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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。
OBJECTIVE: To isolate and purify small molecule antimicrobial peptides from human LAK cells and test the activity of anti-tumor cell line K562. Methods: Human peripheral blood mononuclear cells were isolated by density gradient centrifugation and stimulated with IL 2 and PHA. 5% acetic acid homogenate cells to obtain their acid-soluble extract. Peptide was isolated and purified by preparative acidic polyacrylamide gel electrophoresis and reversed-phase high performance liquid chromatography. The molecular weight was identified by Tricine SDS-PAGE. The antibacterial and antitumor activity was evaluated by agarose diffusion assay and MTT assay. Results: A polypeptide was purified from acid-soluble extracts from human LAK cells. The polypeptide was named as HLP 2b with a molecular weight of 7 9 kD and had anti-Staphylococcus aureus activity and anti-tumor cell K5 6 2 activity. Conclusion: HLP 2b may be a new anti-tumor anti-tumor effector molecule of LAK cells.