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Oral Immunization of Mice With Vaccine of Attenuated Salmonella typhimurium Expressing Helicobacter pylori Urease B Subunit

查看全文 作　　者：XING-LONG [1]YANG;WEN-CHAO [1]LID;Wu-WEI [2]YANG;DONG [3]ZHONG;Yu-Hu [3]LIU;JING-DONG [4]ZHANG;JIAN-HUI [5]JIANG;SHAN-SHAN [2]LI 高影响力作者 机构地区：[1]Department of Oncology, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, Shaanxi, China;[2]Department of Oncology, Affiliated Hospital of the Academy of Military Medical Sciences, Beijing 100039, China;[3]Department of Gastroenterology, Nanfang Hospital Guangzhou 510515, Guangdong, China;[4]Department of Pathology, University of Vermont, 208 South Park, Dr. Suite 2, Colchester, VT05446, U.S.A.;[5]Department of Epidemiology, the Fourth Military Medical University, Xi'an 710032, Shaanxi, China高影响力机构 出　　处：《Biomedical and Environmental Sciences》索引2005年第18卷第6期,共8页高影响力期刊 基　　金：This work was supported by Chinese Postdoctoral Fund (Grant No. 2004035426). 摘　　要：Objective To prepare the live recombinant vaccine of attenuated Salmonella typhimurium SL3261 expressing Helicobacterpylori (H. pylori) B subunit (UreB) and to determine whether it could be used as an oral vaccine against H. pylori infection. Methods Using genomic DNA of H. pylori Sydney strain (SS1) as template, the H. pylori UreB gene fragment was amplified by PCR and subcloned into the expression vector pTC01. The recombinant plasmid pTC01-UreB was then transferred into LB5000 to obtain modified forms, and further conversed into the attenuated Salmonella typhimurium SL3261 to obtain recombinant SL3261/pCT01-UreB as an oral immunization reagent, which was then used to orally immunize Balb/c mice twice at a three-week interval. Twelve weeks later, anti-UreB IgA antibodies in intestinal fluid and IgG antibodies in sera were determined by ELISA. The relating data in control groups (including body weight, gastric inflammation, etc.) were also collected. Results The sequencing analysis showed that the UreB gene fragment amplified by PCR was consistent with the sequence of the H. pylori UreB gene. The restriction enzyme digestion revealed that the correct pTC01-UreB was obtained.SDS-PAGE and Western blot showed that a 61KD protein was expressed in SL3261/pTC01-UreB, which could be recognized by anti-H. pylori UreB antiserum and was absent in the control containing only Salmonella typhimurium SL3261 strain. The multiple oral immunization with SL3261/pTC01-UreB could significantly induce H. pylori specific mucosal IgA response as well as serum IgG responses. IFN-γ and IL-10 levels were significantly increased in SL3261/pTC01-UreB group, and no obvious side effect and change in gastric inflammation were observed. Conclusion The attenuated vaccine of Salmonella typhimurium expressing H. pylori UreB can be used as an oral vaccine against H. pylori infection. 关 键 词：免疫机制 减毒活疫苗 沙门氏菌 幽门螺杆菌
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