Abstract
The use of recombinant DNA technology to study the HAP organisms has greatly improved our ability to characterize the many potential virulence factors of these bacteria. Instead of relying on conventional identification and purification methods based on biochemical, biophysical or immunological techniques, the genes coding for the virulence factors could be isolated for more detailed studies. After isolation and characterization of the genes, the recombinant antigens could be expressed for detailed biochemical and immunological studies. Further, large scale production of the recombinant antigens is also possible by high level expression in a heterologous system. These recombinant antigens could be used in vaccine trial and challenge experiments to investigate their protective capabilities. As a result of these molecular approaches, we now have a better understanding of some of the virulence factors of the HAP organisms. Some of these characterized antigens are being used as potential candidates for the development of an efficacious vaccine against these opportunistic pathogens in animals. In addition to studying the virulence antigens of the bacteria, the accumulation of molecular data is beginning to give us a picture into the biology, genetics and evolution of these bacteria.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
Abdullah, K.M., 1991. Biochemical and genetic characterization of a Pasteurella haemolytica Al glycoprotease. Ph.D. Thesis, University of Guelph.
Abdullah, K.M., Lo, R.Y.C., and Mellors, A., 1991, Cloning. nucleotide sequence, and expression of the Pasteurella haemolyticn AI glycoprotease gene. J. Bacteriol. 173: 5597–5603.
Abdullah. K.M, Udoh, E.A., Shewen, P.E., and Mellors, A., 1992. A neutral glycoprotease of Pasteurella haemolyticn AI specifically cleaves 0-sialoglycoproteins. Infect. Immun. 60: 56–62.
Adhya, S., 1987. The galactose operon. in Escherichia coli and Salmonella typhimurium: Cellular and
Molecular Biology. F.C. Neidhart et al.,. eds., American Society for Microbiology, Wachington, D.C. Braun, V., and Focareta, T., 1991. Pore-forming bacterial protein hemolysins (cytolysins). Crit. Rev. Microhiol. 18: 115–158.
Burrows, L.L., 1993, Molecular characterization of the RTX cytolysin determinants from Gram-negative
pathogens of veterinary significance. Ph.D. Thesis, University of Guelph, Guelph. Ontario, Canada. Burrows, L.L., and Lo, R.Y.C., 1992, Molecular characterization of an RTX toxin determinant from Actinobacillus suis. Infect. immun. 60: 2166–2173.
Burrows, L.L., Olah-Winfield. E., and Lo, R.Y.C., 1993, Molecular analysis of the leukotoxin determinants from Pasteurella haemolytica serotypes 1 to 16. infect. immun. 61: 5001–5007.
Carle, G.F., Frank, M. and Olson, M.V., 1986, Electrophoretic separation of large molecules by periodic inversion of the electric field. Science 232: 65–68.
Cavalieri. S.J., Bohach, G.A., and Snyder, i.S., 1984, Escherichia coli à-haemolysin: characteristics and probable role in pathogenicity. Microhiol. Rev. 48: 326–343.
Chang, Y.-F., Young. R., Moulds. T.L., and Struck, D.K., 1989, Secretion of the Pasteurella leukotoxin by Escherichia coli. FEMS Microhiol. Lett. 60: 169–174.
Chen, L., and Coleman. Jr., W.G., 1993, Cloning and characterization of the Escherichia coli K-12 (rfaC) gene, a gene required for lipopolysaccharide inner core synthesis. J. Bacteriol. 175: 2534–2540.
Cho, H.J. and Jericho, K.W.F., 1986, Induction of immunity against pneumonic pasteurellosis following experimental infection in calves. Can. J. Vet. Res. 50: 27–31.
Chu, G., Vollrath. D., and Davis. R.W., 1986. Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science 234: 1582–1585.
Confer, A.W., and Simons, K.R., 1986, Effects of Pasteurella haemo/vtica lipopolysaccharide on selected functions of bovine leukocytes. Am. J. Vet. Res. 47: 154–157.
Conlon. J.A., Shewen, P.E., and Lo. R.Y.C., 1991. Efficacy of a recombinant leukotoxin in protection against pneumoniae challenge with Pasteurella haemolyticn Al. infect. Immun. 59: 587–591.
Cooney. B.J., and Lo. R.Y.C., 1993, Three contiguous lipoprotein genes in Pasteurella haernolvtica which are homologous to a lipoprotein gene in Haemophilia influen:ae type B. infect. immun. 61: 4682–4688.
Coote, J.G., 1992, Structural and functional relationships among the RTX toxin determinants of Gram-negative bacteria. FEMS Microbiol. Rev. 88: 137–162.
Cope, L.D., Yogev, R., Mertsola. J., Latimer, J.L., Hanson, M.S., McCracken. Jr., G.H., and Hansen. E.J., 1991, Molecular cloning of a gene involved in lipopogosaccharide biosynthesis and virulence expression by Haentophiltts influenzae type B. Molec. Microhiol. 5: 1113–1124.
