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. 1989 Dec;57(12):3808–3815. doi: 10.1128/iai.57.12.3808-3815.1989

Cloning and characterization of Treponema hyodysenteriae antigens and protection in a CF-1 mouse model by immunization with a cloned endoflagellar antigen.

D A Boyden 1, F G Albert 1, C S Robinson 1
PMCID: PMC259909  PMID: 2807548

Abstract

We cloned genes that code for Treponema hyodysenteriae antigens into Escherichia coli with the purpose of identifying protective antigens for vaccine development. Three different genomic libraries were screened with various antisera reactive with T. hyodysenteriae antigens. The cloned antigens and corresponding native T. hyodysenteriae antigens were analyzed for molecular size, serum reactivity, solubility in sarcosine, and segregation during phase partitioning with the nonionic detergent Triton X-114. The results from these analyses suggested that the gene products were components of either the cytoplasmic membrane, periplasm, or endoflagella of T. hyodysenteriae. The cloned antigens were tested as vaccine candidates in a CF-1 mouse model of T. hyodysenteriae infection and immunity. Intraperitoneal injection of crude E. coli extracts containing cloned antigens did not protect mice from challenge. However, serum from mice injected with a crude extract of an E. coli clone which expressed an endoflagellar antigen killed T. hyodysenteriae in vitro. Partially purified preparations of this cloned endoflagellar antigen protected mice against oral challenge with both the homologous serotype (B204) and a heterologous serotype (B234) of T. hyodysenteriae. These results suggest that the endoflagellar proteins could be used as an effective subunit vaccine against T. hyodysenteriae.

