-Antoine Dutertre, Matthieu Allez, Thomas Höllt, Wenjun Ouyang, Regina Stark, Maries van den Broek, Shimon Sakaguchi, Paul K. Wallace, Silvano Sozzani, Francesca LaRosa, Annette Oxenius, Malgorzata J. Podolska, Ivana Marventano, Wilhelm Gerner, Oliver F. Wirz, Britta Frehse, Gevitha Ravichandran, Martin Herrmann, Carl S. Goodyear, Gary Warnes, Helen Ferry, Stefan Frischbutter, Tim R. Radstake, Salomé LeibundGut-Landmann, Yi Zhao, Axel Schulz, Angela Santoni, Pablo Engel, Daniela C. Hernández, Andreas Acs, Cristiano Scottà, Francesco Annunziato, Thomas Weisenburger, Wolfgang Beisker, Sue Chow, Fritz Melchers, Daniel E. Speiser, Immanuel Kwok, Florent Ginhoux, Dominic A. Boardman, Natalie Stanley, Carsten Watzl, Marie Follo, Erik Lubberts, Andreas Krueger, Susanne Ziegler, Göran K. Hansson, David Voehringer, Antonia Niedobitek, Eleni Christakou, Lai Guan Ng, Sabine Baumgart, Nicholas A Gherardin, Antonio Cosma, Orla Maguire, Jolene Bradford, Daniel Schraivogel, Linda Quatrini, Stephen D. Miller, Rheumatology, Università degli Studi di Modena e Reggio Emilia (UNIMORE), Deutsches Rheuma-ForschungsZentrum (DRFZ), Deutsches Rheuma-ForschungsZentrum, Swiss Institute of Allergy and Asthma Research (SIAF), Universität Zürich [Zürich] = University of Zurich (UZH), Institut de Recherche Saint-Louis - Hématologie Immunologie Oncologie (Département de recherche de l’UFR de médecine, ex- Institut Universitaire Hématologie-IUH) (IRSL), Université de Paris (UP), Ecotaxie, microenvironnement et développement lymphocytaire (EMily (UMR_S_1160 / U1160)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Department of Internal Medicine, Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI)-DENOTHE Center, Institute of Clinical Molecular Biology, Kiel University, Department of Life Sciences [Siena, Italy], Università degli Studi di Siena = University of Siena (UNISI), Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP), Dulbecco Telethon Institute/Department of Biology, Caprotec Bioanalytics GmbH, International Occultation Timing Association European Section (IOTA ES), International Occultation Timing Association European Section, European Molecular Biology Laboratory [Heidelberg] (EMBL), VIB-UGent Center for Inflammation Research [Gand, Belgique] (IRC), VIB [Belgium], Fondazione Santa Lucia (IRCCS), Department of Immunology, Chinese Academy of Medical Sciences, FIRC Institute of Molecular Oncology Foundation, IFOM, Istituto FIRC di Oncologia Molecolare (IFOM), Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of Physiopatology and Transplantation, University of Milan (DEPT), University of Milan, Monash University [Clayton], Institut des Maladies Emergentes et des Thérapies Innovantes (IMETI), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Institute of Cellular Pathology, Université Catholique de Louvain = Catholic University of Louvain (UCL), Lymphopoïèse (Lymphopoïèse (UMR_1223 / U1223 / U-Pasteur_4)), Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Experimental Immunology Unit, Dept. of Oncology, DIBIT San Raffaele Scientific Institute, Immunité Innée - Innate Immunity, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Department of Biopharmacy [Bruxelles, Belgium] (Institute for Medical Immunology IMI), Université libre de Bruxelles (ULB), Charité Hospital, Humboldt-Universität zu Berlin, Agency for science, technology and research [Singapore] (A*STAR), Laboratory of Molecular Immunology and the Howard Hughes Institute, Rockefeller University [New York], Kennedy Institute of Rheumatology [Oxford, UK], Imperial College London, Theodor Kocher Institute, University of Bern, Leibniz Research Institute for Environmental Medicine [Düsseldorf, Germany] ( IUF), Université Lumière - Lyon 2 (UL2), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), University of Edinburgh, Integrative Biology Program [Milano], Istituto Nazionale Genetica Molecolare [Milano] (INGM), Singapore Immunology Network (SIgN), Biomedical Sciences Institute (BMSI), Universitat de Barcelona (UB), Rheumatologie, Cell Biology, Department of medicine [Stockholm], Karolinska Institutet [Stockholm]-Karolinska University Hospital [Stockholm], Department for Internal Medicine 3, Institute for Clinical Immunology, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Delft University of Technology (TU Delft), Medical Inflammation Research, Karolinska Institutet [Stockholm], Department of Photonics Engineering [Lyngby], Technical University of Denmark [Lyngby] (DTU), Dpt of Experimental Immunology [Braunschweig], Helmholtz Centre for Infection Research (HZI), Department of Internal Medicine V, Universität Heidelberg [Heidelberg], Department of Histology and Embryology, University of Rijeka, Freiburg University Medical Center, Nuffield Dept of Clinical Medicine, University of Oxford [Oxford]-NIHR Biomedical Research Centre, Institute of Integrative Biology, Molecular Biomedicine, Berlin Institute of Health (BIH), Laboratory for Lymphocyte Differentiation, RIKEN Research Center, Institutes of Molecular Medicine and Experimental Immunology, University of Bonn, Immunité et cancer (U932), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Department of Surgery [Vancouver, BC, Canada] (Child and Family Research Institute), University of British Columbia (UBC)-Child and Family Research Institute [Vancouver, BC, Canada], College of Food Science and Technology [Shangai], Shanghai Ocean University, Institute for Medical Microbiology and Hygiene, University of Marburg, King‘s College London, Erasmus University Medical Center [Rotterdam] (Erasmus MC), Centre d'Immunophénomique (CIPHE), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Brustzentrum Kantonsspital St. Gallen, Immunotechnology Section, Vaccine Research Center, National Institutes of Health [Bethesda] (NIH)-National Institute of Allergy and Infectious Diseases, Heinrich Pette Institute [Hamburg], Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), Department of Immunology and Cell Biology, Mario Negri Institute, Laboratory of Molecular Medicine and Biotechnology, Don C. Gnocchi ONLUS Foundation, Institute of Translational Medicine, Klinik für Dermatologie, Venerologie und Allergologie, School of Biochemistry and Immunology, Department of Medicine Huddinge, Karolinska Institutet [Stockholm]-Karolinska University Hospital [Stockholm]-Lipid Laboratory, Università di Genova, Dipartimento di Medicina Sperimentale, Department of Environmental Microbiology, Helmholtz Zentrum für Umweltforschung = Helmholtz Centre for Environmental Research (UFZ), Department of Radiation Oncology [Munich], Ludwig-Maximilians-Universität München (LMU), Centre de Recherche Publique- Santé, Université du Luxembourg (Uni.lu), William Harvey Research Institute, Barts and the London Medical School, University of Michigan [Ann Arbor], University of Michigan System, Centro de Investigacion del Cancer (CSIC), Universitario de Salamanca, Molecular Pathology [Tartu, Estonia], University of Tartu, Hannover Medical School [Hannover] (MHH), Centre d'Immunologie de Marseille - Luminy (CIML), Monash Biomedicine Discovery Institute, Cytometry Laboratories and School of Veterinary Medicine, Purdue University [West Lafayette], Data Mining and Modelling for Biomedicine [Ghent, Belgium], VIB Center for Inflammation Research [Ghent, Belgium], Laboratory for Cell Signaling, RIKEN Research Center for Allergy and Immunology, RIKEN Research Center for Allergy and Immunology, Osaka University [Osaka], Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP), Institute of Medical Immunology [Berlin, Germany], FACS and Array Core Facility, Johannes Gutenberg - Universität Mainz (JGU), Otto-von-Guericke University [Magdeburg] (OVGU), SUPA School of Physics and Astronomy [University of St Andrews], University of St Andrews [Scotland]-Scottish Universities Physics Alliance (SUPA), Biologie Cellulaire des Lymphocytes - Lymphocyte Cell Biology, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), General Pathology and Immunology (GPI), University of Brescia, Université de Lausanne (UNIL), Terry Fox Laboratory, BC Cancer Agency (BCCRC)-British Columbia Cancer Agency Research Centre, Department of Molecular Immunology, Medizinische Universität Wien = Medical University of Vienna, Dept. Pediatric Cardiology, Universität Leipzig [Leipzig], Universitaetsklinikum Hamburg-Eppendorf = University Medical Center Hamburg-Eppendorf [Hamburg] (UKE), Center for Cardiovascular Sciences, Albany Medical College, Dept Pathol, Div Immunol, University of Cambridge [UK] (CAM), Department of Information Technology [Gent], Universiteit Gent, Department of Plant Systems Biology, Department of Plant Biotechnology and Genetics, Universiteit Gent = Ghent University [Belgium] (UGENT), Division of Molecular Immunology, Institute for Immunology, Department of Geological Sciences, University of Oregon [Eugene], Centers for Disease Control and Prevention [Atlanta] (CDC), Centers for Disease Control and Prevention, University of Colorado [Colorado Springs] (UCCS), FACS laboratory, Cancer Research, London, Cancer Research UK, Regeneration in Hematopoiesis and Animal Models of Hematopoiesis, Faculty of Medicine, Dresden University of Technology, Barbara Davis Center for Childhood Diabetes (BDC), University of Colorado Anschutz [Aurora], School of Computer and Electronic Information [Guangxi University], Guangxi University [Nanning], School of Materials Science and Engineering, Nanyang Technological University [Singapour], Max Planck Institute for Infection Biology (MPIIB), Max-Planck-Gesellschaft, Work in the laboratory of Dieter Adam is supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Projektnummer 125440785 – SFB 877, Project B2.Petra Hoffmann, Andrea Hauser, and Matthias Edinger thank BD Biosciences®, San José, CA, USA, and SKAN AG, Bale, Switzerland for fruitful cooperation during the development, construction, and installation of the GMP‐compliant cell sorting equipment and the Bavarian Immune Therapy Network (BayImmuNet) for financial support.Edwin van der Pol and Paola Lanuti acknowledge Aleksandra Gąsecka M.D. for excellent experimental support and Dr. Rienk Nieuwland for textual suggestions. This work was supported by the Netherlands Organisation for Scientific Research – Domain Applied and Engineering Sciences (NWO‐TTW), research program VENI 15924.Jessica G Borger, Kylie M Quinn, Mairi McGrath, and Regina Stark thank Francesco Siracusa and Patrick Maschmeyer for providing data.Larissa Nogueira Almeida was supported by DFG research grant MA 2273/14‐1. Rudolf A. Manz was supported by the Excellence Cluster 'Inflammation at Interfaces' (EXC 306/2).Susanne Hartmann and Friederike Ebner were supported by the German Research Foundation (GRK 2046).Hans Minderman was supported by NIH R50CA211108.This work was funded by the Deutsche Forschungsgemeinschaft through the grant TRR130 (project P11 and C03) to Thomas H. Winkler.Ramon Bellmàs Sanz, Jenny Kühne, and Christine S. Falk thank Jana Keil and Kerstin Daemen for excellent technical support. The work was funded by the Germany Research Foundation CRC738/B3 (CSF).The work by the Mei laboratory was supported by German Research Foundation Grant ME 3644/5‐1 and TRR130 TP24, the German Rheumatism Research Centre Berlin, European Union Innovative Medicines Initiative ‐ Joint Undertaking ‐ RTCure Grant Agreement 777357, the Else Kröner‐Fresenius‐Foundation, German Federal Ministry of Education and Research e:Med sysINFLAME Program Grant 01ZX1306B and KMU‐innovativ 'InnoCyt', and the Leibniz Science Campus for Chronic Inflammation (http://www.chronische-entzuendung.org).Axel Ronald Schulz, Antonio Cosma, Sabine Baumgart, Brice Gaudilliere, Helen M. McGuire, and Henrik E. Mei thank Michael D. Leipold for critically reading the manuscript.Christian Kukat acknowledges support from the ISAC SRL Emerging Leaders program.John Trowsdale received funding from the European Research Council under the European Union's Horizon 2020 research and innovation program (Grant Agreement 695551)., European Project: 7728036(1978), Università degli Studi di Modena e Reggio Emilia = University of Modena and Reggio Emilia (UNIMORE), Université Paris Cité (UPCité), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Università degli Studi di Firenze = University of Florence (UniFI)-DENOTHE Center, Università degli Studi di Milano = University of Milan (UNIMI), Institut Pasteur [Paris] (IP)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Humboldt University Of Berlin, Leibniz Research Institute for Environmental Medicine [Düsseldorf, Germany] (IUF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Danmarks Tekniske Universitet = Technical University of Denmark (DTU), Universität Heidelberg [Heidelberg] = Heidelberg University, Universitäts Klinikum Freiburg = University Medical Center Freiburg (Uniklinik), University of Oxford-NIHR Biomedical Research Centre, Universität Bonn = University of Bonn, Università degli Studi di Firenze = University of Florence (UniFI), Università degli studi di Genova = University of Genoa (UniGe), Universidad de Salamanca, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome] (UNIROMA), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Johannes Gutenberg - Universität Mainz = Johannes Gutenberg University (JGU), Otto-von-Guericke-Universität Magdeburg = Otto-von-Guericke University [Magdeburg] (OVGU), Université de Lausanne = University of Lausanne (UNIL), Universität Leipzig, Universiteit Gent = Ghent University (UGENT), HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany., Cossarizza, A., Chang, H. -D., Radbruch, A., Acs, A., Adam, D., Adam-Klages, S., Agace, W. W., Aghaeepour, N., Akdis, M., Allez, M., Almeida, L. N., Alvisi, G., Anderson, G., Andra, I., Annunziato, F., Anselmo, A., Bacher, P., Baldari, C. T., Bari, S., Barnaba, V., Barros-Martins, J., Battistini, L., Bauer, W., Baumgart, S., Baumgarth, N., Baumjohann, D., Baying, B., Bebawy, M., Becher, B., Beisker, W., Benes, V., Beyaert, R., Blanco, A., Boardman, D. A., Bogdan, C., Borger, J. G., Borsellino, G., Boulais, P. E., Bradford, J. A., Brenner, D., Brinkman, R. R., Brooks, A. E. S., Busch, D. H., Buscher, M., Bushnell, T. P., Calzetti, F., Cameron, G., Cammarata, I., Cao, X., Cardell, S. L., Casola, S., Cassatella, M. A., Cavani, A., Celada, A., Chatenoud, L., Chattopadhyay, P. K., Chow, S., Christakou, E., Cicin-Sain, L., Clerici, M., Colombo, F. S., Cook, L., Cooke, A., Cooper, A. M., Corbett, A. J., Cosma, A., Cosmi, L., Coulie, P. G., Cumano, A., Cvetkovic, L., Dang, V. D., Dang-Heine, C., Davey, M. S., Davies, D., De