以减毒沙门氏菌为载体的抗贾第虫肠溶疫苗制备及免疫效果检测

发布时间：2018-11-13 20:05

【摘要】：目的以携带α1-giardin DNA的减毒沙门菌为对象,探讨肠溶制剂的制备、免疫剂量和免疫效果。方法携带有贾第虫α1-giardin基因的重组减毒沙门菌经培养扩增冻干后,对各种辅料,包括微分硅胶、分散剂、润滑剂、崩解剂、包衣液等成分与减毒沙门菌的相容性,及包衣条件进行检测。采用最佳配方制备肠溶片剂,用于动物免疫试验。初始免疫后第25d后,分别检测血清和小肠灌洗液中抗rα1-giardin特异性IgG和SIgA的水平;并用106个贾第虫滋养体灌胃,每天检测各组小鼠粪便中的滋养体和(或)包囊的排出率。结果肠溶片剂的处方中微分硅胶和分散剂的用量分别为菌粉用量的10%和2倍;润滑剂和崩解剂的用量分别占处方量的0.8%和2%;包衣液的浓度为20%,包衣增重为处方量的6%。每个包衣片活菌含量在4×106以上。动物试验表明,以SL7207/pVAX1-rα1-giardin肠溶片剂免疫鼠小肠灌洗液中特异性抗rα1-giardin SIgA的水平明显高于对照组,并在攻虫试验具有46.7%的减虫率,明显高于rα1-giardin肌注免疫组(23%)(P0.01)。结论本研究为多种以减毒沙门菌为载体的DNA疫苗的肠溶制剂的研究提供了实验依据,如规模化生产,该处方尚需进一步完善。
[Abstract]:Objective to study the preparation, immune dose and immune effect of enteric lytic preparation with attenuated Salmonella carrying 伪 1-giardin DNA. Methods after lyophilization of recombinant attenuated Salmonella containing the 伪 1-giardin gene of Giardia lamblia, the compatibility of various adjuvants, including differential silica gel, dispersant, lubricant, disintegrating agent, coating solution and so on, with attenuated Salmonella was studied. And the coating conditions are tested. The best formulation was used to prepare enteric tablets for animal immunity test. The levels of anti-r 伪 1-giardin specific IgG and SIgA in serum and small intestinal lavage fluid were measured 25 days after initial immunization, and 106 gibberellism trophozoites were used to gavage the excretion rate of trophozoites and / or cyst in feces of each group of mice. Results the dosage of differential silica gel and dispersant was 10% and 2 times as much as that of bacterial powder respectively, and the dosage of lubricant and disintegrating agent accounted for 0.8% and 2% of the prescriptions, respectively. The concentration of the coating solution was 20 and the weight gain of the coating was 6% of the prescription. The content of live bacteria in each coating tablet was more than 4 脳 10 ~ 6. Animal experiments showed that the level of specific anti-r 伪 1-giardin SIgA in the intestinal lavage fluid of rats immunized with SL7207/pVAX1-r 伪 1-giardin was significantly higher than that in the control group, and the worm reduction rate was 46.7% in the attack test. It was significantly higher than that in r 伪 1-giardin immunized group (23%) (P0.01). Conclusion this study provides an experimental basis for the study of enteric lytic preparations of DNA vaccine with attenuated Salmonella. The formulation needs to be further improved if it is produced on a large scale.
【作者单位】：吉林医药学院;
【基金】：国家自然科学基金(No.81301450,31572262) 吉林省科技厅国际合作项目(No.20130413035GH) 吉林省教育厅科研项目(No.2014367) 吉林市杰出青年项目(No.201464037)联合资助~~
【分类号】：R392

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