Guidelines for the use of flow cytometry and cell sorting in immunological studies
Andrea Cossarizza
(1)
,
Hyun-Dong Chang
(2)
,
Andreas Radbruch
(3)
,
Immanuel Andrä
,
Francesco Annunziato
(4)
,
Petra Bacher
,
Regina Barnaba
,
Luca Battistini
(5)
,
Wolfgang Bauer
(6)
,
Sabine Baumgart
(7)
,
Burkhard Becher
,
Wolfgang Beisker
(8)
,
Claudia Berek
(9)
,
Alfonso Blanco
,
Giovanna Borsellino
(10)
,
David Boulais
,
Ryan R Brinkman
(11)
,
Martin Büscher
,
Dirk Busch
,
Timothy P Bushnell
,
Xuetao Cao
(12)
,
Andrea Cavani
,
Pratip K Chattopadhyay
,
Qingyu Cheng
,
Sue Chow
,
Mario Clerici
(13)
,
Anne Cooke
,
Antonio Cosma
(14)
,
Lorenzo Cosmi
,
van Duc Dang
,
Derek Davies
,
Mor de Biasi
,
Genny del Zotto
,
Silvia Della Bella
,
Paolo Dellabona
(15)
,
Günnur Deniz
,
Mark Dessing
,
Andreas Diefenbach
,
James P. Di Santo
(16)
,
Francesco Dieli
,
Bo Dolf
,
Vera S Donnenberg
,
Thomas Dörner
(17)
,
Götz R A Ehrhardt
,
Bo Endl
,
Pablo Engel
,
Britta Engelhardt
,
Charlotte Esser
,
Bart Everts
,
Christine Falk
,
Todd A Fehniger
,
Andrew Filby
,
Simon Simon
,
Marie Follo
,
Bo Förster
,
John Foster
,
John Foulds
,
Paul S Frenette
,
Ari Galbraith
,
Bo Garbi
,
Maria Dolores García-Godoy
,
Kamran Ghoreschi
,
Mor Gibellini
,
Christoph Goettlinger
,
Carl S Goodyear
,
Andrea Gori
(18)
,
Francisco Grogan
,
Mor Gross
,
Andreas Grützkau
(19)
,
Daryl Grummitt
,
Alexander Hahn
,
Quirin Hammer
,
Anja E Hauser
,
David Houston
,
David Hedley
,
Guadalupe Herrera
,
Alexander Herrmann
(20)
,
Falk Hiepe
,
Bo Holland
,
Pleun Hombrink
,
Jessica P Houston
,
Bimba F Hoyer
,
Ton Huang
,
Christopher A Hunter
,
Anna Iannone
,
Hans-Martin Jäck
,
Beatriz Jávega
,
Stipan Jonjic
(21)
,
Kerstin Juelke
,
Steffen Jung
(22)
,
Toralf Kaiser
,
Tomas Kalina
,
Baerbel Keller
,
Srijit Khan
,
Alexander Kienhöfer
,
Thomas Kroneis
,
Désirée Kunkel
(23)
,
Christian Kurts
(24)
,
Pia Kvistborg
,
Virginia Lannigan
,
Olivier Lantz
(25)
,
Anis A Larbi
(26)
,
Salomé Leibundgut-Landmann
(27)
,
Michael D Leipold
,
Megan K Levings
,
Virginia Litwin
,
Yanling Liu
(28)
,
Michael Lohoff
(29)
,
Giovanna Lombardi
(30)
,
Lilly Lopez
,
Amy Lovett-Racke
,
Erik Lubberts
(31)
,
Burkhard Ludewig
,
Enrico Lugli
(32)
,
Holden T Maecker
,
Gloria Martrus
(33)
,
Giuseppe Matarese
,
Alexander Maueröder
,
Mairi Mcgrath
(34)
,
Iain Mcinnes
,
Henrik E Mei
,
Fritz Melchers
,
Susanne Melzer
,
Dirk Mielenz
,
Kingston Mills
,
Jenny Mjösberg
(35)
,
Jonni Moore
,
Barry Moran
,
Alessandro Moretta
(36)
,
Lorenzo Moretta
(36)
,
David Mosmann
,
Susann Müller
(37)
,
Werner Müller
(38)
,
Christian Münz
(39)
,
Gabriele Multhoff
(40)
,
Alexander Munoz
,
Kenneth M Murphy
,
Toshinori Nakayama
,
Milena Nasi
,
Christine Neudörfl
,
John Nolan
(41)
,
Sussan Nourshargh
(42)
,
José-Enrique O'Connor
,
Francisco Ouyang
,
Annette Oxenius
,
Raghav Palankar
,
Isabel Panse
(2)
,
Peter Peterson
,
Christian Peth
,
Jordi Petriz
,
Daisy Philips
,
Winfried Pickl
,
Regina Piconese
,
Marcello Pinti
,
a Graham Pockley
,
Alexander Podolska
,
Carlo Pucillo
,
David Quataert
,
Timothy R D J Radstake
,
Bartek Rajwa
(43)
,
David Rebhahn
,
Diether Recktenwald
,
Ester B M Remmerswaal
,
Katy Rezvani
,
Laura G Rico
,
J Paul Robinson
,
Chiara Romagnani
,
Anna Rubartelli
,
Jürgen Ruland
,
Shimon Sakaguchi
(44)
,
Francisco Sala-De-Oyanguren
,
Yvonne Samstag
,
Sharon Sanderson
,
Birgit Sawitzki
,
