Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
권호 표지
DOI QR코드

DOI QR Code

Protective efficacy of a novel multivalent vaccine in the prevention of diarrhea induced by enterotoxigenic Escherichia coli in a murine model

  • Zhao, Hong (School of Bioengineering, Dalian University of Technology) ;
  • Xu, Yongping (School of Bioengineering, Dalian University of Technology) ;
  • Li, Gen (School of Bioengineering, Dalian University of Technology) ;
  • Liu, Xin (School of Bioengineering, Dalian University of Technology) ;
  • Li, Xiaoyu (School of Bioengineering, Dalian University of Technology) ;
  • Wang, Lili (School of Bioengineering, Dalian University of Technology)
  • Received : 2021.03.13
  • Accepted : 2021.09.05
  • Published : 2022.01.31
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

Keywords

Acknowledgement

This study was supported by the National Key Research and Development Projects [grant number: 2017YFD0500601], National Natural Science Foundation of China [grant number: 31701719] and the Fundamental Research Funds for the Central Universities [grant number: DUT20LK29].

References

  1. Nagy B, Fekete PZ. Enterotoxigenic Escherichia coli in veterinary medicine. Int J Med Microbiol. 2005;295(6-7):443-454. https://doi.org/10.1016/j.ijmm.2005.07.003
  2. Chen X, Gao S, Jiao X, Liu XF. Prevalence of serogroups and virulence factors of Escherichia coli strains isolated from pigs with postweaning diarrhoea in eastern China. Vet Microbiol. 2004;103(1-2):13-20. https://doi.org/10.1016/j.vetmic.2004.06.014
  3. Mani S, Toapanta FR, McArthur MA, Qadri F, Svennerholm AM, Devriendt B, et al. Role of antigen specific T and B cells in systemic and mucosal immune responses in ETEC and Shigella infections, and their potential to serve as correlates of protection in vaccine development. Vaccine. 2019;37(34):4787-4793. https://doi.org/10.1016/j.vaccine.2019.03.040
  4. Sizemore DR, Roland KL, Ryan US. Enterotoxigenic Escherichia coli virulence factors and vaccine approaches. Expert Rev Vaccines. 2004;3(5):585-595. https://doi.org/10.1586/14760584.3.5.585
  5. Fleckenstein JM, Kuhlmann FM. Enterotoxigenic Escherichia coli Infections. Curr Infect Dis Rep. 2019;21(3):9-24. https://doi.org/10.1007/s11908-019-0665-x
  6. Chu H, Kang S, Ha S, Cho K, Park SM, Han KH, et al. Lactobacillus acidophilus expressing recombinant K99 adhesive fimbriae has an inhibitory effect on adhesion of enterotoxigenic Escherichia coli. Microbiol Immunol. 2005;49(11):941-948. https://doi.org/10.1111/j.1348-0421.2005.tb03687.x
  7. Do KH, Byun JW, Lee WK. Prevalence of O-serogroups, virulence genes, and F18 antigenic variants in Escherichia coli isolated from weaned piglets with diarrhea in Korea during 2008-2016. J Vet Sci. 2019;20(1):43-50. https://doi.org/10.4142/jvs.2019.20.1.43
  8. Zhu Y, Luo Q, Davis SM, Westra C, Vickers TJ, Fleckenstein JM. Molecular determinants of enterotoxigenic Escherichia coli heat-stable toxin secretion and delivery. Infect Immun. 2018;86(11):e00526-e18.
  9. Allen KP, Randolph MM, Fleckenstein JM. Importance of heat-labile enterotoxin in colonization of the adult mouse small intestine by human enterotoxigenic Escherichia coli strains. Infect Immun. 2006;74(2):869-875. https://doi.org/10.1128/IAI.74.2.869-875.2006
  10. Wolf MK. Occurrence, distribution, and associations of O and H serogroups, colonization factor antigens, and toxins of enterotoxigenic Escherichia coli. Clin Microbiol Rev. 1997;10(4):569-584. https://doi.org/10.1128/cmr.10.4.569
  11. Peterson JW, Whipp SC. Comparison of the mechanisms of action of cholera toxin and the heat-stable enterotoxins of Escherichia coli. Infect Immun. 1995;63(4):1452-1461. https://doi.org/10.1128/iai.63.4.1452-1461.1995
  12. Zhu C, Setty P, Boedeker EC. Development of live attenuated bacterial vaccines targeting Escherichia coli heat-labile and heat-stable enterotoxins. Vet Microbiol. 2017;202:72-78. https://doi.org/10.1016/j.vetmic.2017.04.010
  13. You J, Xu Y, He M, McAllister TA, Thacker PA, Li X, et al. Protection of mice against enterotoxigenic E. coli by immunization with a polyvalent enterotoxin comprising a combination of LTB, STa, and STb. Appl Microbiol Biotechnol. 2011;89(6):1885-1893. https://doi.org/10.1007/s00253-010-2991-7
