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Published on: October 27, 2023
Expression of CD34 and L-selectin on human corneal keratocytes
Annie Joseph1, Parwez Hossain, Seema Jham
1Larry A. Donoso Laboratory for Eye Research, Division of Ophthalmology and Visual Sciences, Eye, Ear, Nose, and Throat Centre, University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom.
Insights
Normal human corneal keratocytes express CD34, a hematopoietic stem cell marker. This finding suggests CD34 may anchor keratocytes within the cornea, while L-selectin
Area of Science:
- Ophthalmology
- Cell Biology
- Immunology
Background:
- CD34 is a hematopoietic stem cell marker and adhesion molecule.
- L-selectin is a ligand for CD34.
- The role of CD34 and L-selectin in the cornea is not well understood.
Purpose of the Study:
- To investigate the expression of CD34 and its ligand L-selectin in the human cornea.
- To determine the cellular localization and molecular nature of CD34 and L-selectin expression in corneal tissue.
Main Methods:
- Immunohistochemistry was performed on 17 normal adult human corneal specimens using antibodies against CD34 and L-selectin.
- Western blotting and PCR were used to confirm protein and mRNA expression of CD34 and L-selectin in six additional corneal specimens.
Main Results:
- All three classes of CD34 were expressed by corneal keratocytes, confirmed by immunohistochemistry, Western blotting, and PCR.
- L-selectin showed positive staining around keratocytes via immunohistochemistry, but was not detected by Western blotting or PCR.
Conclusions:
- Normal human corneal keratocytes express CD34, suggesting a role in cell adhesion and anchoring within the corneal stroma.
- The discrepancy in L-selectin detection suggests potential cross-reactivity or the presence of alternative ligands in the cornea.
Purpose:
To investigate the expression of CD34, a hematopoietic stem cell marker and an adhesion molecule, and its ligand L-selectin in the human cornea.
Methods:
Seventeen normal adult human corneal specimens were studied by immunohistochemistry using a panel of monoclonal antibodies against all three classes of the hematopoietic stem cell marker CD34 and its ligand L-selectin. An additional six corneal specimens were used for protein extraction and analysis by Western blotting, using the CD34 and L-selectin antibodies. PCR was used to determine expression of mRNA for CD34 and L-selectin in the corneal specimens.
Results:
Only corneal keratocytes showed positive immunostaining for all three classes of CD34. Western blotting confirmed the expression of CD34 by these cells and mRNA expression for CD34 in the corneal stroma was demonstrated by PCR. For L-selectin, positive staining around keratocytes was noted on immunohistochemistry but L-selectin could not be detected either by Western blotting or PCR.
Conclusions:
Normal human corneal keratocytes express all three classes of CD34. The expression of this adhesion molecule on corneal keratocytes suggests that it may have a role in keeping the keratocytes anchored in their microniche, between the collagen lamellae. The positive staining for L-selectin found by immunohistochemistry but not by Western blotting or PCR would indicate the presence of either another ligand from the selectin family or a cross-reactive epitope on corneal keratocytes.
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