DOI QR코드

DOI QR Code

Construction of a live attenuated Salmonella strain expressing FanC protein to prevent bovine enterotoxigenic Escherichia coli and evaluation of its immunogenicity in mice

  • Won, Gayeon (College of Veterinary Medicine, Chonbuk National University) ;
  • Kim, Hee Jung (College of Veterinary Medicine, Chonbuk National University) ;
  • Lee, John Hwa (College of Veterinary Medicine, Chonbuk National University)
  • Received : 2016.08.24
  • Accepted : 2016.12.20
  • Published : 2017.03.31

Abstract

To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

Keywords

References

  1. Acres SD, Isaacson RE, Babiuk LA, Kapitany RA. Immunization of calves against enterotoxigenic colibacillosis by vaccinating dams with purified K99 antigen and whole cell bacterins. Infect Immun 1979, 25, 121-126.
  2. Ascon MA, Hone DM, Walters N, Pascual DW. Oral immunization with a Salmonella typhimurium vaccine vector expressing recombinant enterotoxigenic Escherichia coli K99 fimbriae elicits elevated antibody titers for protective immunity. Infect Immun 1998, 66, 5470-5476.
  3. Blomfield IC, Calie PJ, Eberhardt KJ, McClain MS, Eisenstein BI. Lrp stimulates phase variation of type 1 fimbriation in Escherichia coli K-12. J Bacteriol 1993, 175, 27-36. https://doi.org/10.1128/jb.175.1.27-36.1993
  4. Chen H, Schifferli DM. Mucosal and systemic immune responses to chimeric fimbriae expressed by Salmonella enterica serovar Typhimurium vaccine strains. Infect Immun 2000, 68, 3129-3139. https://doi.org/10.1128/IAI.68.6.3129-3139.2000
  5. Garg R, Tolbert M, Oakes JL, Clemente TE, Bost KL, Piller KJ. Chloroplast targeting of FanC, the major antigenic subunit of Escherichia coli K99 fimbriae, in transgenic soybean. Plant Cell Rep 2007, 26, 1011-1023. https://doi.org/10.1007/s00299-007-0322-y
  6. Hajishengallis G, Arce S, Gockel CM, Connell TD, Russell MW. Immunomodulation with enterotoxins for the generation of secretory immunity or tolerance: applications for oral infections. J Dent Res 2005, 84, 1104-1116. https://doi.org/10.1177/154405910508401205
  7. Hariharan H, Coles M, Poole D, Page R. Antibiotic resistance among enterotoxigenic Escherichia coli from piglets and calves richia coli with diarrhea. Can Vet J 2004, 45, 605-606.
  8. Harmsen MM, Van Solt CB, Hoogendoorn A, Van Zijderveld FG, Niewold Ta, van der Meulen J. Escherichia coli F4 fimbriae specific llama single-domain antibody fragments effectively inhibit bacterial adhesion in vitro but poorly protect against diarrhoea. Vet Microbiol 2005, 111, 89-98. https://doi.org/10.1016/j.vetmic.2005.09.005
  9. Holmgren J, Czerkinsky C. Mucosal immunity and vaccines. Nat Med 2005, 11 (Suppl 4), S45-53. https://doi.org/10.1038/nm1213
  10. Husseiny MI, Hensel M. Evaluation of Salmonella live vaccines with chromosomal expression cassettes for translocated fusion proteins. Vaccine 2009, 27, 3780-3787. https://doi.org/10.1016/j.vaccine.2009.03.053
  11. Isaacson RE, Dean EA, Morgan RL, Moon HW. Immunization of suckling pigs against enterotoxigenic Escherichia coli-induced diarrheal disease by vaccinating dams with purified K99 or 987P pili: antibody production in response to vaccination. Infect Immun 1980, 29, 824-826.
  12. Kang HY, Srinivasan J, Curtiss R 3rd. Immune responses to recombinant pneumococcal PspA antigen delivered by live attenuated Salmonella enterica serovar Typhimurium vaccine. Infect Immun 2002, 70, 1739-1749. https://doi.org/10.1128/IAI.70.4.1739-1749.2002
  13. Mathers K, Leinonen M, Goldblatt D. Antibody response to outer membrane proteins of Moraxella catarrhalis in children with otitis media. Pediatr Infect Dis J 1999, 18, 982-988. https://doi.org/10.1097/00006454-199911000-00010
  14. Moon HW, Bunn TO. Vaccines for preventing enterotoxigenic Escherichia coli infections in farm animals. Vaccine 1993, 11, 213-220. https://doi.org/10.1016/0264-410X(93)90020-X
  15. Moon HW, Nagy B, Isaacson RE, Orskov I. Occurrence of K99 antigen on Escherichia coli isolated from pigs and colonization of pig ileum by $K99^+$ enterotoxigenic E. coli from calves and pigs. Infect Immun 1977, 15, 614-620.
  16. Morgan RL, Isaacson RE, Moon HW, Brinton CC, To CC. Immunization of suckling pigs against enterotoxigenic Escherichia coli-induced diarrheal disease by vaccinating dams with purified 987 or K99 pili: protection correlates with pilus homology of vaccine and challenge. Infect Immun 1978, 22, 771-777.
  17. Nagy B, Fekete PZ. Enterotoxigenic Escherichia coli (ETEC) in farm animals. Vet Res 1999, 30, 259-284.
  18. Nicolle LE, Ujack E, Brunka J, Bryan LE. Immunoblot analysis of serologic response to outer membrane proteins of Escherichia coli in elderly individuals with urinary tract infections. J Clin Microbiol 1988, 26, 2087-2091.
  19. O'Garra A, Arai N. The molecular basis of T helper 1 and T helper 2 cell differentiation. Trends Cell Biol 2000, 10, 542-550. https://doi.org/10.1016/S0962-8924(00)01856-0
  20. Pathangey L, Kohler JJ, Isoda R, Brown TA. Effect of expression level on immune responses to recombinant oral Salmonella enterica serovar Typhimurium vaccines. Vaccine 2009, 27, 2707-2711. https://doi.org/10.1016/j.vaccine.2009.02.072
  21. Puiprom O, Chantaroj S, Gangnonngiw W, Okada K, Honda T, Taniguchi T, Sawanpanyalert P. Identification of colonization factors of enterotoxigenic Escherichia coli with PCR-based technique. Epidemiol Infect 2010, 138, 519-524. https://doi.org/10.1017/S0950268809990720
  22. van der Woude MW, Arts PA, Bakker D, van Verseveld HW, de Graaf FK. Growth-rate-dependent synthesis of K99 fimbrial subunits is regulated at the level of transcription. J Gen Microbiology 1990, 136, 897-903. https://doi.org/10.1099/00221287-136-5-897
  23. VanCott JL, Staats HF, Pascual DW, Roberts M, Chatfield SN, Yamamoto M, Coste M, Carter PB, Kiyono H, McGhee JR. Regulation of mucosal and systemic antibody responses by T helper cell subsets, macrophages, and derived cytokines following oral immunization with live recombinant Salmonella. J Immunol 1996, 156, 1504-1514.
  24. Verdugo-Rodriguez A, Gam LH, Devi S, Koh CL, Puthucheary SD, Calva E, Pang T. Detection of antibodies against Salmonella typhi outer membrane protein (OMP) preparation in typhoid fever patients. Asians Pac J Allergy Immunol 1993, 11, 45-52.