Applied Microbiology and Biotechnology

, Volume 93, Issue 5, pp 1937–1945 | Cite as

Immunological features and the ability of inhibitory effects on enzymatic activity of an epitope vaccine composed of cholera toxin B subunit and B cell epitope from Helicobacter pylori urease A subunit

  • Le Guo
  • Xiaokang Li
  • Feng Tang
  • Yunmian He
  • Yingying Xing
  • Xuepeng Deng
  • Tao XiEmail author
Biotechnological products and process engineering


Epitope vaccine based on urease of Helicobacter pylori is a promising option for prophylactic and therapeutic vaccination against H. pylori infection. In this study, we constructed an epitope vaccine with mucosal adjuvant cholera toxin B subunit (CTB) and an epitope (UreA183-203) of H. pylori urease A subunit named CTB-UA. The CTB-UA fusion protein was expressed in Escherichia coli, and the purified protein was used for intraperitoneal immunization experiments in BALB/c mice. The experimental results indicated that anti-CTB-UA antibody could recognize both H. pylori urease A subunit (UreA) and urease B subunit (UreB). Besides, the CTB-UA epitope vaccine had good immunogenicity and immunoreactivity and could induce specific neutralizing antibodies which showed effectively inhibitory effect on the enzymatic activity of H. pylori urease. CTB-UA is a promising molecule to be investigated as H. pylori vaccine antigen candidate.


Epitope vaccine Helicobacter pylori Cholera toxin B subunit Urease A subunit Neutralizing antibody 



This work was supported by the Science Foundation of China Pharmaceutical University (grant no, JKY2009023) and Postgraduate Innovation Project of Jiangsu Province (grant no, CXZZ11_0817). We thank especially Professor Wutong Wu for his contributions in implementation of this experiment.