Cornelissen, C.N., Biswas, G.D., Tsai, J., Paruchuri, D.K., Thompson, S.A., and Spading, P.F., 1992. Gonococcal transferrin-binding protein 1 is required for transferrin utilization and is homologous to TonB-dependent outer membrane receptors. J. Bacteriol. 174: 5788–5797.
Frey, J., 1994, RTX-toxins in Actinobacillus pleuropn eu noniae and their potential role in virulence. in “Molecular Mechanisms of Bacterial Virulence”, C. I. Kado and J.H. Crosa, eds., Kluwer Academic Publishers.
Frey, J., Beck. M., and Nicolet, J., 1994. RTX-toxins of Actinobacilhts pleta’opneunonine. In “Bacterial Protein Toxins. Freer et al. eds., Gustav Fisher.
Gauthier, A., Turmel, M., and Lemieux, C., 1991, A group I intron in the chloroplast large subunit rRNA gene of Chlamydarnonas eugametos encodes a double-strand endonuclease that cleaves the homing site of this intros. Cuff. Genet. 19: 43–47.
Gonzalez, G.C., Caamano, D,L., and Schryvers, A.B., 1990, Identification and characterization of a porcinespecific transferrin receptor in Actinobacillus pleuropnetumoniae. Molec. Microbiol. 4: 1173–1179.
Gonzaleze-Rayos, C., Lo, R.Y.C., Shewen, P.S., and Beveridge, T.J., 1986, Cloning of a serotype-specific antigen from Pasteurella haemolytica Al. infect. Immun. 53: 505–510.
Herrington, D.A., and Spading, P.F., 1985, Haemophilus influenzae can use human transferrin as a sole source for required iron. Infect. Immun. 48: 248–251.
High, N.J., Deadman, M•E., and E.R. Moxon, 1993, The role of a repetitive DNA motif (5’-CAAT-3’) in the variable expression of the Haemophilus influenzae lipopolysaccharide epitope 1 Gal(1–4) Gal. Molec. Microbiol. 9: 1275–1282.
Hone,D., Morona, R., Attridge, S., and Hackett, J., 1987, Construction of defined galE mutants of Salmonella for use as vaccines. J. infect. Dis. 156: 167–174.
Houng, H.S,H., Kopecko, D.J., and Baron, L.S., 1990. Molecular cloning and physical and functional characterization of the Salmonella typhimurium and Salmonella typhi galactose utilization opérons. J. Bacteriol. 172: 4392–4398.
Jennings, M.P., van der Ley, P., Wilks, K.E., Maskell, D.J., Poolman, J.T., and Moxon, E.R., 1993, Cloning and molecular analysis of the galE gene of Neisseria meningitidis and its role in lipopolysaccharide biosynthesis. Molec. Microbiol. 10: 361–369.
Kadner, R.J., 1990, Vitamin B 12 transport in Escherichia coli: energy coupling between membranes. Moles. Microbiol. 4: 2027–2033.
Kciss, R.E., Will, D.H., and Collier. J.R., 1964, Skin toxicity and hemodynamic properties of endotoxin derived from Pasteurella haemolytica. Am. J. Vet. Res. 25: 935–942.
Kolodrubetz, D., Dailey. T., Ebersole. J., and Kraig, E., 1989, Cloning and expression of the leukotoxin gene from Actinobacillus actinomycetemcomitcm.s. infect. immun. 57: 1465–1469.
Koronakis, V., Cross, M., Senior, B., Koronakis, E., and Hughes, C., 1987, The secreted hemolysins of Proteus mirabilis, Proteus vul garis, and Morgane/la morganii are genetically related to each other and to the alpha-hemolysin of Escherichia coli. J. Bacteriol. 169: 1509–1515.
Koronakis, V., Stanley, P., Koronakis, E., and Hughes, C., 1992, The HiyB/HiyD-dependent secretion of toxins by Gram-negative bacteria. FEMS Microbiol. Immunol. 105: 45–54.
LaCroix, R.P., Duncan, J.R., Jenkins, R.P., Leitch. R.A., Petry, J.A., and Richards. J.C., 1993. Structural and serological specificities of Pasteurella haemolytica lipolysaccharides. infect. Immun. 61: 170–181.
Legrain, M., Mazarin. V., Irwin, S.W., Bouchon. B., Quentin-Millet. M., Jacobs, E., and Schryvers, A.B., 1993, Cloning and characterization of Neisseria meningitidis genes encoding the transferrin-binding proteins Tbp 1 and Tbp2. Gene, 130: 73–80.
Lemaire, H.-G., and Muller-Hill. B., 1986, Nucleotide sequences of the galE gene and the galT gene of E. coli. Nucl. Acids Res. 14: 7705–7711.
