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Volume 64, Issue 1

Expression of urease B on the surface of spores Free

Abstract

infection is a major risk factor for chronic gastritis, digestive ulcers, gastric adenocarcinoma and lymphoma. Due to the decreasing efficacy of anti- antibiotic therapy in clinical practice, there is renewed interest in the development of anti- vaccines. is non-pathogenic and can produce endospores, which can survive under extreme conditions. These features make the spore an ideal vehicle for delivery of heterologous antigens to extreme environments such as the gastrointestinal tract. In this study, we displayed urease B protein on the spore coat using the spore coat protein CotC as a fusion partner. Western blot analyses were used to verify urease B surface expression on spores. Recombinant spores displaying the urease B antigen were used for oral immunization and were shown to generate humoral response in mice. Urease B-specific secretory IgA in faeces and IgG in serum reached significant levels 2 weeks after oral dosing. In addition, oral immunization of recombinant urease B spores induced a significant reduction (84 %) in the stomach bacterial load (0.25±0.13×10 c.f.u.) compared to that in the non-recombinant spores treated group (1.56±0.3×10 c.f.u.; <0.01). This report shows that urease B expressed on spores was immunogenic, and oral administration of urease B spores can provide protection against infection.

  • Received:
  • Accepted:
  • Published Online:
© 2015 The Authors
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2015-01-01
2024-04-25
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References

