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. 1986 Dec;54(3):751–760. doi: 10.1128/iai.54.3.751-760.1986

Antibody response of swine to outer membrane components of Haemophilus pleuropneumoniae during infection.

V J Rapp, R F Ross
PMCID: PMC260233  PMID: 3536748

Abstract

Sera from pigs infected with Haemophilus (Actinobacillus) pleuropneumoniae were tested for antibodies to outer membrane proteins (OMPs) of the organism by immunoblotting. Convalescent sera were produced in naturally born, colostrum-fed pigs and in cesarean-derived, colostrum-deprived pigs given H. pleuropneumoniae serotype 5 intranasally twice at 5-week intervals. Sera, collected at weekly intervals, were reacted with Sarkosyl-insoluble, OMP-enriched preparations of H. pleuropneumoniae which had been separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose. Antibodies were detected to OMPs with an apparent molecular weight of 16,500 (16.5K OMP); to 29K, 38.5K, 43.5K, 45K, 49.5K, and 66.5K OMPs; and to several high-molecular-weight (greater than or equal to 94,000) OMPs, but not to the major 42K OMP. Antibodies to the heat-modifiable OMP (29K/43.5K) and the 38.5K OMP were detected in sera from noninfected pigs. Antibodies were also detected to two broad 54,000- and 95,000-molecular-weight bands which did not stain with Coomassie blue, stained with silver nitrate, resisted proteinase K digestion, and were eliminated by oxidation with sodium metaperiodate. This indicates that the 54,000- and 95,000-molecular-weight bands represent polysaccharide, possibly capsular or lipopolysaccharide immunogens. Adsorption of sera with cells from the homologous serotype 5 strain removed antibodies to the 45K, 49.5K, 66.5K, and greater than or equal to 94K OMPs and to the two polysaccharide bands, indicating that these antibodies were directed primarily to surface-exposed epitopes. When tested with OMP preparations from other serotype 5 strains, heterogeneity was apparent, both in the reactions with OMPs and with the polysaccharide bands. Silver staining of proteinase K-treated, whole-cell lysates from serotype 5 strains also indicated variable expression of the polysaccharide bands. Sera also reacted with OMPs from H. pleuropneumoniae serotypes 1 and 7; however, several OMPs and the lipopolysaccharide or polysaccharide determinants of these serotypes appeared to be type specific.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  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. Beher M. G., Schnaitman C. A., Pugsley A. P. Major heat-modifiable outer membrane protein in gram-negative bacteria: comparison with the ompA protein of Escherichia coli. J Bacteriol. 1980 Aug;143(2):906–913. doi: 10.1128/jb.143.2.906-913.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blaser M. J., Hopkins J. A., Vasil M. L. Campylobacter jejuni outer membrane proteins are antigenic for humans. Infect Immun. 1984 Mar;43(3):986–993. doi: 10.1128/iai.43.3.986-993.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Erwin A. L., Kenny G. E. Haemophilus influenzae type b isolates show antigenic variation in a major outer membrane protein. Infect Immun. 1984 Nov;46(2):570–577. doi: 10.1128/iai.46.2.570-577.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fenwick B. W., Osburn B. I., Olander H. J. Isolation and biological characterization of two lipopolysaccharides and a capsular-enriched polysaccharide preparation from Haemophilus pleuropneumoniae. Am J Vet Res. 1986 Jul;47(7):1433–1441. [PubMed] [Google Scholar]
  6. Frasch C. E. Status of a group B Neisseria meningitidis vaccine. Eur J Clin Microbiol. 1985 Dec;4(6):533–536. doi: 10.1007/BF02013388. [DOI] [PubMed] [Google Scholar]