Alexander Scheffold
,
Matthias Schiemann
,
Frank Schildberg
,
Esther Schimisky
,
Stephan A Schmid
,
Steffen Schmitt
(45)
,
Kilian Schober
,
Thomas Schüler
(46)
,
Axel Ronald Schulz
,
Ton Schumacher
,
Cristiano Scotta
,
T Vincent Shankey
,
Anat Shemer
,
Anna-Katharina Simon
,
Josef Spidlen
(11)
,
Alan M Stall
,
Regina Stark
,
Christina Stehle
,
Merle Stein
,
Tobit Steinmetz
,
Hannes Stockinger
(47)
,
Yousuke Takahama
,
Attila Tarnok
(48)
,
Zhigang Tian
(49)
,
Gergely Toldi
,
Julia Tornack
,
Elisabetta Traggiai
,
Joe Trotter
,
Paulo Ulrich
,
Marlous van Der Braber
,
René a W van Lier
,
Marcello Veldhoen
,
Salvador Vento-Asturias
,
Paulo Vieira
(50)
,
David Voehringer
(51)
,
Hans-Dieter Volk
(3)
,
Konrad von Volkmann
,
Ari Waisman
(52)
,
Rachael Walker
,
Michael D Ward
,
Klaus Warnatz
,
Sarah Warth
,
James Watson
,
Carsten Watzl
,
Leonie Wegener
,
Annika Wiedemann
,
Jürgen Wienands
,
Gerald Willimsky
,
James Wing
,
Peter Wurst
,
Liping Yu
,
Alice Yue
,
Qianjun Zhang
,
Yi Zhao
(53)
,
Susanne Ziegler
(54)
,
Jakob Zimmermann
1
Dipartimento di Scienze Biomediche
2 Cell Biology
3 Klinik für Dermatologie, Venerologie und Allergologie
4 Department of Internal Medicine
5 Neuroimmunology Unit
6 Inorganic Chemistry II
7 Caprotec Bioanalytics GmbH
8 IOTA ES - International Occultation Timing Association European Section
9 Institut der Leibniz-Gemeinschaft, Berlin
10 IRCCS - Fondazione Santa Lucia
11 Terry Fox Laboratory
12 Department of Immunology
13 Fondazione Don Carlo Gnocchi
14 IMVA - U1184 - Immunologie des Maladies Virales et Autoimmunes
15 Experimental Immunology Unit, Dept. of Oncology
16 Département d'Immunologie - Department of Immunology
17 Charité Hospital
18 UB - Universitat de Barcelona
19 Rheumatologie
20 Department of Internal Medicine I
21 Department of Histology and Embryology
22 Weizmann Institute of Science [Rehovot, Israël]
23 Régulation des Infections Rétrovirales
24 Institutes of Molecular Medicine and Experimental Immunology
25 U932 - Immunité et cancer
26 SIgN - Singapore Immunology Network
27 Institute of Virology [Zürich]
28 College of Food Science and Technology [Shangai]
29 Institute for Medical Microbiology and Hygiene
30 King‘s College London
31 Erasmus MC - Erasmus University Medical Center [Rotterdam]
32 Unit of Clinical and Experimental Immunology
33 Heinrich Pette Institute [Hamburg]
34 Institute of Translational Medicine
35 Department of Medicine Huddinge
36 UniGe - Università degli studi di Genova = University of Genoa
37 Department of Environmental Microbiology
38 Experimental Immunology
39 Viral Immunobiology
40 Department of Radiation Oncology [Munich]
41 Department of Mathematics and Statistics
42 William Harvey Research Institute, Barts and the London Medical School
43 Cytometry Laboratories and School of Veterinary Medicine
44 Osaka University [Osaka]
45 FACS and Array Core Facility
46 Institute for Cognitive Science
47 Department of Molecular Immunology
48 Universität Leipzig
49 Institute of Immunology
50 Lymphopoïèse (UMR_1223 / U1223 / U-Pasteur_4) - Lymphopoïèse
51 Institute for Immunology
52 Department of Internal Medicine
53 PROTEE - Processus de Transfert et d'Echanges dans l'Environnement - EA 3819
54 Heinrich-Pette-Institut
2 Cell Biology
3 Klinik für Dermatologie, Venerologie und Allergologie
4 Department of Internal Medicine
