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. 1997 Jun;65(6):2502–2507. doi: 10.1128/iai.65.6.2502-2507.1997

Oral immunization of mice with attenuated Salmonella typhimurium aroA expressing a recombinant Mycoplasma hyopneumoniae antigen (NrdF).

P K Fagan 1, S P Djordjevic 1, J Chin 1, G J Eamens 1, M J Walker 1
PMCID: PMC175353  PMID: 9169801

Abstract

Mycoplasma hyopneumoniae is the etiological agent of porcine enzootic pneumonia, a commercially expensive respiratory disease of swine. Salmonella typhimurium SL3261 was used as a live carrier of plasmid pKF1, which encodes a 15-kDa recombinant M. hyopneumoniae protein. This expressed recombinant protein consists of the carboxy-terminal 11 kDa of a 42-kDa M. hyopneumoniae NrdF ribonucleotide reductase R2 subunit protein. Rabbit anti-15-kDa serum was able to inhibit the growth of viable M. hyopneumoniae J in vitro. When used as a live oral vaccine, S. typhimurium SL3261(pKF1) induced a significant secretory immunoglobulin A immune response in the lungs of mice orally immunized against the M. hyopneumoniae antigen. Utilization of live oral vaccines expressing potentially protective M. hyopneumoniae proteins, such as the NrdF antigen, which can stimulate a lung mucosal response against surface-accessible proteins may provide a cost-effective alternative to the present control strategies used for porcine enzootic pneumonia.

