人LAK细胞抗菌杀瘤活性多肽HLP-2b的分离纯化

目的 :分离纯化人LAK细胞小分子抗菌多肽 ,并检测其抗肿瘤细胞株K5 6 2活性。方法 :采用密度梯度离心法分离人外周血单个核细胞 ,并用IL 2和PHA刺激培养。 5 %乙酸匀浆细胞获得其酸溶性提取物。应用制备性酸性尿素聚丙烯酰胺凝胶电泳技术和反相高效液相色谱技术分离纯化多肽 ,Tricine SDS PAGE鉴定其分子量 ,并运用琼脂糖弥散法、MTT法鉴定其抗菌、抗肿瘤活性。结果 :从人LAK细胞酸溶性提取物中纯化出一多肽 ,分子量为 7 9kD ,命名为HLP 2b ,具有抗金黄色葡萄球菌活性和抗肿瘤细胞株K5 6 2活性。结论 :HLP 2b可能为LAK细胞一新的抗菌抗瘤效应分子。

Isolation and purification of antibacterial and anti-tumor polypeptide from human LAK cells

Objective:To isolate and purify new peptides with anti-tumor and antibacterial activity from human LAK cells.Methods:Preparative Acid-Urea Polyacrylamide Electrophoresis and Reverse Phase HPLC were performed to isolate and purify polypeptides from acid extract of human LAK cells. The molecule weight was analyzed by Tricine-SDS-PAGE. Radial agar diffusion assay and MTT assay were used to analyze the antibacterial and anti-tumor activities of isolated polypeptides.Results:A 7.9 kD bioactive polypeptide that had anti-tumor and antibacterial activities was purified from human LAK cells, which was named as HLP-2b.Conclusion:HLP-2b may be a new effector molecule of human LAK cells.