Progress 10/01/01 to 09/30/06
Outputs
In the last several years we have studied the effect of the small antibacterial peptide, cecropin B on the nasal flora of cattle with bovine respiratory disease (i.e. pneumonic pasteurellosis). These experiments were designed to deliver and express a cecropin B transgene on bovine nasal mucosa, and to determine the effect on Mannheimia haemolytica serotype 1 (S1) colonization. These results showed that, cecropin B has effective antibacterial activity against M. haemolytica S1, and can prevent colonization of the nasal mucosa after transfection with a plasmid expressing cecropin B. Clinical pneumonic pasteurellosis is inevitable after M. haemolytica S1 colonization and proliferation occurs in the respiratory tract. To provide protection against pneumonic pasteurellosis, M. haemolytica should not be allowed to colonize the nasal passage. The short-lived nature and site specificity of gene therapy makes it an attractive alternative for the prevention of pneumonic
pasteurellosis, because short-term expression of cecropin B would be adequate to prevent respiratory tract infection during transit or in the feedlot. Clearance of the transgene would be beneficial in that no biocontainment would be necessary. Findings of our studies indicate that the antimicrobial peptide cecropin B was useful in inhibiting M. haemolytica colonization. The use of gene therapy will help to eliminate primary M. haemolytica infection in cattle destined for the feedlots.
Impacts
The use of an antimicrobial that is effective in controlling respiratory disease in calves and leaves no unwanted residue is of economic importance to the consumer as well as the grower and meat processor. The value of gene therapy utilizing cecropin B will be greatly enhanced if all of the predominant microbial pathogens in bovine respiratory disease are controlled by this peptide.
Publications
- Corstvet, R.E., F.M. Enright, R.K. Cooper, C.M. Boudreaux NC-107 Annual Report. St. Paul, Minnesota, September 2006.
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Progress 01/01/05 to 12/31/05
Outputs
Utilizing gene therapy as a novel molecular biology method, we found that the calf produces an antimicrobial peptide (cecropin B) locally in the upper respiratory tract. We found that 100ug of the cecropin plasmid was necessary to affect this production, resulting in inhibition of the number one cause of shipping fever pneumonia, Mannheimia haemolytica. Another group of calves was used this year as a repeat of the previous work. The antibacterial results were confirmed as well as the production of cecropin B peptide. We are still in the process of analyzing all of the data. There are factors in the time after exposure to the transgene and amount of cecropin B produced that we plan to study in the future. This research is an important first step in using gene therapy to control shipping fever and bovine respiratory disease in calves. Our research has shown that the antibacterial effect lasts for at least 21 days after exposure to the transgene which is enough time to
protect the calves during shipment and the first critical days in the feedlot.
Impacts
The use of an antimicrobial that is effective in controlling respiratory disease in calves and leaves no unwanted residue is of economic importance to the consumer as well as the grower and meat processor. The value of gene therapy utilizing cecropin B will be greatly enhanced if all of the predominant microbial pathogens in bovine respiratory disease are controlled by this peptide.
Publications
- Boudreaux, C.M, R.E. Corstvet, R.K. Cooper and F.M. Enright. NC-107 Annual Report. Athens, GA, September 2005.
- Boudreaux, C.M, R.E. Corstvet, R.K. Cooper and F.M. Enright. Effects of cecropin B transgene expression on Mannheimia haemolytica serotype 1 colonization of the nasal mucosa of calves. AJVR 66 (No. 11) November 2005, pp 1922-1930.
