【摘要】：目的以攜帶α1-giardin DNA的減毒沙門菌為對象,探討腸溶制劑的制備、免疫劑量和免疫效果。方法攜帶有賈第蟲α1-giardin基因的重組減毒沙門菌經(jīng)培養(yǎng)擴(kuò)增凍干后,對各種輔料,包括微分硅膠、分散劑、潤滑劑、崩解劑、包衣液等成分與減毒沙門菌的相容性,及包衣條件進(jìn)行檢測。采用最佳配方制備腸溶片劑,用于動物免疫試驗(yàn)。初始免疫后第25d后,分別檢測血清和小腸灌洗液中抗rα1-giardin特異性IgG和SIgA的水平;并用106個(gè)賈第蟲滋養(yǎng)體灌胃,每天檢測各組小鼠糞便中的滋養(yǎng)體和(或)包囊的排出率。結(jié)果腸溶片劑的處方中微分硅膠和分散劑的用量分別為菌粉用量的10%和2倍;潤滑劑和崩解劑的用量分別占處方量的0.8%和2%;包衣液的濃度為20%,包衣增重為處方量的6%。每個(gè)包衣片活菌含量在4×106以上。動物試驗(yàn)表明,以SL7207/pVAX1-rα1-giardin腸溶片劑免疫鼠小腸灌洗液中特異性抗rα1-giardin SIgA的水平明顯高于對照組,并在攻蟲試驗(yàn)具有46.7%的減蟲率,明顯高于rα1-giardin肌注免疫組(23%)(P0.01)。結(jié)論本研究為多種以減毒沙門菌為載體的DNA疫苗的腸溶制劑的研究提供了實(shí)驗(yàn)依據(jù),如規(guī)�；a(chǎn),該處方尚需進(jìn)一步完善。
[Abstract]:Objective to study the preparation, immune dose and immune effect of enteric lytic preparation with attenuated Salmonella carrying 偽 1-giardin DNA. Methods after lyophilization of recombinant attenuated Salmonella containing the 偽 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 偽 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 偽 1-giardin SIgA in the intestinal lavage fluid of rats immunized with SL7207/pVAX1-r 偽 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 偽 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.
【作者單位】： 吉林醫(yī)藥學(xué)院;
【基金】：國家自然科學(xué)基金(No.81301450,31572262) 吉林省科技廳國際合作項(xiàng)目(No.20130413035GH) 吉林省教育廳科研項(xiàng)目(No.2014367) 吉林市杰出青年項(xiàng)目(No.201464037)聯(lián)合資助~~
【分類號】：R392

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