Regulation of macrophage functions by cytokine and 1,25-dihydroxyvtamin D_3 on glomerulonephritis

新月体肾小球肾炎是发生在肾小球毛细血管外并涉及足细胞的许多严重病变中的一个代表。关于新月体形成细胞是来源于肾小球上皮细胞还是单核细胞/巨噬细胞的问题存在很大争议，尚未得到澄清。为了研究新月体肾小球肾炎足细胞的病理生理学，我们试图建立一种培养足细胞的方法，特别关注足细胞和巨噬细胞之间的关系。从正常大鼠分离的肾小球培养物中生长的未知来源的非贴壁细胞数量增加，在第11天达到3.5 × 10^5/ml。它们具有巨噬细胞的特征，表达特异性抗原和酶，形态学和产生H_2O_2。它们表达FxlA，但缺乏Thy1.1或因子VIII的表达。 ...更多信息 艾德认为肾小球基底膜（GBM）上有足突的细胞转变为巨噬细胞（MC），并从肾小球中迁移出来。免疫荧光双染显示第8天从肾小球表面迁移的细胞均为足糖萼蛋白和艾德-1阳性。免疫电镜显示艾德1阳性细胞位于GBM上。用抗巨噬细胞抗体和Thy1.1抗体预处理并不能减少MC的数量，而嘌呤霉素氨基糖苷预处理主要减少MC的数量。来自骨髓嵌合体大鼠肾小球培养物的MC表达来自受体的Ia抗原，表明MC不是来自骨髓细胞。巨噬细胞集落刺激因子（macrophagecolonystimulatingfactor，CSF）可加速肾小球足细胞向MC转化的速度，而粒细胞-巨噬细胞集落刺激因子（granulocyte-macrophagecolonystimulatingfactor，GCSF）则可显著促进肾小球足细胞向MC转化的程度。这些结果为新月体形成细胞的起源提供了全新的见解。少

One representative of a number of severe lesions that occur outside the glomerular capillaries and involve podocytes is crescentic glomerulonephritis. The question of whether the crescent-forming cells are derived from glomerular epithelial cells or monocytes/macrophages is highly controversial and has not yet been clarified. To investigate pathophysiology of podocyte in crescentic glomerulonephritis, we attempted to establish methods for culturing cells confirmed to be derived from podocytes, focusing particularly on the relationship between podocytes and macrophages.Non-adherent cells of unknown origin that grew from normal rat isolated glomerular cultures increased in number, reaching a total of 3.5 x 10^5/ml on day 11. They showed several characteristics of macrophages, the expression of specific antigens and enzyme, morphology, and production of H_2O_2. They expressed FxlA,but lacked the expressions of Thy1.1 or factor VIII.A morphological kinetic study on day 3-11 of culture show … More ed that the cells with foot processes on the glomerular basement membrane (GBM) changed into macrophagic cells (MC) and migrated from the glomeruli. Immunofluorescence double staining indicated that the cells migrated from glomerular surface on day 8 were both anti-podocalyxin and ED-1 positive. Furthermore, immunoelectron microscopy revealed that the ED-1 positive cells were located on GBM.Pretreatment with an anti-macrophages and -Thy1.1 antibodies both with complement did not reduce the number of MC,while pretreatment with puromycin aminonucleoside predominantly reduced the number of MC.Predominant decrease in the number of glomerular macrophage by gamma-irradiation did not result in the reduction in the number of MC.MC derived from glomerular cultures of bone marrow chimeric rats expressed the Ia antigen originated from recipient, indicating that MC is not derived from bone marrow cells. Macrophage colony-stimulating factor accelerated the speed of the change into MC and granulocyte-macrophage colony-stimulating factor dramatically enhanced its degree with increase of cell number on day 8.We concluded that podocytes change into MC in normal rat glomerular culture and the change is enhanced by CSFs. The results provide a completely new insight into the origin of crescent forming cells. Less