  1. Blaser MJ (1990) Helicobacter pylori and the pathogenesis of gastroduodenal inflammation. J Infect Dis 161(4):626–633CrossRefGoogle Scholar
  2. Boyanova L, Mentis A, Gubina M, Rozynek E, Gosciniak G, Kalenic S, Goral V, Kupcinskas L, Kantarceken B, Aydin A, Archimandritis A, Dzierzanowska D, Vcev A, Ivanova K, Marina M, Mitov I, Petrov P, Ozden A, Popova M (2002) The status of antimicrobial resistance of Helicobacter pylori in eastern Europe. Clin Microbiol Infect 8(7):388–396CrossRefGoogle Scholar
  3. Bumann D, Metzger WG, Mansouri E, Palme O, Wendland M, Hurwitz R, Haas G, Aebischer T, von Specht BU, Meyer TF (2001) Safety and immunogenicity of live recombinant Salmonella enterica serovar Typhi Ty21a expressing urease A and B from Helicobacter pylori in human volunteers. Vaccine 20(5–6):845–852CrossRefGoogle Scholar
  4. Cheng H, Hu FL (2005) The epidemiology of Helicobacter pylori resistance to antibiotics in Beijing. Zhonghua Yi Xue Za Zhi 85(39):2754–2757Google Scholar
  5. Clements JD (1990) Construction of a nontoxic fusion peptide for immunization against Escherichia coli strains that produce heat-labile and heat-stable enterotoxins. Infect Immun 58(5):1159–1166Google Scholar
  6. DiPetrillo MD, Tibbetts T, Kleanthous H, Killeen KP, Hohmann EL (1999) Safety and immunogenicity of phoP/phoQ-deleted Salmonella typhi expressing Helicobacter pylori urease in adult volunteers. Vaccine 18(5–6):449–459CrossRefGoogle Scholar
  7. Eaton KA, Brooks CL, Morgan DR, Krakowka S (1991) Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun 59(7):2470–2475Google Scholar
  8. Fujii R, Morihara F, Oku T, Hifumi E, Uda T (2004) Epitope mapping and features of the epitope for monoclonal antibodies inhibiting enzymatic activity of Helicobacter pylori urease. Biotechnol Bioeng 86(4):434–444CrossRefGoogle Scholar
  9. Gomez-Duarte OG, Lucas B, Yan ZX, Panthel K, Haas R, Meyer TF (1998) Protection of mice against gastric colonization by Helicobacter pylori by single oral dose immunization with attenuated Salmonella typhimurium producing urease subunits A and B. Vaccine 16(5):460–471CrossRefGoogle Scholar
  10. Hifumi E, Yamada Y, Uda T (2006) A catalytic antibody heavy chain HpU-2 degrading its epitope peptide and H. pylori urease. Immunol Lett 103(1):68–74CrossRefGoogle Scholar
  11. Hirota K, Nagata K, Norose Y, Futagami S, Nakagawa Y, Senpuku H, Kobayashi M, Takahashi H (2001) Identification of an antigenic epitope in Helicobacter pylori urease that induces neutralizing antibody production. Infect Immun 69(11):6597–6603CrossRefGoogle Scholar
  12. Kang SM, Yao Q, Guo L, Compans RW (2003) Mucosal immunization with virus-like particles of simian immunodeficiency virus conjugated with cholera toxin subunit B. J Virol 77(18):9823–9830CrossRefGoogle Scholar
  13. Kwon SB, Lee KL, Kim JS, Lee JK, Kim W, Jung YJ, Jeong JB, Kim JW, Kim BG (2010) Antibiotics-associated diarrhea and other gastrointestinal abnormal responses regarding Helicobacter pylori eradication. Korean J Gastroenterol 56(4):229–235CrossRefGoogle Scholar
  14. Labenz J, Borsch G (1994) Evidence for the essential role of Helicobacter pylori in gastric ulcer disease. Gut 35(1):19–22CrossRefGoogle Scholar
  15. Leal-Herrera Y, Torres J, Perez-Perez G, Gomez A, Monath T, Tapia-Conyer R, Munoz O (1999) Serologic IgG response to urease in Helicobacter pylori-infected persons from Mexico. Am J Trop Med Hyg 60(4):587–592Google Scholar
  16. Liu W, Peng Z, Liu Z, Lu Y, Ding J, Chen YH (2004) High epitope density in a single recombinant protein molecule of the extracellular domain of influenza A virus M2 protein significantly enhances protective immunity. Vaccine 23(3):366–371CrossRefGoogle Scholar
  17. Liu Z, Wang Z, Chen YH (2005) Predefined spacers between epitopes on a recombinant epitope-peptide impacted epitope-specific antibody response. Immunol Lett 97(1):41–45CrossRefGoogle Scholar
