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Hemolytic plaque inhibition by synthetic antigenic peptides of sperm whale myoglobin
Abstract:
We have investigated the systemic antibody response to sperm whale myoglobin (SWMb) antigenic sites in three strains of inbred mice using an inhibition of plaque assay. Sperm whale myoglobin was attached to sheep red blood cells (SRBC) via rabbit anti-SRBC Fab' fragments. Inhibition of lysis was obtained with synthetic peptides representing the purported five antigenic sites but not with a peptide whose sequence was unrelated to SWMb and synthetic peptides of SWMb from outside the antigenic sites gave minimal or no inhibition. The results of our studies show that the pattern of response to the five antigenic sites differs in each strain, but that almost total inhibition is obtained in all strains with these five sites. The antigenic dominance of these sites supports the concept of discrete antigenic sites on soluble proteins. They also suggest a reason for contrary reports in the literature based on hybridoma technology.
Insights
Researchers studied immune responses to sperm whale myoglobin (SWMb) antigenic sites in mice. Different mouse strains showed varied responses to SWMb sites, confirming discrete antigenic sites on proteins.
Area of Science:
- Immunology
- Protein Chemistry
Background:
- Antigenic sites on soluble proteins are crucial for understanding immune responses.
- Previous studies on sperm whale myoglobin (SWMb) have yielded conflicting results regarding its antigenic sites.
Purpose of the Study:
- To investigate the systemic antibody response to specific antigenic sites of SWMb.
- To compare the immune response patterns across different inbred mouse strains.
Main Methods:
- Utilized an inhibition of plaque assay to measure antibody response.
- Attached SWMb to sheep red blood cells (SRBC) using rabbit anti-SRBC Fab' fragments.
- Employed synthetic peptides representing SWMb antigenic sites and control peptides.
Main Results:
- Synthetic peptides corresponding to the five SWMb antigenic sites effectively inhibited lysis.
- Peptides from outside the antigenic sites and unrelated sequences showed minimal to no inhibition.
- Each mouse strain exhibited a distinct antibody response pattern to the five antigenic sites, yet near-complete inhibition was achieved with these sites across all strains.
Conclusions:
- The findings support the concept of discrete, immunodominant antigenic sites on soluble proteins like SWMb.
- The observed strain-specific responses may explain discrepancies reported in prior hybridoma-based studies.