  1. Akhiani A. A., Schön K., Franzén L. E., Pappo J., Lycke N. 2004; Helicobacter pylori-specific antibodies impair the development of gastritis, facilitate bacterial colonization, and counteract resistance against infection. J Immunol 172:5024–5033 [View Article][PubMed]
     [Google Scholar]
  2. Bégué R. E., Sadowska-Krowicka H. 2010; Protective efficacy of recombinant urease B and aluminum hydroxide against Helicobacter pylori infection in a mouse model. FEMS Immunol Med Microbiol 60:142–146 [View Article][PubMed]
     [Google Scholar]
  3. Bhuiyan T. R., Saha A., Lundgren A., Qadri F., Svennerholm A. M. 2010; Immune responses to Helicobacter pylori infection in Bangladeshi children during their first two years of life and the association between maternal antibodies and onset of infection. J Infect Dis 202:1676–1684 [View Article][PubMed]
     [Google Scholar]
  4. Brandtzaeg P. 2010; Function of mucosa-associated lymphoid tissue in antibody formation. Immunol Invest 39:303–355 [View Article][PubMed]
     [Google Scholar]
  5. Czinn S. J., Blanchard T. 2011; Vaccinating against Helicobacter pylori infection. Nat Rev Gastroenterol Hepatol 8:133–140 [View Article][PubMed]
     [Google Scholar]
  6. de Martel C., Ferlay J., Franceschi S., Vignat J., Bray F., Forman D., Plummer M. 2012; Global burden of cancers attributable to infections in 2008: a review and synthetic analysis. Lancet Oncol 13:607–615 [View Article][PubMed]
     [Google Scholar]
  7. DeLyria E. S., Redline R. W., Blanchard T. G. 2009; Vaccination of mice against H pylori induces a strong Th-17 response and immunity that is neutrophil dependent. Gastroenterology 136:247–256 [View Article][PubMed]
     [Google Scholar]
  8. Driks A. 1999; Bacillus subtilis spore coat. Microbiol Mol Biol Rev 63:1–20[PubMed]
     [Google Scholar]
  9. Duc H., Hong H. A., Fairweather N., Ricca E., Cutting S. M. 2003; Bacterial spores as vaccine vehicles. Infect Immun 71:2810–2818 [View Article][PubMed]
     [Google Scholar]
  10. Duc H., Hong H. A., Atkins H. S., Flick-Smith H. C., Durrani Z., Rijpkema S., Titball R. W., Cutting S. M. 2007; Immunization against anthrax using Bacillus subtilis spores expressing the anthrax protective antigen. Vaccine 25:346–355 [View Article][PubMed]
     [Google Scholar]
  11. Eaton K. A., Brooks C. L., Morgan D. R., Krakowka S. 1991; Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun 59:2470–2475[PubMed]
     [Google Scholar]
  12. Ermak T. H., Giannasca P. J., Nichols R., Myers G. A., Nedrud J., Weltzin R., Lee C. K., Kleanthous H., Monath T. P. 1998; Immunization of mice with urease vaccine affords protection against Helicobacter pylori infection in the absence of antibodies and is mediated by MHC class II-restricted responses. J Exp Med 188:2277–2288 [View Article][PubMed]
     [Google Scholar]
  13. Handa O., Naito Y. J., Yoshikawa T. 2011; Redox biology and gastric carcinogenesis: the role of Helicobacter pylori. Redox Rep 16:1–7 [View Article][PubMed]
     [Google Scholar]
  14. Hatzifoti C., Roussel Y., Harris A. G., Wren B. W., Morrow J. W., Bajaj-Elliott M. 2006; Mucosal immunization with a urease B DNA vaccine induces innate and cellular immune responses against Helicobacter pylori. Helicobacter 11:113–122 [View Article][PubMed]
     [Google Scholar]
  15. He B., Xu W., Santini P. A., Polydorides A. D., Chiu A., Estrella J., Shan M., Chadburn A., Villanacci V.& other authors ( 2007; Intestinal bacteria trigger T cell-independent immunoglobulin A(2) class switching by inducing epithelial-cell secretion of the cytokine APRIL. Immunity 26:812–826 [View Article][PubMed]
     [Google Scholar]
  16. Kleanthous H., Myers G. A., Georgakopoulos K. M., Tibbitts T. J., Ingrassia J. W., Gray H. L., Ding R., Zhang Z. Z., Lei W.& other authors ( 1998; Rectal and intranasal immunizations with recombinant urease induce distinct local and serum immune responses in mice and protect against Helicobacter pylori infection. Infect Immun 66:2879–2886[PubMed]
     [Google Scholar]
  17. Kritas S. K., Govaris A., Christodoulopoulos G., Burriel A. R. 2006; Effect of Bacillus licheniformis and Bacillus subtilis supplementation of ewe’s feed on sheep milk production and young lamb mortality. J Vet Med A Physiol Pathol Clin Med 53:170–173 [View Article][PubMed]
     [Google Scholar]
  18. Lamprecht M., Bogner S., Schippinger G., Steinbauer K., Fankhauser F., Hallstroem S., Schuetz B., Greilberger J. F. 2012; Probiotic supplementation affects markers of intestinal barrier, oxidation, and inflammation in trained men; a randomized, double-blinded, placebo-controlled trial. J Int Soc Sports Nutr 9:45 [View Article][PubMed]
     [Google Scholar]
  19. Lee J. S., Cho J. Y., Song H., Kim E. H., Hahm K. B. 2012; Revaprazan, a novel acid pump antagonist, exerts anti-inflammatory action against Helicobacter pylori-induced COX-2 expression by inactivating Akt signaling. J Clin Biochem Nutr 51:77–83 [View Article][PubMed]
     [Google Scholar]
  20. Lee J. W., Kim N., Kim J. M., Nam R. H., Chang H., Kim J. Y., Shin C. M., Park Y. S., Lee D. H., Jung H. C. 2013; Prevalence of primary and secondary antimicrobial resistance of Helicobacter pylori in Korea from 2003 through 2012. Helicobacter 18:206–214 [View Article][PubMed]
     [Google Scholar]
  21. Leighton T. J., Doi R. H. 1971; The stability of messenger ribonucleic acid during sporulation in Bacillus subtilis. J Biol Chem 246:3189–3195[PubMed]
     [Google Scholar]
  22. Li Y., Ning Y., Wang Y., Peng D., Jiang Y., Zhang L., Long M., Luo J., Li M. 2010; Mimotopes selected with a neutralizing antibody against urease B from Helicobacter pylori induce enzyme inhibitory antibodies in mice upon vaccination. BMC Biotechnol 10:84 [View Article][PubMed]
     [Google Scholar]