  7. Gilleland H. E., Jr, Parker M. G., Matthews J. M., Berg R. D. Use of a purified outer membrane protein F (porin) preparation of Pseudomonas aeruginosa as a protective vaccine in mice. Infect Immun. 1984 Apr;44(1):49–54. doi: 10.1128/iai.44.1.49-54.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Granoff D. M., Munson R. S., Jr Prospects for prevention of Haemophilus influenzae type b disease by immunization. J Infect Dis. 1986 Mar;153(3):448–461. doi: 10.1093/infdis/153.3.448. [DOI] [PubMed] [Google Scholar]
  9. Gulig P. A., McCracken G. H., Jr, Frisch C. F., Johnston K. H., Hansen E. J. Antibody response of infants to cell surface-exposed outer membrane proteins of Haemophilus influenzae type b after systemic Haemophilus disease. Infect Immun. 1982 Jul;37(1):82–88. doi: 10.1128/iai.37.1.82-88.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunnarsson A., Biberstein E. L., Hurvell B. Serologic studies on porcine strains of Haemophilus parahaemolyticus (pleuropneumoniae): agglutination reactions. Am J Vet Res. 1977 Aug;38(8):1111–1114. [PubMed] [Google Scholar]
  11. Gunnarsson A. Serologic studies on porcine strains of Haemophilus parahaemolyticus (pleuropneumoniae): extraction of type-specific antigens. Am J Vet Res. 1979 Apr;40(4):469–472. [PubMed] [Google Scholar]
  12. Hancock K., Tsang V. C. India ink staining of proteins on nitrocellulose paper. Anal Biochem. 1983 Aug;133(1):157–162. doi: 10.1016/0003-2697(83)90237-3. [DOI] [PubMed] [Google Scholar]
  13. Hansen M. V., Musher D. M., Baughn R. E. Outer membrane proteins of nontypable Haemophilus influenzae and reactivity of paired sera from infected patients with their homologous isolates. Infect Immun. 1985 Mar;47(3):843–846. doi: 10.1128/iai.47.3.843-846.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hawkes R. Identification of concanavalin A-binding proteins after sodium dodecyl sulfate--gel electrophoresis and protein blotting. Anal Biochem. 1982 Jun;123(1):143–146. doi: 10.1016/0003-2697(82)90634-0. [DOI] [PubMed] [Google Scholar]
  15. Hedstrom R. C., Pavlovskis O. R., Galloway D. R. Antibody response of infected mice to outer membrane proteins of Pseudomonas aeruginosa. Infect Immun. 1984 Jan;43(1):49–53. doi: 10.1128/iai.43.1.49-53.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Higgins R., Larivière S., Mittal K. R., Martineau G. P., Rousseau P., Cameron J. Evaluation of a Killed Vaccine Against Porcine Pleuropneumonia Due to Haemophilus pleuropneumoniae. Can Vet J. 1985 Feb;26(2):86–89. [PMC free article] [PubMed] [Google Scholar]
  17. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hofstra H., Van Tol J. D., Dankert J. Cross-reactivity of major outer membrane proteins of Enterobacteriaceae, studied by crossed immunoelectrophoresis. J Bacteriol. 1980 Jul;143(1):328–337. doi: 10.1128/jb.143.1.328-337.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jensen A. E., Bertram T. A. Morphological and biochemical comparison of virulent and avirulent isolates of Haemophilus pleuropneumoniae serotype 5. Infect Immun. 1986 Feb;51(2):419–424. doi: 10.1128/iai.51.2.419-424.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jessop H. L., Lambert P. A. Immunochemical characterization of the outer membrane complex of Serratia marcescens and identification of the antigens accessible to antibodies on the cell surface. J Gen Microbiol. 1985 Sep;131(9):2343–2348. doi: 10.1099/00221287-131-9-2343. [DOI] [PubMed] [Google Scholar]
  21. Kume K., Nakai T., Sawata A. Efficacy of Haemophilus pleuropneumoniae vaccine in pigs. Nihon Juigaku Zasshi. 1985 Apr;47(2):201–206. doi: 10.1292/jvms1939.47.201. [DOI] [PubMed] [Google Scholar]
