Related Experiment Videos
Distribution of a monoclonal antibody-recognized protective protein immunogen on the outer membranes of Pasteurella
Y S Lu1, L W Gerrity, S J Afendis
1Department of Pathology, University of Texas Southwestern Medical Center, Dallas 75235.
Abstract:
The distribution of a monoclonal antibody (MAb)-recognized protective protein immunogen on the outer membrane of 153 Pasteurella multocida rabbit isolates was determined by dot blot (DB) analysis. MAb 1608 reacted with 36 (24%) of the 153 clinical isolates. The DB-positive clinical isolates expressed capsular antigens A, D, and nontypable and somatic antigens 2, 3, 10, 12, 15, and nontypable. Western blot (immunoblot) analysis with adsorbed and eluted MAb 1608 confirmed that the antigenic determinant identified was located on the cell surface. With MAb 1608 as a probe for antibody-accessible radioimmunoassay, 31 of 36 DB-positive P. multocida rabbit isolates were shown to have surface-exposed and antibody-accessible antigenic determinants, while 44 of 44 DB-negative isolates were negative by antibody-accessible radioimmunoassay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed DB-negative P. multocida isolates both with (6 of 13, 46%) and without (7 of 13, 54%) the 37.5-kilodalton protein. This study establishes that the protective antigenic determinant of the 37.5-kilodalton outer membrane protein is present in 24% of rabbit clinical isolates tested and is detectable in P. multocida strains distributed among the major somatic types (3, 10, 12, and 15) and the capsular types (A and D) commonly isolated from rabbits in North America.
Insights
A protective protein immunogen on Pasteurella multocida outer membranes was detected in 24% of rabbit isolates. This key antigen is present in common Pasteurella multocida types affecting rabbits.
Area of Science:
- Veterinary Microbiology
- Immunology
- Bacteriology
Background:
- Pasteurella multocida is a significant pathogen in rabbits.
- Outer membrane proteins are crucial for bacterial virulence and host immune response.
- Identifying protective antigens is vital for vaccine development.
Purpose of the Study:
- To determine the distribution of a specific protective protein immunogen recognized by a monoclonal antibody (MAb 1608) on Pasteurella multocida outer membranes.
- To assess the presence of this antigen in clinical isolates from rabbits.
Main Methods:
- Dot blot (DB) analysis using MAb 1608 to screen 153 Pasteurella multocida rabbit isolates.
- Western blot (immunoblot) to confirm the location of the antigenic determinant.
- Antibody-accessible radioimmunoassay to verify surface exposure and accessibility of the antigen.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to analyze protein profiles.
Main Results:
- MAb 1608 reacted with 36 (24%) of the 153 clinical isolates.
- The antigenic determinant was confirmed to be on the cell surface via Western blot.
- Antibody-accessible radioimmunoassay showed the antigen was surface-exposed in 31 of 36 DB-positive isolates.
- The 37.5-kilodalton protein was present in both DB-positive and DB-negative isolates, indicating varied expression or accessibility.
Conclusions:
- The protective antigenic determinant of the 37.5-kilodalton outer membrane protein is present in 24% of tested rabbit clinical isolates.
- This antigen is detectable in Pasteurella multocida strains representing common capsular (A, D) and somatic types isolated from rabbits.
- The findings are significant for understanding P. multocida pathogenesis and developing targeted immunotherapies or vaccines.