  14. Altmann K, Pyliotis NA, Mukkur TK. A new method for the extraction and purification of K99 pili from enterotoxigenic Escherichia coli and their characterization. Biochem J. 1982;201(3):505-513. https://doi.org/10.1042/bj2010505
  15. Luiz WB, Rodrigues JF, Crabb JH, Savarino SJ, Ferreira LC. Maternal vaccination with a fimbrial tip adhesin and passive protection of neonatal mice against lethal human enterotoxigenic Escherichia coli challenge. Infect Immun. 2015;83(12):4555-4564. https://doi.org/10.1128/IAI.00858-15
  16. Reese AT, Chadaideh KS, Diggins CE, Schell LD, Beckel M, Callahan P, et al. Effects of domestication on the gut microbiota parallel those of human industrialization. eLife. 2021;10:e60197. https://doi.org/10.7554/eLife.60197
  17. Nagpal R, Wang S, Solberg Woods LC, Seshie O, Chung ST, Shively CA, et al. Comparative microbiome signatures and short-chain fatty acids in mouse, rat, non-human primate, and human feces. Front Microbiol. 2018;9:2897. https://doi.org/10.3389/fmicb.2018.02897
  18. Nadeau E, Fairbrother JM, Zentek J, Belanger L, Tremblay D, Tremblay CL, et al. Efficacy of a single oral dose of a live bivalent E. coli vaccine against post-weaning diarrhea due to F4 and F18-positive enterotoxigenic E. coli. Vet J. 2017;226:32-39. https://doi.org/10.1016/j.tvjl.2017.07.004
  19. Xu C, Wang Y, Sun R, Qiao X, Shang X, Niu W. Modulatory effects of vasoactive intestinal peptide on intestinal mucosal immunity and microbial community of weaned piglets challenged by an enterotoxigenic Escherichia coli (K88). PLoS One. 2014;9(8):e104183. https://doi.org/10.1371/journal.pone.0104183
  20. Chiu CJ, McArdle AH, Brown R, Scott HJ, Gurd FN. Intestinal mucosal lesion in low-flow states. I. A morphological, hemodynamic, and metabolic reappraisal. Arch Surg. 1970;101(4):478-483. https://doi.org/10.1001/archsurg.1970.01340280030009
  21. Duan Q, Xia P, Nandre R, Zhang W, Zhu G. Review of newly identified functions associated with the heatlabile toxin of enterotoxigenic Escherichia coli. Front Cell Infect Microbiol. 2019;9:292. https://doi.org/10.3389/fcimb.2019.00292
  22. Matias J, Berzosa M, Pastor Y, Irache JM, Gamazo C. Maternal vaccination. Immunization of sows during pregnancy against ETEC infections. Vaccines (Basel). 2017;5(4):48. https://doi.org/10.3390/vaccines5040048
  23. Xu C, Wang Y, Sun R, Qiao X, Shang X, Niu W. Modulatory effects of vasoactive intestinal peptide on intestinal mucosal immunity and microbial community of weaned piglets challenged by an enterotoxigenic Escherichia coli (K88). PLoS One. 2014;9(8):e104183. https://doi.org/10.1371/journal.pone.0104183
  24. Duan Q, Pang S, Wu W, Jiang B, Zhang W, Liu S, et al. A multivalent vaccine candidate targeting enterotoxigenic Escherichia coli fimbriae for broadly protecting against porcine post-weaning diarrhea. Vet Res (Faisalabad). 2020;51(1):93. https://doi.org/10.1186/s13567-020-00818-5
  25. Yu J, Cassels F, Scharton-Kersten T, Hammond SA, Hartman A, Angov E, et al. Transcutaneous immunization using colonization factor and heat-labile enterotoxin induces correlates of protective immunity for enterotoxigenic Escherichia coli. Infect Immun. 2002;70(3):1056-1068. https://doi.org/10.1128/iai.70.3.1056-1068.2002
  26. Lin J, Mateo KS, Zhao M, Erickson AK, Garcia N, He D, et al. Protection of piglets against enteric colibacillosis by intranasal immunization with K88ac (F4ac) fimbriae and heat labile enterotoxin of Escherichia coli. Vet Microbiol. 2013;162(2-4):731-739. https://doi.org/10.1016/j.vetmic.2012.09.025
  27. Nguyen UV, Melkebeek V, Devriendt B, Goetstouwers T, Van Poucke M, Peelman L, et al. Maternal immunity enhances systemic recall immune responses upon oral immunization of piglets with F4 fimbriae. Vet Res (Faisalabad). 2015;46(1):72. https://doi.org/10.1186/s13567-015-0210-3
  28. Yang X, Thornburg T, Holderness K, Suo Z, Cao L, Lim T, et al. Serum antibodies protect against intraperitoneal challenge with enterotoxigenic Escherichia coli. J Biomed Biotechnol. 2011;2011:632396. https://doi.org/10.1155/2011/632396