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Progress 01/01/04 to 12/31/04
Outputs
The nasal passages of 6-8 month of age cross-bred beef calves were aerosolized with different amounts of plasmid DNA containing a gene coding for cecropin B. Results indicated that the cecropin B gene or cecropin B mRNA expression can be detected in DNA or RNA extracted from nasal swabs and nasal aspirates. Furthermore the cecropin B gene could be detected for at least 4 weeks post transfection. Detection of the ceropin B gene prior to calves being transfected and in untreated control calves may implicate the possible presence of a native bovine cecropin B or cecropin-like gene which is a new finding in cattle. Further research is necessary to determine the longevity of transfection in cattle. Transfection of the upper respiratory tract of calves with a gene coding for cecropin B does not result in a change in the indigenous and transient nasal flora. Transfection of the upper respiratory tract with 100 ug of DNA per nostril does inhibit colonization and
multiplication of a virulent strain of Mannhemia haemolytica 1:A.
Impacts
Enhancement of local defense mechanisms of the upper respiratory tract using a transiently expressed gene for cecropin B repesents a novel method for prevention of shipping fever in cattle. Characterization of new viral pathogens that may be involved in the pathogenesis of shipping fever of cattle is necessary for eventual control of this disease in cattle.
Publications
- Boudreaux, CM. 2004. A novel strategy of controlling bovine pneumonic pasteurellosis: Transfecting the upper respiratory tract of cattle with a gene coding for the antimicrobial peptide Cecropin B. M.S. Thesis, Louisiana State University Baton Rouge.
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Progress 01/01/03 to 12/31/03
Outputs
In-vitro results indicated that cecropin B is very effective in killing M. haemolytica 1:A. Transfection of the upper respiratory tract of cattle with a gene coding for cecropin B does not result in a change in the indigenous and transient nasal flora.A transfection dose of 100 micrograms of DNA per nostril inhibits colonization of a virulent strain of M. haemolytica 1:A. Over 300 bovine coronavirus strains have been isolated from animals that experienced acute respiratory disease during two different shipping fever epizootics. The entire genome sequence of three different strains, including one of enteric origin have been obtained. Comparing the sequences of the enteric and pulmonary isolates has revealed numerous single nucleotide substitutions.
Impacts
Enhancement of local defense mechanisms of the upper respiratory tract using a transiently expressed gene for cecropin B repesents a novel method for prevention of shipping fever in cattle. Characterization of new viral pathogens that may be involved in the pathogenesis of shipping fever of cattle is necessary for eventual control of shipping fever in cattle.
Publications
- Corstvet, R.E., Enright, F.M., Miller, J.E., Kousoulas, K.G., Cooper, R.K., and McBride, J.M., Boudreaux, C.M., Research Associate. NC-107 Annual Report. Baton Rouge, Louisiana; September 2003.
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Progress 01/01/02 to 12/31/02
Outputs
We have continued to test cattle sera for antibody to the Mannheim haemolytica (MH) antigens we reported as determining infection in the last AD-421 progress report. We still find that the 28kD and 30kD antigens are important in the determination of recent infection with MH. We also began experiments in cattle to determine if a plasmid containing a gene for cecropin B could be transfected into the epithelial cells of the nasal mucosa. In vitro, MH is highly susceptible to the cecropin B peptide. The cattle work has been done this past year. The specimens have to be analyzed. Preliminary results indicate that the cecropin B gene possibly can be transfected. The effect of cecropin B on MH multiplication in the nasal passage of cattle has to be ascertained. These specimens are being processed.
Impacts
If all of our cattle experiments with cecropin B indicate it is effective in vivo against MH; then we have an important tool in combating shipping fever in cattle and pasteurellosis.
Publications
- Corstvet RE, Enright FM, Miller JE and Kousoulas K.G. 2002 NC-107 report given at Michigan State University, East Lansing, MI Sept. 2002. Both written and oral report.