Lo, R.Y.C., 1990, Molecular characterization of cytotoxins produced by Haemophilus, Actinobacillus, Pasteurella. Can. J. Vet. Res. 54: S33 - S35.
Lo, R.Y.C., 1992, An analysis of the codon usage of Pasteurella haemolytica Al. FEMS Microbiol. Lett. 100: 125–132.
Lo, R.Y.C., and Cameron, L.A., 1986. A simple immunological detection method for the direct screening of genes form clone Banks. Can. J. Biochem. Cell Biol. 64: 73–76.
Lo. R.Y.C., Shewen, P.E., Strathdee, C.A., and Greer, C.N., 1985, Cloning and expression of the leukotoxin gene of Pasteurella haemolytica AI in Escherichia coli K-12. Infect. Immun. 50: 667–671.
Lo, R.Y.C., Strathdee, C.A., and Shewen. P.E., 1987. Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A 1. Infect. immun. 55: 1987–1996.
Lo, R.Y.C., Strathdee, C.A., Shewen, P.E., and Cooney, B.J., 1991, Molecular studies of Ssal, a serotypespecific antigen of Pasteurella haemolytica Al. infect. Immun. 59: 3398–3406.
Lo, R.Y.C., Watt, M.-A., Gyorffy, S., and Mellors, A., 1994, Preparation of recombinant glycoprotease of Pasteurella haemolytica Al utilizing the Escherichia coli ahemolysin secretion system. FEMS Microbiol. Lett. 116: 225–230.
Mannheim, W. 1984, In Bergey’s Manual of Systematic Bacteriology, Vol.l. (Kraig, N.R., Ed.). pp. 550–552, Williams and Wilkins Press, Baltimore/London.
Marshall. P., and Lemieux, C., 1991, Cleavage pattern of the homing endonuclease encoded by the fifth intros in the chloroplast large subunit rRNA-encoding gene of Chlamydomonas eugametos. Gene 104: 241–245.
Marsh, J.C.W., Sutherland, D.R., Davidson, J., Mellors, A., and Keating, A., 1992, Retention of progenitor cell fraction in CD34’ cells purified using a nova) 0-sialoglycoprotease. Leukemia 6: 926–934.
Maskell, D.J., Szabo, M.J., Dcadman, M.E., and Moxon, E.R., 1992, The gal locus from Haemophilus influenzae: cloning, sequencing and the use of gal mutants to study lipopolysaccharide. Molec. Microbiol. 6: 3051–3063.
Mickelsen, P.A., and Sparling, P.F., 1981, Ability of Neisseria gonorrhoeae, Neisseria meningitidis and commensal Neisseria species to obtain iron form transferrin and iron compounds. Infect. Immun. 33: 555–564.
Neilands, J.B., 1981, Microbial iron compounds. Annu. Rev. Microbiol. 50: 715–731.
Ogunnariwo, J.A., and Schryvers. A.B., 1990, iron acquisition in Pasteurella haemolytica: expression and identification of bovine-specific transferrin receptor. Infect. Immun. 58: 2091–2097.
Otto. B.R., Verweij-van Vught, A.M.J.J., and MacLaren. D.M., 1992, Transferrins and heme-compounds as iron sources for pathogenic bacteria. Crit. Rev. Microbiol. 13: 217–233.
Paulsen. D.B., Mosier, D.A., Clinkenbeard, K.D., and Confer, A.W., 1989, Direct effects of Pasteurella haemolytica lipopolysaccharide on bovine pulmonary endothelial cells in vitro. Am. J. Vet. Res. 50: 1633–1637.
Paulsen. D.B., Confer, A.W., Clinkenbeard, K.D., and Mosier, D.A., 1990, Pasteurella haemolytica
lipopolysaccharide-induced arachidonic acid release from and neutrophil adherence to bovine pulmonary artery endothelial cells. Am J. Vet. Res. 51: 1635–1639.
Postle, K., 1990, TonB and the Gram-negative dilemma. Molec. Microbiol. 4: 2019–2025.
Potter, M.D. and Lo, R.Y.C., 1993, Genetic characterization of Pasteurella haemolytica Al LPS biosynthetic genes. 93rd Ann. Meet. Am. Soc. Microbiol., 1993. Atlanta, Georgia, Abstr. B354.
Raetz, C.R.H., 1990, Biochemistry of endotoxins. Annu. Rev. Biochem. 59: 129–170.
Rimsay, R.L., Coyle-Dennis. J.E., Lauerman. L.H., and Squire, P.G., 1981. Purification and biological
characterization of endotoxin fractions from Pasteurella haemolytica. Am. J. Vet. Res. 42:2134–2138.