  23. Liu K. Y., Shi Y., Luo P., Yu S., Chen L., Zhao Z., Mao X. H., Guo G., Wu C., Zou Q. M. 2011; Therapeutic efficacy of oral immunization with attenuated Salmonella typhimurium expressing Helicobacter pylori CagA, VacA and UreB fusion proteins in mice model. Vaccine 29:6679–6685 [View Article][PubMed]
     [Google Scholar]
  24. Mauriello E. M., Duc H., Isticato R., Cangiano G., Hong H. A., De Felice M., Ricca E., Cutting S. M. 2004; Display of heterologous antigens on the Bacillus subtilis spore coat using CotC as a fusion partner. Vaccine 22:1177–1187 [View Article][PubMed]
     [Google Scholar]
  25. Nakamura S., Sugiyama T., Matsumoto T., Iijima K., Ono S., Tajika M., Tari A., Kitadai Y., Matsumoto H.& other authors ( 2012; Long-term clinical outcome of gastric MALT lymphoma after eradication of Helicobacter pylori: a multicentre cohort follow-up study of 420 patients in Japan. Gut 61:507–513 [View Article][PubMed]
     [Google Scholar]
  26. Negri A., Potocki W., Iwanicki A., Obuchowski M., Hinc K. 2013; Expression and display of Clostridium difficile protein FliD on the surface of Bacillus subtilis spores. J Med Microbiol 62:1379–1385 [View Article][PubMed]
     [Google Scholar]
  27. Nicholson W. L., Setlow P. 1990; Sporulation, germination and outgrowth. In Molecular Biological Methods for Bacillus pp. 391–450 Edited by Harwood C. R., Cutting S. M. Chichester: Wiley;
     [Google Scholar]
  28. Nicholson W. L., Munakata N., Horneck G., Melosh H. J., Setlow P. 2000; Resistance of Bacillus endospores to extreme terrestrial and extraterrestrial environments. Microbiol Mol Biol Rev 64:548–572 [View Article][PubMed]
     [Google Scholar]
  29. Raghavan S., Ostberg A. K., Flach C. F., Ekman A., Blomquist M., Czerkinsky C., Holmgren J. 2010; Sublingual immunization protects against Helicobacter pylori infection and induces T and B cell responses in the stomach. Infect Immun 78:4251–4260 [View Article][PubMed]
     [Google Scholar]
  30. Robinson K., Chamberlain L. M., Schofield K. M., Wells J. M., Le Page R. W. F. 1997; Oral vaccination of mice against tetanus with recombinant Lactococcus lactis. Nat Biotechnol 15:653–657 [View Article][PubMed]
     [Google Scholar]
  31. Spizizen J. 1958; Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleotide. Proc Natl Acad Sci U S A 44:1072–1078 [View Article][PubMed]
     [Google Scholar]
  32. Su P., Li Y., Li H., Zhang J., Lin L., Wang Q., Guo F., Ji Z., Mao J.& other authors ( 2013; Antibiotic resistance of Helicobacter pylori isolated in the southeast coastal region of China. Helicobacter 18:274–279 [View Article][PubMed]
     [Google Scholar]
  33. Summerton N. A., Welch R. W., Bondoc L., Yang H. H., Pleune B., Ramachandran N., Harris A. M., Bland D., Jackson W. J.& other authors ( 2010; Toward the development of a stable, freeze-dried formulation of Helicobacter pylori killed whole cell vaccine adjuvanted with a novel mutant of Escherichia coli heat-labile toxin. Vaccine 28:1404–1411 [View Article][PubMed]
     [Google Scholar]
  34. Toller I. M., Neelsen K. J., Steger M., Hartung M. L., Hottiger M. O., Stucki M., Kalali B., Gerhard M., Sartori A. A.& other authors ( 2011; Carcinogenic bacterial pathogen Helicobacter pylori triggers DNA double-strand breaks and a DNA damage response in its host cells. Proc Natl Acad Sci U S A 108:14944–14949 [View Article][PubMed]
     [Google Scholar]
  35. Tsuda M., Karita M., Morshed M. G., Okita K., Nakazawa T. 1994; A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach. Infect Immun 62:3586–3589[PubMed]
     [Google Scholar]
  36. Uemura N., Okamoto S., Yamamoto S., Matsumura N., Yamaguchi S., Yamakido M., Taniyama K., Sasaki N., Schlemper R. J. 2001; Helicobacter pylori infection and the development of gastric cancer. N Engl J Med 345:784–789 [View Article][PubMed]
     [Google Scholar]
  37. Vermoote M., Van Steendam K., Flahou B., Smet A., Pasmans F., Glibert P., Ducatelle R., Deforce D., Haesebrouck F. 2012; Immunization with the immunodominant Helicobacter suis urease subunit B induces partial protection against H. suis infection in a mouse model. Vet Res 43:72 [View Article][PubMed]
     [Google Scholar]
  38. You Y., Liu L., Zhang M., Han X., He L., Zhu Y., Ni P., Zhang J. 2012; Genome sequences of three Helicobacter pylori strains isolated from atrophic gastritis and gastric ulcer patients in China. J Bacteriol 194:6314–6315 [View Article][PubMed]
     [Google Scholar]
  39. Zhang X. Z., Cui Z. L., Hong Q., Li S. P. 2005; High-level expression and secretion of methyl parathion hydrolase in Bacillus subtilis WB800. Appl Environ Microbiol 71:4101–4103 [View Article][PubMed]
     [Google Scholar]
  40. Zhao W., Wu W., Xu X. 2007; Oral vaccination with liposome-encapsulated recombinant fusion peptide of urease B epitope and cholera toxin B subunit affords prophylactic and therapeutic effects against H. pylori infection in BALB/c mice. Vaccine 25:7664–7673 [View Article][PubMed]
     [Google Scholar]
  41. Zhou Z. W., Xia H. M., Hu X., Huang Y., Li Y., Li L., Ma C. L., Chen X. X., Hu F. Y.& other authors ( 2008a; Oral administration of a Bacillus subtilis spore-based vaccine expressing Clonorchis sinensis tegumental protein 22.3 kDa confers protection against Clonorchis sinensis. Vaccine 26:1817–1825 [View Article][PubMed]
     [Google Scholar]
  42. Zhou Z. W., Xia H. M., Hu X. C., Huang Y., Ma C. L., Chen X. X., Hu F. Y., Xu J., Lu F. L.& other authors ( 2008b; Immunogenicity of recombinant Bacillus subtilis spores expressing Clonorchis sinensis tegumental protein. Parasitol Res 102:293–297 [View Article][PubMed]
     [Google Scholar]
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Expression of Helicobacter pylori urease B on the surface of Bacillus subtilis spores
J. Med. Microbiol. 64, 104 (2015); https://doi.org/10.1099/jmm.0.076430-0
/content/journal/jmm/10.1099/jmm.0.076430-0
/content/journal/jmm/10.1099/jmm.0.076430-0
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