1887

Abstract

Typhoid fever caused by serovar Typhi has contributed to the global public health burden, particularly in developing countries. In this study, an . Typhi ghost was developed and its capacity as a vaccine candidate against typhoid fever was assessed.

An plasmid pJHL187 harbouring a ghost cassette comprising the PhiX 174 lysis gene tightly controlled under the convergent promotor system was transformed into an gene-deleted mutant Typhi strain (STG). The gene encoding the heat-labile enterotoxin (LTB) protein was subcloned into a foreign antigen delivery cassette of pJHL187 to increase mucosal immunity.

The stringent repression and expression of the lethal lysis gene in the system allowed stable production of the ghost strain and secretion of LTB, which was confirmed by immune blot analysis. The level of IgG and sIgA was significantly increased in the mice subcutaneously immunized with STG-LTB compared to the non-immunized mice (<0.05). The CD3CD4 T cell subpopulation was augmented in the immunized group (<0.05) and showed the increment of immunomodulatory cytokines IL-2, IL-6, IL-12, IL-17 and IFN-γ in restimulated splenocytes isolated from the inoculated mice. The serum bactericidal activity of antibodies generated in the rabbits injected with STG-LTB was proved by the elimination of approximately 87.5 % of wild-type . Typhi in the presence of exogenous complement.

The results demonstrated that the STG-LTB ghost effectively enhanced the immunological responses, meaning that STG-LTB is potentially available as a vaccine candidate against typhoid fever.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000543
2017-08-01
2019-12-15
Loading full text...

Full text loading...

/deliver/fulltext/jmm/66/8/1235.html?itemId=/content/journal/jmm/10.1099/jmm.0.000543&mimeType=html&fmt=ahah