Biasi, S., Del Zotto, G., Dela Cruz, G. V., Delacher, M., Della Bella, S., Dellabona, P., Deniz, G., Dessing, M., Di Santo, J. P., Diefenbach, A., Dieli, F., Dolf, A., Dorner, T., Dress, R. J., Dudziak, D., Dustin, M., Dutertre, C. -A., Ebner, F., Eckle, S. B. G., Edinger, M., Eede, P., Ehrhardt, G. R. A., Eich, M., Engel, P., Engelhardt, B., Erdei, A., Esser, C., Everts, B., Evrard, M., Falk, C. S., Fehniger, T. A., Felipo-Benavent, M., Ferry, H., Feuerer, M., Filby, A., Filkor, K., Fillatreau, S., Follo, M., Forster, I., Foster, J., Foulds, G. A., Frehse, B., Frenette, P. S., Frischbutter, S., Fritzsche, W., Galbraith, D. W., Gangaev, A., Garbi, N., Gaudilliere, B., Gazzinelli, R. T., Geginat, J., Gerner, W., Gherardin, N. A., Ghoreschi, K., Gibellini, L., Ginhoux, F., Goda, K., Godfrey, D. I., Goettlinger, C., Gonzalez-Navajas, J. M., Goodyear, C. S., Gori, A., Grogan, J. L., Grummitt, D., Grutzkau, A., Haftmann, C., Hahn, J., Hammad, H., Hammerling, G., Hansmann, L., Hansson, G., Harpur, C. M., Hartmann, S., Hauser, A., Hauser, A. E., Haviland, D. L., Hedley, D., Hernandez, D. C., Herrera, G., Herrmann, M., Hess, C., Hofer, T., Hoffmann, P., Hogquist, K., Holland, T., Hollt, T., Holmdahl, R., Hombrink, P., Houston, J. P., Hoyer, B. F., Huang, B., Huang, F. -P., Huber, J. E., Huehn, J., Hundemer, M., Hunter, C. A., Hwang, W. Y. K., Iannone, A., Ingelfinger, F., Ivison, S. M., Jack, H. -M., Jani, P. K., Javega, B., Jonjic, S., Kaiser, T., Kalina, T., Kamradt, T., Kaufmann, S. H. E., Keller, B., Ketelaars, S. L. C., Khalilnezhad, A., Khan, S., Kisielow, J., Klenerman, P., Knopf, J., Koay, H. -F., Kobow, K., Kolls, J. K., Kong, W. T., Kopf, M., Korn, T., Kriegsmann, K., Kristyanto, H., Kroneis, T., Krueger, A., Kuhne, J., Kukat, C., Kunkel, D., Kunze-Schumacher, H., Kurosaki, T., Kurts, C., Kvistborg, P., Kwok, I., Landry, J., Lantz, O., Lanuti, P., Larosa, F., Lehuen, A., LeibundGut-Landmann, S., Leipold, M. D., Leung, L. Y. T., Levings, M. K., Lino, A. C., Liotta, F., Litwin, V., Liu, Y., Ljunggren, H. -G., Lohoff, M., Lombardi, G., Lopez, L., Lopez-Botet, M., Lovett-Racke, A. E., Lubberts, E., Luche, H., Ludewig, B., Lugli, E., Lunemann, S., Maecker, H. T., Maggi, L., Maguire, O., Mair, F., Mair, K. H., Mantovani, A., Manz, R. A., Marshall, A. J., Martinez-Romero, A., Martrus, G., Marventano, I., Maslinski, W., Matarese, G., Mattioli, A. V., Maueroder, C., Mazzoni, A., Mccluskey, J., Mcgrath, M., Mcguire, H. M., Mcinnes, I. B., Mei, H. E., Melchers, F., Melzer, S., Mielenz, D., Miller, S. D., Mills, K. H. G., Minderman, H., Mjosberg, J., Moore, J., Moran, B., Moretta, L., Mosmann, T. R., Muller, S., Multhoff, G., Munoz, L. E., Munz, C., Nakayama, T., Nasi, M., Neumann, K., Ng, L. G., Niedobitek, A., Nourshargh, S., Nunez, G., O'Connor, J. -E., Ochel, A., Oja, A., Ordonez, D., Orfao, A., Orlowski-Oliver, E., Ouyang, W., Oxenius, A., Palankar, R., Panse, I., Pattanapanyasat, K., Paulsen, M., Pavlinic, D., Penter, L., Peterson, P., Peth, C., Petriz, J., Piancone, F., Pickl, W. F., Piconese, S., Pinti, M., Pockley, A. G., Podolska, M. J., Poon, Z., Pracht, K., Prinz, I., Pucillo, C. E. M., Quataert, S. A., Quatrini, L., Quinn, K. M., Radbruch, H., Radstake, T. R. D. J., Rahmig, S., Rahn, H. -P., Rajwa, B., Ravichandran, G., Raz, Y., Rebhahn, J. A., Recktenwald, D., Reimer, D., Reis e Sousa, C., Remmerswaal, E. B. M., Richter, L., Rico, L. G., Riddell, A., Rieger, A. M., Robinson, J. P., Romagnani, C., Rubartelli, A., Ruland, J., Saalmuller, A., Saeys, Y., Saito, T., Sakaguchi, S., Sala-de-Oyanguren, F., Samstag, Y., Sanderson, S., Sandrock, I., Santoni, A., Sanz, R. B., Saresella, M., Sautes-Fridman, C., Sawitzki, B., Schadt, L., Scheffold, A., Scherer, H. U., Schiemann, M., Schildberg, F. A., Schimisky, E., Schlitzer, A., Schlosser, J., Schmid, S., Schmitt, S., Schober, K., Schraivogel, D., Schuh, W., Schuler, T., Schulte, R., Schulz, A. R., Schulz, S. R., Scotta, C., Scott-Algara, D., Sester, D. P., Shankey, T. V., Silva-Santos, B., Simon, A. K., Sitnik, K. M., Sozzani, S., Speiser, D. E., Spidlen, J., Stahlberg, A., Stall, A. M., Stanley, N., Stark, R., Stehle, C., Steinmetz, T., Stockinger, H., Takahama, Y., Takeda, K., Tan, L., Tarnok, A., Tiegs, G., Toldi, G., Tornack, J., Traggiai, E., Trebak, M., Tree, T. I. M., Trotter, J., Trowsdale, J., Tsoumakidou, M., Ulrich, H., Urbanczyk, S., van de Veen, W., van den Broek, M., van der Pol, E., Van Gassen, S., Van Isterdael, G., van Lier, R. A. W., Veldhoen, M., Vento-Asturias, S., Vieira, P., Voehringer, D., Volk, H. -D., von Borstel, A., von Volkmann, K., Waisman, A., Walker, R. V., Wallace, P. K., Wang, S. A., Wang, X. M., Ward, M. D., Ward-Hartstonge, K. A., Warnatz, K., Warnes, G., Warth, S., Waskow, C., Watson, J. V., Watzl, C., Wegener, L., Weisenburger, T., Wiedemann, A., Wienands, J., Wilharm, A., Wilkinson, R. J., Willimsky, G., Wing, J. B., Winkelmann, R., Winkler, T. H., Wirz, O. F., Wong, A., Wurst, P., Yang, J. H. M., Yang, J., Yazdanbakhsh, M., Yu, L., Yue, A., Zhang, H., Zhao, Y., Ziegler, S. M., Zielinski, C., Zimmermann, J., Zychlinsky, A., UCL - SSS/DDUV - Institut de Duve, UCL - SSS/DDUV/GECE - Génétique cellulaire, Netherlands Organization for Scientific Research, German Research Foundation, European Commission, European Research Council, Repositório da Universidade de Lisboa, CCA - Imaging and biomarkers, Experimental Immunology, AII - Infectious diseases, AII - Inflammatory diseases, Biomedical Engineering and Physics, ACS - Atherosclerosis & ischemic syndromes, and Landsteiner Laboratory
Subjects: 0301 basic medicine, Consensus, Immunology, Consensu, Cell Separation, Biology, Article, Flow cytometry, 03 medical and health sciences, 0302 clinical medicine, Guidelines, Allergy and Immunology, medicine, Cell separation, Immunology and Allergy, Humans, guidelines, flow cytometry, immunology, medicine.diagnostic_test, BIOMEDICINE AND HEALTHCARE. Basic Medical Sciences, Cell sorting, Flow Cytometry, Cell selection, Data science, 3. Good health, 030104 developmental biology, Phenotype, [SDV.IMM]Life Sciences [q-bio]/Immunology, BIOMEDICINA I ZDRAVSTVO. Temeljne medicinske znanosti, 030215 immunology, Human
Abstract: All authors: Andrea Cossarizza Hyun‐Dong Chang Andreas Radbruch Andreas Acs Dieter Adam Sabine Adam‐Klages William W. Agace Nima Aghaeepour Mübeccel Akdis Matthieu Allez Larissa Nogueira Almeida Giorgia Alvisi Graham Anderson Immanuel Andrä Francesco Annunziato Achille Anselmo Petra Bacher Cosima T. Baldari Sudipto Bari Vincenzo Barnaba Joana Barros‐Martins Luca Battistini Wolfgang Bauer Sabine Baumgart Nicole Baumgarth Dirk Baumjohann Bianka Baying Mary Bebawy Burkhard Becher Wolfgang Beisker Vladimir Benes Rudi Beyaert Alfonso Blanco Dominic A. Boardman Christian Bogdan Jessica G. Borger Giovanna Borsellino Philip E. Boulais Jolene A. Bradford Dirk Brenner Ryan R. Brinkman Anna E. S. Brooks Dirk H. Busch Martin Büscher Timothy P. Bushnell Federica Calzetti Garth Cameron Ilenia Cammarata Xuetao Cao Susanna L. Cardell Stefano Casola Marco A. Cassatella Andrea Cavani Antonio Celada Lucienne Chatenoud Pratip K. Chattopadhyay Sue Chow Eleni Christakou Luka Čičin‐Šain Mario Clerici Federico S. Colombo Laura Cook Anne Cooke Andrea M. Cooper Alexandra J. Corbett Antonio Cosma Lorenzo Cosmi Pierre G. Coulie Ana Cumano Ljiljana Cvetkovic Van Duc Dang Chantip Dang‐Heine Martin S. Davey Derek Davies Sara De Biasi Genny Del Zotto Gelo Victoriano Dela Cruz Michael Delacher Silvia Della Bella Paolo Dellabona Günnur Deniz Mark Dessing James P. Di Santo Andreas Diefenbach Francesco Dieli Andreas Dolf Thomas Dörner Regine J. Dress Diana Dudziak Michael Dustin Charles‐Antoine Dutertre Friederike Ebner Sidonia B. G. Eckle Matthias Edinger Pascale Eede Götz R.A. Ehrhardt Marcus Eich Pablo Engel Britta Engelhardt Anna Erdei Charlotte Esser Bart Everts Maximilien Evrard Christine S. Falk Todd A. Fehniger Mar Felipo‐Benavent Helen Ferry Markus Feuerer Andrew Filby Kata Filkor Simon Fillatreau Marie Follo Irmgard Förster John Foster Gemma A. Foulds Britta Frehse Paul S. Frenette Stefan Frischbutter Wolfgang Fritzsche David W. Galbraith Anastasia Gangaev Natalio Garbi Brice Gaudilliere Ricardo T. Gazzinelli Jens Geginat Wilhelm Gerner Nicholas A. Gherardin Kamran Ghoreschi Lara Gibellini Florent Ginhoux Keisuke Goda Dale I. Godfrey Christoph Goettlinger Jose M. González‐Navajas Carl S. Goodyear Andrea Gori Jane L. Grogan Daryl Grummitt Andreas Grützkau Claudia Haftmann Jonas Hahn Hamida Hammad Günter Hämmerling Leo Hansmann Goran Hansson Christopher M. Harpur Susanne Hartmann Andrea Hauser Anja E. Hauser David L. Haviland David Hedley Daniela C. Hernández Guadalupe Herrera Martin Herrmann Christoph Hess Thomas Höfer Petra Hoffmann Kristin Hogquist Tristan Holland Thomas Höllt Rikard Holmdahl Pleun Hombrink Jessica P. Houston Bimba F. Hoyer Bo Huang Fang‐Ping Huang Johanna E. Huber Jochen Huehn Michael Hundemer Christopher A. Hunter William Y. K. Hwang Anna Iannone Florian Ingelfinger Sabine M Ivison Hans‐Martin Jäck Peter K. Jani Beatriz Jávega Stipan Jonjic Toralf Kaiser Tomas Kalina Thomas Kamradt Stefan H. E. Kaufmann Baerbel Keller Steven L. C. Ketelaars Ahad Khalilnezhad Srijit Khan Jan Kisielow Paul Klenerman Jasmin Knopf Hui‐Fern Koay Katja Kobow Jay K. Kolls Wan Ting Kong Manfred Kopf Thomas Korn Katharina Kriegsmann Hendy Kristyanto Thomas Kroneis Andreas Krueger Jenny Kühne Christian Kukat Désirée Kunkel Heike Kunze‐Schumacher Tomohiro Kurosaki Christian Kurts Pia Kvistborg Immanuel Kwok Jonathan Landry Olivier Lantz Paola Lanuti Francesca LaRosa Agnès Lehuen Salomé LeibundGut‐Landmann Michael D. Leipold Leslie Y.T. Leung Megan K. Levings Andreia C. Lino Francesco Liotta Virginia Litwin Yanling Liu Hans‐Gustaf Ljunggren Michael Lohoff Giovanna Lombardi Lilly Lopez Miguel López‐Botet Amy E. Lovett‐Racke Erik Lubberts Herve Luche Burkhard Ludewig Enrico Lugli Sebastian Lunemann Holden T. Maecker Laura Maggi Orla Maguire Florian Mair Kerstin H. Mair Alberto Mantovani Rudolf A. Manz Aaron J. Marshall Alicia Martínez‐Romero Glòria Martrus Ivana Marventano Wlodzimierz Maslinski Giuseppe Matarese Anna Vittoria Mattioli Christian Maueröder Alessio Mazzoni James McCluskey Mairi McGrath Helen M. McGuire Iain B. McInnes Henrik E. Mei Fritz Melchers Susanne Melzer Dirk Mielenz Stephen D. Miller Kingston H.G. Mills Hans Minderman Jenny Mjösberg Jonni Moore Barry Moran Lorenzo Moretta Tim R. Mosmann Susann Müller Gabriele Multhoff Luis Enrique Muñoz Christian Münz Toshinori Nakayama Milena Nasi Katrin Neumann Lai Guan Ng Antonia Niedobitek Sussan Nourshargh Gabriel Núñez José‐Enrique O'Connor Aaron Ochel Anna Oja Diana Ordonez Alberto Orfao Eva Orlowski‐Oliver Wenjun Ouyang Annette Oxenius Raghavendra Palankar Isabel Panse Kovit Pattanapanyasat Malte Paulsen Dinko Pavlinic Livius Penter Pärt Peterson Christian Peth Jordi Petriz Federica Piancone Winfried F. Pickl Silvia Piconese Marcello Pinti A. Graham Pockley Malgorzata Justyna Podolska Zhiyong Poon Katharina Pracht Immo Prinz Carlo E. M. Pucillo Sally A. Quataert Linda Quatrini Kylie M. Quinn Helena Radbruch Tim R. D. J. Radstake Susann Rahmig Hans‐Peter Rahn Bartek Rajwa Gevitha Ravichandran Yotam Raz Jonathan A. Rebhahn Diether Recktenwald Dorothea Reimer Caetano Reis e Sousa Ester B.M. Remmerswaal Lisa Richter Laura G. Rico Andy Riddell Aja M. Rieger J. Paul Robinson Chiara Romagnani Anna Rubartelli Jürgen Ruland Armin Saalmüller Yvan Saeys Takashi Saito Shimon Sakaguchi Francisco Sala‐de‐Oyanguren Yvonne Samstag Sharon Sanderson Inga Sandrock Angela Santoni Ramon Bellmàs Sanz Marina Saresella Catherine Sautes‐Fridman Birgit Sawitzki Linda Schadt Alexander Scheffold Hans U. Scherer Matthias Schiemann Frank A. Schildberg Esther Schimisky Andreas Schlitzer Josephine Schlosser Stephan Schmid Steffen Schmitt Kilian Schober Daniel Schraivogel Wolfgang Schuh Thomas Schüler Reiner Schulte Axel Ronald Schulz Sebastian R. Schulz Cristiano Scottá Daniel Scott‐Algara David P. Sester T. Vincent Shankey Bruno Silva‐Santos Anna Katharina Simon Katarzyna M. Sitnik Silvano Sozzani Daniel E. Speiser Josef Spidlen Anders Stahlberg Alan M. Stall Natalie Stanley Regina Stark Christina Stehle Tobit Steinmetz Hannes Stockinger Yousuke Takahama Kiyoshi Takeda Leonard Tan Attila Tárnok Gisa Tiegs Gergely Toldi Julia Tornack Elisabetta Traggiai Mohamed Trebak Timothy I.M. Tree Joe Trotter John Trowsdale Maria Tsoumakidou Henning Ulrich Sophia Urbanczyk Willem van de Veen Maries van den Broek Edwin van der Pol Sofie Van Gassen Gert Van Isterdael René A.W. van Lier Marc Veldhoen Salvador Vento‐Asturias Paulo Vieira David Voehringer Hans‐Dieter Volk Anouk von Borstel Konrad von Volkmann Ari Waisman Rachael V. Walker Paul K. Wallace Sa A. Wang Xin M. Wang Michael D. Ward Kirsten A Ward‐Hartstonge Klaus Warnatz Gary Warnes Sarah Warth Claudia Waskow James V. Watson Carsten Watzl Leonie Wegener Thomas Weisenburger Annika Wiedemann Jürgen Wienands Anneke Wilharm Robert John Wilkinson Gerald Willimsky James B. Wing Rieke Winkelmann Thomas H. Winkler Oliver F. Wirz Alicia Wong Peter Wurst Jennie H. M. Yang Juhao Yang Maria Yazdanbakhsh Liping Yu Alice Yue Hanlin Zhang Yi Zhao Susanne Maria Ziegler Christina Zielinski Jakob Zimmermann Arturo Zychlinsky., 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., This work was supported by the Netherlands Organisation for Scientific Research – Domain Applied and Engineering Sciences (NWO-TTW), research program VENI 15924. This work was funded by the Deutsche Forschungsgemeinschaft. European Union Innovative Medicines Initiative - Joint Undertaking - RTCure Grant Agreement 777357 and innovation program (Grant Agreement 695551).
Published: 2019

