Changes in activation states of murine polymorphonuclear leukocytes (PMN) during inflammation: a comparison of bone marrow and peritoneal exudate PMN
Changes in activation states of murine polymorphonuclear leukocytes (PMN) during inflammation: a comparison of bone marrow and peritoneal exudate PMN
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DOI：
10.1128/cvi.13.5.575-583.2006
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发表时间：
2006-05-01
影响因子：
--
通讯作者：
Karlsson, Anna
中科院分区：
文献类型：
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作者：
Itou, Takuya;Collins, L. Vincent;Karlsson, Anna
为了研究小鼠多形核白细胞 (PMN) 的不同激活状态，我们比较了从骨髓获得的小鼠 PMN (BMPMN) 与通过硫代乙醇酸盐 (TGPMN) 或尿酸 (UAPMN) 腹膜内诱导获得的 PMN 的功能。当用趋化肽，例如甲酰基-甲硫氨酰-亮氨酰-苯丙氨酸（fMLF）、WKYMVM或WKYMVm刺激时，与BMPMN相比，TGPMN和UAPMN表现出大大增强的活性氧（ROS）的产生，这表明渗出到腹膜本身会诱导细胞处于启动状态。 WKYMVm 肽是 ROS 生成最有效的刺激剂，并且它对随后的 fMLF 或 WKYMVM 刺激不敏感。添加细胞松弛素 B 可以打破这种脱敏作用。TGPMN 和 UAPMN 似乎已完全启动，因为肿瘤坏死因子 α (TNF-α) 预处理并未诱导反应增加。相比之下，BMPMN 反应增加了 2.5 至 3 倍。脱颗粒研究支持了氧化反应的差异。与 TNF-α 预孵育促进 BMPMN 上 CR3 的表达，并且 WKYMVm 也增强了这种表达水平。相比之下，未经处理的TGPMN和UAPMN上的CR3表达已经与TNF-α引发的BMPMN上的CR3表达相似，并且通过TNF-α处理只能略微增强。综上所述，这些结果表明 BMPMN 处于静息状态并且有能力启动，而腹膜渗出物 PMN 在分离时已经完全启动。这些结果对小鼠中性粒细胞研究具有重大意义，并表明在研究小鼠模型时定义使用哪些 PMN 子集的重要性。
To study different activation states in polymorphonuclear leukocytes (PMN) in mice, we compared the function of murine PMN obtained from the bone marrow (BMPMN) with those of PMN obtained by intraperitoneal induction with thioglycolate (TGPMN) or uric acid (UAPMN). When stimulated with chemotactic peptides, e.g., formyl-methionyl-leucyl-phenylalanine (fMLF), WKYMVM, or WKYMVm, the TGPMN and UAPMN showed greatly enhanced generation of reactive oxygen species (ROS) compared with BMPMN, which suggests that exudation to the peritoneum per se induces a primed state in the cells. The WKYMVm peptide was the most potent stimulant of ROS generation, and it desensitized for subsequent stimulation with fMLF or WKYMVM. This desensitization was broken by the addition of cytochalasin B. The TGPMN and UAPMN appeared to be fully primed, since no increase in response was induced by pretreatment with tumor necrosis factor alpha (TNF-alpha). In contrast, the BMPMN response was increased 2.5- to 3-fold. The differences in oxidative responses were supported by degranulation studies. Preincubation with TNF-alpha promoted CR3 expression on BMPMN, and this level of expression was also enhanced by WKYMVm. In contrast, CR3 expression on untreated TGPMN and UAPMN was already similar to that on TNF-alpha-primed BMPMN and could be only slightly enhanced by TNF-alpha treatment. Taken together, these results indicate that BMPMN are in a resting state and have the capacity to become primed, while peritoneal exudate PMN are already fully primed upon isolation. These results have major implications for murine neutrophil research and show the importance of defining which PMN subsets to use when investigating murine models.