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Solubilization of membrane-associated cross-protection factor(s) of Pasteurella multocida
1National Animal Disease Center, Ames, Iowa 50010.
Abstract:
Pasteurella multocida harvested from the blood of turkeys dying of experimental fowl cholera were purified by centrifugation and lysed. The soluble and membrane-associated components of the bacteria were separated by centrifugation. Nonionic (octylglucoside) and zwitterionic (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate; CHAPS) detergents were tested for their abilities to solubilize the cross-protection factor(s) (CPF) from the membrane-associated component. Protection studies in turkeys showed that optimum solubilization was by 1.0% octylglucoside and 0.5% CHAPS. Antibodies from turkeys made against solubilized membrane-associated CPF passively cross-protected poults against challenge. Ion exchange chromatography of detergent-solubilized CPF resulted in elution of two protein-containing peaks, each of which conferred active immune protection.
Insights
Researchers identified key components of Pasteurella multocida responsible for fowl cholera cross-protection. These components, when solubilized with specific detergents, induced protective immunity in turkeys.
Area of Science:
- Bacteriology
- Immunology
- Veterinary Medicine
Background:
- Fowl cholera, caused by Pasteurella multocida, is a significant disease in turkeys.
- Understanding the immunogenic components of P. multocida is crucial for developing effective vaccines.
- Previous studies suggested the presence of cross-protection factors (CPF) within the bacteria.
Purpose of the Study:
- To identify and characterize the components of Pasteurella multocida responsible for cross-protection against fowl cholera.
- To optimize the solubilization of these cross-protection factors (CPF) from bacterial membranes.
- To evaluate the immunoprotective potential of solubilized CPF and antibodies against them.
Main Methods:
- Purification and lysis of Pasteurella multocida from infected turkey blood.
- Separation of bacterial components into soluble and membrane-associated fractions.
- Solubilization of membrane-associated components using nonionic (octylglucoside) and zwitterionic (CHAPS) detergents.
- Protection studies in turkeys and poults to assess immune response.
- Ion exchange chromatography to isolate protein-containing fractions of CPF.
Main Results:
- Optimal solubilization of CPF was achieved using 1.0% octylglucoside and 0.5% CHAPS.
- Antibodies generated against solubilized membrane-associated CPF conferred passive cross-protection in poults.
- Ion exchange chromatography yielded two distinct protein peaks from detergent-solubilized CPF.
- Both isolated protein peaks demonstrated active immune protection in challenge studies.
Conclusions:
- The cross-protection factors (CPF) of Pasteurella multocida are associated with the bacterial membrane.
- Specific detergents effectively solubilize CPF, enabling further characterization and antibody production.
- Solubilized CPF contains at least two immunoprotective protein components that can elicit protective immunity against fowl cholera.