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Monoclonal antibodies to CD44 epitopes on mouse endothelium
S M Davern1, P K Lankford, L J Foote
1Life Sciences Division, P O Box 2008, MS-6101, Oak Ridge, TN 37831-6101, USA.
Insights
Researchers developed new monoclonal antibodies (MAbs) targeting CD44, a protein crucial in cell adhesion and various biological processes. These novel MAbs offer potential for studying CD44
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- CD44 is a widely expressed plasma membrane glycoprotein involved in critical biological processes like inflammation and cancer metastasis.
- Alternative splicing and glycosylation of CD44 result in diverse molecular forms with varied functions.
- CD44's role as an adhesion molecule links it to angiogenesis, lymphogenesis, wound healing, and cancer metastasis.
Purpose of the Study:
- To develop and characterize a novel panel of rat monoclonal antibodies (MAbs) specifically targeting murine CD44.
- To investigate the distinct epitopes recognized by these new anti-CD44 MAbs.
- To assess the utility of these antibodies in studying CD44's role, particularly in endothelial cells, within disease models.
Main Methods:
- Immunization of rats with mouse lung endothelial cells (LEII cells) to generate MAbs.
- Characterization of MAbs using immunoprecipitation and mass spectrometry.
- Assessment of antibody binding specificity through competition assays and cross Western blotting.
Main Results:
- A new panel of rat MAbs against murine CD44 was successfully developed.
- The MAbs recognize three distinct epitope groups on CD44.
- Specific MAbs were identified that bind to CD44 forms expressed in vivo and those unique to endothelial cells.
Conclusions:
- The novel anti-CD44 MAbs provide valuable tools for immunological research.
- These antibodies can be used to investigate the role of CD44 in various biological processes and disease pathogenesis.
- The panel shows particular promise for studying the involvement of endothelial CD44 in disease models.
Abstract:
CD44 is a widely expressed, plasma membrane protein. Many molecular forms of CD44 are possible as it is encoded by a gene with multiple exons that can be alternatively spliced and its deduced protein sequence contains numerous glycosylation sites. Through its role as an adhesion molecule, CD44 is involved in many and diverse biological processes, including angiogenesis, lymphogenesis, wound healing, inflammation, and cancer metastasis. We have developed a new panel of rat monoclonal antibodies (MAbs) to murine CD44 by immunization with mouse lung endothelial cells (LEII cells). The antibodies were characterized using immunoprecipitation, mass spectrometry, competition binding, and cross Western blot experiments with MAb 133-13A, which recognizes CD44 expressed on tumor cells. The new MAbs recognize three distinct epitope groups. MAbs 531-2A and 531-32A compete for binding with the MAb 133-13A that was described previously. MAb 531-30A identifies a CD44 epitope found on low molecular weight forms expressed in vivo, while MAb 531-22A appears to recognize an epitope specific for endothelial cells. This novel panel of anti-CD44 antibodies has potential for investigating the role of CD44 in disease pathogenesis models in the mouse. They may be particularly useful for examining the role of endothelial cells in these models.
