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. 1991 Dec;59(12):4295–4301. doi: 10.1128/iai.59.12.4295-4301.1991

Characterization of immunodominant surface antigens of Haemophilus somnus.

L B Corbeil 1, S A Kania 1, R P Gogolewski 1
PMCID: PMC259040  PMID: 1937791

Abstract

An immunodominant Haemophilus somnus outer membrane protein with an apparent molecular mass of 40 kDa on Western blots (immunoblots) of gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels was characterized because a monospecific antibody against this antigen was protective. This monospecific antibody was used for immunoaffinity purification of the antigen. The immunoaffinity-purified antigen reacted with a polyclonal antibody to the 40-kDa antigen but not with a monoclonal antibody (3G9) which reacted with the 40-kDa antigen in gradient gels. On 8 or 10% gels, the approximately 40-kDa antigen was resolved as two bands, a 40-kDa band which reacted with the protective monospecific polyclonal antibody (p40) and a band of lower molecular mass which reacted with monoclonal antibody 3G9. The latter antigen was designated p39. Both antigens were conserved in all H. somnus isolates tested. The specific antibodies were also used to detect cross-reacting antigens in other gram-negative bacteria. Antibody to p40 reacted with proteins of 55 to 28 kDa, with the greatest intensity shown among proteins from other members of the family Pasteurellaceae. Antibody to p40 was reduced by absorption with live H. somnus or other members of the family Pasteurellaceae, so the antigen appears to be surface exposed. Antibody to p39 only cross-reacted with a broad band (38 to 40 kDa) in Haemophilus agni. Since H. agni is not a bovine pathogen and since convalescent-phase serum from H. somnus-infected animals did recognize p39, the latter may be a good immunodiagnostic antigen, if the lack of cross-reactivity with antigens in other gram-negative bacteria is confirmed with a polyclonal antibody to p39. The cross-reactivity of antiserum to p40 with antigens of members of the family Pasteurellaceae and the ability of this antiserum to protect against H. somnus pneumonia indicate that p40 may be a useful vaccine antigen for H. somnus disease and perhaps even diseases caused by other members of the family Pasteurellaceae.

