A DNA Vaccine Encoding the Enterohemorragic Escherichia coli Shiga-Like Toxin 2 A2 and B Subunits Confers Protective Immunity to Shiga Toxin Challenge in the Murine Model (original) (raw)

Abstract

Production of verocytotoxin or Shiga-like toxin (Stx), particularly Stx2, is the basis of hemolytic uremic syndrome, a frequently lethal outcome for subjects infected with Stx2-producing enterohemorrhagic Escherichia coli (EHEC) strains. The toxin is formed by a single A subunit, which promotes protein synthesis inhibition in eukaryotic cells, and five B subunits, which bind to globotriaosylceramide at the surface of host cells. Host enzymes cleave the A subunit into the A 1 peptide, endowed with N-glycosidase activity to the 28S rRNA, and the A 2 peptide, which confers stability to the B pentamer. We report the construction of a DNA vaccine (pStx2⌬AB) that expresses a nontoxic Stx2 mutated form consisting of the last 32 amino acids of the A 2 sequence and the complete B subunit as two nonfused polypeptides. Immunization trials carried out with the DNA vaccine in BALB/c mice, alone or in combination with another DNA vaccine encoding granulocytemacrophage colony-stimulating factor, resulted in systemic Stx-specific antibody responses targeting both A and B subunits of the native Stx2. Moreover, anti-Stx2 antibodies raised in mice immunized with pStx2⌬AB showed toxin neutralization activity in vitro and, more importantly, conferred partial protection to Stx2 challenge in vivo. The present vector represents the second DNA vaccine so far reported to induce protective immunity to Stx2 and may contribute, either alone or in combination with other procedures, to the development of prophylactic or therapeutic interventions aiming to ameliorate EHEC infection-associated sequelae.

Loading...

Loading Preview

Sorry, preview is currently unavailable. You can download the paper by clicking the button above.

References (49)