- Boudreaux CM and Corstvet RE. Susceptibility of a virulent strain of Mannheimia haemolytica: A to cecropin B. Phi Zeta Research Emphasis Day, Louisiana State University, Wednesday, September 25, 2002
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Progress 01/01/01 to 12/31/01
Outputs
The NC-107 project was approved for another 5 years. Louisiana has a significant role in this project. A transition team of researchers has been organized by Corstvet. This group will be part of the "calf to market" program in the Ag Center and has also submitted a NRI proposal in conjunction with researchers at Mississippi State University on an aspect of BRD. Last year we said we would work with the antigens expressed in BHI broth and develop a sensitive serological test for detection of pasteurellosis in cattle. Western blots of nasal aspirates, bronchioalvolar lavage fluids, and serum samples were performed using BHI cell free supernatant using a goat anti-bovine IgG (H&L) alkaline phosphate conjugate. Western blots were compared to IgG (H&L) antibody titers against BHI cell free supernatant as determined by ELISA. The BHI cell free supernatant contained 28-kD, 45-kD, 50-kD, 63-kD and 130-kD bands. The 28kD and 30-kD bands apparently are important in determination
of recent exposure to the bacteria.
Impacts
The 28-kD and 30-kD antigens are useful in determination of recent infection with pasteurellae.
Publications
- Corstvet, R.E., Enright, F.M., Miller, J.E., Kousoulas, K.G. and Thompson, A. 2001. NC-107 report given at Iowa State University, Ames, IA. Sept. 2001. Both written and oral repot.
- Corstvet, R.E. and Boudreaux, C.M. 2001. Essential Tools in Pasteurella Research. Mtg. of ASM South Central Branch and the South Central Branch ASM Regional Microbil. Ed. Conf. Oct.12 and 13, LSU, SVM, Baton Rouge, LA. Abstracts p. 37.
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Progress 01/01/00 to 12/31/00
Outputs
Data obtained from aerosol vaccination (using our patented vaccine) of 450 pound calves has reaffirmed that aerosol vaccination at the level of the middle meatus enhances PH-1 clearance from the nasal passages and prevents migration of PH-1 to the lung. Vaccination at the level of the tracheal bifurcation does not affect nasal colonization of PH-1, nor does it affect immunoglobulin production in the nose. Our recently developed BHI antigen preparation for ELISA detection of PH-1 antibodies has made these results possible to obtain. In addition, we can and have ascertained concentrations of the various immunoglobulin classes after various intervals after vaccination. Of the antibody classes, IgA appears to have an adverse effect on colonization of PH-1 in the nasal passage. The BHI antigen preparation of PH-1 is very effective and specific in measuring specific exposure to PH-1. This preparation has been determined to contain the antigen responsible for
hemagglutination activity of PH-1. Further work with the antigens contained in BHI will be done in the development of a sensitive serologic test for detection of infection of cattle. The NC-107 group has completed a research plan for renewel of our funding for 5 more years after year 2002. This research plan is awaiting approval.
Impacts
The findings indicate the importance of aerosol exposure of the respiratory tract in development of effective immunity to PH-1. Future vaccines and host interactions should address the importance of mucosal-pulmonary immune systems to resistance to PH-1. This work will add important facts to the database necessary to plan strategies for control of BRD. The BHI antigen preparation will be useful in determination of exposure to PH-1.
Publications
- Corstvet, R.E., Enright, F.M., et al. 2000. NC-107 report given at Custer State Park, South Dakota State University. September 2000. Both a written and an oral report.
