Daley Jean M, Reichner Jonathan S, Mahoney Eric J, Manfield Laura, Henry William L, Mastrofrancesco Balduino, Albina Jorge E
Department of Surgery, Division of Surgical Research, Rhode Island Hospital and Brown Medical School, Providence, RI 02903, USA.
J Immunol. 2005 Feb 15;174(4):2265-72. doi: 10.4049/jimmunol.174.4.2265.
The regulation of macrophage phenotype by neutrophils was studied in the s.c. polyvinyl alcohol sponge wound model in mice made neutropenic by anti-Gr-1 Ab, as well as in cell culture. Wounds in neutropenic mice contained 100-fold fewer neutrophils than those in nonneutropenic controls 1 day after sponge implantation. Wound fluids from neutropenic mice contained 68% more TNF-alpha, 168% more IL-6, and 61% less TGF-beta1 than those from controls. Wound fluid IL-10 was not different between the two groups, and IL-4 was not detected. Intracellular TNF-alpha staining was greater in cells isolated from neutropenic wounds than in those from control wounds. The hypothesis that wound neutrophil products modulate macrophage phenotype was tested in Transwell cocultures of LPS-stimulated J774A.1 macrophages and day 1 wound cells (84% neutrophils/15% macrophages). Overnight cocultures accumulated 60% less TNF-alpha and IL-6 than cultures of J774A.1 alone. The suppression of cytokine release was mediated by a soluble factor(s), because culture supernatants from wound cells inhibited TNF-alpha and IL-6 release from LPS-stimulated J774A.1 cells. Culture supernatants from purified wound neutrophils equally suppressed TNF-alpha release from LPS-stimulated J774A.1 cells. Wound cell supernatants also suppressed TNF-alpha and superoxide release from murine peritoneal macrophages. The TNF-alpha inhibitory factor has a molecular mass <3000 Da and is neither PGE2 nor adenosine. The present findings confirm a role for neutrophils in the regulation of innate immune responses through modulation of macrophage phenotype.
在通过抗Gr-1抗体使小鼠中性粒细胞减少的皮下聚乙烯醇海绵伤口模型以及细胞培养中，研究了中性粒细胞对巨噬细胞表型的调节作用。海绵植入后1天，中性粒细胞减少的小鼠伤口中的中性粒细胞数量比非中性粒细胞减少的对照组少100倍。中性粒细胞减少的小鼠伤口液中的肿瘤坏死因子-α（TNF-α）比对照组多68%，白细胞介素-6（IL-6）多168%，转化生长因子-β1（TGF-β1）少61%。两组伤口液中的白细胞介素-10（IL-10）无差异，且未检测到白细胞介素-4（IL-4）。从中性粒细胞减少的伤口分离的细胞中细胞内TNF-α染色比从对照伤口分离的细胞中更明显。在LPS刺激的J774A.1巨噬细胞与第1天伤口细胞（84%中性粒细胞/15%巨噬细胞）的Transwell共培养中，测试了伤口中性粒细胞产物调节巨噬细胞表型的假设。过夜共培养积累的TNF-α和IL-6比单独培养J774A.1少60%。细胞因子释放的抑制是由一种可溶性因子介导的，因为伤口细胞的培养上清液抑制了LPS刺激的J774A.1细胞中TNF-α和IL-6的释放。纯化的伤口中性粒细胞的培养上清液同样抑制了LPS刺激的J774A.1细胞中TNF-α的释放。伤口细胞上清液也抑制了小鼠腹腔巨噬细胞中TNF-α和超氧化物的释放。TNF-α抑制因子的分子量<3000 Da，既不是前列腺素E2也不是腺苷。目前的研究结果证实了中性粒细胞在通过调节巨噬细胞表型来调节先天免疫反应中的作用。
