Welle M M, Proske S M, Harvima I T, Schechter N M
Institut für Veterinär Pathologie der Freien Universität Berlin, Germany.
J Histochem Cytochem. 1995 Nov;43(11):1139-44. doi: 10.1177/43.11.7560896.
We examined three tissue samples from each of four cows with non-lesional skin, tissue samples from a cow with multiple cutaneous mast cell tumors, and samples from another cow in which mast cells were infiltrating multiple lymphosarcomas of the skin, for the presence of tryptase and chymase by enzyme cytochemical and immunohistological methods. The enzyme activities of tryptase and chymase were tested using N-carbobenzoxy-glycilglycil-L-arginine-2-naphthylamide (Z-Gly-Gly-Arg-NA) and naphthol-AS-D-chloroacetate (N-AS-D-CA) as substrates, respectively. Tryptase reactivity could be demonstrated in frozen and Carnoy-fixed paraffin sections. Chymase reactivity was seen in neither frozen nor paraffin sections of formalin- or Carnoy-fixed skin tissues. Antibody linkage with a polyclonal rabbit anti-human skin tryptase antibody was highly specific in bovine normal cutaneous, infiltrating, and tumor mast cells. More than 90% of the tumor mast cells were distinctly tryptase-positive. With alcian blue, only slightly more than 10% of the mast cells stained clearly positive and with methylene blue hardly any staining of mast cell granules could be demonstrated. No antibody labeling of mast cell granules in any of the tissue sections was detected by the use of rabbit anti-dog chymase antiserum. These results indicate that there is a striking antigenic similarity of bovine tryptase to its canine and human equivalents. The demonstration of tryptase is an important tool in confirming the diagnosis of undifferentiated mast cell tumors. In contrast to other species, chymase appears to be completely absent in bovine skin mast cells.
我们用酶细胞化学和免疫组织学方法,检测了4头皮肤无病变奶牛的3个组织样本、1头患有多发性皮肤肥大细胞瘤奶牛的组织样本,以及另一头肥大细胞浸润皮肤多发性淋巴肉瘤奶牛的样本,以检测类胰蛋白酶和糜蛋白酶的存在情况。分别使用N-苄氧羰基-甘氨酰甘氨酰-L-精氨酸-2-萘酰胺(Z-Gly-Gly-Arg-NA)和萘酚-AS-D-氯乙酸酯(N-AS-D-CA)作为底物,检测类胰蛋白酶和糜蛋白酶的酶活性。在冷冻和卡诺固定的石蜡切片中均可显示类胰蛋白酶反应性。在福尔马林或卡诺固定的皮肤组织的冷冻切片或石蜡切片中均未观察到糜蛋白酶反应性。与多克隆兔抗人皮肤类胰蛋白酶抗体的抗体连接在牛正常皮肤、浸润性和肿瘤性肥大细胞中具有高度特异性。超过90%的肿瘤肥大细胞明显呈类胰蛋白酶阳性。用阿尔辛蓝染色时,只有略多于10%的肥大细胞染色明显阳性,用亚甲蓝几乎无法显示肥大细胞颗粒的染色。使用兔抗犬糜蛋白酶抗血清未检测到任何组织切片中肥大细胞颗粒的抗体标记。这些结果表明,牛类胰蛋白酶与其犬类和人类同类物存在显著的抗原相似性。类胰蛋白酶的检测是确诊未分化肥大细胞瘤的重要工具。与其他物种不同,牛皮肤肥大细胞中似乎完全不存在糜蛋白酶。