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观察旋毛虫肌肉期幼虫分泌排泄物(TsL1-ES)及其46-58KD蛋白的保护性免疫作用,并作比较研究。方法:通过无血清的RPMI1640培养基培养旋毛虫肌肉期幼虫,获得TsL1-ES;再以常规SDS-PAGE和电洗脱技术相结合纯化TsL1-ES中46-58KD蛋白。用TsL1-ES及其46-58KD蛋白加福氏完全佐剂(FCA)腹腔注射免疫昆明小鼠3次,每鼠再以旋毛虫300条感染攻击,观察感染后第7天鼠肠道成虫数、第30天肌肉幼虫数及血清抗体滴度。结果:46-58KD免疫组鼠的肠道成虫减虫率、肌肉幼虫减虫率及血清抗体的几何平均倒数滴度(GMRT)分别为46.4%、46.1%及2841.2;与TsL1-ES诱导的保护性免疫力接近(48.3%、50.2%及3458.6);明显高于佐剂对照组(4.8%、8.2%及748.5。结论:TsL1-ES及其46-58KD蛋白均可产生明显的保护性免疫,且保护性免疫水平相近。
The protective immune function of TsL1-ES and its 46-58KD protein secreted by muscle larvae of Trichinella were observed and compared. Methods: TsL1-ES was obtained by culturing Trichinella spiralis larvae in serum-free RPMI 1640 medium. The 46-58KD protein in TsL1-ES was purified by conventional SDS-PAGE and electroelution technique. Kunming mice were immunized intraperitoneally with TsL1-ES and its 46-58KD protein plus Freund’s complete adjuvant (FCA) for 3 times. Each mouse was challenged with 300 Trichinella spiralis worms. The number of adult mice in the intestinal tract was observed on the 7th day after infection. Number of muscle larvae on day 30 and serum antibody titer. Results: The intestinal worm reduction rate, muscle worm larval reduction rate and the geometric mean reciprocal titers (GMRT) of 46-58KD immunized mice were 46.4%, 46.1% and 2841.2, respectively. Compared with TsL1- Protective immunity induced by ES was similar (48.3%, 50.2% and 3458.6%) and significantly higher than that of the adjuvant control group (4.8%, 8.2% and 748.5, respectively) .Conclusion: TsL1- ES and its 46-58KD protein can produce significant protective immunity, and protective immunity levels are similar.