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Delivery of Chicken Egg Ovalbumin to Dendritic Cells by Listeriolysin O-Secreting Vegetative Bacillus subtilis

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초록

Listeriolysin O (LLO), one of the most immunogenic proteins of Listeria monocytogenes and its main virulence factor, mediates bacterial escape from the phagosome of the infected cell. Thus, its expression in a nonpathogenic bacterial host may enable effective delivery of heterologous antigens to the host cell cytosol and lead to their processing predominantly through the cytosolic MHC class I presentation pathway. The aim of this project was to characterize the delivery of a model antigen, chicken egg ovalbumin (OVA), to the cytosol of dendritic cells by recombinant Bacillus subtilis vegetative cells expressing LLO. Our work indicated that LLO produced by non-sporulating vegetative bacteria was able to support OVA epitope presentation by MHC I molecules on the surface of antigen presenting cells and consequently influence OVA-specific cytotoxic T cell activation. Additionally, it was proven that the genetic context of the epitope sequence is of great importance, as only the native full-sequence OVA fused to the N-terminal fragment of LLO was sufficient for effective epitope delivery and activation of CD8+ lymphocytes. These results demonstrate the necessity for further verification of the fusion antigen potency of enhancing the MHC I presentation, and they prove that LLO-producing B. subtilis may represent a novel and attractive candidate for a vaccine vector.

Abstract

Listeriolysin O (LLO), one of the most immunogenic proteins of Listeria monocytogenes and its main virulence factor, mediates bacterial escape from the phagosome of the infected cell. Thus, its expression in a nonpathogenic bacterial host may enable effective delivery of heterologous antigens to the host cell cytosol and lead to their processing predominantly through the cytosolic MHC class I presentation pathway. The aim of this project was to characterize the delivery of a model antigen, chicken egg ovalbumin (OVA), to the cytosol of dendritic cells by recombinant Bacillus subtilis vegetative cells expressing LLO. Our work indicated that LLO produced by non-sporulating vegetative bacteria was able to support OVA epitope presentation by MHC I molecules on the surface of antigen presenting cells and consequently influence OVA-specific cytotoxic T cell activation. Additionally, it was proven that the genetic context of the epitope sequence is of great importance, as only the native full-sequence OVA fused to the N-terminal fragment of LLO was sufficient for effective epitope delivery and activation of CD8+ lymphocytes. These results demonstrate the necessity for further verification of the fusion antigen potency of enhancing the MHC I presentation, and they prove that LLO-producing B. subtilis may represent a novel and attractive candidate for a vaccine vector.

참고문헌 (52)

