Citations to this article
Sabine Ivison, … , Ryan R. Brinkman, Megan K. Levings
Published December 6, 2018
Citation Information: JCI Insight. 2018;3(23):e121867. https://doi.org/10.1172/jci.insight.121867.
Citation Information: JCI Insight. 2018;3(23):e121867. https://doi.org/10.1172/jci.insight.121867.
Abstract
The analysis and validation of flow cytometry–based biomarkers in clinical studies are limited by the lack of standardized protocols that are reproducible across multiple centers and suitable for use with either unfractionated blood or cryopreserved PBMCs. Here we report the development of a platform that standardizes a set of flow cytometry panels across multiple centers, with high reproducibility in blood or PBMCs from either healthy subjects or patients 100 days after hematopoietic stem cell transplantation. Inter-center comparisons of replicate samples showed low variation, with interindividual variation exceeding inter-center variation for most populations (coefficients of variability <20% and interclass correlation coefficients >0.75). Exceptions included low-abundance populations defined by markers with indistinct expression boundaries (e.g., plasmablasts, monocyte subsets) or populations defined by markers sensitive to cryopreservation, such as CD62L and CD45RA. Automated gating pipelines were developed and validated on an independent data set, revealing high Spearman’s correlations (rs >0.9) with manual analyses. This workflow, which includes pre-formatted antibody cocktails, standardized protocols for acquisition, and validated automated analysis pipelines, can be readily implemented in multicenter clinical trials. This approach facilitates the collection of robust immune phenotyping data and comparison of data from independent studies.
Authors
Sabine Ivison, Mehrnoush Malek, Rosa V. Garcia, Raewyn Broady, Anne Halpin, Manon Richaud, Rollin F. Brant, Szu-I Wang, Mathieu Goupil, Qingdong Guan, Peter Ashton, Jason Warren, Amr Rajab, Simon Urschel, Deepali Kumar, Mathias Streitz, Birgit Sawitzki, Stephan Schlickeiser, Janetta J. Bijl, Donna A. Wall, Jean-Sebastien Delisle, Lori J. West, Ryan R. Brinkman, Megan K. Levings
Citation information
Citations to this article (22)
| Title and authors | Publication | Year |
|---|---|---|
|
Improving Reliability of Immunological Assays by Defining Minimal Criteria for Cell Fitness.
Ivison S, Boucher G, Zheng G, Garcia RV, Kohen R, Bitton A, Rioux JD, Levings MK |
ImmunoHorizons | 2024 |
|
Next Generation Biobanking: Employing a Robotic System for Automated Mononuclear Cell Isolation
Fuchs YF, Brunner J, Weigelt M, Schieferdecker A, Morgenstern R, Sturm A, Winter B, Jambor H, Stölzel F, Ruhnke L, von Bonin M, Rücker-Braun E, Heidenreich F, Fuchs A, Bonifacio E, Bornhäuser M, Poitz DM, Altmann H |
Biopreservation and Biobanking | 2023 |
|
Strategies for optimizing CITE-seq for human islets and other tissues
Colpitts SJ, Budd MA, Monajemi M, Reid KT, Murphy JM, Ivison S, Verchere CB, Levings MK, Crome SQ |
Frontiers in immunology | 2023 |
|
Assessment of a multisite standardized biospecimen collection protocol for immune phenotyping in neurodevelopmental disorders
Cleary S, Teskey G, Mathews C, Sachachar RJ, Nicolson R, Weksberg R, Anagnostou E, Bowdish DM, Foster JA |
Scientific Reports | 2023 |
|
Deep Immunophenotyping of Human Whole Blood by Standardized Multi-parametric Flow Cytometry Analyses
Gao J, Luo Y, Li H, Zhao Y, Zhao J, Han X, Han J, Lin H, Qian F |
2023 | |
|
A Microfluidics Approach for Ovarian Cancer Immune Monitoring in an Outpatient Setting
Libbrecht S, Vankerckhoven A, de Wijs K, Baert T, Thirion G, Vandenbrande K, Van Gorp T, Timmerman D, Coosemans A, Lagae L |
Cells | 2023 |
|
flowSim: Near duplicate detection for flow cytometry data.
