中华医学杂志（英文版）2002，Vol.115Issue(10)：1513-1516,4.
表达幽门螺杆菌尿素酶B亚单位的减毒鼠伤寒杆菌口服疫苗的制备
Oral immunization of mice with attenuated Salmonella typhimu rium expressing Helicobacter pylori urease B subunit
刘晓峰 1张霞 1胡家露 2樊代明2
作者信息
- 1. 济南军区总医院消化科,济南,250031
- 2. 第四军医大学西京医院全军消化病研究所,西安,710032
- 折叠
摘要
Abstract
Objective To establish attenuated Salmonella typhimu rium producing Helicobacter pylori (H. pylori) urease subunit B (UreB) and determine whether it could be used as an oral vaccine against H. pylori.Methods H. pylori (SS1 strain) UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector pTC01 after sequencing, and then transformed into attenuated Salmonella typhimurium SL32 61 to acquire SL3261/pTC01-UreB. The expression of H. pylori UreB in SL32 61 was detected by Western blot. Twelve weeks after oral immunization of mice, antibody responses were evaluated using serum and intestinal fluid by ELISA assay. Interferon-γ (IFN-γ) and interleukin-10 (IL-10) in the supernatant of spleen cells culture were also assessed by ELISA. Invitro stability of pTC01-UreB plasmid in SL3261 was confirmed by growing in Luria Broth (LB) medi um to 80 generations. Results The UreB gene fragment amplified by PCR was co nsistent with the sequence of the H. pylori UreB as evidenced by sequence analysis. Enzyme digestion revealed that the correct pTC01-UreB was obtained. Western blot showed that a 61kDa protein was expressed in SL3261/pTC01-UreB, wh ich could be recognized by anti-H. pylori UreB antiserum. After 80 genera tions of continuous culture, the recombinant plasmid pTC01-UreB was stable in S L3261 and had no obvious toxicity. Multiple oral immunizations with SL3261/pTC0 1-UreB could significantly induce H. pylori-specific mucosal IgA response as well as serum IgG response. Moreover, there were significant increases of IFN-γand IL-10 in the SL3261/pTC01-UreB group. Finally, no obvious side eff ects for mice and no change in gastric inflammation were observed. Conclusion Attenuated Salmonella typhimurium expre ssing H. pylori UreB may be used as oral vaccine against H. pylori inf ection.关键词
幽门螺杆菌/尿素酶/鼠伤寒杆菌/疫苗Key words
Helicobacter pylori/urease/Sal monella typhimurium/vaccine分类
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刘晓峰,张霞,胡家露,樊代明..表达幽门螺杆菌尿素酶B亚单位的减毒鼠伤寒杆菌口服疫苗的制备[J].中华医学杂志（英文版）,2002,115(10):1513-1516,4.