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Progress 01/01/99 to 12/31/99
Outputs
Data obtained from aerosol vaccination of 450 pound calves has shown the aerosol vaccination at the level of the middle meatus enhances PH-1 clearance from the nasal passage and prevents migration of PH-1 to the lung. Vaccination at the level of the tracheal bifurcation does not affect nasal colonization of PH-1. Vaccination of the middle meatus results in the production IgA type antibodies to various PH-1 whole cell and outer membrane antigens which are detected in the nasal aspirates. The use of a PH-1 cell free culture supernate (BHI) makes an excellent antigen preparation for determination of a specific antibody response to PH-1. One of us (McBride) has found that the initial inflammatory response in the lung probably is caused by complexes of antigen, antibody and complement. This inflammatory response reduces lung clearance of PH-1 and may initate and accentuate the lung injury associated with pneumonic pasteurellosis.The BHI supernate contains a 49 kd protein
(48-50 kd) and a 33 kd protein. There is also an indistinguishable protein at the bottom of the gel. It is lower than the 20 kd, and is possibly closer to 7 kd. All of these proteins can be detected by Western blotting using the whole cell Pasteurella haemolytica antigens as well. The serum samples detect all of the above 3 proteins in BHI supernate. The nasal and lavage samples do not detect the 33 kd protein even when the samples are concentrated. These samples all show a high ELISA titer using the BHI antigen preparation. Western blots using the BHI supernate antigen and negative serum form no bands at all. This is unlike the blots using whole cell antigen and negative serum where 4-5 bands are formed. Antibody to PH-1 in sera, nasal aspirates and lavages are easily distinguishable from negative specimens using the BHI supernate antigen. Different sera samples (from different sources) have been shown to be specific for the different proteins in the BHI supernate (i.e.different
samples develop different combinations of the proteins and some bands are darker or sharper than with other samples). The bands developed with the BHI supernate antigen are comparable when the same sample is developed using the whole cell antigen. However, the use of whole cell antigen results in many more bands on both positive and negative PH-1 antibody specimens. The BHI preparation is very effective in the ELISA test to detect specific antibody to PH-1 in serum, nasal aspirates and lavage specimens.
Impacts
The findings indicate the importance of aerosol exposure of the respiratory tract to immunogens of PH-1. They also indicate that the mucosal-pulmonary immune systems should be considered in resistance to PH-1. This work will add important facts to the data base necessary to plan strategies for control of BRD. The BHI antigen preparation will be useful in determination of exposure to PH-1.
Publications
- McBride, J.W., Wozniak, E.J., Brewer, A.W., Naydan, D.K., and Osburn, B.I. 1999. Evidence of Pasteurella haemolytica linked immune complex disease in natural and experimental models. Microbol. Pathology. 26(4):183-193.
- McBride, J.W., Corstvet, R.E., Taylor, B.C., and Osburn, B.I. 1999. Primary and anamnestic responses of bovine bronchoalveolar peripheral blood lymphocyte subsets to aerosolized Pasteurella haemolytica A1. Vet. Immunol. & Immunopathol. 67(2):161-170.
- Corstvet, R.E., Enright, F.M., et al. 1999. NC-107 report given at Davis, CA, Sept. 1999. Both a written and oral report.
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Progress 01/01/98 to 12/31/98
Outputs
We have continued to explore vaccination of calves with Pasteurella haemolytica (Ph-1). Data obtained from aerosol vaccination of 450 pound calves has shown the aerosol vaccination at the level of the middle meatus enhances Ph-1 clearance from the nasal passage. Vaccination at the level of the tracheal bifurcation does not. Vaccination at the level of the middle meatus results in production of IgA type antibodies to various Ph-1 whole cell and outer membrane antigens which are detected in the nasal aspirates. The use of the 40k outer membrane from Ph-1 is useful in determination of prior exposure of cattle to Ph-1. The antibody response measured by broncheoalveolar lavage and nasal aspirate specimens after nasal aerosol and aerosol at the tracheal bifurcation showed that IgA was formed after nasal aerosol but not after tracheal bifurcation aerosol. After tracheal bifurcation aerosol, antibody concentration did not increase in the nasal aspirates but did in the
bronchoalveolar lavage fluids. In both cases antibody was found in serum specimens. We have used a preparation from broth grown Ph-1 to determine its efficacy in detecting antibodies to Ph-1 after vaccination. Prelimary results indicate that this will be an effective antigen to use in ELISA to measure antibody response to Ph-1 vaccination. It will be evaluated as a diagnostic reagent for detection of Ph-1 infection in cattle. One of us (McBride) has found that the initial inflammatory response in the lung may be caused by complexes of antigen, antibody and complement. This inflammatory response reduces lung clearance of Ph-1 and may accentuate the lung injury associated with pneumonic pasturellosis.
