Resident dendritic cells are the predominant TNF-secreting cell in early renal ischemia-reperfusion injury
Resident dendritic cells are the predominant TNF-secreting cell in early renal ischemia-reperfusion injury
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DOI：
10.1038/sj.ki.5002132
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发表时间：
2007-04-01
影响因子：
19.6
通讯作者：
Griffin, M. D.
中科院分区：
文献类型：
--
作者：
Dong, X.;Swaminathan, S.;Griffin, M. D.
肾缺血再灌注损伤（IRI）迅速诱导炎症介质的产生，尤其包括肿瘤坏死因子（TNF）。可能的来源包括驻留的实质细胞和骨髓来源的细胞以及招募的白细胞。通过细胞分离、体外培养和酶联免疫吸附测定 (ELISA)、免疫过氧化物酶和免疫荧光显微镜以及多色流式细胞术对来自接受 IRI 的肾脏的细胞悬液进行检查，以确定树突状细胞 (DC) 对 TNF 和其他炎症介质早期产生的贡献。与对照肾脏相比，IRI 细胞悬浮液中 TNF、白细胞介素 (IL-6)、单核细胞趋化蛋白 1 (MCP-1) 和活化调节正常 T 细胞表达和分泌 (RANTES) 的分泌增加，并且在富含 DC 的制剂中更高。免疫染色鉴定出共表达 DC 标记 CD11c 的 TNF+ve 细胞。 IRI 后 24 小时对骨髓源性 (CD45(+ve)) 细胞群进行流式细胞术检测，结果表明 F4/80(+ve)/CD11c(+ve) DC 保持比例稳定，并表现出较高水平的 DC 成熟标记物，而 F4/80(-ve) DC、单核细胞、中性粒细胞和 T 细胞的比例则有所增加。 TNF 的细胞内染色证实 F4/80(+ve) DC 是主要的 TNF+ve 细胞，并且表达水平高于其他 TNF+ve 细胞。 IRI后，体内肾DCs的消耗显着减弱了总细胞和CD45(+ve)细胞的TNF分泌。结果揭示了常驻 F4/80(+ve) DC 在 IRI 24 小时内作为 TNF 主要分泌者的作用。
Renal ischemia-reperfusion injury (IRI) rapidly induces production of inflammatory mediators including, and in particular, tumor necrosis factor (TNF). Possible sources include resident parenchymal and bone marrow-derived cells as well as recruited leukocytes. Cell suspensions from kidneys subjected to IRI were examined by cell separation followed by in vitro culture and enzyme-linked immunosorbent assay (ELISA), immunoperoxidase and immunofluorescence microscopy, and multicolor flow cytometry to determine the contribution of dendritic cells (DCs) to early production of TNF and other inflammatory mediators. Secretion of TNF, interleukin (IL-6), monocyte chemoattractant protein-1 (MCP-1), and regulated on activation normal T cell expressed and secreted ( RANTES) was increased in cell suspensions from IRI compared with control kidneys and was higher in DC-enriched preparations. Immunostaining identified TNF+ve cells that coexpressed the DC marker CD11c. Flow cytometry of bone marrow-derived (CD45(+ve)) cell populations at 24h post-IRI demonstrated that F4/80(+ve)/CD11c(+ve) DCs remained proportionately stable and exhibit higher levels of DC maturation markers, whereas the proportion of F4/80(-ve) DCs, monocytes, neutrophils, and T cells increased. Intracellular staining for TNF confirmed that F4/80(+ve) DCs were the predominant TNF+ve cell and expressed higher levels than other TNF+ve cells. In vivo depletion of DCs from the kidney substantially attenuated TNF secretion by total and CD45(+ve) cells following IRI. The results uncover a role for resident F4/80(+ve) DCs as the predominant secretors of TNF within 24h of IRI.