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Serum antibody response to carbohydrate antigens of Pasteurella haemolytica serotype 1: relation to experimentally induced bovine pneumonic pasteurellosis
1989
Confer, A.W. | Simons, K.R. | Panciera, R.J. | Mort, A.J. | Mosier, D.A.
The antibody responses to the capsular carbohydrate (CC) purified from Pasteurella haemolytica serotype 1 were determined by an ELISA using 135 sera from 6 calves vaccinated with phosphate-buffered saline solution, formalin-killed P haemolytica bacterins, live P haemolytica, or an extract of P haemolytica referred to as carbohydrate-protein subunit (CPS). Calves vaccinated with live P haemolytica, bacterins, or CPS developed serum antibodies to CC. Bacterins containing Freund incomplete adjuvant or Freund complete adjuvant induced higher antibody responses than did bacterins containing aluminum hydroxide. In 4 of 6 experiments, high antibody responses to CC were significantly (P less than 0.05) correlated with resistance to transthoracic challenge exposure with P haemolytica. When calves were challenge exposed with a dose of P haemolytica that was 4.5 times greater than the standard challenge exposure dose or when calves that had been vaccinated with CPS were challenge exposed, antibody responses did not significantly (P greater than 0.05) correlate with resistance to challenge exposure. The amount of serum antibodies to CPS increased significantly (P less than 0.05) when calves were vaccinated with live or killed P haemolytica or with CPS, compared with that in calves given saline solution. In 5 of 6 experiments, correlation between high antibody responses and resistance to challenge exposure was significant (P less than 0.05). The correlation between those variables was not significant (P less than 0.07) for CPS-vaccinated calves. In the ELISA, treatment of CPS with sodium m-periodate, to oxidize periodate-sensitive carbohydrate epitopes, failed to markedly alter the antibody response to CPS. However, the correlation between high antibody responses to periodate-treated CPS and resistance was significant (P less than 0.05) for all 6 experiments. In the ELISA, periodate treatment of CC, lipopolysaccharide, and CPS caused average reductions in antibody reactivity of 7.1%, 53.8%, and 34.5%, respectively. Preadsorption of sera with CC or lipopolysaccharide did not markedly reduce antibody reactivity with CPS. Preadsorption of sera with CC and reaction with periodate-treated and nontreated CPS indicatedthat for calves given phosphate-buffered saline solution vaccines, antibody reactivity was reduced 65.4%, whereas for those vaccinated with a bacterin with aluminum hydroxide, a bacterin with Freund incomplete adjuvant, or live P haemolytica, antibody reactivity was reduced 47.1%, 40.5%, and 25.0%, respectively. It was concluded that serum antibodies to CC are of some importance in resistance and that certain epitopes in CPS that are not sensitive to periodate are of importance in resistance to bovine pneumonic pasteurellosis. There are qualitative and quantitative differences among the serum antibody responses to carbohydrate epitopes for calves vaccinated with phosphate-buffered saline solution, bacterins, or live P haemolytica.
Afficher plus [+] Moins [-]Pathogenesis of naturally acquired bovine respiratory syncytial virus infection in calves: evidence for the involvement of complement and mast cell mediators
1989
Kimman, T.G. | Terpstra, G.K. | Daha, M.R. | Westenbrink, F.
Indicators of immune-mediated disease were studied in calves with severe natural bovine respiratory syncytial virus infection. Although antigen and antibody were detected concurrently in most calves, immune complexes were not detected by use of immunofluorescence, ELISA, and binding of the 1q component of complement. Complement component C3, however, was observed by immunofluorescence in the cranioventral, virus-infected portion of the lungs of 19 of 25 calves. Reductions in the amount of histamine and in the numbers of mast cells and mast cell granules in the virus-positive cranioventral and virus-negative caudodorsal portions of the lungs, indicated activation of mast cells and liberation of their granule contents. On the basis of these and previous findings, a model for the pathogenesis of bovine respiratory syncytial virus-induced disease was proposed.
Afficher plus [+] Moins [-]Hemagglutination and hemagglutination inhibition of turkey red blood cells with Mycoplasma hyopneumoniae
1989
Young, T.F. | Erickson, B.Z. | Ross, R.F. | Wannemuehler, Y.