Impacts
(N/A)
Publications
- CORSTVET, R.E.. F.M.ENRIGHT, ET AL. 1998. NC-107 report given at Madison, WI. Sept. 1998. Both a written and oral report.
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Progress 01/01/97 to 12/31/97
Outputs
This year we continued to explore the effect of P. haemolytica serotype A1 (Ph1) exposure on the bovine immune response. 23 calves were used to determine antibody responses to three outer membrane proteins (OMPs) of 31, 43, and 70 kDa. The animals were exposed to Ph1 either intranasally or at the tracheal bifurcation. The intranasal exposure caused a significant increase in the level of sera, nasal and lung Ig to the OMPs. The greatest concentration of IgA was in the nasal fluids. However, exposure at the bifurcation caused no nasal increase of Ig. This indicates that intranasal exposure is a most effective method for stimulating immune response. It was also noted that prior to vaccination animals displayed no antibody to the 40 kDa protein, this would indicate that it is a genus, or species specific protein and this implies the antigen could be used in diagnostic tests. In addition, 10 calves were used to test the effect of repeated intrabronchial exposure to Ph1 by
aerosol. Bronchoalveolar lavages (BAL) were performed and peripheral blood taken. The BAL showed a significant increase in lymphocytes and neutrophils after repeated exposure; the peripheral blood lymphocyte and neutrophil populations remained unchanged. Together these data indicate that the immune responses to Ph1 are compartmentalized in the upper and lower respiratory tract. This knowledge will allow researchers to better design vaccines and implement methods of administration.
Impacts
(N/A)
Publications
- Brennan, R.E., Corstvet, R.E., and D.B. Paulsen. 1998. Detection of antibodies against Pasteurella haemolytica A1 (Ph1) and three outer membrane proteins of Ph1 in serum, nasal secretions, and bronchoalveolar lavages by ELISA. Am. J. Vet. Res. In Press.
- McBride, J.W., Corstvet, R.E., McClure, J.R., Brennan, R.E., Taylor, B.C., and B.I. Osburn. 1997. Phenotypic and differential characterization of broncoalveolar and peripheral blood lymphocytes in calves sequentially exposed to aerosolized Pasteurella haemolytica. Vet. Immunol. Immunopathol. In Press.
- McBride, J.W., Corstvet, R.E., and B.I. Osburn. 1997. Pasteurella haemolytica outer membrane antigenicity is influenced by growth phase and conditions. Vet. Immunol. Immunopathol. In Press.
- Brennan, R.E., Corstvet, R.E., and D.B. Paulsen. 1997. Systemic and Mucosal Antibody Responses to Pasteurella haemolytica A1 (Ph1) and three outer membrane proteins of Ph1 in cattle. Conf. Res. Wrkrs. Animal Dis. Abstract #183.
- Storz, J., Stine, L., Lin, X.Q., Angel, K.L., Corstvet, R.E., and S.S. Nicholson. 1997. Currently Prevailing Virus Infections of Cattle with Acute Respiratory Disease. Conf. Res. Wrkrs. Animal Dis. Abstract #204.
- Brennan, R.E., Corstvet, R.E., and J.W. McBride. 1997. A simplified method for isolating outermembrane protein from Pasteurella haemolytica A1. J. Microbiol. Met. 29:201-206.
- McBride, J.W., Corstvet, R.E., Dietrich, M.A., Berry C., Brennan, R.E., Taylor, B.C., Stott, J.L., and B.I. Osburn. 1997. Memory and CD8+ are the predominant bovine bronchoalveolar lymphocyte phenotypes. Vet. Immunol. Immunopathol. 58:55-62.
- McBride, J.W., Corstvet, R.E., McClure, J.R., Brennan, R.E., Taylor, B.C., and B.I. Osburn. 1997. Phenotypic and differential characterization of bronchoalveolar and peripheral blood lymphocytes in calves sequentially exposed to aerosolized Pasteurella haemolytica A1. Conf. Res. Wrkers. Animal Dis. Abstract #208.