17. 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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18. Differential effects of PD-1 and CTLA-4 blockade on the melanoma-reactive CD8 T cell response

Author: Sanne Patiwael, Christian U. Blank, Ton N. Schumacher, Antoni Ribas, John B. A. G. Haanen, Anastasia Gangaev, Daisy Philips, Dirk Schadendorf, Joost H. van den Berg, Maartje W. Rohaan, Bastian Schilling, Elisa A. Rozeman, Pia Kvistborg, and Olga I. Isaeva
Subjects: Male, Receptors, Antigen, T-Cell, alpha-beta, Programmed Cell Death 1 Receptor, Medizin, Epitopes, T-Lymphocyte, CD8-Positive T-Lymphocytes, Lymphocyte Activation, Epitope, Cohort Studies, Epitopes, melanoma-reactive CD8 T cells, Receptors, PD-1, 80 and over, Cytotoxic T cell, CTLA-4 Antigen, Hepatitis A Virus Cellular Receptor 2, Immune Checkpoint Inhibitors, Melanoma, Cancer, alpha-beta, Aged, 80 and over, Multidisciplinary, Middle Aged, Biological Sciences, medicine.anatomical_structure, 5.1 Pharmaceuticals, Antigen, Female, Development of treatments and therapeutic interventions, medicine.symptom, Melanoma-Specific Antigens, Adult, Receptors, CXCR5, T cell, In Vitro Techniques, Clinical Research, medicine, Humans, Aged, business.industry, Compartment (ship), T-Cell, medicine.disease, CXCR5, Blockade, Kinetics, T-Lymphocyte, Mechanism of action, CTLA-4, Cancer research, business
Abstract: Immune checkpoint inhibitors targeting programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have revolutionized the treatment of melanoma patients. Based on early studies addressing the mechanism of action, it was assumed that PD-1 blockade mostly influences T cell responses at the tumor site. However, recent work has demonstrated that PD-1 blockade can influence the T cell compartment in peripheral blood. If the activation of circulating, tumor-reactive T cells would form an important mechanism of action of PD-1 blockade, it may be predicted that such blockade would alter either the frequency and/or the breadth of the tumor-reactive CD8 T cell response. To address this question, we analyzed CD8 T cell responses toward 71 melanoma-associated epitopes in peripheral blood of 24 melanoma patients. We show that both the frequency and the breadth of the circulating melanoma-reactive CD8 T cell response was unaltered upon PD-1 blockade. In contrast, a broadening of the circulating melanoma-reactive CD8 T cell response was observed upon CTLA-4 blockade, in concordance with our prior data. Based on these results, we conclude that PD-1 and CTLA-4 blockade have distinct mechanisms of action. In addition, the data provide an argument in favor of the hypothesis that anti-PD-1 therapy may primarily act at the tumor site.
Published: 2021