  22. Kuusi N., Nurminen M., Saxen H., Valtonen M., Mäkelä P. H. Immunization with major outer membrane proteins in experimental salmonellosis of mice. Infect Immun. 1979 Sep;25(3):857–862. doi: 10.1128/iai.25.3.857-862.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Loeb M. R. Immunoblot method for identifying surface components, determining their cross-reactivity, and investigating cell topology: results with Haemophilus influenzae type b. Anal Biochem. 1984 Nov 15;143(1):196–204. doi: 10.1016/0003-2697(84)90576-1. [DOI] [PubMed] [Google Scholar]
  24. Loeb M. R., Smith D. H. Human antibody response to individual outer membrane proteins of Haemophilus influenzae type b. Infect Immun. 1982 Sep;37(3):1032–1036. doi: 10.1128/iai.37.3.1032-1036.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maeland J. A. Antigenic properties of various preparations of Neisseria gonorrhoeae endotoxin. Acta Pathol Microbiol Scand. 1968;73(3):413–422. doi: 10.1111/j.1699-0463.1968.tb04610.x. [DOI] [PubMed] [Google Scholar]
  26. Maudsley J. R., Kadis S., Mayberry W. R. Isolation, purification, and partial characterization of a lipopolysaccharide from Haemophilus pleuropneumoniae. Infect Immun. 1986 Feb;51(2):501–506. doi: 10.1128/iai.51.2.501-506.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mills S. D., Bradbury W. C. Human antibody response to outer membrane proteins of Campylobacter jejuni during infection. Infect Immun. 1984 Feb;43(2):739–743. doi: 10.1128/iai.43.2.739-743.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mittal K. R., Higgins R., Larivière S., Leblanc D. A 2-mercaptoethanol tube agglutination test for diagnosis of Haemophilus pleuropneumoniae infection in pigs. Am J Vet Res. 1984 Apr;45(4):715–719. [PubMed] [Google Scholar]
  29. Montaraz J. A., Novotny P., Ivanyi J. Identification of a 68-kilodalton protective protein antigen from Bordetella bronchiseptica. Infect Immun. 1985 Mar;47(3):744–751. doi: 10.1128/iai.47.3.744-751.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nicolet J., Paroz P., Krawinkler M., Baumgartner A. An enzyme-linked immunosorbent assay, using an EDTA-extracted antigen for the serology of Haemophilus pleuropneumoniae. Am J Vet Res. 1981 Dec;42(12):2139–2142. [PubMed] [Google Scholar]
  31. Nicolet J. Sur l'hémophilose du pore. 3. Différenciation sérologique de Haemophilus parahaemolyticus. Zentralbl Bakteriol Orig. 1971;216(4):487–495. [PubMed] [Google Scholar]
  32. Nicolet J., de Meuron P. A., Bachmann P. Sur l'hémophilose du porc. IV. L'épreuve de déviation du complément, un test de dépistage des infections à Haemophilus parahaemolyticus. Schweiz Arch Tierheilkd. 1971 Apr;113(4):191–200. [PubMed] [Google Scholar]
  33. Nielsen R. Haemophilus parahaemolyticus serotypes. Pathogenicity and cross immunity. Nord Vet Med. 1979 Oct;31(10):407–413. [PubMed] [Google Scholar]
  34. Nielsen R. Haemophilus parahaemolyticus serotypes. Serological response. Nord Vet Med. 1979 Oct;31(10):401–406. [PubMed] [Google Scholar]
  35. Nielsen R. Haemophilus pleuropneumoniae (Actinobacillus pleuropneumoniae). Serotypes 8, 3 and 6. Serological response and cross immunity in pigs. Nord Vet Med. 1985 Jul-Aug;37(4):217–227. [PubMed] [Google Scholar]
  36. Nielsen R. Haemophilus pleuropneumoniae serotypes--cross protection experiments. Nord Vet Med. 1984 Jul-Aug;36(7-8):221–234. [PubMed] [Google Scholar]
