Characterization of the Murine Salmonella Typhimurium-Specific Primary B-Cell Repertoire
Abstract
Recent studies suggest that B cells play an important role in the immune response to Salmonella typhimurium which causes a typhoid-like disease in mice. One approach to understanding the role of B cells in the immune response to S. typhimurium would be to analyze the repertoire of salmonella-specific B cells in normal and salmonella-susceptible mice. However, to date no studies of the B-cell repertoire specific for salmonella, nor any other bacterial infectious disease agent, have yet been undertaken. The splenic focus system permits the identification and characterization of individual B cells and their antibody products and an estimate of the size of the antigen-specific repertoire. Therefore, the splenic focus system has been modified to analyze the B-cell repertoire specific for S. typhimurium strain TML (TML). The studies described herein define the conditions for obtaining monoclonal responses to acetone-killed and dried TML (AKD-TML) in salmonella resistant CBA/Ca mice and salmonella-susceptible CBA/N mice. The frequency of primary TML-specific splenic B cells in CBA/Ca mice is approximately 1 per 1 x 10(expn 5) B cells. In contrast, the frequency of memory TML-specific cells is approximately 1 per 5-8 x 10 (expn 3) splenic B cells and the majority of these B cells are specific for LPS. These results suggest that the frequency of primary TML-specific B cells is extremely low and that it expands 15- to 20-fold after antigen exposure. Interestingly, less than 30% of the primary B cells are specific for the LPS molecule which is considered to be the major antigenic determinant on Salmonella organisms. Furthermore, the majority of the LPS-specific anti-TML antibody-producing clones are directed against the LPS O-antigen region. Conversely, less than half of the memory LPS-specific anti-TML B-cell clones are directed against the O-antigen. The remaining half are directed against the KDO or lipid A region of the LPS molecule.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 27, 1984
- Accession Number
- AD1010703
Entities
People
- Lise W. Duran
Organizations
- Uniformed Services University of the Health Sciences