19. Imatinib Regulates miR-483-3p and Mitochondrial Respiratory Complexes in Gastrointestinal Stromal Tumors

Author: Huang, Wen-Kuan, Shi, Hao, Akçakaya, Pinar, Zeljic, Katarina, Gangaev, Anastasia, Caramuta, Stefano, Yeh, Chun-Nan, Bränström, Robert, Larsson, Catharina, and Lui, Weng-Onn
Subjects: Cell Survival, Gastrointestinal Stromal Tumors, QH301-705.5, oxidative phosphorylation, Down-Regulation, Antineoplastic Agents, Transfection, Article, Cell Line, Tumor, hemic and lymphatic diseases, Humans, succinate dehydrogenase B, RNA, Messenger, Biology (General), neoplasms, QD1-999, Gastrointestinal Neoplasms, microRNA, Electron Transport Complex II, gastrointestinal stromal tumor (GIST), Mitochondria, Gene Expression Regulation, Neoplastic, miR-483, MicroRNAs, Chemistry, imatinib, Complex II, Imatinib Mesylate, Signal Transduction
Abstract: Metabolic adaptation to increased oxidative phosphorylation (OXPHOS) has been found in gastrointestinal stromal tumor (GIST) upon imatinib treatment. However, the underlying mechanism of imatinib-induced OXPHOS is unknown. Discovering molecules that mediate imatinib-induced OXPHOS may lead to the development of therapeutic strategies synergizing the efficacy of imatinib. In this study, we explored the role of microRNAs in regulating OXPHOS in GIST upon imatinib treatment. Using a microarray approach, we found that miR-483-3p was one of the most downregulated miRNAs in imatinib-treated tumors compared to untreated tumors. Using an extended series of GIST samples, we further validated the downregulation of miR-483-3p in imatinib-treated GIST samples by RT-qPCR. Using both gain- and loss-of-function experiments, we showed that miR-483-3p could regulate mitochondrial respiratory Complex II expression, suggesting its role in OXPHOS regulation. Functionally, miR-483-3p overexpression could rescue imatinib-induced cell death. These findings provide the molecular link for imatinib-induced OXPHOS expression and the biological role of miR-483-3p in regulating cell viability upon imatinib treatment.
Published: 2021

20. Identification and characterization of a SARS-CoV-2 specific CD8+ T cell response with immunodominant features

Author: Ton N. Schumacher, Giovanni Guaraldi, Pia Kvistborg, Andrea Cossarizza, Cristina Mussini, Kelly Hoefakker, Olga I. Isaeva, Steven L. C. Ketelaars, Lara Gibellini, Anastasia Gangaev, Huib Ovaa, Cami Talavera Ormeño, Robert Balderas, Anna Dopler, Sara De Biasi, Massimo Girardis, Sanne Patiwael, Paul J. M. Hekking, Mireille Toebes, and Neubury M. Lardy
Subjects: 0301 basic medicine, Multidisciplinary, Effector, Science, medicine.medical_treatment, T cell, General Physics and Astronomy, General Chemistry, Human leukocyte antigen, Biology, Acquired immune system, Virology, General Biochemistry, Genetics and Molecular Biology, Epitope, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, Cytokine, medicine.anatomical_structure, 030220 oncology & carcinogenesis, medicine, Cytotoxic T cell, CD8
Abstract: Many viral antigens have been identified in patients with COVID-19 patients, but which of these result in meaningful immune responses is unclear. Here the authors identify a range of SARS-CoV-2 CD8(+) T cell responses across patients including a response targeting an epitope of ORF1ab with immunodominant properties.The COVID-19 pandemic caused by SARS-CoV-2 is a continuous challenge worldwide, and there is an urgent need to map the landscape of immunogenic and immunodominant epitopes recognized by CD8(+) T cells. Here, we analyze samples from 31 patients with COVID-19 for CD8(+) T cell recognition of 500 peptide-HLA class I complexes, restricted by 10 common HLA alleles. We identify 18 CD8(+) T cell recognized SARS-CoV-2 epitopes, including an epitope with immunodominant features derived from ORF1ab and restricted by HLA-A*01:01. In-depth characterization of SARS-CoV-2-specific CD8(+) T cell responses of patients with acute critical and severe disease reveals high expression of NKG2A, lack of cytokine production and a gene expression profile inhibiting T cell re-activation and migration while sustaining survival. SARS-CoV-2-specific CD8(+) T cell responses are detectable up to 5 months after recovery from critical and severe disease, and these responses convert from dysfunctional effector to functional memory CD8(+) T cells during convalescence.
Published: 2021

21. Alternative trafficking of Weibel‐Palade body proteins in CRISPR/Cas9‐engineered von Willebrand factor–deficient blood outgrowth endothelial cells

Author: Benjamin Nota, Jan Voorberg, Anastasia Gangaev, Maartje van den Biggelaar, Martin de Boer, Marije Kat, Maaike Schillemans, Jurjen Westeneng, Menno Hofman, Floris P. J. van Alphen, Ruben Bierings, Coert Margadant, Hematology, ACS - Microcirculation, Graduate School, AII - Inflammatory diseases, Landsteiner Laboratory, and Experimental Vascular Medicine
Subjects: congenital, hereditary, and neonatal diseases and abnormalities, secretory vesicles, von Willebrand factor, Cell morphology, Angiopoietin, Von Willebrand factor, hemic and lymphatic diseases, Weibel–Palade body, Von Willebrand disease, medicine, CRISPR, Secretion, Original Articles: Haemostasis, biology, lcsh:RC633-647.5, gene knockout techniques, lcsh:Diseases of the blood and blood-forming organs, Hematology, medicine.disease, endothelial cells, Cell biology, Endothelial stem cell, cardiovascular system, biology.protein, protein transport, Original Article, circulatory and respiratory physiology
Abstract: textabstractBackground: Synthesis of the hemostatic protein von Willebrand factor (VWF) drives formation of endothelial storage organelles called Weibel‐Palade bodies (WPBs). In the absence of VWF, angiogenic and inflammatory mediators that are costored in WPBs are subject to alternative trafficking routes. In patients with von Willebrand disease (VWD), partial or complete absence of VWF/WPBs may lead to additional bleeding complications, such as angiodysplasia. Studies addressing the role of VWF using VWD patient–derived blood outgrowth endothelial cells (BOECs) have reported conflicting results due to the intrinsic heterogeneity of patient‐derived BOECs. Objective: To generate a VWF‐deficient endothelial cell model using clustered regularly interspaced short palindromic repeats (CRISPR) genome engineering of blood outgrowth endothelial cells. Methods: We used CRISPR/CRISPR‐associated protein 9 editing in single‐donor cord blood–derived BOECs (cbBOECs) to generate clonal VWF−/− cbBOECs. Clones were selected using high‐throughput screening, VWF mutations were validated by sequencing, and cells were phenotypically characterized. Results: Two VWF−/− BOEC clones were obtained and were entirely devoid of WPBs, while their overall cell morphology was unaltered. Several WPB proteins, including CD63, syntaxin‐3 and the cargo proteins angiopoietin (Ang)‐2, interleukin (IL)‐6, and IL‐8 showed alternative trafficking and secretion in the absence of VWF. Interestingly, Ang‐2 was relocated to the cell periphery and colocalized with Tie‐2. Conclusions: CRISPR editing of VWF provides a robust method to create VWF‐ deficient BOECs that can be directly compared to their wild‐type counterparts. Results obtained with our model system confirmed alternative trafficking of several WPB proteins in the absence of VWF and support the theory that increased Ang‐2/Tie‐2 interaction contributes to angiogenic abnormalities in VWD patients.
Published: 2019

22. Identification of presented SARS-CoV-2 HLA class I and HLA class II peptides using HLA peptidomics

Author: Nagler, Adi, primary, Kalaora, Shelly, additional, Barbolin, Chaya, additional, Gangaev, Anastasia, additional, Ketelaars, Steven L.C., additional, Alon, Michal, additional, Pai, Joy, additional, Benedek, Gil, additional, Yahalom-Ronen, Yfat, additional, Erez, Noam, additional, Greenberg, Polina, additional, Yagel, Gal, additional, Peri, Aviyah, additional, Levin, Yishai, additional, Satpathy, Ansuman T., additional, Bar-Haim, Erez, additional, Paran, Nir, additional, Kvistborg, Pia, additional, and Samuels, Yardena, additional
Published: 2021
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23. 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

24. Differential effects of PD-1 and CTLA-4 blockade on the melanoma-reactive CD8 T cell response

Author: Ton N. Schumacher, Elisa A. Rozeman, Bastian Schilling, Maartje W. Rohaan, John B. A. G. Haanen, Pia Kvistborg, Antoni Ribas, Dirk Schadendorf, Sanne Patiwael, Joost H. van den Berg, Anastasia Gangaev, Daisy Philips, and Christian U. Blank
Subjects: business.industry, Melanoma, T cell, medicine.disease, Epitope, Peripheral, Blockade, medicine.anatomical_structure, Mechanism of action, CTLA-4, Cancer research, Medicine, Cytotoxic T cell, medicine.symptom, business
Abstract: Immune checkpoint inhibitors targeting programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) have revolutionized the treatment of melanoma patients. Based on early studies addressing the mechanism of action, it was assumed that PD-1 blockade mostly influences T cell responses at the tumor site. However, recent work has demonstrated that PD-1 blockade can influence the T cell compartment in peripheral. blood. If activation of circulating tumor-reactive T cells would form an important mechanism of action of PD-1 blockade, it may be predicted that such blockade would alter either the frequency and/or the breadth of the tumor-reactive CD8 T cell response. To address this question, we analyzed CD8 T cell responses towards 71 melanoma associated epitopes in peripheral blood of 24 melanoma patients. We show that both the frequency and the breadth of the melanoma-reactive CD8 T cell response in peripheral blood was unaltered upon PD-1 blockade. In contrast, a broadening of the melanoma-reactive CD8 T cell response was observed upon CTLA-4 blockade, in concordance with our prior data. On the basis of these results, we conclude that PD-1 and CTLA-4 blockade impact the tumor-reactive CD8 T cell response in a distinct manner. In addition, the data provide an argument in favor of the hypothesis that anti-PD-1 therapy may primarily act at the tumor site.One sentence summaryWe demonstrate that, contrary to CTLA-4 blockade, PD-1 blockade in melanoma patients does not lead to an increase in the breadth or magnitude of the melanoma-reactive CD8 T cell response in peripheral blood, thereby arguing for a dominant effect of PD-1 blockade at the tumor site.
Published: 2020

