Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Protective Immune Response of Bacterially-Derived Recombinant FaeG in Piglets

  • Yahong, Huang (Department of Biological Science and Technology, School of Life Sciences of Nanjing University) ;
  • Liang, Wanqi (Shanghai Jiao Tong University-Shanghai Institutes for Biological Sciences-Pennsylvania State University Joint Center for Life Sciences, Key Laboratory of Microbial Metabolism, Ministry of Education, School of Life Science and Biotechnology) ;
  • Pan, Aihu (Shanghai Key Laboratory of Agricultural Genetics and Breeding, Agro-biotech Research Center, Shanghai Academy of Agricultural Sciences) ;
  • Zhou, Zhiai (The Institute of Animal and Veterinary Sciences, Shanghai Academy of Agricultural Sciences) ;
  • Wang, Qiang (Department of Biological Science and Technology, School of Life Sciences of Nanjing University) ;
  • Huang, Cheng (Department of Biological Science and Technology, School of Life Sciences of Nanjing University) ;
  • Chen, Jianxiu (Department of Biological Science and Technology, School of Life Sciences of Nanjing University) ;
  • Zhang, Dabing (Shanghai Jiao Tong University-Shanghai Institutes for Biological Sciences-Pennsylvania State University Joint Center for Life Sciences, Key Laboratory of Microbial Metabolism, Ministry of Education, School of Life Science and Biotechnology)
  • Published : 2006.10.31
Download PDF
⟨ Previous Next ⟩

Abstract

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

Keywords

References

  1. Barman, N.N. and D.K. Sarma. 1999. Passive immunization of piglets against enterotoxigenic colibacillosis by vaccinating dams with K88ac pili bearing bacterins. Indian J. Experimen. Biol. 37, 1132-1135
  2. Bakker, D., C.E. Vader, B. Roosendaal, P.R. Mooi, B. Oudega, and F.K. de Graaf. 1991. Structure and function of periplasmic chaperone-like proteins involved in the biosynthesis of K88 and K99 fimbriae in enterotoxigenic Escherichia coli. Mol. Microbiol. 5, 875-886 https://doi.org/10.1111/j.1365-2958.1991.tb00761.x
  3. Furer, E., S.J. Cryz, P. Dorner, J. Nicolet, M. Wanner, and R. Germanier. 1982. Protection against colibacillosis in neonatal piglets by immunization of dams with procholeragenoid. Infect. Immun. 35, 887-894
  4. Girardeau, J.P. 1980. A new in vitro technique for attachment to intestinal villi using enteropathogenic Escherichia coli. Ann. Microbiol. (Paris) 131B, 31-37
  5. Greenwood, P.E., S.J. Clark, A.D. Cahill, J. Trevallyn-Jones, and S. Tzipori. 1988. Development and protective efficacy of a recombinant-DNA derived fimbrial vaccine against enterotoxic colibacillosis in neonatal piglets. Vaccine 6, 389-392 https://doi.org/10.1016/0264-410X(88)90135-1
  6. Hong, M.M., J.L. Zhang, R.Z. Cai, Y..Y Fan, M. Yu, B.X. Wu, and J.P. Xiao. 1985. Cloning and expression of genetic determinants for K88 antigen. Chin. J. Biotechnol. 1, 36-45
  7. Huang, Y.H., W.Q. Liang, A.H. Pan, A.P. Wang, C. Huang, Z.J. Zhao, J.X. Chen, Z.A. Zhou, and D.B. Zhang. 2001. Cloning and expression of the K99 fimbrial antigen gene of Enterotoxingenic Escherichia coli. Chin. J. Vet. Sci. 21, 471-474
  8. Huang, Y.H., W.Q. Liang, A.H. Pan, Z.A. Zhou, C. Huang, J.X. Chen, and D.B. Zhang. 2003. Production of FaeG, the major subunit of K88 fimbriae, in transgenic tobacco plants and its immunogenicity in mice. Infect. Immun. 71, 5436-5439 https://doi.org/10.1128/IAI.71.9.5436-5439.2003
  9. Levine, M.M. 1987. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J. Infect. Dis. 155, 377-389 https://doi.org/10.1093/infdis/155.3.377
  10. Lopez-Vidal, Y. and A.M. Svennerholm. 1990. Svennerholm. Monoclonal antibodies against the different subcomponents of colonization factor antigen II of enterotoxigenic Escherichia coli. J. Clin. Microbiol. 28, 1906-1912
  11. Mol, O. and B. Oudega. 1996. Molecular and structural aspects of fimbriae biosynthesis and assembly in Escherichia coli. FEMS Microbiol. Rev. 19, 25-52 https://doi.org/10.1111/j.1574-6976.1996.tb00252.x
  12. Mooi, P.R., A. Wijfjes, and F.K. de Graaf. 1983. Identification and characterization of precursors in the biosynthesis of the K88ab fimbria of Escherichia coli. J. Bacteriol. 154, 41-49
  13. Mooi, F.R., C. Wouters, A. Wijfjes, and F.K. de Graaf. 1982. Construction and characterization of mutants impaired in the biosynthesis of the K88ab antigen. J. Bacteriol. 150, 512-521
  14. Rutter, J.M. and G.W. Jones. 1973. Protection against enteric disease caused by Escherichia coli; a model for vaccination with a virulence determinant? Nature 242, 531-532 https://doi.org/10.1038/242531a0
  15. Sambrook, J., W.R. David, and J. Sambrook. 2001. Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York, USA
  16. Studier, F.W., A.H. Rosenberg, J.J. Dunn, and J.W. Dubendorff. 1990. Use of T7 RNA polymerase to direct expression of cloned genes, p.60-89. In D.V. Goeddel (ed.), Methods in Enzymology. Academic Press, New York, USA
  17. Turnes, C.G., J.A. Aleixo, A.V. Monteiro, and O.A. Dellagostin. 1999. DNA inoculation with a plasmid vector carrying the faeG adhesin gene of Escherichia coli K88ab induced immune responses in mice and pigs. Vaccine 17, 2089-2095 https://doi.org/10.1016/S0264-410X(98)00384-3
  18. Ubeidat, M. and C.L. Rutherford. 2003. Purification and renaturation of Dictyostelium recombinant alkaline phosphatase by continuous elution electrophoresis. Protein Expr. Purif. 27, 375-383 https://doi.org/10.1016/S1046-5928(02)00613-7
  19. Van den Broeck, W., E. Cox, and B.M. Goddeeris. 1999. Receptor-dependent immune responses in pigs after oral immunization with F4 fimbriae. Infect. Immun. 67, 520-526
  20. Van den Broeck, W., E. Cox, and B.M. Goddeeris. 1999. Induction of immune responses in pigs following oral administration of purified F4 fimbriae. Vaccine 17, 2020-2029 https://doi.org/10.1016/S0264-410X(98)00406-X
  21. Willemsen, P.T. and F.K. de Graaf. 1992. Age and serotype dependent binding of K88 fimbriae to porcine intestinal receptors. Microb. Pathog. 12, 367-375 https://doi.org/10.1016/0882-4010(92)90099-A
  22. Wizemann, T.M., J.E. Adamou, and S. Langermann. 1999. Adhesins as targets for vaccine development. Emerg. Infect. Dis. 5, 395-403 https://doi.org/10.3201/eid0503.990310