5 Neuroimmunology Unit
6 Inorganic Chemistry II
7 Caprotec Bioanalytics GmbH
8 IOTA ES - International Occultation Timing Association European Section
9 Institut der Leibniz-Gemeinschaft, Berlin
10 IRCCS - Fondazione Santa Lucia
11 Terry Fox Laboratory
12 Department of Immunology
13 Fondazione Don Carlo Gnocchi
14 IMVA - U1184 - Immunologie des Maladies Virales et Autoimmunes
15 Experimental Immunology Unit, Dept. of Oncology
16 Département d'Immunologie - Department of Immunology
17 Charité Hospital
18 UB - Universitat de Barcelona
19 Rheumatologie
20 Department of Internal Medicine I
21 Department of Histology and Embryology
22 Weizmann Institute of Science [Rehovot, Israël]
23 Régulation des Infections Rétrovirales
24 Institutes of Molecular Medicine and Experimental Immunology
25 U932 - Immunité et cancer
26 SIgN - Singapore Immunology Network
27 Institute of Virology [Zürich]
28 College of Food Science and Technology [Shangai]
29 Institute for Medical Microbiology and Hygiene
30 King‘s College London
31 Erasmus MC - Erasmus University Medical Center [Rotterdam]
32 Unit of Clinical and Experimental Immunology
33 Heinrich Pette Institute [Hamburg]
34 Institute of Translational Medicine
35 Department of Medicine Huddinge
36 UniGe - Università degli studi di Genova = University of Genoa
37 Department of Environmental Microbiology
38 Experimental Immunology
39 Viral Immunobiology
40 Department of Radiation Oncology [Munich]
41 Department of Mathematics and Statistics
42 William Harvey Research Institute, Barts and the London Medical School
43 Cytometry Laboratories and School of Veterinary Medicine
44 Osaka University [Osaka]
45 FACS and Array Core Facility
46 Institute for Cognitive Science
47 Department of Molecular Immunology
48 Universität Leipzig
49 Institute of Immunology
50 Lymphopoïèse (UMR_1223 / U1223 / U-Pasteur_4) - Lymphopoïèse
51 Institute for Immunology
52 Department of Internal Medicine
53 PROTEE - Processus de Transfert et d'Echanges dans l'Environnement - EA 3819
54 Heinrich-Pette-Institut
Immanuel Andrä
- Function : Author
Petra Bacher
- Function : Author
Regina Barnaba
- Function : Author
Burkhard Becher
- Function : Author
- PersonId : 784536
- ORCID : 0000-0002-1541-7867
- IdRef : 181357747
Alfonso Blanco
- Function : Author
David Boulais
- Function : Author
Martin Büscher
- Function : Author
Dirk Busch
- Function : Author
Timothy P Bushnell
- Function : Author
Andrea Cavani
- Function : Author
Pratip K Chattopadhyay
- Function : Author
Qingyu Cheng
- Function : Author
Sue Chow
- Function : Author
Anne Cooke
- Function : Author
Lorenzo Cosmi
- Function : Author
van Duc Dang
- Function : Author
Derek Davies
- Function : Author
Mor de Biasi
- Function : Author
Genny del Zotto
- Function : Author
- PersonId : 784537
- ORCID : 0000-0001-7272-7776
Silvia Della Bella
- Function : Author
Günnur Deniz
- Function : Author
Mark Dessing
- Function : Author
Andreas Diefenbach
- Function : Author
James P. Di Santo
- Function : Author
- PersonId : 842179
Francesco Dieli
- Function : Author
- PersonId : 784538
- ORCID : 0000-0002-6685-352X
Bo Dolf
- Function : Author
Vera S Donnenberg
- Function : Author
Götz R A Ehrhardt
- Function : Author
Bo Endl
- Function : Author
Pablo Engel
- Function : Author
Britta Engelhardt
- Function : Author
Charlotte Esser
- Function : Author