Montante S, Chen Y, Brinkman RR |
Cytometry Part A | 2023 |
|
Guidelines for standardizing T‐cell cytometry assays to link biomarkers, mechanisms, and disease outcomes in type 1 diabetes
J Yang, K WardHartstonge, D Perry, J Blanchfield, A Posgai, A Wiedeman, K Diggins, A Rahman, T Tree, T Brusko, M Levings, E James, S Kent, C Speake, D Homann, S Long |
European Journal of Immunology | 2022 |
|
Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications
U Blache, R Weiss, A Boldt, M Kapinsky, AR Blaudszun, A Quaiser, A Pohl, T Miloud, M Burgaud, V Vucinic, U Platzbecker, U Sack, S Fricke, U Koehl |
Frontiers in immunology | 2021 |
|
Interrogation of the cellular immunome of cancer patients with regard to the COVID-19 pandemic
RN Donahue, JL Marté, M Goswami, NJ Toney, YT Tsai, JL Gulley, J Schlom |
Journal for ImmunoTherapy of Cancer | 2021 |
|
Current and future biomarkers for outcomes with immunotherapy in non-small cell lung cancer
B Duchemann, J Remon, M Naigeon, L Cassard, JM Jouniaux, L Boselli, J Grivel, E Auclin, A Desnoyer, B Besse, N Chaput |
Translational Lung Cancer Research | 2021 |
|
CXCL13 Is an Indicator of Germinal Center Activity and Alloantibody Formation Following Transplantation
ES Crichton, S Zeng, GM Muraglia, IR Badell |
Transplantation Direct | 2021 |
|
Evaluation of a fit-for-purpose assay to monitor antigen-specific functional CD4+ T cell subpopulations in rheumatoid arthritis using flow-cytometry based peptide-MHC class-II tetramer staining
S Patel, N Ramnoruth, P Wehr, J Rossjohn, HH Reid, K Campbell, HJ Nel, R Thomas |
Clinical & Experimental Immunology | 2021 |
|
Implementing flowDensity for Automated Analysis of Bone Marrow Lymphocyte Population
Ghazaleh Eskandari, Sishir Subedi, Paul Christensen, Randall J Olsen, Youli Zu, Scott W Long |
Journal of Pathology Informatics | 2021 |
|
Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition).
Cossarizza A, Chang HD, Radbruch A, Abrignani S, Addo R, Akdis M, Andrä I, Andreata F, Annunziato F, Arranz E, Bacher P, Bari S, Barnaba V, Barros-Martins J, Baumjohann D, Beccaria CG, Bernardo D, Boardman DA, Borger J, Böttcher C, Brockmann L, Burns M, Busch DH, Cameron G, Cammarata I, Cassotta A, Chang Y, Chirdo FG, Christakou E, Čičin-Šain L, Cook L, Corbett AJ, Cornelis R, Cosmi L, Davey MS, De Biasi S, De Simone G, Del Zotto G, Delacher M, Di Rosa F, Di Santo J, Diefenbach A, Dong J, Dörner T, Dress RJ, Dutertre CA, Eckle SBG, Eede P, Evrard M, Falk CS, Feuerer M, Fillatreau S, Fiz-Lopez A, Follo M, Foulds GA, Fröbel J, Gagliani N, Galletti G, Gangaev A, Garbi N, Garrote JA, Geginat J, Gherardin NA, Gibellini L, Ginhoux F, Godfrey DI, Gruarin P, Haftmann C, Hansmann L, Harpur CM, Hayday AC, Heine G, Hernández DC, Herrmann M, Hoelsken O, Huang Q, Huber S, Huber JE, Huehn J, Hundemer M, Hwang WYK, Iannacone M, Ivison SM, Jäck HM, Jani PK, Keller B, Kessler N, Ketelaars S, Knop L, Knopf J, Koay HF, Kobow K, Kriegsmann K, Kristyanto H, Krueger A, Kuehne