References

  1. Szostak MP, Hensel A, Eko FO, Klein R, Auer T et al. Bacterial ghosts: non-living candidate vaccines. J Biotechnol 1996;44:161–170 [CrossRef][PubMed]
    [Google Scholar]
  2. Haslberger AG, Kohl G, Felnerova D, Mayr UB, Fürst-Ladani S et al. Activation, stimulation and uptake of bacterial ghosts in antigen presenting cells. J Biotechnol 2000;83:57–66 [CrossRef][PubMed]
    [Google Scholar]
  3. Walcher P, Mayr UB, Azimpour-Tabrizi C, Eko FO, Jechlinger W et al. Antigen discovery and delivery of subunit vaccines by nonliving bacterial ghost vectors. Expert Rev Vaccines 2004;3:681–691 [CrossRef][PubMed]
    [Google Scholar]
  4. Chen J, Li N, She F. Helicobacter pylori outer inflammatory protein DNA vaccine-loaded bacterial ghost enhances immune protective efficacy in C57BL/6 mice. Vaccine 2014;32:6054–6060 [CrossRef][PubMed]
    [Google Scholar]
  5. Eko FO, Talin BA, Lubitz W. Development of a Chlamydia trachomatis bacterial ghost vaccine to fight human blindness. Hum Vaccin 2008;4:176–183 [CrossRef][PubMed]
    [Google Scholar]
  6. Mayr UB, Haller C, Haidinger W, Atrasheuskaya A, Bukin E et al. Bacterial ghosts as an oral vaccine: a single dose of Escherichia coli O157: H7 bacterial ghosts protects mice against lethal challenge. Infect Immun 2005;73:4810–4817 [CrossRef][PubMed]
    [Google Scholar]
  7. Eko FO, Witte A, Huter V, Kuen B, Fürst-Ladani S et al. New strategies for combination vaccines based on the extended recombinant bacterial ghost system. Vaccine 1999;17:1643–1649 [CrossRef][PubMed]
    [Google Scholar]
  8. Stevens M. Bacterial ghosts modulation of Innate immunity: immune responses during chlamydia infection. Electronic Theses & Dissertations Collection for Atlanta University & Clark Atlanta University 2015
  9. Crump JA, Luby SP, Mintz ED. The global burden of typhoid fever. Bull World Health Organ 2004;82:346–353[PubMed]
    [Google Scholar]
  10. Lin FY, Vo AH, Phan VB, Nguyen TT, Bryla D et al. The epidemiology of typhoid fever in the dong thap province, Mekong delta region of Vietnam. Am J Trop Med Hyg 2000;62:644–648 [CrossRef][PubMed]
    [Google Scholar]
  11. Garmory HS, Brown KA, Titball RW. Salmonella vaccines for use in humans: present and future perspectives. FEMS Microbiol Rev 2002;26:339–353[PubMed]
    [Google Scholar]
  12. Wain J, Hoa NT, Chinh NT, Vinh H, Everett MJ et al. Quinolone-resistant Salmonella typhi in viet nam: molecular basis of resistance and clinical response to treatment. Clin Infect Dis 1997;25:1404–1410 [CrossRef][PubMed]
    [Google Scholar]
  13. Shanahan PM, Jesudason MV, Thomson CJ, Amyes SG. Molecular analysis of and identification of antibiotic resistance genes in clinical isolates of Salmonella typhi from India. J Clin Microbiol 1998;36:1595–1600[PubMed]
    [Google Scholar]
  14. Ochiai RL, Acosta CJ, Agtini M, Bhattacharya SK, Bhutta ZA et al. The use of typhoid vaccines in Asia: the DOMI experience. Clin Infect Dis 2007;45:S34–S38 [CrossRef][PubMed]
    [Google Scholar]
  15. Fraser A, Paul M, Goldberg E, Acosta CJ, Leibovici L. Typhoid fever vaccines: systematic review and meta-analysis of randomised controlled trials. Vaccine 2007;25:7848–7857 [CrossRef][PubMed]
    [Google Scholar]
  16. Bhutta ZA, Capeding MR, Bavdekar A, Marchetti E, Ariff S et al. Immunogenicity and safety of the Vi-CRM197 conjugate vaccine against typhoid fever in adults, children, and infants in south and southeast Asia: results from two randomised, observer-blind, age de-escalation, phase 2 trials. Lancet Infect Dis 2014;14:119–129 [CrossRef][PubMed]
    [Google Scholar]
  17. Moore SE, Jalil F, Szu SC, Hahn-Zoric M, Prentice AM et al. Revaccination does not improve an observed deficit in antibody responses in Pakistani adults born of a lower birth weight. Vaccine 2008;26:158–165 [CrossRef][PubMed]
    [Google Scholar]
  18. Tapa S, Cvjetanović B. Controlled field trial on the effectiveness of one and two doses of acetone-inactivated and dried typhoid vaccine. Bull World Health Organ 1975;52:75[PubMed]
    [Google Scholar]
  19. Mader HJ, Szostak MP, Hensel A, Lubitz W, Haslberger AG. Endotoxicity does not limit the use of bacterial ghosts as candidate vaccines. Vaccine 1997;15:195–202 [CrossRef][PubMed]
    [Google Scholar]
  20. Lubitz W. Bacterial ghosts as carrier and targeting systems. Expert Opin Biol Ther 2001;1:765–771 [CrossRef][PubMed]
    [Google Scholar]
  21. Huter V, Szostak MP, Gampfer J, Prethaler S, Wanner G et al. Bacterial ghosts as drug carrier and targeting vehicles. J Control Release 1999;61:51–63 [CrossRef][PubMed]
    [Google Scholar]
  22. Ji J, Griffiths KL, Milburn PJ, Hirst TR, O'Neill HC. The B subunit of Escherichia coli heat-labile toxin alters the development and antigen-presenting capacity of dendritic cells. J Cell Mol Med 2015;19:2019–2031 [CrossRef][PubMed]
    [Google Scholar]
  23. Kang HY, Srinivasan J, Curtiss R. Immune responses to recombinant pneumococcal PspA antigen delivered by live attenuated Salmonella enterica serovar Typhimurium vaccine. Infect Immun 2002;70:1739–1749 [CrossRef][PubMed]
    [Google Scholar]
  24. Jawale CV, Kim SW, Lee JH. Tightly regulated bacteriolysis for production of empty Salmonella enteritidis envelope. Vet Microbiol 2014;169:179–187 [CrossRef][PubMed]
    [Google Scholar]
  25. Izant JG, Weintraub H. Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell 1984;36:1007–1015 [CrossRef][PubMed]
    [Google Scholar]
  26. Vinod N, Oh S, Kim S, Choi CW, Kim SC et al. Chemically induced Salmonella enteritidis ghosts as a novel vaccine candidate against virulent challenge in a rat model. Vaccine 2014;32:3249–3255 [CrossRef][PubMed]
    [Google Scholar]
  27. Jawale CV, Lee JH. Development of a biosafety enhanced and immunogenic Salmonella enteritidis ghost using an antibiotic resistance gene free plasmid carrying a bacteriophage lysis system. PLoS One 2013;8:e78193 [CrossRef][PubMed]
    [Google Scholar]
  28. Jawale CV, Lee JH. Salmonella enterica serovar enteritidis ghosts carrying the Escherichia coli heat-labile enterotoxin B subunit are capable of inducing enhanced protective immune responses. Clin Vaccine Immunol 2014;21:799–807 [CrossRef][PubMed]
    [Google Scholar]
  29. Hur J, Kim CS, Eo SK, Park SY, Lee JH. Salmonella ghosts expressing enterotoxigenic Escherichia coli k88ab, k88ac, k99, and fasa fimbrial antigens induce robust immune responses in a mouse model. Vet Q 2015;35:125–132 [CrossRef][PubMed]
    [Google Scholar]
  30. Won G, Kim TH, Lee JH. A novel Salmonella strain inactivated by a regulated autolysis system and expressing the B subunit of Shiga toxin 2e efficiently elicits immune responses and confers protection against virulent Stx2e-producing Escherichia coli. BMC Vet Res 2017;13:40 [CrossRef][PubMed]
    [Google Scholar]
  31. Peirson SN, Butler JN, Foster RG. Experimental validation of novel and conventional approaches to quantitative real-time PCR data analysis. Nucleic Acids Res 2003;31:e73 [CrossRef][PubMed]
    [Google Scholar]
  32. Overbergh L, Valckx D, Waer M, Mathieu C. Quantification of murine cytokine mRNAs using real time quantitative reverse transcriptase PCR. Cytokine 1999;11:305–312 [CrossRef][PubMed]
    [Google Scholar]
  33. Trebicka E, Jacob S, Pirzai W, Hurley BP, Cherayil BJ. Role of antilipopolysaccharide antibodies in serum bactericidal activity against Salmonella enterica serovar Typhimurium in healthy adults and children in the United States. Clin Vaccine Immunol 2013;20:1491–1498 [CrossRef][PubMed]
    [Google Scholar]
  34. Okahashi N, Yamamoto M, Vancott JL, Chatfield SN, Roberts M et al. Oral immunization of interleukin-4 (IL-4) knockout mice with a recombinant Salmonella strain or cholera toxin reveals that CD4+ Th2 cells producing IL-6 and IL-10 are associated with mucosal immunoglobulin A responses. Infect Immun 1996;64:1516–1525[PubMed]
    [Google Scholar]
  35. Amara AA, Salem-Bekhit MM, Alanazi FK. Sponge-like: a new protocol for preparing bacterial ghosts. ScientificWorldJournal 2013;2013:1–7 [CrossRef][PubMed]
    [Google Scholar]
  36. Takahara M, Hibler DW, Barr PJ, Gerlt JA, Inouye M. The ompA signal peptide directed secretion of Staphylococcal nuclease A by Escherichia coli. J Biol Chem 1985;260:2670–2674[PubMed]
    [Google Scholar]
  37. Alone PV, Garg LC. Secretory and GM1 receptor binding role of N-terminal region of LTB in Vibrio cholerae. Biochem Biophys Res Commun 2008;376:770–774 [CrossRef][PubMed]
    [Google Scholar]
  38. de Haan L, Verweij WR, Feil IK, Holtrop M, Hol WG et al. Role of GM1 binding in the mucosal immunogenicity and adjuvant activity of the Escherichia coli heat-labile enterotoxin and its B subunit. Immunology 1998;94:424–430 [CrossRef][PubMed]
    [Google Scholar]
  39. Rappuoli R, Pizza M, Douce G, Dougan G. Structure and mucosal adjuvanticity of cholera and Escherichia coli heat-labile enterotoxins. Immunol Today 1999;20:493–500 [CrossRef][PubMed]
    [Google Scholar]
  40. Mestecky J. The common mucosal immune system and current strategies for induction of immune responses in external secretions. J Clin Immunol 1987;7:265–276 [CrossRef][PubMed]
    [Google Scholar]
  41. Hur J, Lee JH. A new enterotoxigenic Escherichia coli vaccine candidate constructed using a Salmonella ghost delivery system: comparative evaluation with a commercial vaccine for neonatal piglet colibacillosis. Vet Immunol Immunopathol 2015;164:101–109 [CrossRef][PubMed]
    [Google Scholar]
  42. Moon JJ, Mcsorley SJ. Tracking the dynamics of Salmonella specific T cell responses. Visualizing Immunity Berlin, Heidelberg: Springer; 2009; pp.179–198[CrossRef]
    [Google Scholar]
  43. Hornick RB, Greisman SE, Woodward TE, Dupont HL, Dawkins AT et al. Typhoid fever: pathogenesis and immunologic control. N Engl J Med 1970;283:739–746 [CrossRef][PubMed]
    [Google Scholar]
  44. Ohl ME, Miller SI. Salmonella: a model for bacterial pathogenesis. Annu Rev Med 2001;52:259–274 [CrossRef][PubMed]
    [Google Scholar]
  45. Hsieh C-S, Macatonia SE, Tripp CS, Wolf SF, O’Garra A et al. Development of TH1 CD4+ T cells through IL-12. Science 1993;260:547[CrossRef]
    [Google Scholar]
  46. Fitch E, Harper E, Skorcheva I, Kurtz SE, Blauvelt A. Pathophysiology of psoriasis: recent advances on IL-23 and Th17 cytokines. Curr Rheumatol Rep 2007;9:461–467 [CrossRef][PubMed]
    [Google Scholar]
  47. Robbins JB, Schneerson R, Szu SC. Perspective: hypothesis: serum IgG antibody is sufficient to confer protection against infectious diseases by inactivating the inoculum. J Infect Dis 1995;171:1387–1398 [CrossRef][PubMed]
    [Google Scholar]
  48. Boyd MA, Tennant SM, Saague VA, Simon R, Muhsen K et al. Serum bactericidal assays to evaluate typhoidal and nontyphoidal Salmonella vaccines. Clin Vaccine Immunol 2014;21:712–721 [CrossRef][PubMed]
    [Google Scholar]
  49. Borrow R, Balmer P, Miller E. Meningococcal surrogates of protection-serum bactericidal antibody activity. Vaccine 2005;23:2222–2227 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000543
Loading
/content/journal/jmm/10.1099/jmm.0.000543
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error