The ability of Mycoplasma hyopneumoniae to agglutinate RBC was evaluated to develop an in vitro cytadsorption assay. Using swine RBC in a microtitration hemagglutination test, no agglutination or partial agglutination was detected. Comparison of RBC from various other species indicated that improved hemagglutination was obtained with RBC from turkeys. This hemagglutination was detected only when mycoplasma cells used in the assay had been frozen and thawed, heated at 50 C for 30 minutes, or treated with trypsin. Treatment of RBC with trypsin or neuraminidase enhanced hemagglutination. Possible surface lectin activity in M hyopneumoniae was evaluated by use of carbohydrates in a blocking assay; hemagglutination was not inhibited by any of 13 carbohydrates evaluated. Mycoplasma hyopneumoniae convalescent porcine serum and monoclonal antibodies against 2 M hyopneumoniae immunogens of molecular weights of 64,000 and 41,000 inhibited hemagglutination.
Afficher plus [+] Moins [-]Protection of mice against Brucella abortus infection by inoculation with monoclonal antibodies recognizing Brucella O-antigen
1989
Phillips, M. | Deyoe, B.L. | Canning, P.C.
Monoclonal antibodies recognizing the O-polysaccharide portion of Brucella abortus strain 2308 provided BALB/c mice with passive protection against challenge exposure with the homologous strain. Numbers of colony-forming organisms in the spleen were reduced by IgM and IgG monoclonal antibodies. Active immunization of mice, using B abortus 2308S lipopolysaccharide, resulted in production of IgM antibody at 14 days. Clearance of organisms in the actively immunized mice after challenge exposure at 14 days was nearly identical to that in passively immunized mice. Mice either passively or actively immunized were effectively protected from 0 to 28 days. Bacterial colonization of the spleen was observed to increase in both groups of mice at 56 days and indicated that humoral responses were effective in eliminating the organism in the early stages of infection, but other immune mechanisms were necessary for protection of mice in the later stage of infection with virulent strains of B abortus.
Afficher plus [+] Moins [-]Production and characterization of monoclonal antibodies to porcine immunoglobulin gamma, alpha, and light chains
1989
Paul, P.S. | Mengeling, W.L. | Malstrom, C.E. | Van Deusen, R.A.
Monoclonal antibodies (MAB) to porcine immunoglobulins were produced by fusion of SP2/0 cells with splenic lymphocytes of mice that had been immunized with porcine IgG or IgA. Of 16 MAB selected for detailed study, 13 reacted with heavy chains (7 anti-gamma, 6 anti-alpha) and 3 reacted with light chains of native immunoglobulin. Several of the same MAB (3 anti-gamma chain, 1 anti-alpha chain, 3 anti-light chain) also reacted with denatured immunoglobulin by use of immunoblotting analysis. Collective results of competitive ELISA, immunodiffusion, and immunoblotting analysis indicated that the 7 MAB of anti-gamma chain specificity were directed to 3 epitopes, the 6 MAB of anti-alpha chain specificity were directed to at least 2 epitopes, and the 3 MAB of anti-light chain specificity were directed to at least 2 epitopes that may have been located on different types of light chains. When tested by immunodiffusion with 5% polyethylene glycol incorporated in the agar matrix, all anti-gamma chain and antilight chain MAB, but no anti-alpha chain MAB, had precipitating activity. When polyethylene glycol was not used, only 4 MAB (all of the anti-gamma chain specificity and of IgM isotype) had precipitating activity. These MAB, with specificity for gamma and alpha chains and those reported earlier with mu-chain specificity, should be invaluable in the detection and quantification of porcine immunoglobulin isotypes. These MAB have potential applications in delineating the porcine immune response to selected immunogens.
Afficher plus [+] Moins [-]Development and evaluation of an enzyme-linked immunosorbent assay for bovine antibody (IgG) to Pasteurella haemolytica
1989
The sensitivity of an indirect enzyme-linked immunosorbent assay (ELISA) for bovine IgG serum antibody to Pasteurella haemolytica was compared with that of an indirect hemagglutination (IHA) test. Pasteurella haemolytica serotypes were grown in a chemically defined cell culture medium, and soluble antigens released into the growth medium were used in the ELISA and IHA test. An ELISA with serotype-1 antigen consistently detected antibody in sera that were positive by IHA test (correlation, 99%). Sera reacting with serotype-1 ELISA antigens also reacted with ELISA antigens prepared from other serotypes. Although ELISA titers determined by the 2 methods were approximately linear. Titer increases detected in paired serum samples by either test were similar. The ELISA was more sensitive than was the IHA in detecting colostral IgG antibody in serum of newborn calves. The ELISA uses a simple, stable antigen preparation and detects antibody to P haemolytica serotypes that commonly infect cattle.