JF, Kunze-Schumacher H, Kvistborg P, Kwok I, Latorre D, Lenz D, Levings MK, Lino AC, Liotta F, Long HM, Lugli E, MacDonald KN, Maggi L, Maini MK, Mair F, Manta C, Manz RA, Mashreghi MF, Mazzoni A, McCluskey J, Mei HE, Melchers F, Melzer S, Mielenz D, Monin L, Moretta L, Multhoff G, Muñoz LE, Muñoz-Ruiz M, Muscate F, Natalini A, Neumann K, Ng LG, Niedobitek A, Niemz J, Almeida LN, Notarbartolo S, Ostendorf L, Pallett LJ, Patel AA, Percin GI, Peruzzi G, Pinti M, Pockley AG, Pracht K, Prinz I, Pujol-Autonell I, Pulvirenti N, Quatrini L, Quinn KM, Radbruch H, Rhys H, Rodrigo MB, Romagnani C, Saggau C, Sakaguchi S, Sallusto F, Sanderink L, Sandrock I, Schauer C, Scheffold A, Scherer HU, Schiemann M, Schildberg FA, Schober K, Schoen J, Schuh W, Schüler T, Schulz AR, Schulz S, Schulze J, Simonetti S, Singh J, Sitnik KM, Stark R, Starossom S, Stehle C, Szelinski F, Tan L, Tarnok A, Tornack J, Tree TIM, van Beek JJP, van de Veen W, van Gisbergen K, Vasco C, Verheyden NA, von Borstel A, Ward-Hartstonge KA, Warnatz K, Waskow C, Wiedemann A, Wilharm A, Wing J, Wirz O, Wittner J, Yang JHM, Yang J |
European Journal of Immunology | 2021 |
|
SITC cancer immunotherapy resource document: a compass in the land of biomarker discovery
S Hu-Lieskovan, S Bhaumik, K Dhodapkar, JC Grivel, S Gupta, BA Hanks, S Janetzki, TO Kleen, Y Koguchi, AW Lund, C Maccalli, YD Mahnke, RD Novosiadly, SR Selvan, T Sims, Y Zhao, HT Maecker |
Journal for ImmunoTherapy of Cancer | 2020 |
|
Integrating Circulating Biomarkers in the Immune Checkpoint Inhibitor Treatment in Lung Cancer
B Duchemann, J Remon, M Naigeon, L Mezquita, R Ferrara, L Cassard, JM Jouniaux, L Boselli, J Grivel, E Auclin, A Desnoyer, B Besse, N Chaput |
Cancers | 2020 |
|
Multicentre Harmonisation of a Six-Colour Flow Cytometry Panel for Naïve/Memory T Cell Immunomonitoring
I Macchia, VL Sorsa, I Ruspantini, M Sanchez, V Tirelli, M Carollo, G Fedele, P Leone, G Schiavoni, C Buccione, P Rizza, P Nisticò, B Palermo, S Morrone, H Stabile, A Rughetti, M Nuti, IG Zizzari, C Fionda, R Maggio, C Capuano, C Quintarelli, M Sinibaldi, C Agrati, R Casetti, AR Gonzalez, F Iacobone, A Gismondi, F Belardelli, M Biffoni, F Urbani |
Journal of Immunology Research | 2020 |
|
Standardization procedure for flow cytometry data harmonization in prospective multicenter studies
LL Lann, PE Jouve, M Alarcón-Riquelme, C Jamin, JO Pers, M Alvarez, D Alvarez-Errico, N Azevedo, N Barbarroja, A Buttgereit, Q Cheng, C Chizzolini, J Cremer, AD Groof, ED Langhe, J Ducreux, A Dufour, V Gerl, M Hernandez-Fuentes, L Khodadadi, K Kniesch, T Li, C Lopez-Pedrera, Z Makowska, C Marañón, B Muchmore, E Neves, B Rouvière, Q Simon, E Trombetta, N Varela, T Witte, R Aguilar-Quesada, MA Aguirre-Zamorano, I Almeida, N Baerlecken, A Balog, D Belz, L Beretta, RB Alonso, M Bocskai, M Brandão, JL Rubio, A Campar, MC Castro-Villegas, R Cervera, E Collantes, D Cornec, AC Martínez, M Deák, V Devauchelle-Pensec, S Dulic, A Escudero-Contreras, G Espinosa, R Faria, F Farinha, MC Roldán, TG Anjos, MA González-Gay, F Hiepe, N Hunzelmann, S Jousse-Joulin, G Kádár, L Kovács, B Lauwerys, M Lehner, A López-Berrio, R Lories, A Marinho, J Marovac, PL Meroni, B Miranda, IJ Moleón, H Navarro-Linares, R Ortega-Castro, N Ortego, ER Garrido, E Raya, RR Fernández, I Rodríguez-Pintó, A Saraux, G Stummvoll, C Vasconcelos, M Zauner |