  18. Londono-Arcila P, Freeman D, Kleanthous H, O'Dowd AM, Lewis S, Turner AK, Rees EL, Tibbitts TJ, Greenwood J, Monath TP, Darsley MJ (2002) Attenuated Salmonella enterica serovar Typhi expressing urease effectively immunizes mice against Helicobacter pylori challenge as part of a heterologous mucosal priming-parenteral boosting vaccination regimen. Infect Immun 70(9):5096–5106CrossRefGoogle Scholar
  19. Lucas B, Bumann D, Walduck A, Koesling J, Develioglu L, Meyer TF, Aebischer T (2001) Adoptive transfer of CD4+ T cells specific for subunit A of Helicobacter pylori urease reduces H. pylori stomach colonization in mice in the absence of interleukin-4 (IL-4)/IL-13 receptor signaling. Infect Immun 69(3):1714–1721CrossRefGoogle Scholar
  20. Marshall BJ, Barrett LJ, Prakash C, McCallum RW, Guerrant RL (1990) Urea protects Helicobacter (Campylobacter) pylori from the bactericidal effect of acid. Gastroenterology 99(3):697–702Google Scholar
  21. Menezes CA, Amianti J, Harayama HS, Koga PC, Trabulsi LR, Piazza RM (2002) Inhibition of Escherichia coli heat-labile enterotoxin by neoglycoprotein and anti-lectin antibodies which mimic GM1 receptor. FEMS Microbiol Lett 216(1):67–70CrossRefGoogle Scholar
  22. Nagata K, Mizuta T, Tonokatu Y, Fukuda Y, Okamura H, Hayashi T, Shimoyama T, Tamura T (1992) Monoclonal antibodies against the native urease of Helicobacter pylori: synergistic inhibition of urease activity by monoclonal antibody combinations. Infect Immun 60(11):4826–4831Google Scholar
  23. Nystrom J, Raghavan S, Svennerholm AM (2006) Mucosal immune responses are related to reduction of bacterial colonization in the stomach after therapeutic Helicobacter pylori immunization in mice. Microbes Infect 8(2):442–449CrossRefGoogle Scholar
  24. Qiu Y, Wang YC, Tao HX, Zhan DW, Yuan SL, Wang P, Wang LC, Han XP, Li CS, Li JK, Liu CJ (2010) Identification of B-cell epitopes in urease B subunit of Helicobacter pylori bound by neutralizing antibodies. Vaccine 28(32):5220–5227CrossRefGoogle Scholar
  25. Raghavan S, Svennerholm AM, Holmgren J (2002) Effects of oral vaccination and immunomodulation by cholera toxin on experimental Helicobacter pylori infection, reinfection, and gastritis. Infect Immun 70(8):4621–4627CrossRefGoogle Scholar
  26. Rossi G, Ruggiero P, Peppoloni S, Pancotto L, Fortuna D, Lauretti L, Volpini G, Mancianti S, Corazza M, Taccini E, Di Pisa F, Rappuoli R, Del Giudice G (2004) Therapeutic vaccination against Helicobacter pylori in the beagle dog experimental model: safety, immunogenicity, and efficacy. Infect Immun 72(6):3252–3259CrossRefGoogle Scholar
  27. Sun JB, Mielcarek N, Lakew M, Grzych JM, Capron A, Holmgren J, Czerkinsky C (1999) Intranasal administration of a Schistosoma mansoni glutathione S-transferase-cholera toxoid conjugate vaccine evokes antiparasitic and antipathological immunity in mice. J Immunol 163(2):1045–1052Google Scholar
  28. Uemura N, Okamoto S, Yamamoto S, Matsumura N, Yamaguchi S, Yamakido M, Taniyama K, Sasaki N, Schlemper RJ (2001) Helicobacter pylori infection and the development of gastric cancer. N Engl J Med 345(11):784–789CrossRefGoogle Scholar
  29. Wehrle PF, Leedom JM, Portnoy B, Pierce NF, Cowper HH (1963) Safety of sabin oral poliovaccine strains; mass immunization program in Los Angeles County, 1962–1963. JAMA 186:821–826CrossRefGoogle Scholar
  30. Yano A, Onozuka A, Asahi-Ozaki Y, Imai S, Hanada N, Miwa Y, Nisizawa T (2005) An ingenious design for peptide vaccines. Vaccine 23(17–18):2322–2326CrossRefGoogle Scholar
  31. 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(44):7664–7673CrossRefGoogle Scholar
  32. Zhou WY, Shi Y, Wu C, Zhang WJ, Mao XH, Guo G, Li HX, Zou QM (2009) Therapeutic efficacy of a multi-epitope vaccine against Helicobacter pylori infection in BALB/c mice model. Vaccine 27(36):5013–5019CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Le Guo
    • 1
  • Xiaokang Li
    • 1
  • Feng Tang
    • 1
  • Yunmian He
    • 1
  • Yingying Xing
    • 1
  • Xuepeng Deng
    • 1
  • Tao Xi
    • 1
    Email author
  1. 1.Biotechnology Center, School of Life Science and TechnologyChina Pharmaceutical UniversityNanjingChina

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