25. Identification of presented SARS-CoV-2 HLA class I and HLA class II peptides using HLA peptidomics

Author: Yardena Samuels, Adi Nagler, Anastasia Gangaev, Noam Erez, Aviyah Peri, Joy A. Pai, Polina Greenberg, Gil Benedek, Shelly Kalaora, Ansuman T. Satpathy, Yishai Levin, Steven L. C. Ketelaars, Gal Yagel, Michal Alon, Pia Kvistborg, Yfat Yahalom-Ronen, Nir Paran, Erez Bar-Haim, and Chaya Barbolin
Subjects: 0301 basic medicine, COVID-19 Vaccines, QH301-705.5, viruses, T-Lymphocytes, Antigen presentation, Epitopes, T-Lymphocyte, Human leukocyte antigen, Biology, General Biochemistry, Genetics and Molecular Biology, Article, Cell Line, 03 medical and health sciences, 0302 clinical medicine, Antigen, immuno-reactive, Humans, ORFS, Biology (General), Peptidomics, Gene, Antigens, Viral, Antigen Presentation, SARS-CoV-2, HEK 293 cells, Histocompatibility Antigens Class I, Histocompatibility Antigens Class II, COVID-19, Virology, HLA, Open reading frame, 030104 developmental biology, HEK293 Cells, Cell culture, Peptidomimetics, Peptides, out-of-frame-ORFs, 030217 neurology & neurosurgery
Abstract: The human leukocyte antigen (HLA)-bound viral antigens serve as an immunological signature that can be selectively recognized by T cells. As viruses evolve by acquiring mutations, it is essential to identify a range of presented viral antigens. Using HLA peptidomics, we are able to identify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-derived peptides presented by highly prevalent HLA class I (HLA-I) molecules by using infected cells as well as overexpression of SARS-CoV-2 genes. We find 26 HLA-I peptides and 36 HLA class II (HLA-II) peptides. Among the identified peptides, some are shared between different cells and some are derived from out-of-frame open reading frames (ORFs). Seven of these peptides were previously shown to be immunogenic, and we identify two additional immunoreactive peptides by using HLA multimer staining. These results may aid the development of the next generation of SARS-CoV-2 vaccines based on presented viral-specific antigens that span several of the viral genes., Graphical abstract, Using HLA peptidomics, Nagler et al. identify SARS-CoV-2-derived peptides presented by highly prevalent HLA-I and HLA-II. Identified peptides are derived from various canonical and non-canonical viral ORFs, are shared between different cells, and are immunogenic. As the identified peptides are unique to SARS-CoV-2, they could potentially serve as future treatment targets.
Published: 2020

26. LBA73 The ORF1ab of SARS-CoV-2 encodes an immunodominant epitope restricted by HLA-A*01:01

Author: Cristina Mussini, Pia Kvistborg, S. De Biasi, Andrea Cossarizza, Paul J. M. Hekking, Olga I. Isaeva, Steven L. C. Ketelaars, Ton N. Schumacher, Anna Dopler, Anastasia Gangaev, C. Talavera Ormeno, Neubury M. Lardy, Kelly Hoefakker, Giovanni Guaraldi, Massimo Girardis, Huib Ovaa, Robert Balderas, S. Patiweal, and Mireille Toebes
Subjects: Genetics, business.industry, T cell, Human leukocyte antigen, Hematology, Genome, Article, Epitope, HLA-A, medicine.anatomical_structure, Antigen, Oncology, medicine, Cytotoxic T cell, Allele, business
Abstract: Background: A large global effort is ongoing to develop vaccines against SARS-CoV-2, the causative agent of COVID-19. While there is accumulating information on the antibody response against SARS-CoV-2, less is known about the SARS-CoV-2 antigens that are targeted by CD8 T cells. Such knowledge will be of high value to gain fundamental insights into the antigenic landscape of SARS-CoV-2 recognized by CD8 T cells, to develop tool allowing focused analysis of the SARS-CoV-2 specific T cell responses directly ex vivo, and to understand whether current vaccine designs are covering the CD8 T cell recognized antigens. Methods: To address this issue, we have analyzed samples from 18 COVID-19 patients for CD8 T cell recognition of 500 predicted SARS-CoV-2-derived epitopes restricted to 10 common HLA-A and HLA-B alleles. For each HLA allele, the top 50 epitopes were selected based on predicted binding affinity and likelihood of successful proteasomal processing. To probe for CD8 T cell recognition of the selected epitope-HLA complexes, we made use of our in-house technology based on multiplexing of peptide HLA (pHLA) multimers conjugated to fluorescent dyes. Results: In addition to previous studies showing CD8 T cell reactivity towards epitopes derived from the spike protein of SARS-CoV-2, we have identified several CD8 T cell recognized epitopes derived from the ORF1ab, including one epitope displaying clear immunodominant properties across patients positive for HLA-A*01:01. Investigation of the functional status of part of the identified responses (including 4 responses specific for the immunodominant epitope) revealed that the T cell responses were highly dysfunctional. In addition the SARS-CoV-2 specific CD8 T cell responses displayed an increased expression of NKG2A in comparison with bulk CD8 T cells, which may explain their dysfunctional state. Conclusions: Our data suggest that part of the ORF1ab encodes multiple CD8 T cell antigens including one immunodominant epitope. Noteworthy these epitopes were derived from a part of the viral genome that is not included in the majority of vaccine candidates in development, and this may potentially influence their clinical activity and safety profile. Legal entity responsible for the study: The authors. Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.
Published: 2020
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27. Identification and characterization of a SARS-CoV-2 specific CD8

Author: Anastasia, Gangaev, Steven L C, Ketelaars, Olga I, Isaeva, Sanne, Patiwael, Anna, Dopler, Kelly, Hoefakker, Sara, De Biasi, Lara, Gibellini, Cristina, Mussini, Giovanni, Guaraldi, Massimo, Girardis, Cami M P Talavera, Ormeno, Paul J M, Hekking, Neubury M, Lardy, Mireille, Toebes, Robert, Balderas, Ton N, Schumacher, Huib, Ovaa, Andrea, Cossarizza, and Pia, Kvistborg
Subjects: Adult, Aged, 80 and over, Male, Immunodominant Epitopes, SARS-CoV-2, Histocompatibility Antigens Class I, COVID-19, Epitopes, T-Lymphocyte, CD8-Positive T-Lymphocytes, Middle Aged, Lymphocyte Activation, Viral Proteins, Humans, Female, Immunologic Memory, Alleles, Aged, Polyproteins
Abstract: The COVID-19 pandemic caused by SARS-CoV-2 is a continuous challenge worldwide, and there is an urgent need to map the landscape of immunogenic and immunodominant epitopes recognized by CD8
Published: 2020

28. Profound CD8 T cell responses towards the SARS-CoV-2 ORF1ab in COVID-19 patients

Author: Giovanni Guaraldi, Sanne Patiwael, Andrea Cossarizza, Huib Ovaa, Kelly Hoefakker, Pia Kvistborg, Olga I. Isaeva, Cristina Mussini, Paul J. M. Hekking, Steven L. C. Ketelaars, Cami Talavera Ormeño, Anastasia Gangaev, Neubury M. Lardy, Sara De Biasi, Anna Dopler, Robert Balderas, Massimo Girardis, Mireille Toebes, and Ton N. Schumacher
Subjects: 0301 basic medicine, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Human leukocyte antigen, Biology, Genome, Virus, Epitope, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, 030220 oncology & carcinogenesis, Immunology, Cytotoxic T cell, Allele, CD8
Abstract: While there is accumulating evidence on the antibody response against SARS-CoV-2, we are only beginning toacquire knowledge regarding the SARS-CoV-2 specific CD8 T-cell response Therefore, it is an urgent matter to gaina deeper insight into the virus specific CD8 T-cell response to both assist vaccine design and provide tools toevaluate the vaccine-induced T-cell responses To address this issue, we have analyzed samples from 20 COVID-19 patients for CD8 T-cell recognition of 500 predicted SARS-CoV-2-derived epitopes restricted to 10 of the mostcommon HLA-A and HLA-B alleles For each HLA allele, the top 50 epitopes were selected based on predictedbinding affinity and likelihood of successful proteasomal processing In addition, SARS-CoV-2 epitope predictionsshared by the science community were considered To probe for CD8 T-cell recognition of the selected epitopes, wemade use of our in-house technology based on multiplexing of peptide HLA (pHLA) multimers conjugated tofluorescent dyes Our data demonstrated that CD8 T-cell reactivity against SARS-CoV-2 was common Remarkably, a substantial fraction of the observed CD8 T-cell responses were directed towards the ORF1ab polyprotein 1ab These CD8 T-cell responses were frequently of a profound magnitude In particular, a CD8 T-cell response towardsa potentially immunodominant epitope (TTDPSFLGRY) restricted to the HLA-A∗01:01 allele was found in all patientspositive for this allele Interestingly, the fraction of SARS-CoV-2 specific CD8 T cells expressing the inhibitoryreceptor NKG2A was higher as compared to bulk CD8 T cells In conclusion, the fact that a major part of theidentified SARS-CoV-2 specific CD8 T-cell response is directed against a part of the viral genome that is notincluded in the majority of vaccine candidates currently in development may potentially influence their clinical activityand toxicity profile
Published: 2020

