Neutrophil transmigration in renal proximal tubular LLC-PK1 cells
Neutrophil transmigration in renal proximal tubular LLC-PK1 cells
复制标题
DOI：
10.1159/000076931
复制
发表时间：
2004-01-01
影响因子：
--
通讯作者：
Wiedermann, CJ
中科院分区：
文献类型：
--
作者：
Joannidis, M;Truebsbach, S;Wiedermann, CJ
背景/目标：快速进展性肾小球肾炎患者活检中小管内白细胞粘附于近端小管，间质性肾炎患者尿液中出现白细胞，提示肾脏疾病中白细胞与肾小管上皮细胞之间存在相互作用。本研究旨在研究体外细胞因子或内毒素（LPS）孵育肾小管上皮细胞是否会刺激白细胞通过这些上皮细胞的粘附和迁移。研究方法：实验测定了PMN对在组织培养板上培养的LLC-PK细胞的粘附和通过在微孔膜上培养的LLC-PK单层的跨上皮迁移（TEM）。测量跨上皮电阻（TER），细胞因子释放到顶端或基底外侧室和顶端和基底外侧上清液的趋化活性。结果如下：用TNF α或LPS预孵育LLC-PK细胞，可刺激PMN粘附，从而刺激PMN沿顶侧至基底侧和基底侧至顶侧方向通过LLC-PK单层迁移。TEM与TER降低无关。虽然发现LLC-PK细胞主要分泌顶端IL-8，但顶侧至基底侧的迁移发生在IL-8的浓度梯度上，并且不能被IL-8抗体抑制。通过TEM，上清液的趋化活性略有增加，但没有显示出任何显着的差异，独立于PMN迁移的方向，顶侧和基底侧室。在基底外侧至顶端方向上的TEM的效率是在顶端至基底外侧方向上的效率的大约两倍（迁移指数分别= 3.92 +/-0.55和2.28 +/-.21）。只有基底外侧到顶端TEM可以部分抑制预孵育的基底外侧膜与IL-8抗体。结论：炎症介质刺激PMN粘附LLC-PK细胞，随后TEM。细胞因子或内毒素刺激TEM的机制似乎是LLC-PK细胞表面受体性质的变化，而不是趋化刺激。基底外侧至顶端TEM似乎是最有可能通过IL-8相关的趋触性PMN刺激增强的有利方向。版权所有（C）2004 S. Karger AG，巴塞尔。
Background/Aims: Adhesion of intratubular leukocytes to proximal tubules in biopsies of patients with rapidly progressive glomerulonephritis and the appearance of leukocytes in the urine in interstitial nephritis suggest interactions between leukocytes and tubular epithelia in renal disease. The present study was performed to investigate whether incubation of tubular epithelia with cytokines or endotoxin (LPS) does stimulate adhesion and migration of leukocytes through these epithelia in vitro. Methods: Experiments determined adhesion of PMN to LLC-PK cells cultured on tissue culture plates and transepithelial migration (TEM) through LLC-PK monolayers cultured on microporous membranes. Measurements of transepithelial electrical resistance (TER), cytokine release into apical or basolateral compartments and chemotactic activities of apical and basolateral supernatants were performed. Results: Preincubation of LLC-PK cells with either TNFalpha or LPS resulted in stimulation of PMN adhesion and consequently PMN migration through LLC-PK monolayers in both apical-to-basolateral and basolateral-to-apical direction. TEM was not associated with a reduction of TER. Although largely apical IL-8 secretion by LLC-PK cells was found, apical-to-basolateral migration occurred against a concentration gradient of IL-8 and could not be inhibited by IL-8 antibodies. Chemotactic activities of supernatants were slightly increased by TEM but did not show any significant differences between apical and basolateral compartments independent of the direction of PMN migration. TEM in basolateral-to-apical direction was about twice as efficient as in apical-to-basolateral direction (Transmigration Index = 3,92 +/- 0,55 and 2,28 +/-,21, respectively). Only basolateral-to-apical TEM could be partly inhibited by preincubation of basolateral membrane with IL-8 antibodies. Conclusion: Inflammatory mediators stimulate PMN adherence to LLC-PK cells and subsequently TEM. The mechanisms involved in TEM stimulated by cytokines or endotoxin appear to be rather changes in surface receptor properties of LLC-PK cells than chemotactic stimuli. Basolateral-to-apical TEM appears to be the favored direction most likely augmented by IL-8 associated haptotactic PMN stimulation. Copyright (C) 2004 S. Karger AG, Basel.