Mouse glomerular epithelial cells in culture with features of podocytes in vivo express aminopeptidase A and angiotensinogen but not other components of the renin-angiotensin system.
Mouse glomerular epithelial cells in culture with features of podocytes in vivo express aminopeptidase A and angiotensinogen but not other components of the renin-angiotensin system.
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发表时间：
1997-05
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通讯作者：
S. Mentzel;J. V. Son;A. S. D. Jong;H. Dijkman;R.A.P. Koene;J. F. Wetzels;K. Assmann
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作者：
S. Mentzel;J. V. Son;A. S. D. Jong;H. Dijkman;R.A.P. Koene;J. F. Wetzels;K. Assmann
与足细胞抗原如Heymann抗原或氨基肽酶A的抗体结合可导致几个物种的膜性肾小球肾炎。为了研究抗体与这些足细胞上的抗原未来可能的相互作用，我们培养了分离的小鼠肾小球上皮细胞。通过间接免疫荧光，细胞角蛋白、波形蛋白、结蛋白和紧密连接复合体的一种成分ZO-1蛋白呈阳性。当使用鼠单抗时，这些细胞也对水解酶氨基肽酶A和二肽基肽酶IV呈阳性，并用ASD-33染色，ASD-33是一种识别只存在于小鼠足细胞细胞膜上的表位的单抗。它们的von Willebrand因子均为阴性，并且不与与肾小球和肾小管周围毛细血管内皮细胞结合的单抗(ASD-13)染色。在电子显微镜下，细胞连接紧密，但缺少韦氏小体(内皮)、桥粒和纤毛(壁上皮)。此外，还检测了肾素-血管紧张素系统的几种成分的mRNA表达，并通过RT-PCR分析了一些与肾素-血管紧张素系统成分血管紧张素II间接偶联的因子。发现血管紧张素II降解水解酶氨基肽酶A和血管紧张素原的mRNA表达，但没有发现该系统的任何其他成分，如肾素、血管紧张素转换酶或血管紧张素II受体AT1a、AT1b和AT2。二肽基肽酶IV的mRNA表达水平较低。此外，还观察到转化生长因子-β和白介素7的生长因子以及细胞外基质成分纤维连接蛋白、层粘连蛋白B2、Perlecan和IV型胶原α1的表达。鉴于这些特点，建立了一种具有体内足细胞特征的肾小球上皮细胞培养方法，该方法将为进一步研究抗氨基肽酶A单抗和血管紧张素II与氨基肽酶A的相互作用奠定基础。这是有趣的，因为观察到注射抗氨基肽酶A抗体的小鼠会引起急性蛋白尿。
The binding of antibodies to podocytic antigens such as the Heymann antigen or aminopeptidase A may lead to the induction of a membranous glomerulonephritis in several species. To study the possible future interactions of antibodies with antigens on these podocytes, epithelial cells from isolated mouse glomeruli were cultured. By indirect immunofluorescence, the cells were positive for cytokeratin, vimentin, desmin, and the ZO-1 protein, a component of the tight junction complex. When rat monoclonal antibodies were used, the cells were also positive for the hydrolases aminopeptidase A and dipeptidyl peptidase IV, and they stained with ASD-33, a monoclonal antibody that recognized an epitope only present on the cell membranes of mouse podocytes. They were negative for the von Willebrand factor and did not stain with a monoclonal antibody (ASD-13) that binds to endothelial cells of glomeruli and peritubular capillaries. By electron microscopy, the cells showed tight junctions but lacked Weibel Palade bodies (endothelium), desmosomes, and cilia (parietal epithelium). The mRNA expression of several components of the renin-angiotensin system was also examined, and some factors indirectly coupled to the renin-angiotensin system component angiotensin II in this podocytic culture by RT-PCR analysis. mRNA Expression for the angiotensin II degrading hydrolase aminopeptidase A and angiotensinogen was found, but this was not found for any other component of this system, such as renin, angiotensin-converting enzyme, or the angiotensin II receptors AT1a, AT1b, and AT2. Low mRNA expression for dipeptidyl peptidase IV was observed. In addition, expression of the growth factors transforming growth factor-beta and interleukin-7, and the extracellular matrix components fibronectin, laminin B2, perlecan, and collagen IV alpha 1, was observed. Given these characteristics, a glomerular epithelial cell culture with features of podocytes in vivo that will allow future studies on the interaction of anti-aminopeptidase A monoclonal antibodies and angiotensin II with aminopeptidase A was established. This is of interest in light of the observation that injection of mice with anti-aminopeptidase A antibodies causes an acute albuminuria.