  1. Acheson, D. W., S. A. De Breucker, M. Jacewicz, L. L. Lincicome, A. Dono- hue-Rolfe, A. V. Kane, and G. T. Keusch. 1995. Expression and purification of Shiga-like toxin II B subunits. Infect. Immun. 63:301-308.
  2. Acheson, D. W., M. M. Levine, J. B. Kaper, and G. T. Keusch. 1996. Pro- tective immunity to Shiga-like toxin I following oral immunization with Shiga-like toxin I B-subunit-producing Vibrio cholerae CVD 103-HgR. Infect. Immun. 64:355-357.
  3. Amara, R. R., F. Villinger, J. D. Altman, S. L. Lydy, S. P. O'Neil, S. I. Staprans, D. C. Montefiori, Y. Xu, J. G. Herndon, L. S. Wyatt, M. A. Candido, N. L. Kozyr, P. L. Earl, J. M. Smith, H. L. Ma, B. D. Grimm, M. L. Hulsey, J. Miller, H. M. McClure, J. M. McNicholl, B. Moss, and H. L. Robinson. 2001. Control of a mucosal challenge and prevention of AIDS by a multiprotein DNA/MVA vaccine. Science 692:69-74.
  4. Andersen, P. H., M. Nielsen, and O. Lund. 2006. Prediction of residues in discontinuous B cell epitopes using protein 3D structures. Protein Sci. 15: 2558-2567.
  5. Bielaszewska, M., I. Clarke, M. A. Karmali, and M. Petric. 1997. Localiza- tion of intravenously administered verocytotoxins (Shiga-like toxins) 1 and 2 in rabbits immunized with homologous and heterologous toxoids and toxin subunits. Infect. Immun. 65:2509-2516.
  6. Bosworth, B. T., J. E. Samuel, H. W. Moon, A. D. OBrien, V. M. Gordon, and S. C. Whipp. 1995. Vaccination with genetically modified Shiga-like toxin IIe prevents edema disease in swine. Infect. Immun. 64:55-60.
  7. Capozzo, A. V., V. Pistone Creydt, G. Dran, G. Fernandez, S. Gomez, L. V. Bentancor, C. Rubel, C. Ibarra, M. Isturiz, and M. S. Palermo. 2003. De- velopment of DNA vaccines against hemolytic-uremic syndrome in a murine model. Infect. Immun. 71:3971-3978.
  8. Caprioli, A., S. Morabito, H. Brugere, and E. Oswald. 2005. Enterohaemor- rhagic Escherichia coli: emerging issues on virulence and modes of transmis- sion. Vet. Res. 36:289-311.
  9. Donohue-Rolfe, A., D. W. Acheson, and G. T. Keusch. 1991. Shiga toxin: purification, structure, and function. Rev. Infect. Dis. 13(Suppl. 4):S293- S297.
  10. Elango, N., J. Vivekananda, R. Strong, and M. S. Katz. 1997. Nuclei isolation from bone cells for nuclear run-on assays. BioTechniques 23:422-424.
  11. Ferna ´ndez, G. C., C. Rubel, G. Dran, S. Go ´mez, M. A. Isturiz, and M. S. Palermo. 2000. Shiga toxin-2 induces neutrophilia and neutrophil activation in a murine model of hemolytic uremic syndrome. Clin. Immunol. 95:227- 234.
  12. Friedrich, A. W., M. Bielaszewska, W. L. Zhang, M. Pulz, T. Kuczius, A. Ammon, and H. Karch. 2002. Escherichia coli harboring Shiga toxin 2 gene variants: frequency and association with clinical symptoms. J. Infect. Dis. 185:74-84.
  13. Gordon, V. M., S. C. Whipp, H. W. Moon, A. D. O'Brien, and J. E. Samuel. 1992. An enzymatic mutant of Shiga-like toxin II variant is a vaccine candi- date for edema disease of swine. Infect. Immun. 60:485-490.
  14. Stx2 DNA VACCINE AND PROTECTIVE IMMUNITY IN MICE 717
  15. Griffin, P. M., and R. V. Tauxe. 1991. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the asso- ciated hemolytic uremic syndrome. Epidemiol. Rev. 13:60-98.
  16. Harari, I., and R. Arnon. 1990. Carboxy-terminal peptides from the B sub- unit of Shiga toxin induce a local and parenteral protective effect. Mol. Immunol. 27:613-621.
  17. Ishikawa, S., K. Kawahara, Y. Kagami, Y. Isshiki, A. Kaneko, H. Matsui, N. Okada, and H. Danbara. 2003. Protection against Shiga toxin 1 challenge by immunization of mice with purified mutant Shiga toxin 1. Infect. Immun. 71:3235-3239.
