Zhang Rongguang, Wang Chen, Cheng Wenbin, Duan Guangcai, Shi Qingfeng, Chen Shuaiyin, Fan Qingtang
Department of Epidemiology and Statistics, College of Public Health, Zhengzhou University, Kexue Avenue, Zhengzhou, 450001, People's Republic of China.
Henan Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang Medical University, Xinxiang, 453003, People's Republic of China.
Biotechnol Lett. 2018 Mar;40(3):585-590. doi: 10.1007/s10529-017-2502-3. Epub 2018 Jan 3.
To develop a safe and effective oral vaccine against Helicobacter pylori using its HpaA protein expressed in Lactococcus lactis.
The gene encoding HpaA was obtained by PCR and ligated to pNZ8110-lysM following digestion with NaeI + SphI. The recombinant plasmid was transferred into E. coli for multiplication, and then into L. lactis. The recombinant L. lactis was induced to express HpaA, resulting in two products of 29 and 25 kDa, both of which yielded positive immunoreaction with mouse antisera against H. pylori, as confirmed by immunoblot assays. The 29 kDa product constituted 12% of the cell lysates. Oral inoculation with the engineered L. lactis evoked significantly elevated serum IgG level in mice (P < 0.05).
A novel engineered L. lactis strain was developed that efficiently produces whole HpaA protein with desired antigenicity and potent immunogenicity. It provides a basis for approaches to L. lactis-delivered anti-H. pylori vaccination.
利用在乳酸乳球菌中表达的幽门螺杆菌HpaA蛋白开发一种安全有效的口服疫苗。
通过PCR获得编码HpaA的基因,并用NaeI + SphI消化后连接到pNZ8110-lysM上。重组质粒先转入大肠杆菌进行扩增,然后转入乳酸乳球菌。诱导重组乳酸乳球菌表达HpaA,产生两种分子量分别为29 kDa和25 kDa的产物,免疫印迹分析证实这两种产物与抗幽门螺杆菌小鼠抗血清均产生阳性免疫反应。29 kDa的产物占细胞裂解物的12%。用工程化乳酸乳球菌口服接种可显著提高小鼠血清IgG水平(P < 0.05)。
开发出一种新型工程化乳酸乳球菌菌株,该菌株能高效产生具有所需抗原性和强免疫原性的完整HpaA蛋白。这为利用乳酸乳球菌进行抗幽门螺杆菌疫苗接种的方法提供了依据。