细粒棘球蚴细胞系培育及其免疫研究

从细粒棘球蚴病人体内获取生发层和原头节作为培养材料，采用改良DMEM培养液和鼠胶原蛋白包被的培养瓶进行细胞培养，建立了适合人源细粒棘球蚴细胞体外培养的方法，并探讨棘球蚴细胞培养的制约因素。首次成功培育出一株人源细粒棘球蚴细胞系(13G-5)，至1997年3月1日为止该细胞系已培养了140d，其间传了21代。该细胞系主要以成纤维型细胞为主，呈贴壁生长；用该细胞系细胞接种于昆明(Km)小鼠腹腔内，能形成具包囊样结构的类似包囊;接种鼠产生对囊液抗原的特异性抗体；ELISA能够检测出细胞系细胞代谢物中特异性抗原；该细胞系细胞和E.granulosus原头节、包囊液具有相同的EST酶带；使用该细胞系细胞的代谢物作为粗制抗原进行免疫预防接种，能使60%的Km小鼠获得完全抵抗E.granulosus感染的能力；该抗原能使43.33%～77.42%的包虫病人诊断出来。此外，检定了人源细粒棘球蚴染色体，染色体数为14～18条，并首次对其进行了G-、C-带分析。

The Cell Line Establishment and Immunogenic Study of Echinococcus granulosus

The germinal layer and protoscoleces of larval Echinococcusgranulosus excised surgically from a patient with liver hydatid disease were isolated and grew in the modified DMEM in collagen-coated culture flasks. The optimal condition for larval E.granulosus growth in vitro was established, and effect factors for larval E.granulosus growth were analyzed. The cells were continuously grew in the modified DMEM over 140 days and past 21 passages. The cells grew with the fibroblast-like cells dominating and attached to the glass surface. The results indicated that the cell line of larval E.granulosus, 13G-5, was established. The Km mice were inoculated intraperitoneally with cells of 13G-5 and cyst-like structures which proved to be cyst under microscope. The mice produced specific antibody against hydatid cyst-fluid antigens. The specific antigen was found in the excretory products of 13G-5 by ELISA. The same EST isoenzyme band as that of E.granulosus protoscoleces was identified in 13G-5. After the mice were vaccinated with the crude antigen, the excretory products of 13G-5, 60% Km mice have no cyst formed when infected with E.granulosus. When excretory products of 13G-5 were used as diagnostic antigen, the sensitivity is 43.33%～77.42%. The chromosome of larval E.granulosus were numbered 14 to 18 and G-band and C-band were analyzed for the first time.