Bart Everts
- Function : Author
Christine Falk
- Function : Author
Todd A Fehniger
- Function : Author
Andrew Filby
- Function : Author
Simon Simon
- Function : Author
Marie Follo
- Function : Author
Bo Förster
- Function : Author
John Foster
- Function : Author
John Foulds
- Function : Author
Paul S Frenette
- Function : Author
Ari Galbraith
- Function : Author
Bo Garbi
- Function : Author
Maria Dolores García-Godoy
- Function : Author
Kamran Ghoreschi
- Function : Author
Mor Gibellini
- Function : Author
Christoph Goettlinger
- Function : Author
Carl S Goodyear
- Function : Author
Francisco Grogan
- Function : Author
Mor Gross
- Function : Author
Daryl Grummitt
- Function : Author
Alexander Hahn
- Function : Author
Quirin Hammer
- Function : Author
Anja E Hauser
- Function : Author
David Houston
- Function : Author
David Hedley
- Function : Author
Guadalupe Herrera
- Function : Author
Falk Hiepe
- Function : Author
Bo Holland
- Function : Author
Pleun Hombrink
- Function : Author
Jessica P Houston
- Function : Author
Bimba F Hoyer
- Function : Author
Ton Huang
- Function : Author
Christopher A Hunter
- Function : Author
Anna Iannone
- Function : Author
Hans-Martin Jäck
- Function : Author
Beatriz Jávega
- Function : Author
Kerstin Juelke
- Function : Author
Toralf Kaiser
- Function : Author
Tomas Kalina
- Function : Author
Baerbel Keller
- Function : Author
Srijit Khan
- Function : Author
Alexander Kienhöfer
- Function : Author
Thomas Kroneis
- Function : Author
Pia Kvistborg
- Function : Author
Virginia Lannigan
- Function : Author
Olivier Lantz
- Function : Author
- PersonId : 756941
- ORCID : 0000-0003-3161-7719
- IdRef : 092426565
Michael D Leipold
- Function : Author
Megan K Levings
- Function : Author
Virginia Litwin
- Function : Author
Lilly Lopez
- Function : Author
Amy Lovett-Racke
- Function : Author
Burkhard Ludewig
- Function : Author
Holden T Maecker
- Function : Author
Giuseppe Matarese
- Function : Author
Alexander Maueröder
- Function : Author
Iain Mcinnes
- Function : Author
Henrik E Mei
- Function : Author
Fritz Melchers
- Function : Author
Susanne Melzer
- Function : Author
Dirk Mielenz
- Function : Author
Kingston Mills
- Function : Author
Jonni Moore
- Function : Author
Barry Moran
- Function : Author
David Mosmann
- Function : Author
Christian Münz
- Function : Author
- PersonId : 766963
- ORCID : 0000-0001-6419-1940
- IdRef : 19061742X
Alexander Munoz
- Function : Author
Kenneth M Murphy
- Function : Author
Toshinori Nakayama
- Function : Author
Milena Nasi
- Function : Author
Christine Neudörfl
- Function : Author
Sussan Nourshargh
- Function : Author
- PersonId : 916650
José-Enrique O'Connor
- Function : Author
Francisco Ouyang
- Function : Author
Annette Oxenius
- Function : Author
Raghav Palankar
- Function : Author
Peter Peterson
- Function : Author
Christian Peth
- Function : Author
Jordi Petriz
- Function : Author
Daisy Philips
- Function : Author
Winfried Pickl
- Function : Author
Regina Piconese
- Function : Author
Marcello Pinti
- Function : Author
a Graham Pockley
- Function : Author
Alexander Podolska
- Function : Author
Carlo Pucillo
- Function : Author
David Quataert
- Function : Author
Timothy R D J Radstake
- Function : Author
David Rebhahn
- Function : Author
Diether Recktenwald
- Function : Author
Ester B M Remmerswaal