29. Weibel-Palade Body Localized Syntaxin-3 Modulates Von Willebrand Factor Secretion From Endothelial Cells

Author: Johanna C. Escher, Jan Voorberg, Bart L. van den Eshof, Rüdiger Adam, Hans Janssen, Tom Carter, Maartje van den Biggelaar, Anastasia Gangaev, Aat A. Mulder, Ellie Karampini, Henriet Meems, Maaike Schillemans, Menno Hofman, Ruben Bierings, Abraham J. Koster, Dorothee van Breevoort, Carolina R. Jost, Pediatrics, Hematology, Graduate School, Landsteiner Laboratory, and ACS - Microcirculation
Subjects: 0301 basic medicine, 030204 cardiovascular system & hematology, von Willebrand factor, Exocytosis, R-SNARE Proteins, 03 medical and health sciences, 0302 clinical medicine, Von Willebrand factor, Malabsorption Syndromes, Mucolipidoses, Organelle, Weibel–Palade body, Cyclic AMP, Human Umbilical Vein Endothelial Cells, Syntaxin, Humans, Weibel-Palade body, Secretion, Cells, Cultured, Secretory Pathway, biology, Microvilli, Weibel-Palade Bodies, Chemistry, Basic Sciences, Qa-SNARE Proteins, SNARE protein, Syntaxin 3, endothelial cells, Cell biology, 030104 developmental biology, Mutation, biology.protein, ComputingMethodologies_DOCUMENTANDTEXTPROCESSING, Calcium, Rab, biological phenomena, cell phenomena, and immunity, Cardiology and Cardiovascular Medicine, syntaxin, Signal Transduction
Abstract: Supplemental Digital Content is available in the text., Objective— Endothelial cells store VWF (von Willebrand factor) in rod-shaped secretory organelles, called Weibel-Palade bodies (WPBs). WPB exocytosis is coordinated by a complex network of Rab GTPases, Rab effectors, and SNARE (soluble NSF attachment protein receptor) proteins. We have previously identified STXBP1 as the link between the Rab27A-Slp4-a complex on WPBs and the SNARE proteins syntaxin-2 and -3. In this study, we investigate the function of syntaxin-3 in VWF secretion. Approach and Results— In human umbilical vein endothelial cells and in blood outgrowth endothelial cells (BOECs) from healthy controls, endogenous syntaxin-3 immunolocalized to WPBs. A detailed analysis of BOECs isolated from a patient with variant microvillus inclusion disease, carrying a homozygous mutation in STX3(STX3−/−), showed a loss of syntaxin-3 protein and absence of WPB-associated syntaxin-3 immunoreactivity. Ultrastructural analysis revealed no detectable differences in morphology or prevalence of immature or mature WPBs in control versus STX3−/− BOECs. VWF multimer analysis showed normal patterns in plasma of the microvillus inclusion disease patient, and media from STX3−/− BOECs, together indicating WPB formation and maturation are unaffected by absence of syntaxin-3. However, a defect in basal as well as Ca2+- and cAMP-mediated VWF secretion was found in the STX3−/− BOECs. We also show that syntaxin-3 interacts with the WPB-associated SNARE protein VAMP8 (vesicle-associated membrane protein-8). Conclusions— Our data reveal syntaxin-3 as a novel WPB-associated SNARE protein that controls WPB exocytosis.
Published: 2018

30. Differential effects of PD-1 and CTLA-4 blockade on the melanoma-reactive CD8 T cell response

Author: Gangaev, Anastasia, primary, Rozeman, Elisa A., additional, Rohaan, Maartje W., additional, Philips, Daisy, additional, Patiwael, Sanne, additional, van den Berg, Joost H., additional, Ribas, Antoni, additional, Schadendorf, Dirk, additional, Schilling, Bastian, additional, Schumacher, Ton N., additional, Blank, Christian U., additional, Haanen, John B.A.G., additional, and Kvistborg, Pia, additional
Published: 2020
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31. 863 Identification and characterization of an immunodominant SARS-CoV-2-specific CD8 T cell response

Author: Gangaev, Anastasia, primary and Kvistborg, Pia, additional
Published: 2020
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32. Abstract S05-01: Profound CD8 T-cell responses towards SARS-CoV-2 OFR1ab in COVID-19 patients

Author: Gangaev, Anastasia, primary, Ketelaars, Steven L., additional, Patiwael, Sanne, additional, Dopler, Anna, additional, Isaeva, Olga I., additional, Hoefakker, Kelly, additional, De Biasi, Sara, additional, Mussini, Cristina, additional, Guaraldi, Giovanni, additional, Girardis, Massimo, additional, Ormeno, Cami M.P. Talavera, additional, Hekking, Paul J.M., additional, Lardy, Neubury M., additional, Toebes, Mireille, additional, Balderas, Robert, additional, Schumacher, Ton N., additional, Ovaa, Huib, additional, Cossarizza, Andrea, additional, and Kvistborg, Pia, additional
Published: 2020
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33. Identification and characterization of an immunodominant SARS-CoV-2-specific CD8 T cell response

Author: Gangaev, Anastasia, primary, Ketelaars, Steven L. C., additional, Isaeva, Olga I, additional, Patiwael, Sanne, additional, Dopler, Anna, additional, Hoefakker, Kelly, additional, Biasi, Sara De, additional, Gibellini, Lara, additional, Mussini, Cristina, additional, Guaraldi, Giovanni, additional, Girardis, Massimo, additional, Ormeno, Cami M. P. Talavera, additional, Hekking, Paul J. M., additional, Lardy, Neubury M., additional, Toebes, Mireille, additional, Balderas, Robert, additional, Schumacher, Ton N., additional, Ovaa, Huib, additional, Cossarizza, Andrea, additional, and Kvistborg, Pia, additional
Published: 2020
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34. LBA73 The ORF1ab of SARS-CoV-2 encodes an immunodominant epitope restricted by HLA-A*01:01

Author: Kvistborg, P., primary, Gangaev, A., additional, Ketelaars, S., additional, Patiweal, S., additional, Dopler, A., additional, Isaeva, O., additional, Hoefakker, K., additional, De Biasi, S., additional, Mussini, C., additional, Guaraldi, G., additional, Girardis, M., additional, Talavera Ormeno, C., additional, Hekking, P., additional, Lardy, N., additional, Toebes, M., additional, Balderas, R., additional, Schumacher, T.N., additional, Ovaa, H., additional, and Cossarizza, A., additional
Published: 2020
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35. Profound CD8 T cell responses towards the SARS-CoV-2 ORF1ab in COVID-19 patients

Author: Gangaev, Anastasia, primary, Ketelaars, Steven L. C., additional, Patiwael, Sanne, additional, Dopler, Anna, additional, Isaeva, Olga I., additional, Hoefakker, Kelly, additional, Biasi, Sara De, additional, Mussini, Cristina, additional, Guaraldi, Giovanni, additional, Girardis, Massimo, additional, Ormeno, Cami M. P. Talavera, additional, Hekking, Paul J. M., additional, Lardy, Neubury M., additional, Toebes, Mireille, additional, Balderas, Robert, additional, Schumacher, Ton N., additional, Ovaa, Huib, additional, Cossarizza, Andrea, additional, and Kvistborg, Pia, additional
Published: 2020
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36. Alternative trafficking of Weibel-Palade body proteins in CRISPR/Cas9-engineered von Willebrand factor-deficient blood outgrowth endothelial cells

Author: Schillemans, M., Kat, M., Westeneng, J., Gangaev, A., Hofman, M., Nota, B, van Alphen, F.P.J., Boer, M.L. (Monique) den, van den Biggelaar, M., Margadant, C., Voorberg, J., Bierings, R., Schillemans, M., Kat, M., Westeneng, J., Gangaev, A., Hofman, M., Nota, B, van Alphen, F.P.J., Boer, M.L. (Monique) den, van den Biggelaar, M., Margadant, C., Voorberg, J., and Bierings, R.
Abstract: Background: Synthesis of the hemostatic protein von Willebrand factor (VWF) drives formation of endothelial storage organelles called Weibel‐Palade bodies (WPBs). In the absence of VWF, angiogenic and inflammatory mediators that are costored in WPBs are subject to alternative trafficking routes. In patients with von Willebrand disease (VWD), partial or complete absence of VWF/WPBs may lead to additional bleeding complications, such as angiodysplasia. Studies addressing the role of VWF using VWD patient–derived blood outgrowth endothelial cells (BOECs) have reported conflicting results due to the intrinsic heterogeneity of patient‐derived BOECs. Objective: To generate a VWF‐deficient endothelial cell model using clustered regularly interspaced short palindromic repeats (CRISPR) genome engineering of blood outgrowth endothelial cells. Methods: We used CRISPR/CRISPR‐associated protein 9 editing in single‐donor cord blood–derived BOECs (cbBOECs) to generate clonal VWF−/− cbBOECs. Clones were selected using high‐throughput screening, VWF mutations were validated by sequencing, and cells were phenotypically characterized. Results: Two VWF−/− BOEC clones were obtained and were entirely devoid of WPBs, while their overall cell morphology was unaltered. Several WPB proteins, including CD63, syntaxin‐3 and the cargo proteins angiopoietin (Ang)‐2, interleukin (IL)‐6, and IL‐8 showed alternative trafficking and secretion in the absence of VWF. Interestingly, Ang‐2 was relocated to the cell periphery and colocalized with Tie‐2. Conclusions: CRISPR editing of VWF provides a robust method to create VWF‐ deficient BOECs that can be directly compared to their wild‐type counterparts. Results obtained with our model system confirmed alternative trafficking of several WPB proteins in the absence of VWF and support the theory that increased Ang‐2/Tie‐2 interaction contributes to angiogenic abnormalities in VWD patients.
Published: 2019
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37. 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

