人淋巴因子激活的杀伤细胞抗生素肽的分离纯化

Isolation and Purification of Antibacterial Polypeptides from Human LAK cells

目的 分离纯化人淋巴因子激活的杀伤细胞 (lym phokine activated killer,L AK)内源性抗生素肽。方法 采用密度梯度离心法分离人外周血单核细胞 ,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鉴定其抗菌活性。结果 从人 L AK细胞酸溶性提取物中纯化出多个多肽 ,其中 HL P- 2 b、HL P- 3a完全纯化 ,分子量分别为 7.9× 10 3u和 4× 10 3u。HL P- 2 a、HL P- 2 c、HL P- 3b和 HL P- 3c基本纯化 ,主带分子量分别为 7.2× 10 3u、10 .4× 10 3u、6 .4× 10 3u、6 .4× 10 3u。 HL P- 3a具有抗金黄色葡萄球菌活性 ,HL P- 2 a、HL P- 2 b、HL P- 2 c、HL P- 3b、HL P- 3c具有抗金黄色葡萄球菌活性和抗白色念珠菌活性。结论 人 L

Objective To isolate and purify new antibiotic peptides from human lymphokine acfivated killer (LAK) cells. Methods Preparative Acid Urea Polyacrylamide Gel Electrophoresis and Reverse Phase HPLC were performed to isolate and purify polypeptides from the acid extract of human LAK cells. The molecule weight was analyzed by Tricine SDS PAGE. Radial agar diffusion assay was used to analyze the antibacterial activities. Results Several antibiotic peptides were isolated. Two peptides were purified from fractions HLP 2, HLP 3, which had molecular weight of around 7.9×10 3 u and 4×10 3 u and were named HLP 2b and HLP 3a, respectively. Four peptides with molecular weight of 7.2×10 3 u, 10.4×10 3 u, 6.2×10 3 u and 6.2×10 3 u were almost purified and were named HLP 2a, HLP 2c, HLP 3b and HLP 3c, respectively. HLP 2b, HLP 2a, HLP 2c, HLP 3b and HLP 3c all had antimicrobial activities against S. Aureus and C.Albicans, and HLP 3a against S. Aureus only. Conclusion Human LAK cells contained a variety of antimicrobial peptides.