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Selected References

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  1. Barenkamp S. J., Munson R. S., Jr, Granoff D. M. Subtyping isolates of Haemophilus influenzae type b by outer-membrane protein profiles. J Infect Dis. 1981 May;143(5):668–676. doi: 10.1093/infdis/143.5.668. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Canto J., Biberstein E. L., Schulte T. A., Behymer D. Cross-reactivity of Haemophilus somnus antibody in agglutination and complement fixation tests and in the enzyme-linked immunosorbent assay. J Clin Microbiol. 1983 Mar;17(3):500–506. doi: 10.1128/jcm.17.3.500-506.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cardella M. A., Adviento M. A., Nervig R. M. Vaccination studies against experimental bovine Pasteurella pneumonia. Can J Vet Res. 1987 Apr;51(2):204–211. [PMC free article] [PubMed] [Google Scholar]
  5. Confer A. W., Panciera R. J., Gentry M. J., Fulton R. W. Immunologic response to Pasteurella haemolytica and resistance against experimental bovine pneumonic pasteurellosis, induced by bacterins in oil adjuvants. Am J Vet Res. 1987 Feb;48(2):163–168. [PubMed] [Google Scholar]
  6. Confer A. W., Panciera R. J., Mosier D. A. Bovine pneumonic pasteurellosis: immunity to Pasteurella haemolytica. J Am Vet Med Assoc. 1988 Nov 15;193(10):1308–1316. [PubMed] [Google Scholar]
  7. Corbeil L. B., Arthur J. E., Widders P. R., Smith J. W., Barbet A. F. Antigenic specificity of convalescent serum from cattle with haemophilus somnus-induced experimental abortion. Infect Immun. 1987 Jun;55(6):1381–1386. doi: 10.1128/iai.55.6.1381-1386.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Corbeil L. B., Blau K., Prieur D. J., Ward A. C. Serum susceptibility of Haemophilus somnus from bovine clinical cases and carriers. J Clin Microbiol. 1985 Aug;22(2):192–198. doi: 10.1128/jcm.22.2.192-198.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Corbeil L. B., Widders P. R., Gogolewski R., Arthur J., Inzana T. J., Ward A. C. Haemophilus somnus: Bovine Reproductive and Respiratory Disease. Can Vet J. 1986 Feb;27(2):90–93. [PMC free article] [PubMed] [Google Scholar]
  10. Crandell R. A., Smith A. R., Kissil M. Colonization and transmission of Haemophilus somnus in cattle. Am J Vet Res. 1977 Nov;38(11):1749–1751. [PubMed] [Google Scholar]
  11. Gogolewski R. P., Kania S. A., Inzana T. J., Widders P. R., Liggitt H. D., Corbeil L. B. Protective ability and specificity of convalescent serum from calves with Haemophilus somnus pneumonia. Infect Immun. 1987 Jun;55(6):1403–1411. doi: 10.1128/iai.55.6.1403-1411.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gogolewski R. P., Kania S. A., Liggitt H. D., Corbeil L. B. Protective ability of antibodies against 78- and 40-kilodalton outer membrane antigens of Haemophilus somnus. Infect Immun. 1988 Sep;56(9):2307–2316. doi: 10.1128/iai.56.9.2307-2316.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gogolewski R. P., Leathers C. W., Liggitt H. D., Corbeil L. B. Experimental Haemophilus somnus pneumonia in calves and immunoperoxidase localization of bacteria. Vet Pathol. 1987 May;24(3):250–256. doi: 10.1177/030098588702400309. [DOI] [PubMed] [Google Scholar]
  14. Gogolewski R. P., Schaefer D. C., Wasson S. K., Corbeil R. R., Corbeil L. B. Pulmonary persistence of Haemophilus somnus in the presence of specific antibody. J Clin Microbiol. 1989 Aug;27(8):1767–1774. doi: 10.1128/jcm.27.8.1767-1774.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gonzales H. F., Bingham D. P. Genetic relatedness of Haemophilus somnus to select genera of bacteria. Am J Vet Res. 1983 Sep;44(9):1793–1795. [PubMed] [Google Scholar]
  16. Groom S. C., Little P. B. Effects of vaccination of calves against induced Haemophilus somnus pneumonia. Am J Vet Res. 1988 Jun;49(6):793–800. [PubMed] [Google Scholar]
  17. Groom S. C., Little P. B., Rosendal S. Virulence differences among three strains of Haemophilus somnus following intratracheal inoculation of calves. Can J Vet Res. 1988 Jul;52(3):349–354. [PMC free article] [PubMed] [Google Scholar]
