Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis

Abstract

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.

Molecular studies of Ssa1, a serotype-specific antigen of Pasteurella haemolytica A1

“…Previous results have demonstrated the surface location of this protein in the E. coli clones (15), and the present data support that observation. There have been a number of reports in the literature on the characterization of outer membrane proteins from P. haemolytica Al and their possible role(s) as protective antigens (6,27). These data were obtained primarily by SDS-PAGE analysis of outer membrane proteins extracted by various methods or by the use of Western immunoblot analysis of the bacterial antigens with immune serum from P. haemolytica Al-infected ani- mals (6,27).…”

“…There have been a number of reports in the literature on the characterization of outer membrane proteins from P. haemolytica Al and their possible role(s) as protective antigens (6,27). These data were obtained primarily by SDS-PAGE analysis of outer membrane proteins extracted by various methods or by the use of Western immunoblot analysis of the bacterial antigens with immune serum from P. haemolytica Al-infected ani- mals (6,27). However, the identity of these proteins or antigens is often unknown, making it difficult for follow-up and detailed characterization.…”

Proteomic and bioinformatic analyses of putative Mannheimia haemolytica secretome by liquid chromatography and tandem mass spectrometry

“…Shewen and Wilkie (1988) demonstrated that immunity against M. haemolytica requires leukotoxin (LKT) neutralizing antibodies as well as antibodies against bacterial cell surface antigens. Studies in our and others' laboratories demonstrated that high antibody responses to OMPs, ranging from 16 to 86 kDa, correlated with resistance to challenge with virulent M. haemolytica S1, and vaccination of cattle with OMP-enriched cellular fractions from M. haemolytica S1 enhanced resistance of cattle against experimental challenge (Ayalew et al, 2010(Ayalew et al, , 2011aConfer et al, 2003Confer et al, , 2006Morton et al, 1995;Mosier et al, 1989;Orouji et al, 2012). In this regard, we used immunoproteomics, which uses a combination of 2D-gel electrophoresis (2-DE) and Western blotting with bovine convalescent sera, and identified 132 immunoreactive proteins in outer membrane preparations of M. haemolytica using LC-MS/MS.…”

Characterization of a fourth lipoprotein fromPasteurella haemolyticaA1 and its homology to the OmpA family of outer membrane proteins

“…A number of P. haemolytica A1 antigens in the 28–31‐kDa range have been suggested to be protective antigens by their correlation with resistance to pneumonic pasteurellosis [18]. Previously we have reported the characterization of three 30‐kDa lipoproteins Plp1, 2 and 3 from P. haemolytica A1.…”