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Selected References

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  1. Achtman M., Manning P. A., Edelbluth C., Herrlich P. Export without proteolytic processing of inner and outer membrane proteins encoded by F sex factor tra cistrons in Escherichia coli minicells. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4837–4841. doi: 10.1073/pnas.76.10.4837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baum D. H., Joens L. A. Serotypes of beta-hemolytic Treponema hyodysenteriae. Infect Immun. 1979 Sep;25(3):792–796. doi: 10.1128/iai.25.3.792-796.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beall J. A., Mitchell G. F. Identification of a particular antigen from a parasite cDNA library using antibodies affinity purified from selected portions of Western blots. J Immunol Methods. 1986 Feb 12;86(2):217–223. doi: 10.1016/0022-1759(86)90456-4. [DOI] [PubMed] [Google Scholar]
  4. Blanco D. R., Radolf J. D., Lovett M. A., Miller J. N. The antigenic interrelationship between the endoflagella of Treponema phagedenis biotype Reiter and Treponema pallidum Nichols strain. I. Treponemicidal activity of cross-reactive endoflagellar antibodies against T. pallidum. J Immunol. 1986 Nov 1;137(9):2973–2979. [PubMed] [Google Scholar]
  5. Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
  6. Chatfield S. N., Fernie D. S., Penn C., Dougan G. Identification of the major antigens of Treponema hyodysenteriae and comparison with those of Treponema innocens. Infect Immun. 1988 May;56(5):1070–1075. doi: 10.1128/iai.56.5.1070-1075.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cunningham T. M., Walker E. M., Miller J. N., Lovett M. A. Selective release of the Treponema pallidum outer membrane and associated polypeptides with Triton X-114. J Bacteriol. 1988 Dec;170(12):5789–5796. doi: 10.1128/jb.170.12.5789-5796.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fernie D. S., Ripley P. H., Walker P. D. Swine dysentery: protection against experimental challenge following single dose parenteral immunisation with inactivated Treponema hyodysenteriae. Res Vet Sci. 1983 Sep;35(2):217–221. [PubMed] [Google Scholar]
  9. Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerl L., Sumper M. Halobacterial flagellins are encoded by a multigene family. Characterization of five flagellin genes. J Biol Chem. 1988 Sep 15;263(26):13246–13251. [PubMed] [Google Scholar]
  11. Glock R. D., Schwartz K. J., Harris D. L. Parenteral immunization of pigs against infection with Treponema hyodysenteriae. Am J Vet Res. 1978 Apr;39(4):639–642. [PubMed] [Google Scholar]
  12. Harris D. L., Glock R. D., Christensen C. R., Kinyon J. M. Inoculation of pigs with Treponema hyodysenteriae (new species) and reproduction f the disease. Vet Med Small Anim Clin. 1972 Jan;67(1):61–64. [PubMed] [Google Scholar]
  13. Hindersson P., Petersen C. S., Axelsen N. H. Purified flagella from Treponema phagedenis biotype Reiter does not induce protective immunity against experimental syphilis in rabbits. Sex Transm Dis. 1985 Jul-Sep;12(3):124–127. doi: 10.1097/00007435-198507000-00006. [DOI] [PubMed] [Google Scholar]
  14. Joens L. A., Glock R. D. Experimental infection in mice with Treponema hyodysenteriae. Infect Immun. 1979 Aug;25(2):757–760. doi: 10.1128/iai.25.2.757-760.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Joens L. A., Harris D. L., Baum D. H. Immunity to Swine dysentery in recovered pigs. Am J Vet Res. 1979 Oct;40(10):1352–1354. [PubMed] [Google Scholar]
  16. Joens L. A., Marquez R. B. Molecular characterization of proteins from porcine spirochetes. Infect Immun. 1986 Dec;54(3):893–896. doi: 10.1128/iai.54.3.893-896.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Joens L. A., Nuessen M. E. Bactericidal effect of normal swine sera on Treponema hyodysenteriae. Infect Immun. 1986 Jan;51(1):282–285. doi: 10.1128/iai.51.1.282-285.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Joens L. A., Whipp S. C., Glock R. D., Neussen M. E. Serotype-specific protection against Treponema hyodysenteriae infection in ligated colonic loops of pigs recovered from swine dysentery. Infect Immun. 1983 Jan;39(1):460–462. doi: 10.1128/iai.39.1.460-462.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kinyon J. M., Harris D. L. Growth in Treponema hyodysenteriae in liquid medium. Vet Rec. 1974 Sep 7;95(10):219–220. doi: 10.1136/vr.95.10.219. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. 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]
  22. Limberger R. J., Charon N. W. Antiserum to the 33,000-dalton periplasmic-flagellum protein of "Treponema phagedenis" reacts with other treponemes and Spirochaeta aurantia. J Bacteriol. 1986 Nov;168(2):1030–1032. doi: 10.1128/jb.168.2.1030-1032.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lin W., Kasamatsu H. On the electrotransfer of polypeptides from gels to nitrocellulose membranes. Anal Biochem. 1983 Feb 1;128(2):302–311. doi: 10.1016/0003-2697(83)90379-2. [DOI] [PubMed] [Google Scholar]
  24. Mapother M. E., Joens L. A. New serotypes of Treponema hyodysenteriae. J Clin Microbiol. 1985 Aug;22(2):161–164. doi: 10.1128/jcm.22.2.161-164.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mestecky J. The common mucosal immune system and current strategies for induction of immune responses in external secretions. J Clin Immunol. 1987 Jul;7(4):265–276. doi: 10.1007/BF00915547. [DOI] [PubMed] [Google Scholar]
  26. Miller D. P., Toivio-Kinnucan M., Wu G., Wilt G. R. Ultrastructural and electrophoretic analysis of Treponema hyodysenteriae axial filaments. Am J Vet Res. 1988 Jun;49(6):786–789. [PubMed] [Google Scholar]
  27. Minnich S. A., Newton A. Promoter mapping and cell cycle regulation of flagellin gene transcription in Caulobacter crescentus. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1142–1146. doi: 10.1073/pnas.84.5.1142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moriyon I., Berman D. T. Effects of nonionic, ionic, and dipolar ionic detergents and EDTA on the Brucella cell envelope. J Bacteriol. 1982 Nov;152(2):822–828. doi: 10.1128/jb.152.2.822-828.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Norris S. J., Charon N. W., Cook R. G., Fuentes M. D., Limberger R. J. Antigenic relatedness and N-terminal sequence homology define two classes of periplasmic flagellar proteins of Treponema pallidum subsp. pallidum and Treponema phagedenis. J Bacteriol. 1988 Sep;170(9):4072–4082. doi: 10.1128/jb.170.9.4072-4082.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pierce N. F., Koster F. T. Priming and suppression of the intestinal immune response to cholera toxoid/toxin by parenteral toxoid in rats. J Immunol. 1980 Jan;124(1):307–311. [PubMed] [Google Scholar]
  31. Pleier E., Schmitt R. Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti. J Bacteriol. 1989 Mar;171(3):1467–1475. doi: 10.1128/jb.171.3.1467-1475.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Queen C. A vector that uses phage signals for efficient synthesis of proteins in Escherichia coli. J Mol Appl Genet. 1983;2(1):1–10. [PubMed] [Google Scholar]
  33. Radolf J. D., Blanco D. R., Miller J. N., Lovett M. A. Antigenic interrelationship between endoflagella of Treponema phagedenis biotype Reiter and Treponema pallidum (Nichols): molecular characterization of endoflagellar proteins. Infect Immun. 1986 Dec;54(3):626–634. doi: 10.1128/iai.54.3.626-634.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Radolf J. D., Chamberlain N. R., Clausell A., Norgard M. V. Identification and localization of integral membrane proteins of virulent Treponema pallidum subsp. pallidum by phase partitioning with the nonionic detergent triton X-114. Infect Immun. 1988 Feb;56(2):490–498. doi: 10.1128/iai.56.2.490-498.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rosenberg A. H., Lade B. N., Chui D. S., Lin S. W., Dunn J. J., Studier F. W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene. 1987;56(1):125–135. doi: 10.1016/0378-1119(87)90165-x. [DOI] [PubMed] [Google Scholar]
  36. Taylor D. J., Alexander T. J. The production of dysentery in swine by feeding cultures containing a spirochaete. Br Vet J. 1971 Nov;127(11):58–61. doi: 10.1016/s0007-1935(17)37282-2. [DOI] [PubMed] [Google Scholar]
  37. Wannemuehler M. J., Hubbard R. D., Greer J. M. Characterization of the major outer membrane antigens of Treponema hyodysenteriae. Infect Immun. 1988 Dec;56(12):3032–3039. doi: 10.1128/iai.56.12.3032-3039.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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