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Progress 01/01/96 to 12/30/96
Outputs
The relationship of colonization of the respiratory tract and immunization to P.haemolytica was repeated this year. The groups consisted of 6 calves that were nonvaccinates and 6 that were vaccinates. The vaccine strain of PH-1 did not persist in the respiratory tract of any calves. The challenge strain of PH-1 persisted in the respiratory tract of non vaccinates 1 day and 1 week post challenge; in the vaccinates PH-1 was isolated from the nasal passage in 3 of 6 animals (nonvaccinates 6/6) 1 day post challenge and 1 of 6 animals (nonvaccinates 3/6) 1 week later, at 2 weeks post challenge PH-1 was isolated from the nasal passage of 1/6 of the non vaccinates. Two and a half months after challenge no PH-1 was isolated from any of the calves. PH-1 was isolated from more areas of the respiratory tract in the nonvaccinates than in the vaccinates. It was found, as it was last year, that when night temperatures dropped to 58-60F and daytime temperatures were in the 70's,
PH-1 persisted for a longer period of time in more of the nonvaccinates than in the vaccinates. The nasal passage appears to indicate persistence better than the tonsil or any other part of the respiratory tract. The clearance from the trachea on down apparently is quite formidable in vaccinates and nonvaccinates. Data concerning phenotypic differences observed between bronchoalveolar and peripheral blood T lymphocyte subpopulation showed that these differences may indicate selective lymphocyte migration to the bovine lung.
Impacts
(N/A)
Publications
- PAULSEN, D. B., M.K. LOPEZ AND ET AL. Characterization of the plasma cell infiltrate in experimental bovine pneumonic pasteurellosis. CRWAD (Abstract), 77th Mtg., Nov. 11-12, 1996. Abst. #207.
- McBRIDE, J. W., R.E. CORSTVET AND ET AL. 1996. Memory and CD8-+ are the Predominant Bovine Bronchoalveolar Lymphocyte Phenotypes. Cell Immunol. (Accepted for Publication).
- McBRIDE, J. W., R.E.CORSTVET AND ET AL.. 1996. Characterization of Bovine Pulmonary and Serum Antibody Responses After Parenteral or Intrapulmonary Vaccination with live Pasteurella haemolytica. Comp. Immun. Microbiol. Infect. Dis. 19(2):99- CORSTVET, R.
- E., D.B. PAULSON AND ET AL. NC-107 Annual Meeting, Baton Rouge, LA.,Sept. 1996.
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Progress 01/01/95 to 12/30/95
Outputs
1. A highly portable apparatus for aerosol vaccination of calves using P. haemolytica was assembled and tested in calves. 2. P. haemolytica colonization of the bovine respiratory tract was ascertained. The organism did not persist. Climatic conditions adversely influenced the resistance to colonization and clearance even in vaccinated calves. 3. Intrapulmonary vaccination with P. haemolytica resulted in the lavage fluids from these calves having IgG antibodies with different patterns of antigen specificity compared with IgG antibodies in analogous sera. 4. Anti-PH-1 IgA and IgG antibody concentrations were significantly higher (P,0.05) in lung lavage fluids from the intrapulmonary vaccinated group before and after challenge compared to the subcutaneous and sham groups. 5. The intrapulmonary and subcutaneous vaccinated groups developed IgA, IgG and IgM antibody in nonvaccinated or nonchallenged lung lobes. (In dogs antibody is only associated with the treated lobe.) 6.
Pulmonary IgG and IgA antibodies were associated with increased lung protection after challenge. 7. Developed monoclonals and technologies to determine lymphocyte markers in lung lavage fluids from cattle. 8. Assisted in the writing of the next 5 year proposal for the NC-107. 9. Prepared an NRI proposal that was submitted for 1996 funding.