Robertson. B.D., Frosch, M., and van Pulten. J.P.M., 1993. The role of galE in the biosynthesis and function of gonococcal lipopolysaccharide. Molec. Microbiol. 8: 891–901.
Schnaitman. C.A., and Klena, J.D., 1993, Genetics of lipopolysaccharide biosynthesis in enteric bacteria. Microbiol. Rev. 57: 655–682.
Schryvers, A.B., and Morris, L.J., 1988, Identification and characterization of the transferrin receptor from Neisseria meningitidis. Molec. Microbiol. 2: 281–288.
Severn, W.B., and Richards, J.C., 1993, Characterization of the O-polysaccharide of Pasteurella haemolytica serotype Al. Carbohydr. Res. 240: 277–285.
Shewen. P.E., and Wilkie, B.N., 1988. Vaccination of calves with leukotoxic culture supernatant from Pasteurella haemolytica. Can. J. Vet. Res. 52: 30–36.
Shewen, P.E., Sharpe, A., and Wilkie, B.N., 1988. Efficacy testing of a Pasteurella haemolytica extract vaccine. Vet. Med. 10: 1078–1083.
Sirisena, D.M., Brozek, K.A., MacLachlan, P.R., Sanderson, K.E., and Raetz, C.R.H., 1992, The rfaC gene of Salmonella typhimurium. J. Biol. Chem. 267: 18874–18884.
Slocombe, R.F., Mulks, M., Killingsworth, C.R., Derksen, F.J., and Robinson. N.E., 1990. Effect of Pasteurella haemolytica-derived endotoxin on pulmonary structure and function in calves. Am. J. Vet. Res. 51: 433–438.
Smits, M.A., Briaire, J., Jansen, R., Smith. H.E., Kamp, E.M., and Gielkens, A.L.J., 1991, Cytolysins of Actinobacillus pleuropnetunoniae serotype 9. Infect. Immun. 59: 4497–4504.
Sneath, P.H.A. and Stevens, M., 1990, Actinobacillus rossi sp. nov., Actinobacillus seminis sp. nov., nom. rev., Pasteurella bettii sp. nov. Pasteurella lymphangitidis sp. nov., Pasteurella main’ sp. nov., and Pasteurella trehalosi sp. Nov. Int. J. Syst. Bact. 40: 148–153.
Strathdee, C.A., 1989, Molecular characterization of the Pasteurella haemolytica leukotoxin determinant. Ph.D. Thesis, University of Guelph, Guelph. Ontario, Canada.
Strathdee, C.A., and Lo, R.Y.C., 1987, Extensive homology between the leukotoxin of Pasteurella haemolytica Al and the alpha-hemolysin of Escherichia coli. Infect. Immun. 55: 3233–3236.
Strathdee. C.A., and Lo. R.Y.C., 1989. Cloning, nucleotide sequence, and characteriztion of genes encoding the secretion function of the Pasteurella haemolytica leukotoxin determinant. J. Bacteriol. 171: 916–928.
Sutherland, A.D., and Donachie, W., 1986, Cytotoxic effect of serotypes of Pasteurella haemolytica on sheep bronchoalveolar macrophages. Vet. Microbiol. 11: 331–336.
Welch, R.A., 1991, Pore-forming cytolysins of Gram-negative bacteria. Molec. Microbiol. 5: 521–528.
Welch, R.A., Forestier, C., Lobo, A., Pellett, S., Thomas, Jr., W., and Rowe, G., 1992, The synthesis and function of the Escherichia coli hemolysin and related RTX exotoxins. FEMS Microbiol. Immunol. 105: 29–36.
Williams, P., and Griffiths, E., 1992, Bacterial transferrin receptors - structure, function and contribution to virulence. Med. Microbial. Immunol. 181: 301–322.
Whiteley, L.O., Maheswaran, S.K., Weiss, D.J., and Ames, T.R., 1990, Immunohistochemical localization of Pasteurella haemolytica A 1-derived endotoxin, leukotoxin, and capsular polysaccharide in experimental bovine pasteurella pneumoniae. Vet Pathol. 27: 150–161.
Woo, T.K.W., and Lo, R.Y.C., 1993, The transferrin binding protein of Pasteurella haemolytica is homologous to the gonococcal transferrin binding protein. 93rd Ann. Meet. American Society for Microbiol., Atlanta Georgia, Abstr. B294.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Springer Science+Business Media New York
About this chapter
Cite this chapter
Lo, R.Y.C. (1995). Molecular Studies of Antigens in HAP Organisms. In: Donachie, W., Lainson, F.A., Hodgson, J.C. (eds) Haemophilus, Actinobacillus, and Pasteurella. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-0978-7_11
Download citation
DOI: https://doi.org/10.1007/978-1-4899-0978-7_11
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4899-0980-0
Online ISBN: 978-1-4899-0978-7
eBook Packages: Springer Book Archive