Afficher plus [+] Moins [-]Total and antigen-specific serum immunoglobulin isotype concentrations in hyperimmunized cattle that have undergone plasmapheresis
1989
McVey, D.S. | Loan, R.W.
The effects of prolonged plasmapheresis of cattle on total and antigen-specific immunoglobulin production were evaluated. Five adult cows were hyperimmunized by repeated IV administration of live, logarithmic-phase Pasteurella haemolytica A1 organisms. Three of the cows underwent plasmapheresis daily for 3 weeks. From 2 cows, serum was only obtained periodically. Anti-P haemolytica antibody was assayed by indirect hemagglutination and a kinetic-augmented, antigen-capture ELISA for capsular polysaccharide and lipopolysaccharide/outer membrane protein antigens. Total serum immunoglobulin concentration was determined for IgM, IgG1, and IgG2 by primary radial immunodiffusion. Anti-P haemolytica A1 activity increased rapidly after immunization. After beginning plasmapheresis, the antigen-specific antibody activities remained nearly constant. In general, antilipopolysaccharide/outer membrane protein activity (in terms of concentration) was higher than anti-capsular polysaccharide activity and was not affected as much by the plasmapheresis. Total serum Ig concentration decreased transiently by a small amount after beginning plasmapheresis.
Afficher plus [+] Moins [-]Interaction of turkey complement with Escherichia coli isolated from turkeys
1989
Ellis, M.G. | Arp, L.H. | Lamont S.J.
The role of turkey complement in a serum bactericidal reaction was determined using serum-sensitive and serum resistant Escherichia coli isolated from turkeys. Inactivation of complement by heating serum (56 C for 40 minutes) or by treating serum with 10mM EDTA eliminated bactericidal activity. Serum sensitive E coli organisms were killed by turkey serum treated with 10 mM ethylene glycol-bis-beta-(aminoethyl ether)-N, N, N,' N,'-tetraacetic acid and 5 mM MgCl2. Exposure of normal turkey serum to serum-sensitive or serum resistant E coli resulted in equivalent reductions in hemolytic activity of serum. Treatment of serum-resistant E coli with antibody rendered the bacteria sensitive to bactericidal effects of normal turkey serum. Serum-sensitive E coli organisms were readily killed by an alternative complement pathway, serum-sensitive and serum-resistant E coli activated the complement system equally well, and antibody was required for complement-mediated killing of certain serum-resistant E coli organisms from turkeys.
Afficher plus [+] Moins [-]Oral vaccination of dogs fed canine adenovirus in baits
1989
Baer, G.M. | Brooks, R.C. | Foggin, C.M.
Six groups of 5 dogs each were fed dilutions of canine adenovirus-2, either as raw liquid or after insertion into cornmeal baits. By the fourth week after vaccination, 29 of the 30 dogs developed high titers of serum-neutralizing antibodies to the virus.
Afficher plus [+] Moins [-]Evaluation of the specificity of Pasteurella multocida somatic antigen-typing antisera prepared in chickens, using ribosome-lipopolysaccharide complexes as inocula
1989
Rimler, R.B. | Angus, R.D. | Phillips, M.
Purified lipopolysaccharides (LPS) from 16 serotypes of Pasteurella multocida were complexed with Aspergillus fumigatus ribosomes. The complexes were used as inocula to prepare antisera, in chickens, for somatic antigen typing by the gel diffusion precipitin test (GDPT). Antisera made against 15 of 16 LPS reacted with their respective specific heat-stable antigens in the GDPT and homologous LPS in the passive hemagglutination test. Antisera could not be made against serotype 15 LPS. Correlation was not observed between intensity of the precipitin reaction in the GDPT and titer to homologous LPS in the passive hemagglutination test. Most antisera cross-related with other heat-stable antigens of other serotypes in the GDPT. Many of these cross-reactions were eliminated by dilution. Cross-reactions that occurred in the GDPT with antisera made against LPS of serotypes 2, 5, 7 and 8 could not be eliminated by dilution.
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