Scientific Reports | 2020 |
|
A composite immune signature parallels disease progression across T1D subjects
Cate Speake, Samuel O. Skinner, Dror Berel, Elizabeth Whalen, Matthew J. Dufort, William Chad Young, Jared M. Odegard, Anne M Pesenacker, Frans K. Gorus, Eddie A. James, Megan K Levings, Peter S Linsley, Eitan M. Akirav, Alberto Pugliese, Martin J. Hessner, Gerald T Nepom, Raphael Gottardo, S. Alice Long |
JCI Insight | 2019 |
|
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
A Cossarizza, HD Chang, A Radbruch, A Acs, D Adam, S AdamKlages, WW Agace, N Aghaeepour, M Akdis, M Allez, LN Almeida, G Alvisi, G Anderson, I Andrä, F Annunziato, A Anselmo, P Bacher, CT Baldari, S Bari, V Barnaba, J BarrosMartins, L Battistini, W Bauer, S Baumgart, N Baumgarth, D Baumjohann, B Baying, M Bebawy, B Becher, W Beisker, V Benes, R Beyaert, A Blanco, DA Boardman, C Bogdan, JG Borger, G Borsellino, PE Boulais, JA Bradford, D Brenner, RR Brinkman, AE Brooks, DH Busch, M Büscher, TP Bushnell, F Calzetti, G Cameron, I Cammarata, X Cao, SL Cardell, S Casola, MA Cassatella, A Cavani, A Celada, L Chatenoud, PK Chattopadhyay, S Chow, E Christakou, L ČičinŠain, M Clerici, FS Colombo, L Cook, A Cooke, AM Cooper, AJ Corbett, A Cosma, L Cosmi, PG Coulie, A Cumano, L Cvetkovic, VD Dang, C DangHeine, MS Davey, D Davies, SD Biasi, GD Zotto, GV Cruz, M Delacher, SD Bella, P Dellabona, G Deniz, M Dessing, JP Santo, A Diefenbach, F Dieli, A Dolf, T Dörner, RJ Dress, D Dudziak, M Dustin, CA Dutertre, F Ebner, SB Eckle, M Edinger, P Eede, GR Ehrhardt, M Eich, P Engel, B Engelhardt, A Erdei, C Esser, B Everts, M Evrard, CS Falk, TA Fehniger, M FelipoBenavent, H Ferry, M Feuerer, A Filby, K Filkor, S Fillatreau, M Follo, I Förster, J Foster, GA Foulds, B Frehse, PS Frenette, S Frischbutter, W Fritzsche, DW Galbraith, A Gangaev, N Garbi, B Gaudilliere, RT Gazzinelli, J Geginat, W Gerner, NA Gherardin, K Ghoreschi, L Gibellini, F Ginhoux, K Goda, DI Godfrey, C Goettlinger, JM GonzálezNavajas, CS Goodyear, A Gori, JL Grogan, D Grummitt, A Grützkau, C Haftmann, J Hahn, H Hammad, G Hämmerling, L Hansmann, G Hansson, CM Harpur, S Hartmann, A Hauser, AE Hauser, DL Haviland, D Hedley, DC Hernández, G Herrera, M Herrmann, C Hess, T Höfer, P Hoffmann, K Hogquist, T Holland, T Höllt, R Holmdahl, P Hombrink, JP Houston, BF Hoyer, B Huang, FP Huang, JE Huber, J Huehn, M Hundemer, CA Hunter, WY Hwang, A Iannone, F Ingelfinger, SM Ivison, HM Jäck, PK Jani, B Jávega, S Jonjic, T Kaiser, T Kalina, T Kamradt, SH Kaufmann, B Keller, SL Ketelaars, A Khalilnezhad, S Khan, J Kisielow, P Klenerman, J Knopf, HF