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Selected References

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  1. Amass S. F., Clark L. K., van Alstine W. G., Bowersock T. L., Murphy D. A., Knox K. E., Albregts S. R. Interaction of Mycoplasma hyopneumoniae and Pasteurella multocida infections in swine. J Am Vet Med Assoc. 1994 Jan 1;204(1):102–107. [PubMed] [Google Scholar]
  2. Baskerville A. Development of the early lesions in experimental enzootic pneumonia of pigs: an ultrastructural and histological study. Res Vet Sci. 1972 Nov;13(6):570–578. [PubMed] [Google Scholar]
  3. Brandtzaeg P. Overview of the mucosal immune system. Curr Top Microbiol Immunol. 1989;146:13–25. doi: 10.1007/978-3-642-74529-4_2. [DOI] [PubMed] [Google Scholar]
  4. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  5. Carter P. B., Collins F. M. The route of enteric infection in normal mice. J Exp Med. 1974 May 1;139(5):1189–1203. doi: 10.1084/jem.139.5.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chatfield S. N., Roberts M., Dougan G., Hormaeche C., Khan C. M. The development of oral vaccines against parasitic diseases utilizing live attenuated Salmonella. Parasitology. 1995;110 (Suppl):S17–S24. doi: 10.1017/s0031182000001451. [DOI] [PubMed] [Google Scholar]
  7. Ciprián A., Pijoan C., Cruz T., Camacho J., Tórtora J., Colmenares G., López-Revilla R., de la Garza M. Mycoplasma hyopneumoniae increases the susceptibility of pigs to experimental Pasteurella multocida pneumonia. Can J Vet Res. 1988 Oct;52(4):434–438. [PMC free article] [PubMed] [Google Scholar]
  8. Climent I., Sjöberg B. M., Huang C. Y. Carboxyl-terminal peptides as probes for Escherichia coli ribonucleotide reductase subunit interaction: kinetic analysis of inhibition studies. Biochemistry. 1991 May 28;30(21):5164–5171. doi: 10.1021/bi00235a008. [DOI] [PubMed] [Google Scholar]
  9. Cárdenas L., Clements J. D. Oral immunization using live attenuated Salmonella spp. as carriers of foreign antigens. Clin Microbiol Rev. 1992 Jul;5(3):328–342. doi: 10.1128/cmr.5.3.328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fagan P. K., Djordjevic S. P., Eamens G. J., Chin J., Walker M. J. Molecular characterization of a ribonucleotide reductase (nrdF) gene fragment of Mycoplasma hyopneumoniae and assessment of the recombinant product as an experimental vaccine for enzootic pneumonia. Infect Immun. 1996 Mar;64(3):1060–1064. doi: 10.1128/iai.64.3.1060-1064.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galán J. E. Molecular genetic bases of Salmonella entry into host cells. Mol Microbiol. 1996 Apr;20(2):263–271. doi: 10.1111/j.1365-2958.1996.tb02615.x. [DOI] [PubMed] [Google Scholar]
  12. Guzmán C. A., Brownlie R. M., Kadurugamuwa J., Walker M. J., Timmis K. N. Antibody responses in the lungs of mice following oral immunization with Salmonella typhimurium aroA and invasive Escherichia coli strains expressing the filamentous hemagglutinin of Bordetella pertussis. Infect Immun. 1991 Dec;59(12):4391–4397. doi: 10.1128/iai.59.12.4391-4397.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hoiseth S. K., Stocker B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature. 1981 May 21;291(5812):238–239. doi: 10.1038/291238a0. [DOI] [PubMed] [Google Scholar]
  14. Kobisch M., Blanchard B., Le Potier M. F. Mycoplasma hyopneumoniae infection in pigs: duration of the disease and resistance to reinfection. Vet Res. 1993;24(1):67–77. [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Livingston C. W., Jr, Stair E. L., Underdahl N. R., Mebus C. A. Pathogenesis of mycoplasmal pneumonia in swine. Am J Vet Res. 1972 Nov;33(11):2249–2258. [PubMed] [Google Scholar]
  17. Lumsden J. S., Wilkie B. N. Immune response of pigs to parenteral vaccination with an aromatic-dependent mutant of Salmonella typhimurium. Can J Vet Res. 1992 Oct;56(4):296–302. [PMC free article] [PubMed] [Google Scholar]
  18. Mukkur T. K., Walker K. H. Development and duration of protection against salmonellosis in mice and sheep immunised with live aromatic-dependent Salmonella typhimurium. Res Vet Sci. 1992 Mar;52(2):147–153. doi: 10.1016/0034-5288(92)90003-k. [DOI] [PubMed] [Google Scholar]
  19. Pointon A. M., Byrt D., Heap P. Effect of enzootic pneumonia of pigs on growth performance. Aust Vet J. 1985 Jan;62(1):13–18. doi: 10.1111/j.1751-0813.1985.tb06032.x. [DOI] [PubMed] [Google Scholar]
  20. Roth J. A. Characterization of protective antigens and the protective immune response. Vet Microbiol. 1993 Nov;37(3-4):193–199. doi: 10.1016/0378-1135(93)90022-y. [DOI] [PubMed] [Google Scholar]
  21. Schorr J., Knapp B., Hundt E., Küpper H. A., Amann E. Surface expression of malarial antigens in Salmonella typhimurium: induction of serum antibody response upon oral vaccination of mice. Vaccine. 1991 Sep;9(9):675–681. doi: 10.1016/0264-410x(91)90194-b. [DOI] [PubMed] [Google Scholar]
  22. Stanbridge E., Hayflick L. Growth inhibition test for identification of Mycoplasma species utilizing dried antiserum-impregnated paper discs. J Bacteriol. 1967 Apr;93(4):1392–1396. doi: 10.1128/jb.93.4.1392-1396.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tajima M., Yagihashi T. Interaction of Mycoplasma hyopneumoniae with the porcine respiratory epithelium as observed by electron microscopy. Infect Immun. 1982 Sep;37(3):1162–1169. doi: 10.1128/iai.37.3.1162-1169.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Takeshita S., Sato M., Toba M., Masahashi W., Hashimoto-Gotoh T. High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection. Gene. 1987;61(1):63–74. doi: 10.1016/0378-1119(87)90365-9. [DOI] [PubMed] [Google Scholar]
  25. Telford E., Lankinen H., Marsden H. Inhibition of equine herpesvirus type 1 subtype 1-induced ribonucleotide reductase by the nonapeptide YAGAVVNDL. J Gen Virol. 1990 Jun;71(Pt 6):1373–1378. doi: 10.1099/0022-1317-71-6-1373. [DOI] [PubMed] [Google Scholar]
  26. Thelander L., Reichard P. Reduction of ribonucleotides. Annu Rev Biochem. 1979;48:133–158. doi: 10.1146/annurev.bi.48.070179.001025. [DOI] [PubMed] [Google Scholar]
  27. Walker M. J., Rohde M., Timmis K. N., Guzmán C. A. Specific lung mucosal and systemic immune responses after oral immunization of mice with Salmonella typhimurium aroA, Salmonella typhi Ty21a, and invasive Escherichia coli expressing recombinant pertussis toxin S1 subunit. Infect Immun. 1992 Oct;60(10):4260–4268. doi: 10.1128/iai.60.10.4260-4268.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wannemuehler M. J., Galvin J. E. Bacterial immunogens and protective immunity in swine. Vet Immunol Immunopathol. 1994 Oct;43(1-3):117–126. doi: 10.1016/0165-2427(94)90127-9. [DOI] [PubMed] [Google Scholar]
  29. Yagihashi T., Nunoya T., Mitui T., Tajima M. Effect of Mycoplasma hyopneumoniae infection on the development of Haemophilus pleuropneumoniae pneumonia in pigs. Nihon Juigaku Zasshi. 1984 Oct;46(5):705–713. doi: 10.1292/jvms1939.46.705. [DOI] [PubMed] [Google Scholar]

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