Schleicher E D, Wagner E M, Olgemöller B, Nerlich A G, Gerbitz K D
Klinisch-Chemisches Institut, Universität München, Federal Republic of Germany.
Lab Invest. 1989 Sep;61(3):323-32.
A polyclonal antiserum was raised in rabbits against basement membrane heparan sulfate proteoglycan (HSPG) purified from extracts of isolated porcine glomeruli. The antiserum was characterized by enzyme immunoassay, immunoprecipitation, and immunohistological methods. It was shown to recognize specifically the core protein of HSPG from porcine, rat, bovine, and human glomerular basement membrane, but it did not bind to HSPG from guinea pig or rabbit kidney. The affinity-purified antiserum did not cross-react with other basement membrane proteins like laminin, fibronectin, or collagen type IV. Immunohistochemical studies on tissue sections from several human organs revealed specific basement membrane staining, although the intensity of the reaction differed among the organs tested. Whereas glomerular and other capillary basement membranes showed prominent staining with antibody to the core protein of the proteoglycan, those from nerve, skeletal, cardiac, and smooth muscle reacted only weakly. Adipocytes and liver sinusoid walls were not stained. Independent of the extent of HSPG staining the basement membranes of all different tissues tested reacted strongly with an antiserum against type IV collagen.
用从分离的猪肾小球提取物中纯化的基底膜硫酸乙酰肝素蛋白聚糖(HSPG)在兔体内制备了多克隆抗血清。通过酶免疫测定、免疫沉淀和免疫组织学方法对抗血清进行了表征。结果表明,该抗血清能特异性识别来自猪、大鼠、牛和人肾小球基底膜的HSPG核心蛋白,但不与豚鼠或兔肾的HSPG结合。亲和纯化的抗血清不与层粘连蛋白、纤连蛋白或IV型胶原等其他基底膜蛋白发生交叉反应。对来自几个人体器官的组织切片进行的免疫组织化学研究显示出特异性的基底膜染色,尽管在所测试的器官中反应强度有所不同。肾小球和其他毛细血管基底膜用针对蛋白聚糖核心蛋白的抗体染色明显,而神经、骨骼、心脏和平滑肌的基底膜反应较弱。脂肪细胞和肝血窦壁未染色。与HSPG染色程度无关,所有测试的不同组织的基底膜都与抗IV型胶原抗血清发生强烈反应。