  18. Islam, M. S., and W. H. Stimson. 1990. Production and characterization of monoclonal antibodies with therapeutic potential against Shiga toxin. J. Clin. Lab. Immunol. 33:11-16.
  19. Kaper, J. B. 1998. Enterohemorrhagic Escherichia coli. Curr. Opin. Micro- biol. 1:103-108.
  20. Karmali, M. A. 1989. Infection by verotoxin-producing Escherichia coli. Clin. Microbiol. Rev. 2:15-38.
  21. Karmali, M. A., M. Petric, C. Lim, P. C. Fleming, and B. T. Steele. 1983. Escherichia coli cytotoxin, haemolytic-uraemic syndrome, and haemorragic colitis. Lancet ii:1299-1300.
  22. Konadu, E., A. Donohue-Rolfe, S. B. Calderwood, V. Pozsgay, J. Shiloach, J. B. Robbins, and S. C. Szu. 1999. Syntheses and immunologic properties of Escherichia coli O157 O-specific polysaccharide and Shiga toxin 1 B subunit conjugates in mice. Infect. Immun. 67:6191-6193.
  23. Krautz-Peterson, G., S. Chapman-Bonofiglio, K. Boisvert, H. Feng, I. M. Herman, S. Tzipori, and A. S. Sheoran. 2008. Intracellular neutralization of Shiga toxin 2 by an A subunit-specific human monoclonal antibody. Infect. Immun. 76:1931-1939.
  24. Kusakabe, K., K. O. Xin, H. Katoh, K. Sumino, E. Hagiwara, S. Kawamoto, K. Okuda, Y. Miyagi, I. Aoki, K. Nishioka, D. Klinman, and K. Okuda. 2000. The timing of GM-CSF expression plasmid administration influences the Th1/Th2 response induced by an HIV-1-specific DNA vaccine. J. Immunol. 164:3102-3111.
  25. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T7. Nature 227:680-685.
  26. Lasaro, M. O., W. B. Luiz, M. E. Sbrogio-Almeida, L. S. Nishimura, B. E. C. Guth, and L. C. S. Ferreira. 2004. Combined vaccine regimen based on parenteral priming with a DNA vaccine and administration of an oral booster consisting of a recombinant Salmonella enterica Serovar Typhi- murium vaccine strain for immunization against human-derived enterotoxi- genic Escherichia coli strains. Infect. Immun. 72:6480-6491.
  27. Le, T. P., K. M. Coonan, R. C. Hedstrom, Y. Charoenvit, M. Sedegah, J. E. Epstein, S. Kumar, R. Wang, D. L. Doolan, J. D. Maguire, S. E. Parker, P. Hobart, J. Norman, and S. L. Hoffman. 2000. Safety, tolerability and hu- moral immune responses after intramuscular administration of a malaria DNA vaccine to healthy adult volunteers. Vaccine 18:1893-1901.
  28. Lindgren, S. W., J. E. Samuel, C. K. Schmitt, and A. D. O'Brien. 1994. The specific activities of Shiga-like toxin type II (SLT-II) and SLT-II-related toxins of enterohemorrhagic Escherichia coli differ when measured by Vero cell cytotoxicity but not by mouse lethality. Infect. Immun. 62:623-631.
  29. Lingwood, C. A. 1996. Role of verotoxin receptors in pathogenesis. Trends Microbiol. 4:147-153.
  30. Lo ´pez, E. L., V. Prado-Jime ´nez, M. O'Ryan-Gallardo, and M. M. Contrini. 2000. Shigella and Shiga toxin-producing Escherichia coli causing bloody diarrhea in Latin America. Infect. Dis. Clin. North Am. 14:41-65.
  31. Luiz, W. B., R. C. M. Cavalcante, J. D. Paccez, R. D. Souza, M. E. Sbrogio- Almeida, R. C. C. Ferreira, and L. C. S. Ferreira. 2008. 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. Vaccine 26:3998-4005.
  32. MacGregor, R. R., J. D. Boyer, K. E. Ugen, K. E. Lacy, S. J. Gluckman, M. L. Bagarazzi, M. A. Chattergoon, Y. Baine, T. J. Higgins, R. B. Ciccarelli, L. R. Coney, R. S. Ginsberg, and D. B. Weiner. 1998. First human trial of a DNA-based vaccine for treatment of human immunodeficiency virus type 1 infection: safety and host response. J. Infect. Dis. 178:92-100.
  33. Makino, S., M. Watarai, H. Tabuchi, T. Shirahata, H. Furuoka, Y. Koba- yashi, and Y. Takeda. 2001. Genetically modified Shiga toxin 2e (Stx2e) producing Escherichia coli is a vaccine candidate for porcine edema disease. Microb. Pathog. 31:1-8.
