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A Method For Production of Recombinant mCD1d Protein in Insect Cells.
Published on: December 11, 2007
A feline homologue of CD1 is defined using a feline-specific monoclonal antibody
1Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, USA.
Insights
Researchers identified a feline CD1 homologue, a protein similar to human CD1, primarily in the thymus and antigen-presenting cells. Its expression decreases with age, suggesting a role in immune function and thymic involution.
Area of Science:
- Immunology
- Cell Biology
- Veterinary Science
Background:
- CD1 molecules are crucial for antigen presentation in the immune system.
- Characterizing CD1 homologues in different species aids in understanding immune system evolution and function.
Purpose of the Study:
- To describe the characteristics of a feline CD1 homologue.
- To determine its tissue distribution and biochemical properties.
- To compare it with known human CD1 molecules.
Main Methods:
- Utilized a murine monoclonal antibody (Fe1.5F4) to identify and characterize the feline CD1 homologue.
- Employed immunoprecipitation and SDS-PAGE to analyze protein characteristics.
- Used flow cytometry to assess tissue distribution and expression levels on feline thymocytes.
Main Results:
- A feline CD1 homologue was identified, precipitating a 49 kDa protein and beta-2 microglobulin.
- The antigen was predominantly found in thymocytes and antigen-presenting cells (Langerhans cells, dendritic cells).
- Expression decreased with age and was highest on CD4+CD8+ thymocytes, distinct from CD1c.
Conclusions:
- The feline CD1 homologue shares similarities with the classic thymic CD1 family.
- Its biochemical and tissue expression patterns resemble human CD1a.
- This finding contributes to the comparative immunology of CD1 molecules across species.
Abstract:
The characteristics of a feline homologue of CD1, defined by a murine monoclonal antibody, Fe1.5F4 (IgG1), are described. This antibody precipitated a 49 kDa protein from biotinylated feline thymocyte extract in conjunction with a 14 kDa protein, consistent with beta 2 microglobulin subunit. The tissue distribution of this antigen was restricted to cortical thymocytes and antigen presenting cells of the thymic medulla, epidermis (Langerhans cells), dermis and occasional dendritic cells in the mantle and periarteriolar lymphoid areas of the spleen. Although flow cytometry demonstrated a continuous distribution of antigen expression on thymocytes, antigen density was found to decrease with age, consistent with physiological thymic involution. Thymocytes with high density expression of this antigen were predominantly restricted to cells with dual expression of CD4 and CD8 as defined by feline specific murine monoclonal antibodies Fe1.7B12 (IgG1) and Fe1.10E9 (IgG1) respectively. The tissue distribution of this CD1 homologue indicates that it is a member of the classic thymic CD1 family. This feline homologue of CD1 was distinct from CD1c by virtue of its lack of expression in peripheral blood and splenic mantle zone B cells. An unequivocal distinction could not be made between CD1a and CD1b based on tissue distribution due to species variation in expression of these CD1 molecules. Although the biochemical characteristics of this feline CD1 homologue more closely match with CD1a. The pattern of tissue expression and biochemical characteristics of the feline CD1 antigen appear largely similar to those described for human and other species.
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