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. 1991 Sep;59(9):3079–3085. doi: 10.1128/iai.59.9.3079-3085.1991

Identification of hemolytic and cytotoxic proteins of Actinobacillus pleuropneumoniae by use of monoclonal antibodies.

E M Kamp 1, J K Popma 1, J Anakotta 1, M A Smits 1
PMCID: PMC258137  PMID: 1879932

Abstract

Monoclonal antibodies (MAbs) were raised against extracellular hemolytic and cytotoxic proteins of Actinobacillus pleuropneumoniae serotypes 2 and 9. MAbs were tested for inhibition of hemolytic and cytotoxic activities of the reference strains of A. pleuropneumoniae serotypes 1 to 12. Five MAbs inhibited hemolytic or cytotoxic activities of one or more serotypes and were used to probe Western blots (immunoblots) of cytotoxic and hemolytic culture filtrates of serotypes 1 to 12. The MAbs recognized three different proteins: (i) a protein of approximately 103 kDa that was associated with a weak hemolytic activity and a moderate cytotoxic activity, (ii) a protein of approximately 105 kDa that was associated with a strong hemolytic activity and a strong cytotoxic activity, and (iii) a protein of approximately 120 kDa that was associated with a strong cytotoxic activity but not with hemolytic activity. Serotypes 6, 7, and 12 produced only the 103-kDa protein, and serotype 10 produced only the 105-kDa protein. The other serotypes produced two proteins: serotypes 1, 5, 9, and 11 produced the 103- and 105-kDa proteins, and serotypes 2, 3, 4, and 8 produced the 103- and 120-kDa proteins.

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Selected References

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

  1. Bhakdi S., Mackman N., Nicaud J. M., Holland I. B. Escherichia coli hemolysin may damage target cell membranes by generating transmembrane pores. Infect Immun. 1986 Apr;52(1):63–69. doi: 10.1128/iai.52.1.63-69.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang Y. F., Young R., Struck D. K. Cloning and characterization of a hemolysin gene from Actinobacillus (Haemophilus) pleuropneumoniae. DNA. 1989 Nov;8(9):635–647. doi: 10.1089/dna.1.1989.8.635. [DOI] [PubMed] [Google Scholar]
  3. Devenish J., Rosendal S., Bossé J. T., Wilkie B. N., Johnson R. Prevalence of seroreactors to the 104-kilodalton hemolysin of Actinobacillus pleuropneumoniae in swine herds. J Clin Microbiol. 1990 Apr;28(4):789–791. doi: 10.1128/jcm.28.4.789-791.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devenish J., Rosendal S. Identification of the heat-labile hemolysin of Actinobacillus pleuropneumoniae serotype 1. Can J Vet Res. 1989 Apr;53(2):251–254. [PMC free article] [PubMed] [Google Scholar]
  5. Devenish J., Rosendal S., Johnson R., Hubler S. Immunoserological comparison of 104-kilodalton proteins associated with hemolysis and cytolysis in Actinobacillus pleuropneumoniae, Actinobacillus suis, Pasteurella haemolytica, and Escherichia coli. Infect Immun. 1989 Oct;57(10):3210–3213. doi: 10.1128/iai.57.10.3210-3213.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frey J., Nicolet J. Hemolysin patterns of Actinobacillus pleuropneumoniae. J Clin Microbiol. 1990 Feb;28(2):232–236. doi: 10.1128/jcm.28.2.232-236.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gygi D., Nicolet J., Frey J., Cross M., Koronakis V., Hughes C. Isolation of the Actinobacillus pleuropneumoniae haemolysin gene and the activation and secretion of the prohaemolysin by the HlyC, HlyB and HlyD proteins of Escherichia coli. Mol Microbiol. 1990 Jan;4(1):123–128. doi: 10.1111/j.1365-2958.1990.tb02021.x. [DOI] [PubMed] [Google Scholar]
  8. Kamp E. M., Popma J. K., Van Leengoed L. A. Serotyping of Haemophilus pleuropneumoniae in the Netherlands: with emphasis on heterogeneity within serotype 1 and (proposed) serotype 9. Vet Microbiol. 1987 Mar;13(3):249–257. doi: 10.1016/0378-1135(87)90087-3. [DOI] [PubMed] [Google Scholar]
  9. Kamp E. M., van Leengoed L. A. Serotype-related differences in production and type of heat-labile hemolysin and heat-labile cytotoxin of Actinobacillus (Haemophilus) pleuropneumoniae. J Clin Microbiol. 1989 Jun;27(6):1187–1191. doi: 10.1128/jcm.27.6.1187-1191.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lalonde G., McDonald T. V., Gardner P., O'Hanley P. D. Identification of a hemolysin from Actinobacillus pleuropneumoniae and characterization of its channel properties in planar phospholipid bilayers. J Biol Chem. 1989 Aug 15;264(23):13559–13564. [PubMed] [Google Scholar]
  11. Ma J. N., Inzana T. J. Indirect enzyme-linked immunosorbent assay for detection of antibody to a 110,000-molecular-weight hemolysin of Actinobacillus pleuropneumoniae. J Clin Microbiol. 1990 Jun;28(6):1356–1361. doi: 10.1128/jcm.28.6.1356-1361.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rosendal S., Devenish J., MacInnes J. I., Lumsden J. H., Watson S., Xun H. Evaluation of heat-sensitive, neutrophil-toxic, and hemolytic activity of Haemophilus (Actinobacillus) pleuropneumoniae. Am J Vet Res. 1988 Jul;49(7):1053–1058. [PubMed] [Google Scholar]
  13. Sebunya T. N., Saunders J. R. Haemophilus pleuropneumoniae infection in swine: a review. J Am Vet Med Assoc. 1983 Jun 15;182(12):1331–1337. [PubMed] [Google Scholar]
  14. Strathdee C. A., Lo R. Y. Cloning, nucleotide sequence, and characterization of genes encoding the secretion function of the Pasteurella haemolytica leukotoxin determinant. J Bacteriol. 1989 Feb;171(2):916–928. doi: 10.1128/jb.171.2.916-928.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Van Leengoed L. A., Kamp E. M., Pol J. M. Toxicity of Haemophilus pleuropneumoniae to porcine lung macrophages. Vet Microbiol. 1989 Apr;19(4):337–349. doi: 10.1016/0378-1135(89)90099-0. [DOI] [PubMed] [Google Scholar]
  16. van Zijderveld F. G., Westenbrink F., Anakotta J., Brouwers R. A., van Zijderveld A. M. Characterization of the F41 fimbrial antigen of enterotoxigenic Escherichia coli by using monoclonal antibodies. Infect Immun. 1989 Apr;57(4):1192–1199. doi: 10.1128/iai.57.4.1192-1199.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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