  34. Marcato, P., G. Mulvey, R. J. Read, K. Vander Helm, P. N. Nation, and G. D. Armstrong. 2001. Immunoprophylactic potential of cloned Shiga toxin 2B subunit. J. Infect. Dis. 183:435-443.
  35. Mukherjee, J., K. Chios, D. Fishwild, D. Hudson, S. O'Donnell, S. M. Rich, A. Donohue-Rolfe, and S. Tzipori. 2002. Human Stx2-specific monoclonal antibodies prevent systemic complications of Escherichia coli O157:H7 in- fection. Infect. Immun. 70:612-619.
  36. National Institutes of Health. 1998. NIH policy manual. Chapter 3035, working safely with hazardous biological materials. National Institutes of Health, Bethesda, MD.
  37. National Research Council. 1996. Guide for the care and use of laboratory animals. National Academy Press, Washington, DC.
  38. Ohmura, M., S. Yamasaki, H. Kurazono, K. Kashiwagi, K. Igarashi, and Y. Takeda. 1993. Characterization of non-toxic mutant toxins of Vero toxin 1 that were constructed by replacing amino acids in the A subunit. Microb. Pathog. 15:169-176.
  39. Palermo, M. S., C. Rubel, M. F. Alves Rosa, G. Ferna ´ndez, F. G. Ferna ´ndez Alonso, M. Rivas, and M. A. Isturiz. 1999. Pretreatment of mice with LPS or IL-1␤ exerts opposite effects on Shiga-toxin-2 lethality. Clin. Exp. Immunol. 119:77-83.
  40. Pistone Creydt, V., M. Fernandez Miyakawa, F. Martín, E. Zotta, C. Silber- stein, and C. Ibarra. 2004. The Shiga toxin 2 B subunit inhibits net fluid absorption in human colon and elicits fluid accumulation in rat colon loops. Braz J. Med. Biol. Res. 37:799-808.
  41. Rivas, M., E. Miliwebsky, I. Chinen, N. Deza, and G. A. Leotta. 2006. The epidemiology of hemolytic uremic syndrome in Argentina. Diagnosis of the etiologic agent, reservoirs and routes of transmission. Medicina (Buenos Aires) 66:27-32.
  42. Ru ¨ssmann, H., H. Schmidt, J. Heesemann, A. Caprioli, and H. Karch. 1994. Variants of Shiga-like toxin II constitute a major toxin component in Esch- erichia coli O157 strains from patients with haemolytic uraemic syndrome. J. Med. Microbiol. 40:338-343.
  43. Sharon, N. 2006. Carbohydrates as future anti-adhesion drugs for infectious diseases. Biochim. Biophys. Acta 1760:527-537.
  44. Sheoran, A. S., S. Chapman, P. Singh, A. Donohue-Rolfe, and S. Tzipori. 2003. Stx2-specific human monoclonal antibodies protect mice against lethal infection with Escherichia coli expressing Stx2 variants. Infect. Immun. 71: 3125-3130.
  45. Slutsker, L., A. A. Ries, K. D. Greene, J. G. Wells, L. Hutwagner, and P. M. Griffin. 1997. Escherichia coli O157:H7 diarrhea in the United States: clinical and epidemiologic features. Ann. Intern. Med. 126:505-513.
  46. Smith, M. J., L. D. Teel, H. M. Carvalho, A. R. Melton-Celsa, and A. D. O'Brien. 2006. Development of a hybrid Shiga holotoxoid vaccine to elicit heterologous protection against Shiga toxins types 1 and 2. Vaccine 24:4122- 4129.
  47. Tzipori, S., A. Sheoran, D. Akiyoshi, A. Donohue-Rolfe, and H. Trachtman. 2004. Antibody therapy in the management of Shiga toxin-induced hemolytic uremic syndrome. Clin. Microbiol. Rev. 17:926-941.
  48. Wen, S. X., L. D. Teel, N. A. Judge, and A. D. O'Brien. 2006. Genetic toxoids of Shiga toxin types 1 and 2 protect mice against homologous but not heterologous toxin challenge. Vaccine 24:1142-1148.
  49. Zhu, C., J. Yu, Z. Yang, K. Davis, H. Rios, B. Wang, G. Glenn, and E. C. Boedeker. 2007. Protection against Shiga toxin-producing Escherichia coli infection by transcutaneous immunization with Shiga toxin subunit B. Clin. Vaccine Immunol. 15:359-366.