  1. Cohn L, "Dendritic cell-targeted vaccines", Front. Immunol., 5, pp.255~ (2014)
  2. Rudolph MG, "How TCRs bind MHCs, peptides, and coreceptors", Annu. Rev. Immunol., 24, pp.419~466 (2006)
  3. Daniels MA, "TCR signaling in T cell memory", Front. Immunol., 6, pp.617~ (2015)
  4. Trovato M, "Novel antigen delivery systems", World J. Virol., 4, pp.156~168 (2015)[DOI]
  5. VanCott JL, "Regulation of host immune responses by modification of Salmonella virulence genes", Nat. Med., 4, pp.1247~1252 (1998)[DOI]
  6. McBride BW, "Protective efficacy of a recombinant protective antigen against Bacillus anthracis challenge and assessment of immunological markers", Vaccine, 16, pp.810~817 (1998)[DOI]
  7. Airaksinen U, "Production of Chlamydia pneumoniae proteins in Bacillus subtilis and their use in characterizing immune responses in the experimental infection model", Clin. Diagn. Lab. Immunol., 10, pp.367~375 (2003)
  8. Olmos-Soto J, "Genetic system constructed to overproduce and secrete proinsulin in Bacillus subtilis", Appl. Microbiol. Biotechnol., 62, pp.369~373 (2003)
  9. Kakeshita H, "Secretion of biologically-active human interferonbeta by Bacillus subtilis", Biotechnol. Lett., 33, pp.1847~1852 (2011)[DOI]
  10. Westers L, "Secretion of functional human interleukin-3 from Bacillus subtilis", J. Biotechnol., 123, pp.211~224 (2006)[DOI]
  11. Negri A, "Expression and display of Clostridium difficile protein FliD on the surface of Bacillus subtilis spores", J. Med. Microbiol., 62, pp.1379~1385 (2013)[DOI]
  12. Hinc K, "Expression and display of UreA of Helicobacter acinonychis on the surface of Bacillus subtilis spores", Microb. Cell Factor., 9, pp.2~ (2010)[DOI]
  13. Ning D, "Surface-displayed VP28 on Bacillus subtilis spores induce protection against white spot syndrome virus in crayfish by oral administration", J. Appl. Microbiol., 111, pp.1327~1336 (2011)[DOI]
  14. Duc le H, "Bacterial spores as vaccine vehicles", Infect. Immun., 71, pp.2810~2818 (2003)
  15. Mauriello EM, "Display of heterologous antigens on the Bacillus subtilis spore coat using CotC as a fusion partner", Vaccine, 22, pp.1177~1187 (2004)[DOI]
  16. Gomes PA, "Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7", Braz. J. Microbiol., 40, pp.333~338 (2009)[DOI]
  17. Titball RW, "Vaccines against intracellular bacterial pathogens", Drug Discov. Today, 13, pp.596~600 (2008)[DOI]
  18. Vazquez-Boland JA, "Listeria pathogenesis and molecular virulence determinants", Clin. Microbiol. Rev., 14, pp.584~640 (2001)[DOI]
  19. Pamer EG, "Immune responses to Listeria monocytogenes", Nat. Rev. Immunol., 4, pp.812~823 (2004)[DOI]
  20. Bahey-El-Din M, "Expression of two Listeria monocytogenes antigens (P60 and LLO) in Lactococcus lactis and examination for use as live vaccine vectors", J. Med. Microbiol., 59, pp.904~912 (2010)[DOI]
  21. Radford KJ, "A recombinant E. coli v accine t o promote MHC class I-dependent antigen presentation: application to cancer immunotherapy", Gene Ther., 9, pp.1455~1463 (2002)[DOI]
  22. Huang JM, "Cytoplasmic delivery of antigens, by Bacillus subtilis enhances Th1 responses", Vaccine, 26, pp.6043~6052 (2008)[DOI]
  23. Carrero JA, "Listeriolysin O is strongly immunogenic independently of its cytotoxic activity", PLoS One, 7, pp.e32310~ (2012)[DOI]
  24. Sewell DA, "Recombinant Listeria vaccines containing PEST sequences are potent immune adjuvants for the tumorassociated antigen human papillomavirus-16 E7", Cancer Res, 64, pp.8821~8825 (2004)
  25. Decatur AL, "A PEST-like sequence in listeriolysin O essential for Listeria monocytogenes pathogenicity", Science, 290, pp.992~995 (2000)[DOI]
  26. Schnupf P, "Phosphorylation, ubiquitination and degradation of listeriolysin O in mammalian cells: role of the PEST-like sequence", Cell. Microbiol., 8, pp.353~364 (2006)
  27. Carreno LJ, "The half-life of the T-cell receptor", peptide-major histocompatibility complex interaction can modulate T-cell activation in response to bacterial challenge, 2007, pp.121, 227~237
  28. Albaghdadi H, "Selectively reduced intracellular proliferation of Salmonella enterica serovar Typhimurium within APCs limits antigen presentation and development of a rapid CD8 T cell response", J. Immunol., 183, pp.3778~3787 (2009)[DOI]
  29. Radford KJ, "Recombinant E. coli efficiently delivers antigen and maturation signals to human dendritic cells: presentation of MART1 to CD8+ T cells", Int. J. Cancer, 105, pp.811~819 (2003)[DOI]
  30. Youngman P, "Methods for genetic manipulation, cloning, and functional analysis of sporulation genes in Bacillus subtilis;Regulation of Prokaryotic Development", pp.65~87 (1989)
  31. Stachowiak, R., "Cytotoxicity of Listeriolysin O Produced by Membrane-Encapsulated Bacillus subtilis on Leukemia Cells", Journal of Microbiology and Biotechnology, 21(11), pp.1193~1198 (2011), 한국미생물·생명공학회
  32. Stachowiak R, "Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells", BMC Biotechnol, 14, pp.77~ (2014)[DOI]
  33. Antelmann H, "The extracellular proteome of Bacillus subtilis under secretion stress conditions", Mol. Microbiol., 49, pp.143~156 (2003)[DOI]
  34. Sojka L, "YbxF, a protein associated with exponential-phase ribosomes in Bacillus subtilis", J. Bacteriol., 189, pp.4809~4814 (2007)[DOI]
  35. Stachowiak R, "Cytotoxicity of bacterial metabolic products, including listeriolysin O, on leukocyte targets", J. Biomed. Biotechnol., 2012, pp.954375~ (2012)
  36. Sette A, "Epitope-based vaccines: an update on epitope identification, vaccine design and delivery", Curr. Opin. Immunol., 15, pp.461~470 (2003)[DOI]
  37. Li Pira G, "High throughput T epitope mapping and vaccine development", J. Biomed. Biotechnol., 2010, pp.325720~ (2010)
  38. Jawa V, "Immunogenicity of Therapeutic Fusion Proteins: Contributory Factors and Clinical Experience;Fusion Protein Technologies for Biopharmaceuticals", pp.75~90 (2013), John Wiley & Sons, Inc
  39. Schulke S, "A fusion protein consisting of the vaccine adjuvant monophosphoryl lipid A and the allergen ovalbumin boosts allergen-specific Th1, Th2, and Th17 responses in vitro", J. Immunol. Res., 2016, pp.4156456~ (2016)
  40. Yang E, "Recombinant BCG coexpressing Ag85B, ESAT-6 and Rv3620c elicits specific Th1 immune responses in C57BL", 6 mice, 2014, pp.69-70, 53~59
  41. Gupta S, "Vaccination with a fusion protein that introduces HIV-1 Gag antigen into a multitrimer CD40L construct results in enhanced CD8+ T cell responses and protection from viral challenge by vaccinia-Gag", J. Virol., 88, pp.1492~1501 (2014)[DOI]
  42. Cheng WF, "Fusion protein vaccines targeting two tumor antigens generate synergistic anti-tumor effects", PLoS One, 8, pp.e71216~ (2013)[DOI]
  43. Kohda C, "Dissociated linkage of cytokine-inducing activity and cytotoxicity to different domains of listeriolysin O from Listeria monocytogenes", Infect. Immun., 70, pp.1334~1341 (2002)[DOI]
  44. Singh R, "Fusion to listeriolysin O and delivery by Listeria monocytogenes enhances the immunogenicity of HER-2", neu and reveals subdominant epitopes in the FVB, J. Immunol., pp.2005, 175[DOI]
  45. Pohl S, "Proteomic analysis of Bacillus subtilis strains engineered for improved production of heterologous proteins", Proteomics, 13, pp.3298~3308 (2013)
  46. Mader U, "Bacillus subtilis functional genomics: genomewide analysis of the DegS-DegU regulon by transcriptomics and proteomics", Mol. Genet. Genomics, 268, pp.455~467 (2002)[DOI]
  47. Savina A, "Phagocytosis and antigen presentation in dendritic cells", Immunol. Rev., 219, pp.143~156 (2007)[DOI]
  48. Wei CH, "N-terminal trimer extension of nominal CD8 T cell epitopes is sufficient to promote crosspresentation to cognate CD8 T cells in vivo", J. Immunol., 179, pp.8280~8286 (2007)
  49. Luiz WB, "Boosting systemic and secreted antibody responses in mice orally immunized with recombinant Bacillus subtilis strains following parenteral priming with a DNA vaccine encoding the enterotoxigenic Escherichia coli (ETEC) CFA", I fimbriae B subunit, 2008, pp.26, 3998~4005
  50. Lee S, "Development of a Bacillus subtilisbased rotavirus vaccine", Clin. Vaccine Immunol., 17, pp.1647~1655 (2010)[DOI]
  51. Lin IY, "Live-attenuated bacterial vectors: tools for vaccine and therapeutic agent delivery", Vaccines, 3, pp.940~972 (2015)[DOI]
  52. Becavin C, "Comparison of widely used Listeria monocytogenes strains EGD, 10403S, and EGD-e highlights genomic variations underlying differences in pathogenicity", mBio, 5, pp.e00969~e00914 (2014)

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