Koay, K Kobow, JK Kolls, WT Kong, M Kopf, T Korn, K Kriegsmann, H Kristyanto, T Kroneis, A Krueger, J Kühne, C Kukat, D Kunkel, H KunzeSchumacher, T Kurosaki, C Kurts, P Kvistborg, I Kwok, J Landry, O Lantz, P Lanuti, F LaRosa, A Lehuen, S LeibundGutLandmann, MD Leipold, LY Leung, MK Levings, AC Lino, F Liotta, V Litwin, Y Liu, HG Ljunggren, M Lohoff, G Lombardi, L Lopez, M LópezBotet, AE LovettRacke, E Lubberts, H Luche, B Ludewig, E Lugli, S Lunemann, HT Maecker, L Maggi, O Maguire, F Mair, KH Mair, A Mantovani, RA Manz, AJ Marshall, A MartínezRomero, G Martrus, I Marventano, W Maslinski, G Matarese, AV Mattioli, C Maueröder, A Mazzoni, J McCluskey, M McGrath, HM McGuire, IB McInnes, HE Mei, F Melchers, S Melzer, D Mielenz, SD Miller, KH Mills, H Minderman, J Mjösberg, J Moore, B Moran, L Moretta, TR Mosmann, S Müller, G Multhoff, LE Muñoz, C Münz, T Nakayama, M Nasi, K Neumann, LG Ng, A Niedobitek, S Nourshargh, G Núñez, JE O'Connor, A Ochel, A Oja, D Ordonez, A Orfao, E OrlowskiOliver, W Ouyang, A Oxenius, R Palankar, I Panse, K Pattanapanyasat, M Paulsen, D Pavlinic, L Penter, P Peterson, C Peth, J Petriz, F Piancone, WF Pickl, S Piconese, M Pinti, AG Pockley, MJ Podolska, Z Poon, K Pracht, I Prinz, CE Pucillo, SA Quataert, L Quatrini, KM Quinn, H Radbruch, TR Radstake, S Rahmig, HP Rahn, B Rajwa, G Ravichandran, Y Raz, JA Rebhahn, D Recktenwald, D Reimer, CR e Sousa, EB Remmerswaal, L Richter, LG Rico, A Riddell, AM Rieger, JP Robinson, C Romagnani, A Rubartelli, J Ruland, A Saalmüller, Y Saeys, T Saito, S Sakaguchi, F SaladeOyanguren, Y Samstag, S Sanderson, I Sandrock, A Santoni, RB Sanz, M Saresella, C SautesFridman, B Sawitzki, L Schadt, A Scheffold, HU Scherer, M Schiemann, FA Schildberg, E Schimisky, A Schlitzer, J Schlosser, S Schmid, S Schmitt, K Schober, D Schraivogel, W Schuh, T Schüler, R Schulte, AR Schulz, SR Schulz, C Scottá, D ScottAlgara, DP Sester, TV Shankey, B SilvaSantos, AK Simon, KM Sitnik, S Sozzani, DE Speiser, J Spidlen, A Stahlberg, AM Stall, N Stanley, R Stark, C Stehle, T Steinmetz, H Stockinger, Y Takahama, K Takeda, L Tan, A Tárnok, G Tiegs, G Toldi, J Tornack, E Traggiai, M Trebak, TI Tree, J Trotter, J Trowsdale, M Tsoumakidou, H Ulrich, S Urbanczyk, W Veen, M Broek, E Pol, SV Gassen, GV Isterdael, RA Lier, M Veldhoen, S VentoAsturias, P Vieira, D Voehringer, HD Volk, A Borstel, K Volkmann, A Waisman, RV Walker, PK Wallace, SA Wang, XM Wang, MD Ward, KA WardHartstonge, K Warnatz, G Warnes, S Warth, C Waskow, JV Watson, C Watzl, L Wegener, T Weisenburger, A Wiedemann, J Wienands, A Wilharm, RJ Wilkinson, G Willimsky, JB Wing, R Winkelmann, TH Winkler, OF Wirz, A Wong, P Wurst, JH Yang, J Yang, M Yazdanbakhsh, L Yu, A Yue, H Zhang, Y Zhao, SM Ziegler, C Zielinski, J Zimmermann, A Zychlinsky |
European Journal of Immunology | 2019 |
|
Improving the Rigor and Reproducibility of Flow Cytometry‐Based Clinical Research and Trials Through Automated Data Analysis
RR Brinkman |
Cytometry Part A | 2019 |
