Sakashita N, Takeya M, Kishida T, Stackhouse T M, Zbar B, Takahashi K
Second Department of Pathology, Kumamoto University School of Medicine, Japan.
Histochem J. 1999 Feb;31(2):133-44. doi: 10.1023/a:1003554712386.
To examine the localization of von Hippel-Lindau (VHL) protein in human tissues, we produced four novel monoclonal antibodies against human VHL protein. Western blot analysis revealed that two of these antibodies recognized the epitope in amino acid sequence 60-89 of the VHL protein and the others recognized sequences 54-60 and 189-213. In a wild-type VHL gene-transfected cell line, immunocytochemistry and immunoelectron microscopy demonstrated the intracytoplasmic localization of VHL protein, particularly in mitotic cells. In normal human tissues, VHL protein was detected immunohistochemically in epithelial cells covering the body surface and the alimentary, respiratory, and genitourinary tracts; in secretory epithelial cells of exocrine and endocrine organs; in parenchymal cells of visceral organs; in cardiomyocytes; in neurons in nervous tissue; in lymphocytes in lymphoid tissue; and in macrophages. In pathological specimens, VHL protein was expressed in VHL-related tumor, as well as in endothelial cells, fibroblasts, and pericytes, all of which are involved in active angiogenesis. These findings suggest that these monoclonal antibodies can be useful for various immunological assays and that the VHL protein plays fundamental roles in physiological and pathological situations, especially in neovascularization.
为了研究血管母细胞瘤 - 林道（VHL）蛋白在人体组织中的定位，我们制备了四种针对人VHL蛋白的新型单克隆抗体。蛋白质印迹分析表明，其中两种抗体识别VHL蛋白氨基酸序列60 - 89中的表位，其他抗体识别序列54 - 60和189 - 213。在野生型VHL基因转染的细胞系中，免疫细胞化学和免疫电子显微镜显示VHL蛋白定位于细胞质内，特别是在有丝分裂细胞中。在正常人体组织中，通过免疫组织化学检测到VHL蛋白存在于覆盖体表以及消化道、呼吸道和泌尿生殖道的上皮细胞中；存在于外分泌和内分泌器官的分泌上皮细胞中；存在于内脏器官的实质细胞中；存在于心肌细胞中；存在于神经组织的神经元中；存在于淋巴组织的淋巴细胞中；以及存在于巨噬细胞中。在病理标本中，VHL蛋白在VHL相关肿瘤中表达，也在内皮细胞、成纤维细胞和周细胞中表达，所有这些细胞都参与活跃的血管生成。这些发现表明，这些单克隆抗体可用于各种免疫测定，并且VHL蛋白在生理和病理情况下发挥着重要作用，尤其是在新生血管形成方面。
