Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1993 Oct;61(10):4153–4157. doi: 10.1128/iai.61.10.4153-4157.1993

Characterization of an immunoreactive 17.5-kilodalton outer membrane protein of Haemophilus somnus by using a monoclonal antibody.

Y Tagawa 1, M Haritani 1, N Yuasa 1
PMCID: PMC281138  PMID: 7691744

Abstract

A single outer membrane protein (OMP) of Haemophilus somnus, with an apparent molecular mass of 17.5 kDa, was identified in the sodium dodecyl sulfate (SDS)-insoluble fraction after extraction with 1% SDS-0.5 M NaCl-0.1% beta-mercaptoethanol. A hybridoma derived from mice immunized with H. somnus OMP fractions produced a monoclonal antibody (MAb), designated 20-3-5, that bound to the 17.5-kDa OMP of H. somnus. The MAb 20-3-5 epitope was present on 45 of 45 strains of H. somnus tested. MAb 20-3-5 cross-reacted with Haemophilus agni, Histophilus ovis, and Haemophilus haemoglobinophilus but not with 13 other species and subspecies of gram-negative bacteria. Immunoelectron-microscopic and antibody absorption studies revealed that the MAb 20-3-5 epitope is exposed on the surface of bacteria. In an immunoblot analysis, convalescent-phase sera obtained from calves with experimental H. somnus pneumonia contained antibodies to the 17.5-kDa OMP of H. somnus. Future studies will be directed toward examining the role of the 17.5-kDa OMP in immunity to H. somnus infections.

Full text

PDF
4153

Images in this article

4154Image
on p.4154
4154Image
on p.4154
4155Image
on p.4155
4155Image
on p.4155
4155Image
on p.4155
4155Image
on p.4155

Selected References

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

  1. 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]
  2. Corbeil L. B., Kania S. A., Gogolewski R. P. Characterization of immunodominant surface antigens of Haemophilus somnus. Infect Immun. 1991 Dec;59(12):4295–4301. doi: 10.1128/iai.59.12.4295-4301.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Ley J., Mannheim W., Mutters R., Piechulla K., Tytgat R., Segers P., Bisgaard M., Frederiksen W., Hinz K. H., Vanhoucke M. Inter- and intrafamilial similarities of rRNA cistrons of the Pasteurellaceae. Int J Syst Bacteriol. 1990 Apr;40(2):126–137. doi: 10.1099/00207713-40-2-126. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  7. Munson R. S., Jr, Granoff D. M. Purification and partial characterization of outer membrane proteins P5 and P6 from Haemophilus influenzae type b. Infect Immun. 1985 Sep;49(3):544–549. doi: 10.1128/iai.49.3.544-549.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Murphy T. F., Bartos L. C., Campagnari A. A., Nelson M. B., Apicella M. A. Antigenic characterization of the P6 protein of nontypable Haemophilus influenzae. Infect Immun. 1986 Dec;54(3):774–779. doi: 10.1128/iai.54.3.774-779.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ogunnariwo J. A., Cheng C., Ford J., Schryvers A. B. Response of Haemophilus somnus to iron limitation: expression and identification of a bovine-specific transferrin receptor. Microb Pathog. 1990 Dec;9(6):397–406. doi: 10.1016/0882-4010(90)90058-x. [DOI] [PubMed] [Google Scholar]
  10. Osbaldiston G. W. Vaginitis in a bitch associated with Haemophilus sp. Am J Vet Res. 1971 Dec;32(12):2067–2069. [PubMed] [Google Scholar]
  11. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Tagawa Y., Haritani M., Ishikawa H., Yuasa N. Antigenic analysis of the major outer membrane protein of Haemophilus somnus with monoclonal antibodies. Infect Immun. 1993 May;61(5):2257–2259. doi: 10.1128/iai.61.5.2257-2259.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tagawa Y., Haritani M., Ishikawa H., Yuasa N. Characterization of a heat-modifiable outer membrane protein of Haemophilus somnus. Infect Immun. 1993 May;61(5):1750–1755. doi: 10.1128/iai.61.5.1750-1755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tagawa Y., Ishikawa H., Yuasa N. Purification and partial characterization of the major outer membrane protein of Haemophilus somnus. Infect Immun. 1993 Jan;61(1):91–96. doi: 10.1128/iai.61.1.91-96.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Theisen M., Rioux C. R., Potter A. A. Molecular cloning, nucleotide sequence, and characterization of a 40,000-molecular-weight lipoprotein of Haemophilus somnus. Infect Immun. 1992 Mar;60(3):826–831. doi: 10.1128/iai.60.3.826-831.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Yarnall M., Gogolewski R. P., Corbeil L. B. Characterization of two Haemophilus somnus Fc receptors. J Gen Microbiol. 1988 Jul;134(7):1993–1999. doi: 10.1099/00221287-134-7-1993. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES