Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development

Yan Cheng; Youjun Feng; Ping Luo; Jiang Gu; Shu Yu; Wei-jun Zhang; Yan-qing Liu; Qing-xu Wang; Quan-ming Zou; Xu-hu Mao

doi:10.1007/s12275-009-0116-8

Articles

Page Path: HOME > J. Microbiol > Volume 47(4); 2009 > Article

Research Support, Non-U.S. Gov't
Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development: Yan Cheng , Youjun Feng , Ping Luo , Jiang Gu , Shu Yu , Wei-jun Zhang , Yan-qing Liu , Qing-xu Wang , Quan-ming Zou , Xu-hu Mao; Journal of Microbiology 2009;47(4):498-505.
DOI: https://doi.org/10.1007/s12275-009-0116-8
Published online: September 9, 2009

Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University, Chongqing 400038, P. R. China

Corresponding author: Xu-hu Mao , Tel: 86-23-68752315,

Received: 14 April 2009 • Accepted: 5 May 2009

prev next

56 Views
0 Download
20 Crossref
29 Scopus

Full Article

Abstract

Shiga toxin 2 (Stx2) is a major virulence factor for enterohemorrhagic Escherichia coli (EHEC), which is encoded by λ lysogenic phage integrated into EHEC chromosome. Stx2A1, A1 subunit of Stx2 toxin has gathered extensive concerns due to its potential of being developed into a vaccine candidate. However, the substantial progress is hampered in part for the lack of a suitable in vitro expression system. Here we report use of the prokaryotic system pET-28a::espA-Stx2A1/BL21 to carry out the fusion expression of Stx2A1 which is linked to E. coli secreted protein A (EspA) at its N-terminus. Under the IPTG induction, EspA- Stx2A1 fusion protein in the form of inclusion body was obtained successfully, whose expression level can reach about 40% of total bacterial protein at 25°C, much higher than that at 37°C. Western blot test suggested the refolded fusion protein is of excellent immuno-reactivity with both monoclonal antibodies, which are specific to EspA and Stx2A1, respectively. Anti-sera from Balb/c mice immunized with the EspA-Stx2A1 fusion protein were found to exhibit strong neutralization activity and protection capability in vitro and in vivo. These data have provided a novel feasible method to produce Stx2A1 in large scale in vitro, which is implicated for the development of multivalent subunit vaccines candidate against EHEC O157:H7 infections.

