植物乳杆菌凝集素的纯化及其体外抗HIV活性

目的纯化植物乳杆菌(L.plantarum PCL)凝集素,并检测其体外抗HIV活性。方法采用Sephadex G-25凝胶脱盐层析和HITrap Q Sepharose XL离子交换层析纯化L.plantarum PCL凝集素;测定纯化产物的兔红细胞凝集素活性、纯度及荧光光谱;采用MTT法检测其对C8166淋巴细胞的毒性作用,以光镜检查其对HIV-1ⅢB诱导C8166细胞形成合胞体的抑制作用,并计算TI值。结果纯化的L.plantarum PCL凝集素经尿素-SDS-PAGE,可见相对分子质量约85000的单一条带;荧光光谱可见在380nm附近有一个发射强度较大的凝集素特征荧光发射峰;L.plantarum PCL凝集素对C8166细胞的IC50为2.239μg/ml,对HIV-1ⅢB诱导C8166细胞合胞体形成的EC50为0.079μg/ml,TI值为28.34。结论已成功纯化了L.plantarum PCL凝集素,并具有抗HIV活性。

Purification and In Vitro Anti-HIV Activity of L. plantarum PCL Lectin

Objective To purify L. plantarum PCL lectin and study its in vitro anti-HIV activity. Methods L. plantarum PCL lectin was purified from L. plantarum PCL by Sephadex G-25 gel desalination chromatography and HITrap Q Sepharose XL ion exchange chromatography and determined for rabbit erythrocyte agglutination activity, purity and fluorescence spectrum. Test the toxicity of purified protein to C8166 lymphocytes from L. plantarum by MTT method, and observe its inhibiting effect on synplasm formation in C8166 lymphocytes induced by HIV-1ⅢB by light microscopy. Calculate the TI value. Results The purified protein showed a single band with relative molecular mass of about 85 000 on urea-SDS-PAGE profile and a characteristic fluorescent emission peak with high intensity at wavelength of about 380 nm on fluorescence spectrum. The IC50 value of purified protein to C8166 lymphocytes was 2. 239 μg / ml, and its EC50 to synplasm formation in C8166 lymphocytes induced by HIV-1ⅢB was 0. 079 μg / ml. The TI value was 28. 34. Conclusion L. plantarum PCL lectin was successfully purified and showed anti-HIV activity.