  18. Hackstadt T., Todd W. J., Caldwell H. D. Disulfide-mediated interactions of the chlamydial major outer membrane protein: role in the differentiation of chlamydiae? J Bacteriol. 1985 Jan;161(1):25–31. doi: 10.1128/jb.161.1.25-31.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hall R. F., Williams J. M., Smith G. L. Field evaluation of Haemophilus somnus bacterin. Vet Med Small Anim Clin. 1977 Aug;72(8):1368–1370. [PubMed] [Google Scholar]
  20. Harris F. W., Janzen E. D. The Haemophilus somnus disease complex (Hemophilosis): A review. Can Vet J. 1989 Oct;30(10):816–822. [PMC free article] [PubMed] [Google Scholar]
  21. Inzana T. J., Corbeil L. B. Development of a defined medium for Haemophilus somnus isolated from cattle. Am J Vet Res. 1987 Mar;48(3):366–369. [PubMed] [Google Scholar]
  22. Inzana T. J., Iritani B., Gogolewski R. P., Kania S. A., Corbeil L. B. Purification and characterization of lipooligosaccharides from four strains of "Haemophilus somnus". Infect Immun. 1988 Nov;56(11):2830–2837. doi: 10.1128/iai.56.11.2830-2837.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kania S. A., Gogolewski R. P., Corbeil L. B. Characterization of a 78-kilodalton outer membrane protein of Haemophilus somnus. Infect Immun. 1990 Jan;58(1):237–244. doi: 10.1128/iai.58.1.237-244.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Martin W., Acres S., Janzen E., Willson P., Allen B. A field trial of preshipment vaccination of calves. Can Vet J. 1984 Mar;25(3):145–147. [PMC free article] [PubMed] [Google Scholar]
  26. McGillivery D. J., Webber J. J., Dean H. F. Characterisation of Histophilus ovis and related organisms by restriction endonuclease analysis. Aust Vet J. 1986 Dec;63(12):389–393. [PubMed] [Google Scholar]
  27. Miller R. J., Renshaw H. W., Evans J. A. Haemophilus somnus complex: antigenicity and specificity of fractions of Haemophilus somnus. Am J Vet Res. 1975 Aug;36(08):1123–1128. [PubMed] [Google Scholar]
  28. Mosier D. A., Simons K. R., Confer A. W., Panciera R. J., Clinkenbeard K. D. Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis. Infect Immun. 1989 Mar;57(3):711–716. doi: 10.1128/iai.57.3.711-716.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ribble C. S., Jim G. K., Janzen E. D. Efficacy of immunization of feedlot calves with a commercial Haemophilus somnus bacterin. Can J Vet Res. 1988 Apr;52(2):191–198. [PMC free article] [PubMed] [Google Scholar]
  30. Saunders J. R., Thiessen W. A., Janzen E. D. Haemophilus somnus infections I. A ten year (1969-1978) retrospective study of losses in cattle herds in Western Canada. Can Vet J. 1980 Apr;21(4):119–123. [PMC free article] [PubMed] [Google Scholar]
  31. Shewen P. E., Wilkie B. N. Vaccination of calves with leukotoxic culture supernatant from Pasteurella haemolytica. Can J Vet Res. 1988 Jan;52(1):30–36. [PMC free article] [PubMed] [Google Scholar]
  32. Stephens L. R., Humphrey J. D., Little P. B., Barnum D. A. Morphological, biochemical, antigenic, and cytochemical relationships among Haemophilus somnus, Haemophilus agni, Haemophilus haemoglobinophilus, Histophilus ovis, and Actinobacillus seminis. J Clin Microbiol. 1983 May;17(5):728–737. doi: 10.1128/jcm.17.5.728-737.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stephens L. R., Little P. B., Humphrey J. D., Wilkie B. N., Barnum D. A. Vaccination of cattle against experimentally induced thromboembolic meningoencephalitis with a Haemophilus somnus bacterin. Am J Vet Res. 1982 Aug;43(8):1339–1342. [PubMed] [Google Scholar]
  34. Stephens L. R., Little P. B., Wilkie B. N., Barnum D. A. Isolation of Haemophilus somnus antigens and their use as vaccines for prevention of bovine thromboembolic meningoencephalitis. Am J Vet Res. 1984 Feb;45(2):234–239. [PubMed] [Google Scholar]
  35. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Widders P. R., Dorrance L. A., Yarnall M., Corbeil L. B. Immunoglobulin-binding activity among pathogenic and carrier isolates of Haemophilus somnus. Infect Immun. 1989 Feb;57(2):639–642. doi: 10.1128/iai.57.2.639-642.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Widders P. R., Paisley L. G., Gogolewski R. P., Evermann J. F., Smith J. W., Corbeil L. B. Experimental abortion and the systemic immune response to "Haemophilus somnus" in cattle. Infect Immun. 1986 Nov;54(2):555–560. doi: 10.1128/iai.54.2.555-560.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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