Preliminary study on immunotherapy of an oral recombinant DNA vaccine of Helic obacter pylori neutrophil activating protein
- VernacularTitle:幽门螺杆菌中性粒细胞激活蛋白核酸疫苗实验研究
- Author:
Bo SUN
;
Hua YANG
;
Zhaoshen LI
- Publication Type:Journal Article
- Keywords:
Helicobacter pylori;
Neutrophil activating protein;
DNA vaccine
- From:
Chinese Journal of Digestion
2001;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an oral recombinant DNA vaccine of Helicobacter pylori(H. pylori) neutrophil activating protein (Hp-NAP), and to evaluate its immunotherapeutic effects. Methods The napA gene (encoding Hp-NAP) was amplified by poly mera se chain reaction(PCR) and cloned into TA cloning vector pBT. After nucleotide s equencing and sequence analysis, the target sequence was subcloned into an eukar yot ic expression vector pIRES. Then the identified recombinant plasmid, pIRES-napA , was transformed into a live attenuated Salmonella typhimurium(S. typhimurium ) strain SL7207, and lavaged into a long-term(30 weeks) model of BALB/c mice infected by Sydney strain(SS1) of H. pylori. Results A 435 bp target gene of napA was amplified by PCR. Seq uenci ng and BLAST analysis showed that most of the cloned napA sequence was homologou s with that of SS1 strain of H. pylori. provided by GenBank, and the homolog y of neucleotide and protein was over 98%, respectively. PCR and restriction enzyme digestion id entification indicated that a recombinant live attenuated S. typhimurium DNA vaccine strain carrying Hp-napA gene was successfully constructed. After 4 wee ks of oral immunization, 75% of mice treated with DNA vaccine were rapid urease test negative, while those with vacant plasmid or normal saline alone were all p ositive (P= 0.0476). The titer of serum Hp-NAP antibody was signific antly elevated in treatment group. Conclusions The successful construction of an effective oral recom binant DNA vaccine of Hp-NAP may be helpful for the further development of polyvalent DNA vaccine against H. pylori infection.