- Function : Author
Katy Rezvani
- Function : Author
Laura G Rico
- Function : Author
J Paul Robinson
- Function : Author
Chiara Romagnani
- Function : Author
Anna Rubartelli
- Function : Author
Jürgen Ruland
- Function : Author
Francisco Sala-De-Oyanguren
- Function : Author
Yvonne Samstag
- Function : Author
Sharon Sanderson
- Function : Author
Birgit Sawitzki
- Function : Author
- PersonId : 784539
- ORCID : 0000-0001-8166-8579
Alexander Scheffold
- Function : Author
Matthias Schiemann
- Function : Author
Frank Schildberg
- Function : Author
Esther Schimisky
- Function : Author
Stephan A Schmid
- Function : Author
Kilian Schober
- Function : Author
Axel Ronald Schulz
- Function : Author
Ton Schumacher
- Function : Author
Cristiano Scotta
- Function : Author
T Vincent Shankey
- Function : Author
Anat Shemer
- Function : Author
Anna-Katharina Simon
- Function : Author
Alan M Stall
- Function : Author
Regina Stark
- Function : Author
Christina Stehle
- Function : Author
Merle Stein
- Function : Author
Tobit Steinmetz
- Function : Author
- PersonId : 784540
- ORCID : 0000-0002-6541-9369
Yousuke Takahama
- Function : Author
Gergely Toldi
- Function : Author
Julia Tornack
- Function : Author
Elisabetta Traggiai
- Function : Author
Joe Trotter
- Function : Author
Paulo Ulrich
- Function : Author
Marlous van Der Braber
- Function : Author
René a W van Lier
- Function : Author
Marcello Veldhoen
- Function : Author
Salvador Vento-Asturias
- Function : Author
Konrad von Volkmann
- Function : Author
Ari Waisman
- Function : Author
- PersonId : 763005
- ORCID : 0000-0003-4304-8234
Rachael Walker
- Function : Author
Michael D Ward
- Function : Author
Klaus Warnatz
- Function : Author
Sarah Warth
- Function : Author
James Watson
- Function : Author
- PersonId : 770013
- ORCID : 0000-0003-4942-1984
Carsten Watzl
- Function : Author
Leonie Wegener
- Function : Author
Annika Wiedemann
- Function : Author
Jürgen Wienands
- Function : Author
Gerald Willimsky
- Function : Author
- PersonId : 784541
- ORCID : 0000-0002-9693-948X
James Wing
- Function : Author
Peter Wurst
- Function : Author
Liping Yu
- Function : Author
Alice Yue
- Function : Author
Qianjun Zhang
- Function : Author
Jakob Zimmermann
- Function : Author
Abstract
The classical model of hematopoiesis established in the mouse postulates that lymphoid cells originate from a founder population of common lymphoid progenitors. Here, using a modeling approach in humanized mice, we showed that human lymphoid development stemmed from distinct populations of CD127(-) and CD127(+) early lymphoid progenitors (ELPs). Combining molecular analyses with in vitro and in vivo functional assays, we demonstrated that CD127(-) and CD127(+) ELPs emerged independently from lympho-mono-dendritic progenitors, responded differently to Notch1 signals, underwent divergent modes of lineage restriction, and displayed both common and specific differentiation potentials. Whereas CD127(-) ELPs comprised precursors of T cells, marginal zone B cells, and natural killer (NK) and innate lymphoid cells (ILCs), CD127(+) ELPs supported production of all NK cell, ILC, and B cell populations but lacked T potential. On the basis of these results, we propose a "two-family" model of human lymphoid development that differs from the prevailing model of hematopoiesis.