钉螺凝集素的分离纯化及免疫活性的研究

Study on purification and immunological activity of agglutinin from Oncomelania hupensis

目的:探讨湖北钉螺体内凝集素的提取纯化方法及其免疫活性。方法:钉螺原浆用30%硫酸铵进行粗提,Sepharose-4B亲和层析,用十二烷基磺酸钠-聚丙烯酸胺(SDS-PAGE)凝胶电泳对纯化后的凝集素进行浓度测定,并计算相对分子质量。用纯化后的凝集素行促血淋巴细胞吞噬杀菌实验、抑菌实验,并检测2.5 mg.ml-1的D-葡萄糖、蔗糖、D-半乳糖、EDTA对凝集素免疫活性的影响。结果:Sepharose-4B亲和层析对钉螺凝集素的纯化具有特异性,钉螺亚基相对分子质量约为57 kD;凝集素能促使血淋巴细胞吞噬和杀灭细菌,对细菌生长有抑制作用,上述免疫活性能被D-半乳糖和EDTA抑制。结论:钉螺体内存在半乳糖凝集素,并有促噬杀、抑菌活性,能为EDTA所抑制。

Objective： To explore the extraction and and study its immune activity. Methods： Protein of saturation of ammonium sulfate. The extraction of the purification methods of agglutinin from Oncomelania hupensis Oncomelania hupensis was preliminary extracted with 30% Oncomelania hupensis agglutinin was purified by Sepharose- 4B. The purity of the agglutinin was identified by SDS-PAGE and the subunit molecular weight was calculated. The purity was applied for the tests of promoting phagocytosis and bacteriostasis. The immune activity of purity was detected with 2.5 mg ·ml^-1 D- glucose, saccharose, D- galactose, EDTA. Results： Sepharose-4B was specific on purification of agglutinin. The subunit molecular weigh was 57 kD. The agglutinin could promote haemocytes phagocytosis and killing bacteria. It also could inhibit the growth of bacteria. The above immunological activity could be inhibited by D-galactose and EDTA. Conclusion： Galactose specific agglutitin exists in Oncomelania hupensis, it could promote haemocytes phagocytosis and killing bacteria, also could inhibit the growth of bacteria. The above activity is inhibited by EDTA.