Skip to main content

Advertisement

SpringerLink
Log in

Immunogenicity of cholera toxin B epitope inserted in Salmonella flagellin expressed on bacteria and administered as DNA vaccine

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Salmonella vaccine strains have been previously reported to evoke immune response against heterologous antigen cloned in the flagellin gene. A non-toxic cholera toxin subunit B epitope was selected by using computer-based program and genetically fused in single and double copy in Salmonella typhimurium flagellin gene. The chimeric flagellin functioned normally as demonstrated by motility assay. Cholera toxin B epitope cloned in flagellin was expressed at the flagellar surface. The expression was verified by Western blotting. Mice administered orally and subcutaneously with aroA flagellin-negative strain of S. dublin expressing the chimeric flagellin gene resulted in generation of antibody against cholera toxin. Mice administered intramuscularly and subcutaneously with naked mammalian expression vector containing the same cholera toxin epitope could also evoke the antibody response though it was less than the chimeric flagellin. (Mol Cell Biochem 276: 1–6, 2005)

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

Similar content being viewed by others

Abbreviations

CTP1:

cholera toxin B subunit epitope

ELISA:

enzyme linked immunoassay

Ig:

immunoglobulin

References

  1. Svennerholm L, Lange S, Holmgren J: Correlation between intestinal synthesis of specific immunglobulin A and protection against experimental cholera in mice. Infect Immun 21: 1–6, 1978

    PubMed  Google Scholar 

  2. Elson CO, Ealding W: Generalized systemic and mucosal immunity in mice after mucosal stimulation with cholera toxin. J Immunol 132: 27–36, 1984

    Google Scholar 

  3. Formal SB, Baron LS, Kopecko DJ, Washington O, Powell C, Life CA: Construction of a potential bivalant vaccine strain:Introduction of Shigella sonni from 1 antigen genes into the galE Salmonella typhi Ty21a typhoid vaccine strain. Infect Immun 34: 746–750, 1981

    PubMed  Google Scholar 

  4. Fairweather NF, Chatfield SN, Makoff AJ: Oral vaccianation of mice againts tetnus by use of a live attenuated Salmonella carrier. Infect Immun 58: 1323–1326, 1990

    PubMed  Google Scholar 

  5. Newton SMC, Jacob CO, Stocker BAD: Immune response to cholera toxin epitope inserted in Salmonella flagella. Science 244: 70–72, 1989

    PubMed  Google Scholar 

  6. Curtiss RI, Kelly SM, Gulig PA, Nakayama K: Selective delivery of antigens by recombinanat bactaria. Curr Top Microbiol Immunol 146: 35–49, 1989

    PubMed  Google Scholar 

  7. Gonzalez C, Hone D, Noriega F: Salmonella typhi vaccine strain CVD 908 expressing the circumsporozoite protein of plasmodium falciparum: Strain construction and safety and immunogenicity in hamans. J Infect Dis 169: 927–931, 1994

    PubMed  Google Scholar 

  8. Khan CMA, Villareal-Ramos B, Pierce RJ: Construction expression and immunogenicity of the Schistosoma mansoni P28 gluthione S-transferase as a genetic fusion to tetanus toxin fragmaent C in a live Aro attenuated vaccine strain of Salmonella. Proc Natl Acad Sci USA 91: 11261–11265, 1994

    PubMed  Google Scholar 

  9. Clements JD, Lyon FL, Lowe KL, Farrand, AL, El-Morshidy S: Oral immunization of mice with attenuated Salmonella enteretiditis cointaining a recombinanat plasmid which encodes for production of the B subunit of heat labile Escherichia coli enterotoxin. Infect Immun 53: 685–692, 1986

    PubMed  Google Scholar 

  10. Sztein M, Wasserman SS, Tacket CO: Cytokine production patterns and lymproliferative responses in volunteers orally immunized with attenuated vaccine strains of Salmonella typhi. J Infect Dis 170: 1508–1517, 1994

    PubMed  Google Scholar 

  11. Robbins AA: Progress towards vaccines we need and we do not have. Lancet 335: 1436–1438, 1990

    Article  PubMed  Google Scholar 

  12. Chatfield SN, Charles IG, Makoff A.: Use of the nirB promoter to direct the stable expression of heterologous antigens in Salmonella oral vaccine strains: development of a single-dose oral tetanus vaccine. Biotechnology (NY) 10: 888–892, 1992

    Article  Google Scholar 

  13. Carmenes RS, Freije JP, Molina MM, Martin JM: Predict7, a program for protein structure prediction. Biochem Biophys Res Commun 159: 687–693, 1989