  37. Nielsen R., O'Connor P. J. Serological characterization of 8 Haemophilus pleuropneumoniae strains and proposal of a new serotype: serotype 8. Acta Vet Scand. 1984;25(1):96–106. doi: 10.1186/BF03547283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nielsen R. Pleuropneumonia of swine caused by Haemophilus parahaemolyticus. Studies on the protection obtained by vaccination. Nord Vet Med. 1976 Jul-Aug;28(7-8):337–348. [PubMed] [Google Scholar]
  39. Nielsen R. Serological and immunological studies of pleuropneumonia of swine caused by Haemophilus parahaemolyticus. Acta Vet Scand. 1974;15(1):80–89. doi: 10.1186/BF03547495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nielsen R. Serological characterization of Haemophilus pleuropneumoniae (Actinobacillus pleuropneumoniae) strains and proposal of a new serotype: serotype 9. Acta Vet Scand. 1985;26(4):501–512. doi: 10.1186/BF03546522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Poolman J. T., Hopman C. T., Zanen H. C. Immunogenicity of meningococcal antigens as detected in patient sera. Infect Immun. 1983 Apr;40(1):398–406. doi: 10.1128/iai.40.1.398-406.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rapp V. J., Munson R. S., Jr, Ross R. F. Outer membrane protein profiles of Haemophilus pleuropneumoniae. Infect Immun. 1986 May;52(2):414–420. doi: 10.1128/iai.52.2.414-420.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Rapp V. J., Ross R. F., Erickson B. Z. Serotyping of Haemophilus pleuropneumoniae by rapid slide agglutination and indirect fluorescent antibody tests in swine. Am J Vet Res. 1985 Jan;46(1):185–192. [PubMed] [Google Scholar]
  44. Redhead K. Serum antibody responses to the outer membrane proteins of Bordetella pertussis. Infect Immun. 1984 Jun;44(3):724–729. doi: 10.1128/iai.44.3.724-729.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Rosendal S., Boyd D. A. Haemophilus pleuropneumoniae serotyping. J Clin Microbiol. 1982 Nov;16(5):840–843. doi: 10.1128/jcm.16.5.840-843.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rosendal S., Carpenter D. S., Mitchell W. R., Wilson M. R. Vaccination against pleuropneumonia of pigs caused by Haemophilus pleuropneumoniae. Can Vet J. 1981 Feb;22(2):34–35. [PMC free article] [PubMed] [Google Scholar]
  47. Schultz R. A., Young T. F., Ross R. F., Jeske D. R. Prevalence of antibodies to Haemophilus pleuropneumoniae in Iowa swine. Am J Vet Res. 1982 Oct;43(10):1848–1851. [PubMed] [Google Scholar]
  48. Sugasawara R. J. Recognition of serogroup A Neisseria meningitidis serotype antigens by human antisera. Infect Immun. 1985 Apr;48(1):23–28. doi: 10.1128/iai.48.1.23-28.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Swanson J., Barrera O. Immunological characteristics of gonococcal outer membrane protein II assessed by immunoprecipitation, immunoblotting, and coagglutination. J Exp Med. 1983 May 1;157(5):1405–1420. doi: 10.1084/jem.157.5.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Thalhamer J., Freund J. Passive immunization: a method of enhancing the immune response against antigen mixtures. J Immunol Methods. 1985 Jun 12;80(1):7–13. doi: 10.1016/0022-1759(85)90158-9. [DOI] [PubMed] [Google Scholar]
  51. 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]
  52. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  53. Wedege E., Frøholm L. O. Human antibody response to a group B serotype 2a meningococcal vaccine determined by immunoblotting. Infect Immun. 1986 Feb;51(2):571–578. doi: 10.1128/iai.51.2.571-578.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. van Alphen L., Riemens T., Poolman J., Zanen H. C. Characteristics of major outer membrane proteins of Haemophilus influenzae. J Bacteriol. 1983 Aug;155(2):878–885. doi: 10.1128/jb.155.2.878-885.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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