title: Wild immunology assessed by multidimensional mass cytometry.
abstract: A great part of our knowledge on mammalian immunology has been established in laboratory settings. The use of inbred mouse strains enabled controlled studies of immune cell and molecule functions in defined settings. These studies were usually performed in specific-pathogen free (SPF) environments providing standardized conditions. In contrast, mammalians including humans living in their natural habitat are continuously facing pathogen encounters throughout their life. The influences of environmental conditions on the signatures of the immune system and on experimental outcomes are yet not well defined. Thus, the transferability of results obtained in current experimental systems to the physiological human situation has always been a matter of debate. Studies elucidating the diversity of "wild immunology" imprintings in detail and comparing it with those of "clean" lab mice are sparse. Here, we applied multidimensional mass cytometry to dissect phenotypic and functional differences between distinct groups of laboratory and pet shop mice as a source for "wild mice". For this purpose, we developed a 31-antibody panel for murine leukocyte subsets identification and a 35-antibody panel assessing various cytokines. Established murine leukocyte populations were easily identified and diverse immune signatures indicative of numerous pathogen encounters were classified particularly in pet shop mice and to a lesser extent in quarantine and non-SPF mice as compared to SPF mice. In addition, unsupervised analysis identified distinct clusters that associated strongly with the degree of pathogenic priming, including increased frequencies of activated NK cells and antigen-experienced B- and T-cell subsets. Our study unravels the complexity of immune signatures altered under physiological pathogen challenges and highlights the importance of carefully adapting laboratory settings for immunological studies in mice, including drug and therapy testing. © 2016 International Society for Advancement of Cytometry.
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Alberto Sada
authors.initials: AS
authors.lastname: Japp
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Hoffmann
authors.affiliation: BCRT Flow Cytometry Lab (BCRT-FCL), BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.|Institute for Medical Immunology, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Schlickeiser
authors.affiliation: Medical Department I (Gastroenterology, Rheumatology, Infectious Diseases), Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Glauben
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Christos
authors.initials: C
authors.lastname: Nikolaou
authors.affiliation: Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, California.
authors.forename: Holden T
authors.initials: HT
authors.lastname: Maecker
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Julian
authors.initials: J
authors.lastname: Braun
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.|Federal Office of Consumer Protection and Food Safety (BVL), Berlin, Germany.
authors.forename: Nadine
authors.initials: N
authors.lastname: Matzmohr
authors.affiliation: Transplantation Tolerance, Institute for Medical Immunology, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Sawitzki
authors.affiliation: Medical Department I (Gastroenterology, Rheumatology, Infectious Diseases), Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Britta
authors.initials: B
authors.lastname: Siegmund
authors.affiliation: Deutsches Rheuma-Forschungszentrum, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Radbruch
authors.affiliation: Institute for Medical Immunology, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Hans-Dieter
authors.initials: HD
authors.lastname: Volk
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Frentsch
authors.affiliation: BCRT Flow Cytometry Lab (BCRT-FCL), BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Desiree
authors.initials: D
authors.lastname: Kunkel
authors.affiliation: Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Thiel
mesh_terms: D000818:Animals; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D007694:Killer Cells, Natural / Q000276:immunology*; D007962:Leukocytes / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*
keywords: Key terms: wild immunology; adaptive immune system; innate immune system; mass cytometry