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福尔马林固定石蜡包埋组织中VIII因子/血管性血友病因子抗原免疫组化检测的特异性和敏感性

Specificity and sensitivity of immunohistochemical detection of factor VIII/von Willebrand factor antigen in formalin-fixed paraffin-embedded tissue.

作者信息

McComb R D, Jones T R, Pizzo S V, Bigner D D

出版信息

J Histochem Cytochem. 1982 Apr;30(4):371-7. doi: 10.1177/30.4.6801111.

Abstract

The immunohistochemical detection of factor VIII/von Willebrand factor antigen (FVIII/vWF-AG) in formalin-fixed paraffin-embedded tissues was investigated using the peroxidase-antiperoxidase (PAP) method. Highly purified human FVIII/vWF was used to raise rabbit anti-FVIII/vWF-AG serum. In addition to anti-FVIII/vWF-AG activity, the unabsorbed antiserum had anti-IgG, anti-IgM, and anti-alpha2-macroglobulin specificities. Following exhaustive absorption with these proteins, the antiserum reacted monospecifically for FVIII/vWF-AG in immunodiffusion, immunoelectrophoresis, and PAP immunohistochemistry. Sections of normal tissues from six patients and a total of 43 neoplasms were examined. Treatment of the tissue sections with trypsin prior to application of the antiserum markedly increased the sensitivity of FVIII/vWF-AG detection. The positive staining for FVIII/vWF-AG was restricted to endothelial cells in both neoplastic and nonneoplastic tissue. In general, the hyperplastic endothelia in neoplastic and reactive tissues stained more intensely than those in normal tissues. Expression of FVIII/vWF-AG by nonendothelial neoplastic cells was not observed. FVIII/vWF-AG is a reliable marker for endothelial cells.

摘要

采用过氧化物酶抗过氧化物酶(PAP)法研究了福尔马林固定石蜡包埋组织中因子VIII/血管性血友病因子抗原(FVIII/vWF-AG)的免疫组化检测。使用高度纯化的人FVIII/vWF来制备兔抗FVIII/vWF-AG血清。除了抗FVIII/vWF-AG活性外,未吸收的抗血清还具有抗IgG、抗IgM和抗α2-巨球蛋白特异性。用这些蛋白质进行彻底吸收后,抗血清在免疫扩散、免疫电泳和PAP免疫组化中对FVIII/vWF-AG呈单特异性反应。检查了6例患者的正常组织切片和总共43个肿瘤。在应用抗血清之前用胰蛋白酶处理组织切片显著提高了FVIII/vWF-AG检测的灵敏度。FVIII/vWF-AG的阳性染色在肿瘤组织和非肿瘤组织中均局限于内皮细胞。一般来说,肿瘤组织和反应性组织中的增生性内皮细胞比正常组织中的染色更强烈。未观察到非内皮肿瘤细胞表达FVIII/vWF-AG。FVIII/vWF-AG是内皮细胞的可靠标志物。

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