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AIM:To construct and select antigen epitopes of vacuolatingcytotoxin A (VacA) for nontoxic VacA vaccine againstHelicobacter pylori (H pylori) infection.METHODS:Eleven VacA epitopes were predicted accordingto VacA antigenic bioinformatics.Three candidates ofVacA epitope were constructed through differentcombined epitopes.The candidate was linked with E.coliheat-labile enterotoxin B (LTB) by a linker of 7 aminoacids,and cloned into plasmid pQE-60 in which fusionLTB-VacA epitope was efficiently expressed.To test theantigencity of the candidate,6 BALB/c mice were treatedwith the fusion LTB-VacA epitope through intraperitonealinjection.To explore the ability of inhibiting the toxicityof VacA,cantiserum against the candidate was used tocounteract VacA that induced HeLa cells to produce cellvacuoles in vitro.RESULTS:Serum IgG against the candidate was inducedin the BALB/c mice.In vitro,the three antisera against thecandidate efficiently counteracted the toxicity of VacA,anddecreased the number of cell vacuoles by 14.17%,20.20%and 30.41% respectively.CONCLUSION:Two of the three candidates,LZ-VacAlandLZ-VacA2,can be used to further study the mechanism ofvacuolating toxicity of VacA,and to construct nontoxic VacAvaccine against H pylori infection.
AIM: To construct and select antigen epitopes of vacuolating cytotoxin A (VacA) for nontoxic VacA vaccine againstHelicobacter pylori (H pylori) infection. METHODS: Eleven VacA epitopes were evaluated according to VacA antigenic bioinformatics. Three candidates of VacA epitope were constructed through differentcombined epitopes. was linked with E.coliheat-labile enterotoxin B (LTB) by a linker of 7 aminoacids, and cloned into plasmid pQE-60 in which fusion LTB-VacA epitope was highly expressed. Test of the antigenicity of the candidate, 6 BALB / c mice were treated with the fusion LTB-VacA epitope through intraperitoneal injection. To explore the ability of inhibiting the toxicity of VacA, cantiserum against the candidate was used tocounteract VacA that induced HeLa cells to produce cellvacuoles in vitro .RESULTS: Serum IgG against the candidate was inducedin the BALB / c mice. vitro, the three antisera against thecandidate ever effective counteracted the toxicity of VacA, anddecreased the numbe r of cell vacuoles by 14.17%, 20.20% and 30.41% respectively. CONCLUSION: Two of the three candidates, LZ-VacAlandLZ-VacA2, can be used to further study the mechanism of vacuolating toxicity of VacA, and to construct nontoxic VacAvaccine against H pylori infection.