Saline-extracted antigens of : Separation by chromatofocusing, preliminary characterization, and evaluation of immunogenicity.

Lessley, Bruce A.; Confer, Anthony W.; Mosier, Derek A.; Gentry, Martha J.; Durham, J.A.; Rummage, John A.

doi:10.1016/0165-2427(85)90053-4

Cited by 26 publications

(11 citation statements)

References 16 publications

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“…Serotype specificity may be determined by the many different components of the cell surface of the bacteria. The cell surface of P. haemolytica has been postulated to consist of a polysaccharide capsular material (1,5); however, other macromolecules such as lipopolysaccharides or proteins may also be present (8,16). Since the antiserum used in the initial screening experiment was raised against serotype 1 soluble antigens, it is probable that soluble antigens may also be involved in determining serotype specificity for this bacterium.…”

Section: Discussionmentioning

confidence: 99%

Cloning of a serotype-specific antigen from Pasteurella haemolytica A1

Gonzalez-Rayos¹,

Lo²,

Shewen³

et al. 1986

Infect Immun

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Recombinant plasmids coding for a soluble (or surface) antigen ofPasteurella haemolytica Al were identified. Two plasmids, both containing the same 5.4 kilobase pairs of insert DNA, were recovered independently by screening a clone bank of P. haemolytica Al genomic DNA in Escherichia coli for the expression of P. haemolytica Al soluble antigens (R. Y. C. Lo and L. A. Cameron, Can. J. Biochem. Cell Biol. 64: [73][74][75][76] 1986). E. coli cells carrying the plasmids were found to be agglutinated by an antiserum raised against the P. haemolytica Al soluble antigens. Analysis of the E. coli clones by electron microscopy revealed patches of amorphous material on the surface of the cells which were not present on the controls. Further characterization with protein A-colloidal gold labeled both these patches and the outer membranes of these cloned cells pretreated with the specific antiserum. These results indicated that the cloned antigen was expressed on the surface of the E. coli cells. The cloned antigen was found to be specific for serotype 1 when tested by slide agglutination against a collection of P. haemolytica typing antisera. Southern blot hybridization, using the cloned DNA as a probe, labeled the genomic DNA from P. haemolytica serotype 1 as well as the cross-agglutinating serotypes 2 and 7, but not DNA from the non-cross-agglutinating serotypes 3 and 4 and PasteureUa multocida. These results demonstrated that serotype specificity could be attributed to the particular antigenic determinants in the genome of the organism.

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Section: Discussionmentioning

confidence: 99%

Cloning of a serotype-specific antigen from Pasteurella haemolytica A1

Gonzalez-Rayos¹,

Lo²,

Shewen³

et al. 1986

Infect Immun

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“…The limitations of current vaccines have prompted a search for a variety of components in P. haemolytica which may serve as immunogens. These include bacterial cell walls and capsular materials (11), a carbohydrate-protein subunit (17), inner and outer membrane proteins (31), and a lipopolysaccharide with endotoxic properties (8). Some of these have been found to possess immunogenic properties of limited protective value.…”

mentioning

confidence: 99%

Iron acquisition in Pasteurella haemolytica: expression and identification of a bovine-specific transferrin receptor

1990

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Seven type 1 field isolates of Pasteurella haemolytica were screened for their ability to use different transferrins as a source of iron for growth. All seven strains were capable of using bovine but not human, porcine, avian, or equine transferrin. A screening assay failed to detect siderophore production in any of the strains tested. Iron-deficient cells from these strains expressed a binding activity, specific for bovine transferrin, that was regulated by the level of iron in the medium. Inhibition of expression by translation and transcription inhibitors suggested that iron regulation was occurring at the gene level. Affinity isolation of receptor proteins from all seven strains with biotinylated bovine transferrin identified a 100-kilodalton iron-regulated outer membrane protein as the bovine transferrin receptor. Iron-regulated outer membrane proteins of 71 and 77 kilodaltons were isolated along with the 100-kilodalton protein when less stringent washing procedures were employed in the affinity isolation procedure.

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“…Fresh 18-to 22-h-old cultures of P. haemolytica serotype 1 suspended to a concentration of 109 CFU in PBS were utilized as the whole-bacteria somatic antigen (WB). The CPS was prepared by warm saline extraction of 6-h-old cultures of P. haemolytica followed by chromatofocusing of the extract as previously described (16). LKT was prepared by inoculation of brain heart infusion broth with 18to 22-h-old cultures of P. haemolytica.…”

Section: Methodsmentioning

confidence: 99%

“…Antigen extracts were subjected to preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Samples were adjusted to a final protein concentration of 1 mg/ml, denatured, reduced, and separated on vertical slab gels by discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis with 2% stacking and 10% resolving gels (16). Proteins from the gels were then electrophoretically transferred onto a nitrocellulose membrane (30).…”

Section: Methodsmentioning

confidence: 99%

“…Bacterial cell wall and capsular materials have been extracted with saline, sodium salicylate, and potassium thiocyanate (11). A carbohydrate-protein subunit (CPS) has been further separated by chromatography of a saline extract (16). Capsular carbohydrates and inner and outer membrane proteins of P. haemolytica have also been purified and characterized (1,29).…”

mentioning

confidence: 99%

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Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis

et al. 1989

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Antigens associated with whole Pasteurella haemolytica biotype A serotype 1, a capsular carbohydrateprotein extract of the organism, and P. haemolytica leukotoxin were separated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Antigens of the electrophoresed preparations were detected by Western blotting (immunoblotting) with sera from cattle which were either nonvaccinated or vaccinated with live or killed P. haemolytica vaccines and had variable degrees of resistance to experimental pneumonic pasteurellosis. Distinct, easily recognizable antigens of these preparations were identified, and the antibody responses to these antigens were quantified by densitometry. To determine their importance to disease resistance, we then compared antibody responses with experimental lesion scores. Antibody reactivity to surface antigens which were significantly correlated with resistance and present in two or more of the preparations were detected at 86, 66, 51, 49, 34,31, and 16 kilodaltons (kDa). Of these, antibody responses to antigens at 86, 49, and 31 kDa appeared most important based on their concentration and signfficance levels. Antibody reactivity to leukotoxin antigens which were significantly correlated with resistance and common with important surface antigens were detected at 86, 66, and 49 kDa. Antibody responses to unique leukotoxin antigens which were significantly correlated with resistance were present at 92 and 58 kDa. * Corresponding author. t Journal article no. 5314 of the Oklahoma Agricultural Experiment Station.

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Saline-extracted antigens of : Separation by chromatofocusing, preliminary characterization, and evaluation of immunogenicity.

Cited by 26 publications

References 16 publications

Cloning of a serotype-specific antigen from Pasteurella haemolytica A1

Cloning of a serotype-specific antigen from Pasteurella haemolytica A1

Iron acquisition in Pasteurella haemolytica: expression and identification of a bovine-specific transferrin receptor

Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis

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