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表达幽门螺杆菌过氧化氢酶的减毒鼠伤寒沙门氏菌疫苗株的构建及其保护性免疫观察

[Construction of attenuated Salmonella typhimurium vaccine strain expressing Helicobacter pylori catalase and observation on its protective immunity].

作者信息

Liao W, Chen M, Zhu S

机构信息

Department of Gastroenterology, First Affiliated Hospital, Sun Yat-Sen University of Medical Sciences, Guangzhou 510080, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2001 May 25;81(10):613-6.

Abstract

OBJECTIVE

To investigated the effect of attenuated Salmonella typhimurium vaccine strain expressing Helicobacter pylori (Hp) catalase (KatA) on protection against Hp infection.

METHODS

Recombinant plasmid expressing KatA was constructed. Expression of KatA was induced with IPTG, and analyzed by SDS-PAGE and Western blot. The recombinant plasmid were introduced into attenuated Salmonella typhimurium SL3261 strain to construct live oral vaccine strain. C57BL/6 mice was orally immunized with this vaccine strain and then given orogastric challenge with live Hp Sydney strain. The stomach samples were submitted to a rapid urease test and quantitative culture.

RESULTS

The SDS-PAGE showed a dominant additional protein with molecular weight of 79kDa which could specially react with antibody against GST and accounted for 19% of total bacteria proteins. Animal experiment indicated that this vaccine strain could protect mice against Hp infection.

CONCLUSION

Attenuated Salmonella typhimurium vaccine strain expressing KatA could induce effective immune response in protection against Hp infection,which may play an important role in preventing and treating Hp infection and related diseases.

摘要

目的

研究表达幽门螺杆菌(Hp)过氧化氢酶(KatA)的减毒鼠伤寒沙门氏菌疫苗株对预防Hp感染的作用。

方法

构建表达KatA的重组质粒。用异丙基-β-D-硫代半乳糖苷(IPTG)诱导KatA表达,并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹法(Western blot)进行分析。将重组质粒导入减毒鼠伤寒沙门氏菌SL3261株构建口服活疫苗株。用该疫苗株对C57BL/6小鼠进行口服免疫,然后用活的Hp悉尼株进行灌胃攻击。取胃样本进行快速尿素酶试验和定量培养。

结果

SDS-PAGE显示有一条分子量为79kDa的主要额外蛋白带,它能与抗谷胱甘肽S-转移酶(GST)抗体特异性反应,占细菌总蛋白的19%。动物实验表明该疫苗株能保护小鼠免受Hp感染。

结论

表达KatA的减毒鼠伤寒沙门氏菌疫苗株能诱导有效的免疫反应以预防Hp感染,这可能在预防和治疗Hp感染及相关疾病中发挥重要作用。

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