    Article  PubMed  Google Scholar 

  14. Jacob CO, Sella M, Arnon R: Antibodies against synthetic peptides of the B subunit of cholera toxin: Crossreaction and neutralization of the toxin. Proc Natl Acad Sci USA 80: 7611–7615, 1983

    PubMed  Google Scholar 

  15. Ibrahim GF, Fleet GH, Lyons MJ, Walker R: Method for the isolation of highly purified Salmonella flagellins. J Clin Microbiol 22: 1040–1044, 1985

    PubMed  Google Scholar 

  16. Lowry OH, Rosenbrough NJ, Farr AC, Randall RJ: Protein measuurement with folin phenol reagent. J Biol Chem 193: 265–275, 1951

    PubMed  Google Scholar 

  17. Laemmli UK: Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 277: 680–685, 1970

    Article  Google Scholar 

  18. Finkelstein RA: Vibriocidal antibody inhibition analysis: A technique for the identification of predominant vibriocidal antibodies in erum and for the recognition and identification of Vibrio cholerae antigens. J Immunol 89: 264, 1962

    Google Scholar 

  19. Harlow E, Lane DA: A Laboratory Manual. Cold Spring Harbour Laboratory, Cold Spring Harbour, New York, 1988, pp 283–612

    Google Scholar 

  20. Hopp TP, Woods KR: Prediction of protein antigenic determinants from amino acids sequences. Proc Natl Acad Sci USA 78: 3824–3828, 1981

    PubMed  Google Scholar 

  21. Kyte J, Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132, 1982

    Article  PubMed  Google Scholar 

  22. Maskell DJ, Sweeney, KJ, O’Callaghan D, Hormaeche CE, Liew FY, Dougan G: Salmonella tyohinurium aroA mutants as carrier of the Esherichia coli heat labile enterotoxin b subunit to the murine secretory and systemic immune systems. Microb Pathog 2: 211, 1987

    Article  PubMed  Google Scholar 

  23. Leclerc C, Charbit A, Molla A, Hofnung M: Antibody response to a foreign epitope expressed at the surface of recombinant bacteria: Importance of the route of immunisation. Vaccine 7: 242, 1989

    Article  PubMed  Google Scholar 

  24. Agterberg M, Adriaanse H, Lankhof H, Meloen R, Tommasen J: Outer membrane phoE protein of Escherichia coli as carrier of foreign antigenic determinants: Immunogenicity of epitopes of foot and mouth disease virus. Vaccine 8: 85, 1990

    Article  PubMed  Google Scholar 

  25. Wei LN, Joys, TM: The nucleotide sequence of the H-1 gene of Salmonella rubishaw. Nucleic Acids Res 14: 8227, 1986

    PubMed  Google Scholar 

  26. Boyle CM, Morin M, Webster RG, Robinson HL: Role of different lymphoid tissues in the initiation and maitanance of DNA raised antibody responses to the influenza virus h1 glycoprotein. J Virol 70: 9076–9078, 1996

    Google Scholar 

  27. Donnely JJ, Ulmer, JB, Liu MA: Immunization with DNA. J Immun Meth 176: 145–152, 1994

    Article  Google Scholar 

  28. Xiang ZQ, Spitlanik SL, Cheng J, Erickson J, Wozczyk B, Ertl HC: Immune responses to nucleic acid vaccines to rabies virus. Virology 209: 569–579, 1995

    Article  PubMed  Google Scholar 

  29. Zarozinski CC, Gynan EF, Selin LK, Robinson HL, Welsh RM: Protective CTL dependent immunity and enhanced immunopathology in mice immunized by particle bombardment with DNA encoding an internal viron protein. J Immunol 154: 4010–4017, 1995

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Ravindra Kumar

    Present address: Pennsylvania State University, 403 Wartik Laboratory/104 Chemistry Building, University Park, PA, USA

Authors and Affiliations

  1. Department of Microbiology and Immunology, Georgetown University School of Medicine, Washington, DC, USA

    Neeraj Chauhan

  2. Applied Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, India

    Neeraj Chauhan, Ravindra Kumar, Jitendra Badhai, Anju Preet & Pramod Kumar Yadava

  3. Applied Molecular Biology Laboratory, School of Life Sciences,, Jawaharlal Nehru University, New Delhi, 110067, India

    Pramod Kumar Yadava

Authors
  1. Neeraj Chauhan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ravindra Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jitendra Badhai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anju Preet
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Pramod Kumar Yadava
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pramod Kumar Yadava.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chauhan, N., Kumar, R., Badhai, J. et al. Immunogenicity of cholera toxin B epitope inserted in Salmonella flagellin expressed on bacteria and administered as DNA vaccine. Mol Cell Biochem 276, 1–6 (2005). https://doi.org/10.1007/s11010-005-2240-z

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-005-2240-z

Keywords

Search

Navigation