Neutrophils are quickly recruited to tissues in response to proinflammatory cues; however, little is known about tissue neutrophil phenotypes in health. We employ a multicolor flow cytometric approach to assess surface markers of activation on neutrophils from the bone marrow, blood, peritoneum, spleen, liver, fat, colon, and oral cavity of healthy mice. Cell preparations were promptly fixed to preserve native surface marker expression levels. Peritoneal, colonic, and oral neutrophils were also assessed in the setting of pHrodo-induced peritonitis, dextran sodium sulfate-induced colitis, and ligature-induced periodontal disease, respectively. Our results demonstrate consistent detectable neutrophil populations in various sterile and nonsterile tissues of healthy mice, and these cells had tissue-specific neutrophil immunophenotypes. Neutrophils derived from biofilm-associated mucosal tissues had 2- to 3-fold higher expression of surface markers of activation, including CD66a, CD11b, and CD62L, compared to neutrophils derived from both sterile healthy tissues as well as tissues in animals treated with broad-spectrum antibiotics. Furthermore, the unique cluster of differentiation (CD) marker activation signatures of tissue-specific neutrophils from the peritoneum, colon, and oral cavity were altered to a proinflammatory immunophenotype with the presence of an inflammatory stimulus. Based on our results, we propose a model whereby a hierarchy of tissue neutrophil immunophenotypes, based on the differential expression of CD markers of activation, correlates with sterile, healthy commensal biofilm-associated and inflamed tissue states.

中文翻译：

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健康和炎症中的组织特异性鼠中性粒细胞活化状态

中性粒细胞响应促炎信号被迅速募集到组织中；然而，人们对健康中的组织中性粒细胞表型知之甚少。我们采用多色流式细胞术方法来评估来自健康小鼠的骨髓、血液、腹膜、脾脏、肝脏、脂肪、结肠和口腔的中性粒细胞的表面活化标记。细胞制剂被迅速固定以保持天然表面标志物的表达水平。还分别在 pHrodo 诱发的腹膜炎、葡聚糖硫酸钠诱发的结肠炎和结扎诱发的牙周病的情况下评估了腹膜、结肠和口腔中性粒细胞。我们的结果表明，在健康小鼠的各种无菌和非无菌组织中可检测到一致的中性粒细胞群，并且这些细胞具有组织特异性中性粒细胞免疫表型。与来自无菌健康组织以及用广泛治疗的动物组织中的中性粒细胞相比，来自生物膜相关粘膜组织的中性粒细胞的表面活化标志物（包括 CD66a、CD11b 和 CD62L）的表达高出 2 到 3 倍。谱抗生素。此外，来自腹膜、结肠和口腔的组织特异性嗜中性粒细胞的独特的分化簇 (CD) 标记激活特征在炎症刺激的存在下被改变为促炎免疫表型。基于我们的结果，我们提出了一个模型，其中基于 CD 激活标记物的差异表达，组织中性粒细胞免疫表型的层次结构与无菌、健康的共生生物膜相关和发炎组织状态相关。