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

Author: Cossarizza, A, Chang, H-D, Radbruch, A, Acs, A, Adam, D, Adam-Klages, S, Agace, WW, Aghaeepour, N, Akdis, M, Allez, M, Almeida, LN, Alvisi, G, Anderson, G, Andrae, I, Annunziato, F, Anselmo, A, Bacher, P, Baldari, CT, Bari, S, Barnaba, V, Barros-Martins, J, Battistini, L, Bauer, W, Baumgart, S, Baumgarth, N, Baumjohann, D, Baying, B, Bebawy, M, Becher, B, Beisker, W, Benes, V, Beyaert, R, Blanco, A, Boardman, DA, Bogdan, C, Borger, JG, Borsellino, G, Boulais, PE, Bradford, JA, Brenner, D, Brinkman, RR, Brooks, AES, Busch, DH, Buescher, M, Bushnell, TP, Calzetti, F, Cameron, G, Cammarata, I, Cao, X, Cardell, SL, Casola, S, Cassatella, MA, Cavani, A, Celada, A, Chatenoud, L, Chattopadhyay, PK, Chow, S, Christakou, E, Cicin-Sain, L, Clerici, M, Colombo, FS, Cook, L, Cooke, A, Cooper, AM, Corbett, AJ, Cosma, A, Cosmi, L, Coulie, PG, Cumano, A, Cvetkovic, L, Dang, VD, Dang-Heine, C, Davey, MS, Davies, D, De Biasi, S, Del Zotto, G, Dela Cruz, GV, Delacher, M, Della Bella, S, Dellabona, P, Deniz, G, Dessing, M, Di Santo, JP, Diefenbach, A, Dieli, F, Dolf, A, Doerner, T, Dress, RJ, Dudziak, D, Dustin, M, Dutertre, C-A, Ebner, F, Eckle, SBG, Edinger, M, Eede, P, Ehrhardt, GRA, Eich, M, Engel, P, Engelhardt, B, Erdei, A, Esser, C, Everts, B, Evrard, M, Falk, CS, Fehniger, TA, Felipo-Benavent, M, Ferry, H, Feuerer, M, Filby, A, Filkor, K, Fillatreau, S, Follo, M, Foerster, I, Foster, J, Foulds, GA, Frehse, B, Frenette, PS, Frischbutter, S, Fritzsche, W, Galbraith, DW, Gangaev, A, Garbi, N, Gaudilliere, B, Gazzinelli, RT, Geginat, J, Gerner, W, Gherardin, NA, Ghoreschi, K, Gibellini, L, Ginhoux, F, Goda, K, Godfrey, DI, Goettlinger, C, Gonzalez-Navajas, JM, Goodyear, CS, Gori, A, Grogan, JL, Grummitt, D, Gruetzkau, A, Haftmann, C, Hahn, J, Hammad, H, Haemmerling, G, Hansmann, L, Hansson, G, Harpur, CM, Hartmann, S, Hauser, A, Hauser, AE, Haviland, DL, Hedley, D, Hernandez, DC, Herrera, G, Herrmann, M, Hess, C, Hoefer, T, Hoffmann, P, Hogquist, K, Holland, T, Hollt, T, Holmdahl, R, Hombrink, P, Houston, JP, Hoyer, BF, Huang, B, Huang, F-P, Huber, JE, Huehn, J, Hundemer, M, Hunter, CA, Hwang, WYK, Iannone, A, Ingelfinger, F, Ivison, SM, Jaeck, H-M, Jani, PK, Javega, B, Jonjic, S, Kaiser, T, Kalina, T, Kamradt, T, Kaufmann, SHE, Keller, B, Ketelaars, SLC, Khalilnezhad, A, Khan, S, Kisielow, J, Klenerman, P, Knopf, J, Koay, H-F, Kobow, K, Kolls, JK, Kong, WT, Kopf, M, Korn, T, Kriegsmann, K, Kristyanto, H, Kroneis, T, Krueger, A, Kuehne, J, Kukat, C, Kunkel, D, Kunze-Schumacher, H, Kurosaki, T, Kurts, C, Kvistborg, P, Kwok, I, Landry, J, Lantz, O, Lanuti, P, LaRosa, F, Lehuen, A, LeibundGut-Landmann, S, Leipold, MD, Leung, LYT, Levings, MK, Lino, AC, Liotta, F, Litwin, V, Liu, Y, Ljunggren, H-G, Lohoff, M, Lombardi, G, Lopez, L, Lopez-Botet, M, Lovett-Racke, AE, Lubberts, E, Luche, H, Ludewig, B, Lugli, E, Lunemann, S, Maecker, HT, Maggi, L, Maguire, O, Mair, F, Mair, KH, Mantovani, A, Manz, RA, Marshall, AJ, Martinez-Romero, A, Martrus, G, Marventano, I, Maslinski, W, Matarese, G, Mattioli, AV, Maueroder, C, Mazzoni, A, McCluskey, J, McGrath, M, McGuire, HM, McInnes, IB, Mei, HE, Melchers, F, Melzer, S, Mielenz, D, Miller, SD, Mills, KHG, Minderman, H, Mjosberg, J, Moore, J, Moran, B, Moretta, L, Mosmann, TR, Mueller, S, Multhoff, G, Munoz, LE, Munz, C, Nakayama, T, Nasi, M, Neumann, K, Ng, LG, Niedobitek, A, Nourshargh, S, Nunez, G, O'Connor, J-E, Ochel, A, Oja, A, Ordonez, D, Orfao, A, Orlowski-Oliver, E, Ouyang, W, Oxenius, A, Palankar, R, Panse, I, Pattanapanyasat, K, Paulsen, M, Pavlinic, D, Penter, L, Peterson, P, Peth, C, Petriz, J, Piancone, F, Pickl, WF, Piconese, S, Pinti, M, Pockley, AG, Podolska, MJ, Poon, Z, Pracht, K, Prinz, I, Pucillo, CEM, Quataert, SA, Quatrini, L, Quinn, KM, Radbruch, H, Radstake, TRDJ, Rahmig, S, Rahn, H-P, Rajwa, B, Ravichandran, G, Raz, Y, Rebhahn, JA, Recktenwald, D, Reimer, D, Reis e Sousa, C, Remmerswaal, EBM, Richter, L, Rico, LG, Riddell, A, Rieger, AM, Robinson, JP, Romagnani, C, Rubartelli, A, Ruland, J, Saalmueller, A, Saeys, Y, Saito, T, Sakaguchi, S, Sala-de-Oyanguren, F, Samstag, Y, Sanderson, S, Sandrock, I, Santoni, A, Sanz, RB, Saresella, M, Sautes-Fridman, C, Sawitzki, B, Schadt, L, Scheffold, A, Scherer, HU, Schiemann, M, Schildberg, FA, Schimisky, E, Schlitzer, A, Schlosser, J, Schmid, S, Schmitt, S, Schober, K, Schraivogel, D, Schuh, W, Schueler, T, Schulte, R, Schulz, AR, Schulz, SR, Scotta, C, Scott-Algara, D, Sester, DP, Shankey, TV, Silva-Santos, B, Simon, AK, Sitnik, KM, Sozzani, S, Speiser, DE, Spidlen, J, Stahlberg, A, Stall, AM, Stanley, N, Stark, R, Stehle, C, Steinmetz, T, Stockinger, H, Takahama, Y, Takeda, K, Tan, L, Tarnok, A, Tiegs, G, Toldi, G, Tornack, J, Traggiai, E, Trebak, M, Tree, TIM, Trotter, J, Trowsdale, J, Tsoumakidou, M, Ulrich, H, Urbanczyk, S, van de Veen, W, van den Broek, M, van der Pol, E, Van Gassen, S, Van Isterdael, G, van Lier, RAW, Veldhoen, M, Vento-Asturias, S, Vieira, P, Voehringer, D, Volk, H-D, von Borstel, A, von Volkmann, K, Waisman, A, Walker, RV, Wallace, PK, Wang, SA, Wang, XM, Ward, MD, Ward-Hartstonge, KA, Warnatz, K, Warnes, G, Warth, S, Waskow, C, Watson, JV, Watzl, C, Wegener, L, Weisenburger, T, Wiedemann, A, Wienands, J, Wilharm, A, Wilkinson, RJ, Willimsky, G, Wing, JB, Winkelmann, R, Winkler, TH, Wirz, OF, Wong, A, Wurst, P, Yang, JHM, Yang, J, Yazdanbakhsh, M, Yu, L, Yue, A, Zhang, H, Zhao, Y, Ziegler, SM, Zielinski, C, Zimmermann, J, Zychlinsky, A, Cossarizza, A, Chang, H-D, Radbruch, A, Acs, A, Adam, D, Adam-Klages, S, Agace, WW, Aghaeepour, N, Akdis, M, Allez, M, Almeida, LN, Alvisi, G, Anderson, G, Andrae, I, Annunziato, F, Anselmo, A, Bacher, P, Baldari, CT, Bari, S, Barnaba, V, Barros-Martins, J, Battistini, L, Bauer, W, Baumgart, S, Baumgarth, N, Baumjohann, D, Baying, B, Bebawy, M, Becher, B, Beisker, W, Benes, V, Beyaert, R, Blanco, A, Boardman, DA, Bogdan, C, Borger, JG, Borsellino, G, Boulais, PE, Bradford, JA, Brenner, D, Brinkman, RR, Brooks, AES, Busch, DH, Buescher, M, Bushnell, TP, Calzetti, F, Cameron, G, Cammarata, I, Cao, X, Cardell, SL, Casola, S, Cassatella, MA, Cavani, A, Celada, A, Chatenoud, L, Chattopadhyay, PK, Chow, S, Christakou, E, Cicin-Sain, L, Clerici, M, Colombo, FS, Cook, L, Cooke, A, Cooper, AM, Corbett, AJ, Cosma, A, Cosmi, L, Coulie, PG, Cumano, A, Cvetkovic, L, Dang, VD, Dang-Heine, C, Davey, MS, Davies, D, De Biasi, S, Del Zotto, G, Dela Cruz, GV, Delacher, M, Della Bella, S, Dellabona, P, Deniz, G, Dessing, M, Di Santo, JP, Diefenbach, A, Dieli, F, Dolf, A, Doerner, T, Dress, RJ, Dudziak, D, Dustin, M, Dutertre, C-A, Ebner, F, Eckle, SBG, Edinger, M, Eede, P, Ehrhardt, GRA, Eich, M, Engel, P, Engelhardt, B, Erdei, A, Esser, C, Everts, B, Evrard, M, Falk, CS, Fehniger, TA, Felipo-Benavent, M, Ferry, H, Feuerer, M, Filby, A, Filkor, K, Fillatreau, S, Follo, M, Foerster, I, Foster, J, Foulds, GA, Frehse, B, Frenette, PS, Frischbutter, S, Fritzsche, W, Galbraith, DW, Gangaev, A, Garbi, N, Gaudilliere, B, Gazzinelli, RT, Geginat, J, Gerner, W, Gherardin, NA, Ghoreschi, K, Gibellini, L, Ginhoux, F, Goda, K, Godfrey, DI, Goettlinger, C, Gonzalez-Navajas, JM, Goodyear, CS, Gori, A, Grogan, JL, Grummitt, D, Gruetzkau, A, Haftmann, C, Hahn, J, Hammad, H, Haemmerling, G, Hansmann, L, Hansson, G, Harpur, CM, Hartmann, S, Hauser, A, Hauser, AE, Haviland, DL, Hedley, D, Hernandez, DC, Herrera, G, Herrmann, M, Hess, C, Hoefer, T, Hoffmann, P, Hogquist, K, Holland, T, Hollt, T, Holmdahl, R, Hombrink, P, Houston, JP, Hoyer, BF, Huang, B, Huang, F-P, Huber, JE, Huehn, J, Hundemer, M, Hunter, CA, Hwang, WYK, Iannone, A, Ingelfinger, F, Ivison, SM, Jaeck, H-M, Jani, PK, Javega, B, Jonjic, S, Kaiser, T, Kalina, T, Kamradt, T, Kaufmann, SHE, Keller, B, Ketelaars, SLC, Khalilnezhad, A, Khan, S, Kisielow, J, Klenerman, P, Knopf, J, Koay, H-F, Kobow, K, Kolls, JK, Kong, WT, Kopf, M, Korn, T, Kriegsmann, K, Kristyanto, H, Kroneis, T, Krueger, A, Kuehne, J, Kukat, C, Kunkel, D, Kunze-Schumacher, H, Kurosaki, T, Kurts, C, Kvistborg, P, Kwok, I, Landry, J, Lantz, O, Lanuti, P, LaRosa, F, Lehuen, A, LeibundGut-Landmann, S, Leipold, MD, Leung, LYT, Levings, MK, Lino, AC, Liotta, F, Litwin, V, Liu, Y, Ljunggren, H-G, Lohoff, M, Lombardi, G, Lopez, L, Lopez-Botet, M, Lovett-Racke, AE, Lubberts, E, Luche, H, Ludewig, B, Lugli, E, Lunemann, S, Maecker, HT, Maggi, L, Maguire, O, Mair, F, Mair, KH, Mantovani, A, Manz, RA, Marshall, AJ, Martinez-Romero, A, Martrus, G, Marventano, I, Maslinski, W, Matarese, G, Mattioli, AV, Maueroder, C, Mazzoni, A, McCluskey, J, McGrath, M, McGuire, HM, McInnes, IB, Mei, HE, Melchers, F, Melzer, S, Mielenz, D, Miller, SD, Mills, KHG, Minderman, H, Mjosberg, J, Moore, J, Moran, B, Moretta, L, Mosmann, TR, Mueller, S, Multhoff, G, Munoz, LE, Munz, C, Nakayama, T, Nasi, M, Neumann, K, Ng, LG, Niedobitek, A, Nourshargh, S, Nunez, G, O'Connor, J-E, Ochel, A, Oja, A, Ordonez, D, Orfao, A, Orlowski-Oliver, E, Ouyang, W, Oxenius, A, Palankar, R, Panse, I, Pattanapanyasat, K, Paulsen, M, Pavlinic, D, Penter, L, Peterson, P, Peth, C, Petriz, J, Piancone, F, Pickl, WF, Piconese, S, Pinti, M, Pockley, AG, Podolska, MJ, Poon, Z, Pracht, K, Prinz, I, Pucillo, CEM, Quataert, SA, Quatrini, L, Quinn, KM, Radbruch, H, Radstake, TRDJ, Rahmig, S, Rahn, H-P, Rajwa, B, Ravichandran, G, Raz, Y, Rebhahn, JA, Recktenwald, D, Reimer, D, Reis e Sousa, C, Remmerswaal, EBM, Richter, L, Rico, LG, Riddell, A, Rieger, AM, Robinson, JP, Romagnani, C, Rubartelli, A, Ruland, J, Saalmueller, A, Saeys, Y, Saito, T, Sakaguchi, S, Sala-de-Oyanguren, F, Samstag, Y, Sanderson, S, Sandrock, I, Santoni, A, Sanz, RB, Saresella, M, Sautes-Fridman, C, Sawitzki, B, Schadt, L, Scheffold, A, Scherer, HU, Schiemann, M, Schildberg, FA, Schimisky, E, Schlitzer, A, Schlosser, J, Schmid, S, Schmitt, S, Schober, K, Schraivogel, D, Schuh, W, Schueler, T, Schulte, R, Schulz, AR, Schulz, SR, Scotta, C, Scott-Algara, D, Sester, DP, Shankey, TV, Silva-Santos, B, Simon, AK, Sitnik, KM, Sozzani, S, Speiser, DE, Spidlen, J, Stahlberg, A, Stall, AM, Stanley, N, Stark, R, Stehle, C, Steinmetz, T, Stockinger, H, Takahama, Y, Takeda, K, Tan, L, Tarnok, A, Tiegs, G, Toldi, G, Tornack, J, Traggiai, E, Trebak, M, Tree, TIM, Trotter, J, Trowsdale, J, Tsoumakidou, M, Ulrich, H, Urbanczyk, S, van de Veen, W, van den Broek, M, van der Pol, E, Van Gassen, S, Van Isterdael, G, van Lier, RAW, Veldhoen, M, Vento-Asturias, S, Vieira, P, Voehringer, D, Volk, H-D, von Borstel, A, von Volkmann, K, Waisman, A, Walker, RV, Wallace, PK, Wang, SA, Wang, XM, Ward, MD, Ward-Hartstonge, KA, Warnatz, K, Warnes, G, Warth, S, Waskow, C, Watson, JV, Watzl, C, Wegener, L, Weisenburger, T, Wiedemann, A, Wienands, J, Wilharm, A, Wilkinson, RJ, Willimsky, G, Wing, JB, Winkelmann, R, Winkler, TH, Wirz, OF, Wong, A, Wurst, P, Yang, JHM, Yang, J, Yazdanbakhsh, M, Yu, L, Yue, A, Zhang, H, Zhao, Y, Ziegler, SM, Zielinski, C, Zimmermann, J, and Zychlinsky, A
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