Impacts
(N/A)
Publications
- MCBRIDE, J.W., R.E. CORSTVET, D.B. PAULSEN, J.R. MCCLURE AND F.M. ENRIGHT. 1995. Pulmonary and serum immunoglobulin responses and pulmonary immunity against Pasteurella haemolytica A1 in calves. Comp. Immun., Microbiol. and Infect. Dis. (In
- PAULSEN, D.B., A.W. CONFER, K.D. CLINKENBEARD AND D.A. MOSIER. 1995. Pasteurella haemolytica lipopolysaccharide-induced cytotoxicity in bovine pulmonary artery endothelial monolayers: Inhibition by indomethacin. Vet. Path., 32:173-183. CORSTVET, R.E., D.B. PAULSEN AND ET AL. NC-107 Report Annual Meeting, Columbia, MO, Sept. 1995.
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Progress 01/01/94 to 12/30/94
Outputs
Different routes of immunization with Ph-1 were used to stimulate the mucosal immune system. A priming dose was given followed by an intranasal and intratracheal booster 14 days later. We used criteria, such as, colonization of various levels of the respiratory tract, serum and respiratory antibody responses, and antibody production by cells from lavage fluids as measures of resistance. Intraperitoneal inoculation gave protection against colonization of the lung one day after challenge and trucking. Six days after challenge colonization of the nasal passage was found in the nonvaccinated animals only. Seven days later colonization of the nasal passage only was found again in the nonvaccinated animals and also in the intratracheal vaccinated animal This data has been added to by additional experiments which have just been concluded. The study on immune response of the respiratory system will enable us to clearly define important immunogens and isotype class in lavage
fluids involved in resistance of the upper and lower respiratory tract to Pasteurella haemolytica infection.
Impacts
(N/A)
Publications
- MCBRIDE, J.W., R.E. CORSTVET, D.B. PAULSEN, J.R. MCCLURE AND F.M. ENRIGHT. 1994. Differences in antigen recognition by systemic and pulmonary antibodies induced by Pasteurella haemolytica A1. Infect. Immun. (Submitted).
- PACE, L.W., J.R. TURK, R.E. CORSTVET, F.M. ENRIGHT AND W. HENK. 1994. Induction of acute bronchopneumonia in mice by intrabronchial inoculation of Pasteurella haemolytica Serotype I. Can. J. Vet. Res. 58:79-82.
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Progress 01/01/93 to 12/30/93
Outputs
Identifying antigens that are important for immunity is necessary to produce effective vaccines. The purpose of our experiments was to determine if pulmonary induced and systemic induced antibodies recognize the same antigens. P. haemolytica-specific IgG and IgA antibodies contained in lung lavage fluids from pulmonary vaccinates recognized a different array of antigens than the same isotype in serum. We also confirmed that leukotoxin, an antigen which is often correlated with resistance to disease, was not consistently detected in lung lavage fluids, and calves with pulmonary antibody to leukotoxin did not have significantly smaller lesions. The unique pattern of antigen recognition by antibodies in lavage fluids indicates that a unique local production of antibody in the respiratory tract may be occurring. We have developed an in vitro antibody generation assay using lymphocytes isolated from bronchoalveolar lavage fluids and from blood. Results, thus far, indicate
that we only have significant antibody formation by the lung lavage derived lymphocytes. The highest level of antibody formation has been from the intraperitoneal priming of GALT followed by a intranasal/intratracheal booster. Disclosure of Patent: Corstvet, R.E., F.M. Enright and J.R. McClure, U.S. patent No.5,256,415; Vaccine against bovine respiratory disease; issued 10/26/93; file: 6451.044.
Impacts
(N/A)
Publications
- CORSTVET, R.E., D. PAULSEN, ET AL. 1993. Annual Progess Report, NC-107 Technical Committee on Respiratory Diseases of Cattle, University of Tennessee, Gatlingburg, TN (Written report and oral presentation).