Keywords: EHEC O157:H7;EspA;Stx2A1;fusion protein;vaccine

Figure & Data

References

Citations

Citations to this article as recorded by

The Diversity of Escherichia coli Pathotypes and Vaccination Strategies against This Versatile Bacterial Pathogen
Pravil Pokharel, Sabin Dhakal, Charles M. Dozois
Microorganisms.2023; 11(2): 344. CrossRef
In silico design, production and immunization evaluation of a recombinant bivalent fusion protein candidate vaccine against E. coli O157:H7
Hossein Samiei, Shahram Nazarian, Abass Hajizade, Emad Kordbacheh
International Immunopharmacology.2023; 114: 109464. CrossRef
Oral Administration with Live Attenuated Citrobacter rodentium Protects Immunocompromised Mice from Lethal Infection
Shuyu Wang, Xue Xia, Yue Liu, Fengyi Wan, Guy H. Palmer
Infection and Immunity.2022;[Epub] CrossRef
Diagnosis and Treatment for Shiga Toxin-Producing Escherichia coli Associated Hemolytic Uremic Syndrome
Yang Liu, Hatim Thaker, Chunyan Wang, Zhonggao Xu, Min Dong
Toxins.2022; 15(1): 10. CrossRef
Immunization of mice with chimeric antigens displaying selected epitopes confers protection against intestinal colonization and renal damage caused by Shiga toxin-producing Escherichia coli
David A. Montero, Felipe Del Canto, Juan C. Salazar, Sandra Céspedes, Leandro Cádiz, Mauricio Arenas-Salinas, José Reyes, Ángel Oñate, Roberto M. Vidal
npj Vaccines.2020;[Epub] CrossRef
Treatment Strategies for Infections With Shiga Toxin-Producing Escherichia coli
Sabrina Mühlen, Petra Dersch
Frontiers in Cellular and Infection Microbiology.2020;[Epub] CrossRef
Oral immunization of mice with Lactococcus lactis expressing Shiga toxin truncate confers enhanced protection against Shiga toxins of Escherichia coli O157:H7 and Shigella dysenteriae
Sagi Sreerohini, Konduru Balakrishna, Manmohan Parida
APMIS.2019; 127(10): 671. CrossRef
Mucosal and systemic responses of immunogenic vaccines candidates against enteric Escherichia coli infections in ruminants: A review
A. Lawan, F.F.A. Jesse, U.H. Idris, M.N. Odhah, M. Arsalan, N.A. Muhammad, K.R. Bhutto, I.D. Peter, G.A. Abraham, A.H. Wahid, M.L. Mohd-Azmi, M. Zamri-Saad
Microbial Pathogenesis.2018; 117: 175. CrossRef
Intestinal Pathogenic Escherichia coli: Insights for Vaccine Development
Maricarmen Rojas-Lopez, Ricardo Monterio, Mariagrazia Pizza, Mickaël Desvaux, Roberto Rosini
Frontiers in Microbiology.2018;[Epub] CrossRef
Current pathogenic Escherichia coli foodborne outbreak cases and therapy development
Shih-Chun Yang, Chih-Hung Lin, Ibrahim A. Aljuffali, Jia-You Fang
Archives of Microbiology.2017; 199(6): 811. CrossRef
In silico analysis of Shiga toxins (Stxs) to identify new potential vaccine targets for Shiga toxin-producing Escherichia coli
Maryam Golshani, Mana Oloomi, Saeid Bouzari
In Silico Pharmacology.2017;[Epub] CrossRef
Production and purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro) with high-purity and low endotoxin content
Douglas B. Figueiredo, Eneas Carvalho, Mauricio P. Santos, Stefanie Kraschowetz, Rafaela T. Zanardo, Gilson Campani, Gabriel G. Silva, Cíntia R. Sargo, Antonio Carlos L. Horta, Roberto de C. Giordano, Eliane N. Miyaji, Teresa C. Zangirolami, Joaquin Cabre
Applied Microbiology and Biotechnology.2017; 101(6): 2305. CrossRef
Escherichia coli outer membrane protein F (OmpF): an immunogenic protein induces cross-reactive antibodies against Escherichia coli and Shigella
Xiao Wang, Da Teng, Qingfeng Guan, Ruoyu Mao, Ya Hao, Xiumin Wang, Junhu Yao, Jianhua Wang
AMB Express.2017;[Epub] CrossRef
Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for Shigella , Salmonella, enterotoxigenic E. coli (ETEC) enterohemorragic
Miguel O’Ryan, Roberto Vidal, Felipe del Canto, Juan Carlos Salazar, David Montero
Human Vaccines & Immunotherapeutics.2015; 11(3): 601. CrossRef
Recombinant outer membrane protein A induces a protective immune response against Escherichia coli infection in mice
Qingfeng Guan, Xiao Wang, Xiumin Wang, Da Teng, Ruoyu Mao, Yong Zhang, Jianhua Wang
Applied Microbiology and Biotechnology.2015; 99(13): 5451. CrossRef
Paving the way to construct a new vaccine against Escherichia coli from its recombinant outer membrane protein C via a murine model
Xiao Wang, Qingfeng Guan, Xiumin Wang, Da Teng, Ruoyu Mao, Junhu Yao, Jianhua Wang
Process Biochemistry.2015; 50(8): 1194. CrossRef
Comparative Genomics and Immunoinformatics Approach for the Identification of Vaccine Candidates for Enterohemorrhagic Escherichia coli O157:H7
Víctor A. García-Angulo, Anjana Kalita, Mridul Kalita, Luis Lozano, Alfredo G. Torres, S. M. Payne
Infection and Immunity.2014; 82(5): 2016. CrossRef
Advances in the development of enterohemorrhagic Escherichia coli vaccines using murine models of infection
Victor A. Garcia-Angulo, Anjana Kalita, Alfredo G. Torres
Vaccine.2013; 31(32): 3229. CrossRef
Evaluation of hha and hha sepB mutant strains of Escherichia coli O157:H7 as bacterins for reducing E. coli O157:H7 shedding in cattle
Vijay K. Sharma, Evelyn A. Dean-Nystrom, Thomas A. Casey
Vaccine.2011; 29(31): 5078. CrossRef
Vaccination of attenuated EIS-producing Salmonella induces protective immunity against enterohemorrhagic Escherichia coli in mice
Jiang Gu, Yuanyuan Ning, Haiguang Wang, Daping Xiao, Bin Tang, Ping Luo, Yan Cheng, Mingming Jiang, Na Li, Quanming Zou, Xuhu Mao
Vaccine.2011; 29(43): 7395. CrossRef

Cite this Article

Cite this Article: export Copy Download Format; Close

Download Citation

Download a citation file in RIS format that can be imported by all major citation management software, including EndNote, ProCite, RefWorks, and Reference Manager.

Format:

RIS — For EndNote, ProCite, RefWorks, and most other reference management software
BibTeX — For JabRef, BibDesk, and other BibTeX-specific software

Include:

Citation for the content below
Citation and abstract for the content below

Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development

J. Microbiol. 2009;47(4):498-505. Published online September 9, 2009

DOI: https://doi.org/10.1007/s12275-009-0116-8

Related articles