减毒鼠伤寒沙门氏菌防龋疫苗免疫定菌鼠的实验研究  

Research of active immunization with recombinant avirulence Salmonella typhimurium vaccine against dental caries in gnotobiotic rats

作　　者：胡晓莉[1] 凌均棨[1] 陈罕[1] 

机构地区：[1]中山医科大学口腔医学院光华口腔医院,广东广州510060

出　　处：《牙体牙髓牙周病学杂志》2002年第8期416-420,共5页Chinese Journal of Conservative Dentistry

基　　金：广东省自然科学基金项目 (970 10 5 )

摘　　要：目的 :本研究通过建立定菌鼠模型 ,采用携带变形链球菌PAcA区片段的减毒鼠伤寒沙门氏菌S .typhimuriumSL32 6 1(pET - 17b/A)防龋疫苗 ,经不同途径免疫定菌大鼠 ,观察其在大鼠肠道定居的能力和诱发的特异性免疫应答反应。方法 :采用携带变形链球菌表面蛋白基因片段的重组减毒鼠伤寒沙门氏菌S .typhimuriumSL32 6 1(pET - 17b/A) ,通过口服、皮下、粘膜下等途径免疫 5 0只定菌SD大白鼠 ,以酶联免疫吸附试验对鼠血清及唾液中抗PAc-IgG、SIgA水平进行测定。采集不同时间大鼠粪便样本 ,观察减毒鼠伤寒沙门氏菌在肠道的定居情况 ,并进行质粒抽提鉴定。结果 :口服活菌苗组免疫后血清PAc -IgG及唾液SIgA水平皆明显高于免疫前 (P <0 .0 1)。各组免疫水平比较 ,口服活菌苗组免疫水平也明显高于其它 4组 (P <0 .0 1)。其大鼠粪便中存在大量氨苄抗性的沙门氏菌 ,即沙门氏菌SL32 6 1(pET - 17b/A)。结论 :用基因重组减毒鼠伤寒沙门氏菌免疫定菌大鼠 ,能刺激其产生免疫反应。重组减毒鼠伤寒沙门氏菌S .typhimuriumSL32 6 1(pET - 17b/A)可在大鼠肠道较持久的寄生 ,并能较稳定地携带质粒。AIM:To evaluate the effectiveness of gene recombinant attenuated Salmonella typhimurium SL3261(pET-17b/A)in inducing protective immune responses against S.mutans-induced dental caries in experimental model of caries and observe the ability of . typhimurium SL3261(pET-17b/A)parasiting in intestinal trace. METHODS:50 Sprague-Dawley rats were immunized with S. typhimurium SL3261(pET-17b/A) expressing surface protein PAc A region of 35×10 3 by oral, subcutaneous injection, and submucous injection. S. mutans were inoculated onto rat teeth and the rats were fed with caries -inducing diet 2000 # . Saliva and serum samples were collected and assayed by enzyme-linked immunosorbent assay (ELISA). The colonization of S.typhimurium SL3261 (pET-17b/A) in rats' guts was examined by selected culture medium LB-Amp. RESULTS:In orally immunized with recombinant attenuated live of rats typhimurium SL3261(pET-17b/A) the levels of serum anti-PAc IgG and saliva IgA were higher than before, and also higher than that of rats immunized by subcutaneous injection, submucous injection, and with S.typhimurium SL3261 by oral immunization. The feces of rats orally immunized by S.typhimurium SL3261 (pET-17b/A) had a large numbers of S.typhimurium SL3261 (pET-17b/A). CONCLUSION:This study suggested that the effective use of recombinant attenuated live Salmonella typhimurium as oral vaccine induces the protective immune response and may confer protection against dental caries. The recombinant attenuated S.typhimurium can parasite in the intestinal tract for a long time and be safe for the rats.

关 键 词：减毒鼠伤寒沙门氏菌防龋疫苗 基因重组 免疫反应 龋病 口服接种 皮下接种 粘膜下接种 

分 类 号：R780.1[医药卫生—口腔医学]