Skip to main content
SpringerLink
Log in

An engineered Lactococcus lactis strain exerts significant immune responses through efficient expression and delivery of Helicobacter pylori Lpp20 antigen

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Objectives

To produce and deliver Helicobacter pylori lipoprotein Lpp20 via using Lactococcus lactis with aim of developing an efficient way to use this protective antigen in vaccine formulation.

Results

An engineered L. lactis strain carrying the lpp20 gene from H. pylori was constructed. The inducible expression of Lpp20 in L. lactis was detected as a 20 kDa intracellular protein by SDS-PAGE. Lpp20 constituted 10 % of the L. lactis cellular proteins. The expression product was highly immunoreactive, as demonstrated by western blot assays using mouse anti-H. pylori sera. Animal experimentation showed that oral vaccination with the engineered strain excited significantly elevated levels of serum Lpp20-specific IgG antibodies in BALB/c mice (P < 0.05).

Conclusions

This report presents the first efficient expression and delivery of whole Lpp20 protein to the immunization sites by using L. lactis, demonstrating an efficient utilization mode of Lpp20 in anti-H. pylori vaccination.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Chen SY, Zhang RG, Duan GC, Shi JX (2011) Food-grade expression of Helicobacter pylori ureB subunit in Lactococcus lactis and its immunoreactivity. Curr Microbiol 62:1726–1731

    Article  CAS  PubMed  Google Scholar 

  • Gu Q, Song D, Zhu M (2009) Oral vaccination of mice against Helicobacter pylori with recombinant Lactococcus lactis expressing urease subunit B. FEMS Immunol Med Microbiol 56:197–203

    Article  CAS  PubMed  Google Scholar 

  • Keenan J, Oliaro J, Domigan N, Potter H, Aitken G, Allardyce R, Roake J (2000) Immune response to an 18-kilodalton outer membrane antigen identifies lipoprotein 20 as a Helicobacter pylori vaccine candidate. Infect Immun 68:3337–3343

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Keenan J, Neal S, Allardyce R, Roake J (2002) Serum-derived IgG1-mediated immune exclusion as a mechanism of protection against H. pylori infection. Vaccine 20:2981–2988

    Article  CAS  PubMed  Google Scholar 

  • Keenan JI, Rijpkema SG, Durrani Z, Roake JA (2003) Differences in immunogenicity and protection in mice and guinea pigs following intranasal immunization with Helicobacter pylori outer membrane antigens. FEMS Immunol Med Microbiol 36:199–205

    Article  CAS  PubMed  Google Scholar 

  • Kostrzynska M, O’Toole PW, Taylor DE, Trust TJ (1994) Molecular characterization of a conserved 20-kilodalton membrane-associated lipoprotein antigen of Helicobacter pylori. J Bacteriol 176:5938–5948

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lee MH, Roussel Y, Wilks M, Tabaqchali S (2001) Expression of Helicobacter pylori urease subunit B gene in Lactococcus lactis MG1363 and its use as a vaccine delivery system against H. pylori infection in mice. Vaccine 19:3927–3935

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Jiang Y, Xi Y, Zhang L, Luo J, He D, Zeng S, Ning Y (2012) Identification and characterization of H-2d restricted CD4+ T cell epitopes on Lpp20 of Helicobacter pylori. BMC Immunol 13:68

    Article  PubMed  PubMed Central  Google Scholar 

  • Li Y, Chen Z, Ye J, Ning L, Luo J, Zhang L, Jiang Y, Xi Y, Ning Y (2016) Antibody production and Th1-biased response induced by an epitope vaccine composed of cholera toxin B unit and Helicobacter pylori Lpp20 epitopes. Helicobacter 21:234–248

    Article  CAS  PubMed  Google Scholar 

  • Mierau I, Kleerebezem M (2005) 10 years of the nisin-controlled gene expression system (NICE) in Lactococcus lactis. Appl Microbiol Biotechnol 68:1–13

    Article  Google Scholar 

  • Yu W, Zhang Y, Jing J, Liu Z (2010) Construction of Helicobacter pylori Lpp20-IL2 DNA vaccine and evaluation of its immunocompetence in C57BL/6 mice. Wei Sheng Wu Xue Bao 50:554–559

    CAS  PubMed  Google Scholar 

  • Zeng M, Mao XH, Li JX et al (2015) Efficacy, safety, and immunogenicity of an oral recombinant Helicobacter pylori vaccine in children in China: a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet 386:1457–1464

    Article  CAS  PubMed  Google Scholar 

  • Zhang XJ, Duan GC, Zhang RG, Fan QT (2009) Optimized expression of Helicobacter pylori ureB gene in Lactococcus lactis NICE system and experimental study on its immunoreactivity. Current Microbiol 58:308–314

    Article  CAS  PubMed  Google Scholar 

  • Zhang XJ, Feng SY, Li ZT, Feng YM (2015) Expression of Helicobacter pylori hspA gene in Lactococcus lactis NICE system and experimental study on its immunoreactivity. Gastroenterol Res Pract 2015:750932

    PubMed  PubMed Central  Google Scholar 

  • Zhang R, Duan G, Shi Q, Chen S, Fan Q, Sun N, Xi Y (2016a) Construction of a recombinant Lactococcus lactis strain expressing a fusion protein of Omp22 and HpaA from Helicobacter pylori for oral vaccine development. Biotechnol Lett. doi:10.1007/s10529-016-2173-5 Epub ahead of print

    Google Scholar 

  • Zhang RG, Duan GC, Fan QT, Chen SY (2016b) Role of Helicobacter pylori infection in pathogenesis of gastric carcinoma. World J Gastrointest Pathophysiol 7:97–107

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This study was funded by the Henan Innovation Center of Molecular Diagnosis and Laboratory Medicine (XTCX-2015-ZD2) and the China Postdoctoral Science Foundation (No. 200801273).

Supporting information

Supplementary Table 1—Plasmid vectors and bacterial strains used.

Author information

Authors and Affiliations

  1. Department of Epidemiology, College of Public Health, Zhengzhou University, No. 100 Kexue Dadao, Zhengzhou, 450001, China

    Rongguang Zhang, Xiaoyan Peng, Guangcai Duan, Qingfeng Shi, Shuaiyin Chen, Chen Wang, Qingtang Fan & Yuanlin Xi

  2. Henan Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang Medical University, Xinxiang, 453003, China

    Rongguang Zhang & Guangcai Duan

Authors
  1. Rongguang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoyan Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guangcai Duan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qingfeng Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shuaiyin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Qingtang Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yuanlin Xi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guangcai Duan.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 37 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, R., Peng, X., Duan, G. et al. An engineered Lactococcus lactis strain exerts significant immune responses through efficient expression and delivery of Helicobacter pylori Lpp20 antigen. Biotechnol Lett 38, 2169–2175 (2016). https://doi.org/10.1007/s10529-016-2209-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-016-2209-x

Keywords

Search

Navigation