39. Alternative trafficking of Weibel-Palade body proteins in CRISPR/Cas9-engineered von Willebrand factor-deficient blood outgrowth endothelial cells

Author: Schillemans, M, Kat, M, Westeneng, J, Gangaev, A, Hofman, M, Nota, B, van Alphen, FPJ, Boer, ML, van den Biggelaar, M, Margadant, C, Voorberg, J, Bierings, Ruben, Schillemans, M, Kat, M, Westeneng, J, Gangaev, A, Hofman, M, Nota, B, van Alphen, FPJ, Boer, ML, van den Biggelaar, M, Margadant, C, Voorberg, J, and Bierings, Ruben
Published: 2019

40. miR-125a-5p regulation increases phosphorylation of FAK that contributes to imatinib resistance in gastrointestinal stromal tumors

Author: Mehran Ghaderi, Erik Berglund, Anastasia Gangaev, Jan Åhlén, Katarina Zeljic, Linkiat Lee, Catharina Larsson, Praveensingh Hajeri, Pinar Akcakaya, Weng-Onn Lui, Robert Bränström, and Wen-Kuan Huang
Subjects: 0301 basic medicine, STAT3 Transcription Factor, Small interfering RNA, Stromal cell, Cell Survival, Gastrointestinal Stromal Tumors, Antineoplastic Agents, Apoptosis, Biology, Small hairpin RNA, 03 medical and health sciences, 0302 clinical medicine, hemic and lymphatic diseases, Cell Line, Tumor, medicine, Gene silencing, Humans, Viability assay, RNA, Messenger, Phosphorylation, RNA, Small Interfering, neoplasms, Protein Kinase Inhibitors, Cell Proliferation, Gastrointestinal Neoplasms, Oligoribonucleotides, GiST, Imatinib, Cell Biology, Prognosis, Protein Tyrosine Phosphatases, Non-Receptor, Survival Analysis, Gene Expression Regulation, Neoplastic, MicroRNAs, 030104 developmental biology, Imatinib mesylate, Drug Resistance, Neoplasm, 030220 oncology & carcinogenesis, Focal Adhesion Kinase 1, Cancer research, Imatinib Mesylate, medicine.drug, Signal Transduction
Abstract: The use of imatinib mesylate has greatly improved the clinical outcome for gastrointestinal stromal tumor (GIST) patients. However, imatinib resistance is still a major clinical challenge, and the molecular mechanisms are not fully understood. We have previously shown that miR-125a-5p and its mRNA target PTPN18 modulate imatinib response in GIST cells. Herein, we evaluated phosphorylated FAK (pFAK) as a candidate downstream target of PTPN18 and the possible association of this regulation with imatinib resistance in GIST. FAK and pFAK expressions were evaluated in GIST882 cells transfected with short hairpin RNA or short interfering RNA targeting PTPN18 or miR-125a-5p mimic, imatinib-resistant GIST882R subclones and clinical samples using Western blot analyses. FAK phosphorylation was blocked using the FAK inhibitor 14 (FAKi) and the effects on cell viability and apoptosis were evaluated using WST-1 assay and cleaved PARP expression. Clinical associations of FAK and pFAK expression with imatinib resistance, KIT mutation and patient outcome were assessed by Fisher's exact test or log-rank test. Over-expression of miR-125a-5p and silencing of PTPN18 increased pFAK, but not FAK, expression in GIST cells. Higher pFAK expression was observed in the GIST882R subclones with acquired imatinib resistance compared to their imatinib-sensitive parental cells. Treatment with FAKi in imatinib-resistant GIST882R cells reduced cell viability and increased apoptosis upon imatinib treatment. Additionally, FAKi could rescue the imatinib resistance effect mediated by miR-125a-5p over-expression. In clinical samples, high FAK and pFAK expressions were associated with KIT mutation status, and high FAK expression was also associated with metastasis in GIST. Higher pFAK was found in cases with shorter overall survival. Our findings highlight an important role for miR-125a-5p regulation and its downstream target pFAK for imatinib resistance in GIST. pFAK and FAK may have prognostic values in GIST.
Published: 2018

41. 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. 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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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42. Weibel-Palade Body Localized Syntaxin-3 Modulates Von Willebrand Factor Secretion From Endothelial Cells

Author: Schillemans, M, Karampini, E, van den Eshof, BL, Gangaev, A, Hofman, M, van Breevoort, D, Meems, H, Janssen, H, Mulder, AA, Jost, CR, Escher, Hankje, Adam, R, Carter, T, Koster, AJ, van den Biggelaar, M, Voorberg, J, Bierings, R, Schillemans, M, Karampini, E, van den Eshof, BL, Gangaev, A, Hofman, M, van Breevoort, D, Meems, H, Janssen, H, Mulder, AA, Jost, CR, Escher, Hankje, Adam, R, Carter, T, Koster, AJ, van den Biggelaar, M, Voorberg, J, and Bierings, R
Published: 2018

43. miR-125a-5p regulation increases phosphorylation of FAK that contributes to imatinib resistance in gastrointestinal stromal tumors

Author: Huang, Wen-Kuan, primary, Akçakaya, Pinar, additional, Gangaev, Anastasia, additional, Lee, Linkiat, additional, Zeljic, Katarina, additional, Hajeri, Praveensingh, additional, Berglund, Erik, additional, Ghaderi, Mehran, additional, Åhlén, Jan, additional, Bränström, Robert, additional, Larsson, Catharina, additional, and Lui, Weng-Onn, additional
Published: 2018
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44. miR-125a-5p regulation increases phosphorylation of FAK that contributes to imatinib resistance in gastrointestinal stromal tumors.

Author: Akçakaya, Pinar, Gangaev, Anastasia, Lee, Linkiat, Hajeri, Praveensingh, Ghaderi, Mehran, Larsson, Catharina, Lui, Weng-Onn, Huang, Wen-Kuan, Zeljic, Katarina, Berglund, Erik, Åhlén, Jan, and Bränström, Robert
Subjects: *MICRORNA genetics, *GENETIC regulation, *FOCAL adhesion kinase, *PHOSPHORYLATION, *IMATINIB, *GASTROINTESTINAL stromal tumors, *GENETICS
Abstract: Abstract The use of imatinib mesylate has greatly improved the clinical outcome for gastrointestinal stromal tumor (GIST) patients. However, imatinib resistance is still a major clinical challenge, and the molecular mechanisms are not fully understood. We have previously shown that miR-125a-5p and its mRNA target PTPN18 modulate imatinib response in GIST cells. Herein, we evaluated phosphorylated FAK (pFAK) as a candidate downstream target of PTPN18 and the possible association of this regulation with imatinib resistance in GIST. FAK and pFAK expressions were evaluated in GIST882 cells transfected with short hairpin RNA or short interfering RNA targeting PTPN18 or miR-125a-5p mimic, imatinib-resistant GIST882R subclones and clinical samples using Western blot analyses. FAK phosphorylation was blocked using the FAK inhibitor 14 (FAKi) and the effects on cell viability and apoptosis were evaluated using WST-1 assay and cleaved PARP expression. Clinical associations of FAK and pFAK expression with imatinib resistance, KIT mutation and patient outcome were assessed by Fisher's exact test or log-rank test. Over-expression of miR-125a-5p and silencing of PTPN18 increased pFAK, but not FAK, expression in GIST cells. Higher pFAK expression was observed in the GIST882R subclones with acquired imatinib resistance compared to their imatinib-sensitive parental cells. Treatment with FAKi in imatinib-resistant GIST882R cells reduced cell viability and increased apoptosis upon imatinib treatment. Additionally, FAKi could rescue the imatinib resistance effect mediated by miR-125a-5p over-expression. In clinical samples, high FAK and pFAK expressions were associated with KIT mutation status, and high FAK expression was also associated with metastasis in GIST. Higher pFAK was found in cases with shorter overall survival. Our findings highlight an important role for miR-125a-5p regulation and its downstream target pFAK for imatinib resistance in GIST. pFAK and FAK may have prognostic values in GIST. Graphical abstract fx1 Highlights • Phosphorylation of FAK is regulated by PTPN18 and miR-125a-5p. • pFAK level is higher in imatinib-resistant than imatinib-sensitive GIST882 cells. • FAK inihibitor treatment in imatinib-resistant cells increases imatinib-induced apoptosis. • Blocking FAK phosphorylation rescues miR-125a-5p mediated imatinib resistance. [ABSTRACT FROM AUTHOR]
Published: 2018
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45. Differential effects of PD-1 and CTLA-4 blockade on the melanoma-reactive CD8 T cell response

Author: Gangaev, A, primary, Rozeman, E, additional, Rohaan, M, additional, Isaeva, O, additional, Philips, D, additional, Patiwael, S, additional, van den Berg, J, additional, Ribas, A, additional, Schadendorf, D, additional, Schilling, B, additional, Schumacher, T, additional, Blank, C, additional, Haanen, J, additional, and Kvistborg, P, additional
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