Guidelines for the use of flow cytometry and cell sorting in immunological studies

• Published in: European journal of immunology 2017-10, Vol.47 (10), p.1584-1797
• Main Authors: Akdis, Mübeccel, Bacher, Petra, Battistini, Luca, Bauer, Wolfgang M., Becher, Burkhard, Beisker, Wolfgang, Berek, Claudia, Borsellino, Giovanna, Brinkman, Ryan R., Büscher, Martin, Chow, Sue, Clerici, Mario, Cosmi, Lorenzo, Davies, Derek, Biasi, Sara, Del Zotto, Genny, Della Bella, Silvia, Dessing, Mark, Dieli, Francesco, Ehrhardt, Götz R. A., Engel, Pablo, Engelhardt, Britta, Esser, Charlotte, Fehniger, Todd A., Fillatreau, Simon, Follo, Marie, Foster, John, Frenette, Paul S., García‐Godoy, Maria Dolores, Geginat, Jens, Grogan, Jane, Grummitt, Daryl, Hauser, Anja E., Hiepe, Falk, Holland, Tristan, Hombrink, Pleun, Iannone, Anna, Jonjic, Stipan, Juelke, Kerstin, Jung, Steffen, Kaiser, Toralf, Kurts, Christian, Lannigan, Joanne, Liu, Yanling, Lohoff, Michael, Lombardi, Giovanna, Lovett‐Racke, Amy, Ludewig, Burkhard, Lugli, Enrico, McGrath, Mairi, McInnes, Iain, Mei, Henrik E., Mirrer, David, Mjösberg, Jenny, Müller, Susann, Münz, Christian, Nakayama, Toshinori, O'Connor, José‐Enrique, Oxenius, Annette, Pickl, Winfried, Pinti, Marcello, Pucillo, Carlo, Quataert, Sally A., Radstake, Timothy R. D. J., Rebhahn, Jonathan A., Rico, Laura G., Robinson, J. Paul, Rubartelli, Anna, Ruland, Jürgen, Sanderson, Sharon, Sawitzki, Birgit, Schildberg, Frank, Schimisky, Esther, Schmid, Stephan A, Schmitt, Steffen, Schober, Kilian, Schüler, Thomas, Schumacher, Ton, Scotta, Cristiano, Shemer, Anat, Simon, Anna‐Katharina, Stehle, Christina, Stockinger, Hannes, Tarnok, Attila, Tian, ZhiGang, Tornack, Julia, Trotter, Joe, Voehringer, David, Volk, Hans‐Dieter, Walker, Rachael, Warnatz, Klaus, Warth, Sarah, Wegener, Leonie, Wiedemann, Annika, Willimsky, Gerald, Wurst, Peter, Yu, Liping, Yue, Alice, Zhao, Yi, Ziegler, Susanne
• Format: Article
• Language: English
• Subjects: Animals; Cell Proliferation; Cell Separation; DNA - analysis; False Positive Reactions; Flow Cytometry; Guidelines as Topic; Humans; Immunologic Techniques; Immunology; Immunophenotyping; Life Sciences; Medicin och hälsovetenskap; Quality Control; Research Design; RNA - analysis; Software; T-Lymphocytes - cytology
• ISSN: 0014-2980; 1521-4141; 1521-4141
• DOI: 10.1002/eji.201646632

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.