抗角蛋白单克隆抗体5G5对培养角质形成细胞增殖的影响

目的 研究抗角蛋白单克隆抗体5G5在无血清培养的角质形成细胞中的免疫学活性,观察其对角质形成细胞增殖的影响。方法 免疫组化观察5G5对角质形成细胞的反应性;从培养的角质形成细胞中提取角蛋白,免疫印迹法检测5G5所识别的角蛋白区带;四甲基偶氮唑蓝比色法(MTT法)测定5G5对细胞增殖的影响。结果 5G5仅识别角质形成细胞中相对分子质量为50000的角蛋白区带,呈现为一条很强的染色带,而未与其它分子质量的角蛋白反应;免疫组化染色明显呈胞浆阳性;该单克隆抗体明显抑制培养的角质形成细胞增殖,并且存在剂量依赖关系。结论 特异性抗相对分子质量为50000角蛋白的抗体可能是角蛋白抗体家族中影响角质形成细胞增殖的有效成分。

Effect of an Anti-Keratin Monoclonal Antibody 5G5 on the Proliferation of Cultured Keratinocytes

Objective To investigate the immunological activity and the effect of an anti-keratin monoclonal antibody 5G5 on the proliferation of serum-free cultured keratinocytes. Methods The reactivity of 5G5 with cultured keratinocytes was observed by immunohistohemistry technique. The keratins extracted from the cultured keratinocytes was identified by SDS-PAGE and their reactivities with 5G5 were assessed by immunoblot analysis. MTT was used to study the influences of 5G5 on the proliferation of cultured keratinocytes. Results 5G5 recognized the band of keratin with molecular weight of 50 000 only. Positive staining of 5G5 was found in the cytoplasm of cultured keratinocytes. The strong potential of 5G5 to inhibit the proliferation of cultured keratinocytes was shown by MTT assay with dose-dependent manner. Conclusions The antibody which specifically recognizes the keratin with molecular weight of 50 000 may be the effective component to inhibit the proliferation of cultured keratinocytes.