肾上腺髓质素单克隆抗体制备和在人体器官组织中的表达

Preparation of Monoclonal Antibody Against Adrenomedullin and Distribution in Human Organ and Tissues

目的 自制特异性肾上腺髓质素 (AM)单克隆抗体 ,观察AM在人正常组织中的表达。方法 以AM为抗原免疫BALB/c小鼠 ,取其脾细胞与SP2 / 0骨髓瘤细胞株融合 ,建立阳性杂交瘤细胞克隆 ,制备AM单克隆抗体 ;以此为第一抗体 ,采用ABC免疫组化法检测其在人体心、肾、肝、肺和脾组织的表达。结果 AM在上述器官中的分布较广泛 ,表达于这些组织中的血管平滑肌细胞和内皮细胞 ,以及心肌细胞、肾小球上皮细胞和内皮细胞 ,肾小管和集合管上皮细胞 ,肝胆管上皮细胞 ,肺支气管及其黏液腺上皮细胞、平滑肌细胞和肺泡Ⅱ型上皮细胞胞质。结论 AM在人器官组织中分布广泛 ,提示其生物学作用可能是多方面的。

Purpose: To prepare the specific monoclonal antibody against adrenomedullin and to observe the distribution of adrenomedullin(AM) in some normal human organs. Methods: BALB/c mice were immunized with AM protein. Splenocytes derived from the immunized mouse were fused with the SP2/0 myeloma cells. The positive hybridoma clones producing monoclonal antibody against AM were established. With the use of the monoclonal antibody as primary antibody, immunohistochemical assay revealed the expression of AM in normal human organs, including heart, kidney, liver, lung, and spleen. Results: AM was widely expressed on vascular smooth muscle cells and endothelial cells in the detected organs. The immunoreactivity was observed in the cytoplasms of cardiac myocytes, glomerular epithelial and endothelial cells, renal tubules and collecting tubules, biliary ductal epithelia, bronchial mucosa and smooth muscle cells, and type II pneumocytes of the lung. Conclusions: The wide distribution of AM in some organs of human being indicates its multiple biological roles.