- AUSTIN, F.W. AND R.E. CORSTVET. 1993. Purification of a Pasteurella haemolytica serotype 1 specific polysaccaride epitope using monoclonal antibody immunoaffinity. Am. J. Vet. Res. 54(5):695-700.
- MCBRIDE, J.W.. 1993. Pulmonary and systemic immunoglobin response of calves to Pasteurella haemolytica A1. M.S. thesis; Louisiana State University, Baton Rouge, LA.
- MCBRIDE, J.W., R.E. CORSTVET, ET AL. 1993. Differences in antigen recognition by systemic and pulmonary antibodies induced by Pasteurella haemolytica Al. Am. J. Vet. Res. (Submitted).
- MCBRIDE, J.W., R.E. CORSTVET, ET AL. 1993. Pulmonary and systemic antibody responses and pulmonary immunity of calves to Pasteurella haemolytica A1 after prolonged systemic or pulmonary stimulation. Vet. Microbial. (Submitted).
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Progress 01/01/92 to 12/30/92
Outputs
Determined the systemic and pulmonary antibody response of calves vaccinated with Pasteurella haemolytica. Response consisted of IgM, IgG and IgA in the serum and lavage fluids. IgG was the predominant response in the serum, while IgG and IgA were in ratio of 2.5:1 in pulmonary lavage fluids. Compared systemic pulmonary routes of vaccination and correlated pulmonary lesion size. Pulmonary vaccination appears to provide better protection by inducing mucosal immune response, particularly IgA, and a more localized response in the lung which is quicker and more intense, especially for the IgA isotype. The intensity of the antibody response in the left diaphyramatic lobe, right cardiac lobe and left caudal lobe appear to be similar to that of the vaccinated right diaphyramatic lobe. Antigens recognized by antibodies in lavage fluids and serum from calves vaccinated systemically and pulmonary were identified. The antibodies in lavage fluids and serum recognize different
antigens regardless of route of vaccination. IgA and IgG isotypes in lavage fluids recognize the same antigens which are 60-86,77,49,40 kDa proteins, while serum antibodies recognized antigens more independently. These included 105,92,77,60,49 and 30kDa. It was also demonstrated that leukotoxin is not consistently recognized by antibody in lavage fluids, and protection did not correlate with presence of antibody to leukotoxin.
Impacts
(N/A)
Publications
- PAULSEN, D.B., R.E. CORSTVET, ET AL. 1992. Use of an indwelling bronchial catheter model of bovine pneumonic pasteurellosis for evaluation of therapeutic efficacy of various compounds. Am. J. Vet. Res. 53(5):679-683.
- MCBRIDE, J.W., R.E. CORSTVET, D.B. PAULSEN, AND R.J. MCCLURE. 1992. Systemic and pulmonary antibody responses of calves to Pasteurella haemolytica after intralpulmonary inoculation. Am. J. Vet. Res. 53:1889-1894.
- AUSTIN, F.W., R.E. CORSTVET, K.L. SCHNORR AND W.J. TODD. 1992. A characterization of monoclonal antibodies prepared against Pasteurella haemolytica serotype 1 surface antigens. Veterinary Microbiology 32:327-342.
- CORSTVET, R.E., D. PAULSEN, ET AL. 1992. Annual Progress Report, NC-107 Technical Committee on Respiratory Diseases of Cattle, Minnesota State University, St. Paul, MN. (Written report and oral presentation).
- AUSTIN, F.W. AND R.E. CORSTVET. 1992. Purification of a Pasteurella haemolytica serotype 1 specific polysaccharide epitope using monoclonal antibody immunoaffinity. Am. J. Vet. Res. (in-press).
- MCBRIDE, J.W., R.E. CORSTVET, ET AL. 1992. Pulmonary immunoglobulin response of calves against Pasteurella haemolytica after pulmonary or systemic stimulation. Conf. Res. Workers Animal Disease, 73rd Annual Mtg.:44.
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