Burger M, Almanasreh N, Bauermeister K, Knopf H P, Schollmeyer P, Dobos G J
Department of Nephrology, University Hospital Freiburg, Germany.
Immunobiology. 1999 Feb;200(1):62-76. doi: 10.1016/S0171-2985(99)80033-3.
Studies on human macrophages are restricted due to difficulties in isolating significant numbers of human macrophages. High numbers of monocytes/macrophages can be obtained from peritonitis effluents of patients treated with peritoneal dialysis. To determine whether these cells might be useful for functional studies, we characterized peritoneal macrophages (PM) immediately after isolation from the dialysate effluents and their subsequent differentiation. During a 10 days culture period they differentiated morphologically and phenotypically (FACS-analysis) from monocyte-like cells to macrophages. Reflecting the intraperitoneal inflammation we found protein- and mRNA-synthesis of IL-8 and monocyte-chemoattractant-protein-1 (MCP-1) to be upregulated in PM after isolation from the effluents. In contrast, TNF-alpha was downregulated and could not be stimulated by LPS and/or IFN-gamma, reflecting the phenomenon of desensitization. After 10 days in culture, cytokine production normalized to a constitutive level and the TNF-alpha responsiveness to LPS was restored. These data suggest the recovery of PM from the inflammatory prestimulation. Therefore PM harvested from peritoneal dialysis effluents might provide a useful tool for further studies on the role of human macrophages in inflammation.
由于难以分离出大量的人类巨噬细胞，对人类巨噬细胞的研究受到限制。通过腹膜透析治疗的患者的腹膜炎流出液可获得大量的单核细胞/巨噬细胞。为了确定这些细胞是否可用于功能研究，我们在从透析液流出物中分离出腹膜巨噬细胞（PM）后立即对其进行了表征，并观察了它们随后的分化情况。在为期10天的培养期内，它们在形态和表型上（通过流式细胞术分析）从单核细胞样细胞分化为巨噬细胞。反映腹膜内炎症情况，我们发现从流出物中分离出的PM中，白细胞介素-8（IL-8）和单核细胞趋化蛋白-1（MCP-1）的蛋白质和mRNA合成上调。相反，肿瘤坏死因子-α（TNF-α）下调，且不能被脂多糖（LPS）和/或干扰素-γ（IFN-γ）刺激，这反映了脱敏现象。培养10天后，细胞因子产生恢复到组成水平，并且TNF-α对LPS的反应性得以恢复。这些数据表明PM从炎症预刺激中恢复。因此，从腹膜透析流出物中收获的PM可能为进一步研究人类巨噬细胞在炎症中的作用提供有用的工具。
