Japp Alberto Sada, Hoffmann Kerstin, Schlickeiser Stephan, Glauben Rainer, Nikolaou Christos, Maecker Holden T, Braun Julian, Matzmohr Nadine, Sawitzki Birgit, Siegmund Britta, Radbruch Andreas, Volk Hans-Dieter, Frentsch Marco, Kunkel Desiree, Thiel Andreas
Regenerative Immunology and Aging, BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
BCRT Flow Cytometry Lab (BCRT-FCL), BCRT, Charité Universitätsmedizin Berlin, Berlin, Germany.
Cytometry A. 2017 Jan;91(1):85-95. doi: 10.1002/cyto.a.22906. Epub 2016 Jul 12.
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.
我们关于哺乳动物免疫学的大部分知识都是在实验室环境中建立的。近交系小鼠品系的使用使得在特定环境下对免疫细胞和分子功能进行对照研究成为可能。这些研究通常在提供标准化条件的无特定病原体（SPF）环境中进行。相比之下，包括人类在内生活在自然栖息地的哺乳动物在其一生中不断面临病原体接触。环境条件对免疫系统特征和实验结果的影响尚未得到明确界定。因此，当前实验系统中获得的结果能否转化为人类生理状况一直是一个有争议的问题。详细阐明“野生免疫学”印记多样性并将其与“纯净”实验室小鼠的印记进行比较的研究很少。在这里，我们应用多维质谱流式细胞术剖析实验室小鼠和宠物店小鼠不同组之间的表型和功能差异，宠物店小鼠可作为“野生小鼠”的来源。为此，我们开发了一个用于鉴定小鼠白细胞亚群的31抗体面板和一个评估各种细胞因子的35抗体面板。与SPF小鼠相比，已建立的小鼠白细胞群体很容易识别，并且在宠物店小鼠中尤其在隔离和非SPF小鼠中程度较轻地分类出了指示大量病原体接触的多种免疫特征。此外，无监督分析确定了与致病引发程度密切相关的不同簇，包括活化NK细胞以及经历过抗原刺激的B细胞和T细胞亚群频率增加。我们的研究揭示了在生理病原体挑战下改变的免疫特征的复杂性，并强调了为小鼠免疫学研究（包括药物和疗法测试）仔细调整实验室环境的重要性。© 2016国际细胞计量学促进协会。
