Vivanco Gonzalez Nora, Oliveria John-Paul, Tebaykin Dmitry, Ivison Geoffrey T, Mukai Kaori, Tsai Mindy M, Borges Luciene, Nadeau Kari C, Galli Stephen J, Tsai Albert G, Bendall Sean C
Department of Pathology, School of Medicine, Stanford University, Stanford Blood Center, 3373 Hillview Avenue Room 230A, Palo Alto, CA 94305, USA.
Department of Medicine, Division of Respirology, McMaster University, Hamilton, ON, L8S4K1, Canada.
iScience. 2020 Oct 22;23(11):101724. doi: 10.1016/j.isci.2020.101724. eCollection 2020 Nov 20.
Basophils, the rarest granulocyte, play critical roles in parasite- and allergen-induced inflammation. We applied mass cytometry (CyTOF) to simultaneously asses 44 proteins to phenotype and functionally characterize neutrophils, eosinophils, and basophils from 19 healthy donors. There was minimal heterogeneity seen in eosinophils and neutrophils, but data-driven analyses revealed four unique subpopulations within phenotypically basophilic granulocytes (PBG; CD45HLA-DRCD123). Through CyTOF and fluorescence-activated cell sorting (FACS), we classified these four PBG subpopulations as (I) CD16FcεRICD244 (88.5 ± 1.2%), (II) CD16FcεRICD244 (9.1 ± 0.4%), (III) CD16FcεRICD244 (2.3 ± 1.3), and (IV) CD16FcεRICD244 (0.4 ± 0.1%). Prospective isolation confirmed basophilic-morphology of PBG I-III, but neutrophilic-morphology of PBG IV. Functional interrogation via IgE-crosslinking or IL-3 stimulation demonstrated that PBG I-II had significant increases in CD203c expression, whereas PBG III-IV remained unchanged compared with media-alone conditions. Thus, PBG III-IV could serve roles in non-IgE-mediated immunity. Our findings offer new perspectives in human basophil heterogeneity and the varying functional potential of these new subsets in health and disease.
嗜碱性粒细胞是最罕见的粒细胞，在寄生虫和过敏原诱导的炎症中起关键作用。我们应用质谱流式细胞术（CyTOF）同时评估44种蛋白质，以对来自19名健康供体的中性粒细胞、嗜酸性粒细胞和嗜碱性粒细胞进行表型分析和功能表征。在嗜酸性粒细胞和中性粒细胞中观察到的异质性最小，但数据驱动分析揭示了表型嗜碱性粒细胞（PBG；CD45HLA-DRCD123）内的四个独特亚群。通过CyTOF和荧光激活细胞分选（FACS），我们将这四个PBG亚群分类为（I）CD16FcεRICD244（88.5±1.2%）、（II）CD16FcεRICD244（9.1±0.4%）、（III）CD16FcεRICD244（2.3±1.3）和（IV）CD16FcεRICD244（0.4±0.1%）。前瞻性分离证实PBG I-III具有嗜碱性形态，但PBG IV具有中性粒细胞形态。通过IgE交联或IL-3刺激进行功能询问表明，与单独培养基条件相比，PBG I-II的CD203c表达显著增加，而PBG III-IV保持不变。因此，PBG III-IV可能在非IgE介导的免疫中发挥作用。我们的研究结果为人类嗜碱性粒细胞异质性以及这些新亚群在健康和疾病中的不同功能潜力提供了新的视角。
