PURPOSE: To examine the normal murine corneal stroma for the presence of bone marrow-derived leukocytes. METHODS: Wholemounts of paraformaldehyde-fixed corneal stroma from normal mice at 5 to 16 weeks of age were examined in single- and double-color immunomorphologic studies performed with confocal microscopy. The phenotype, morphology, distribution, and density of immunopositive cells were determined. RESULTS: Numerous CD45(+) cells with pleomorphic and dendriform morphology were found within the pericentral and central region of the corneal stroma (200-300 cells/mm(2)). Dual-color immunostaining demonstrated that 100% of the CD45(+) cells coexpressed CD11b and 50% coexpressed F4/80. Approximately 30% of the total cells and 50% of the F4/80(+) cells coexpressed major histocompatibility complex (MHC) class II antigens. Very small to negligible numbers of cells expressed markers of dendritic cells (CD11c) or granulocytes (Ly6G). Markers for T-cells and NK cells were absent from the corneal stroma, indicating that all the cells identified in the stroma were of the myeloid lineage. CONCLUSIONS: The normal murine corneal stroma contains a significant number of CD45(+) leukocytes. Most these cells express the CD11b marker, but not other dendrite, granulocyte, T-cell, or NK markers, placing them in the monocyte/macrophage lineage.
PURPOSE: To examine the normal murinecorneal stroma for the presence of bone marrow-derived leukocytes. METHODS: Wholemounts of paraformaldehyde-fixed corneal stroma from normal mice at 5 to 16 weeks of age were examined in single- and double-color immunomorphologic studies performed with confocal microscopy. The phenotype, morphology, distribution, and density of immunopositive cells were determined. RESULTS: Numerous CD45(+) cells with pleomorphic and dendriform morphology were found within the pericentral and central region of the corneal stroma (200-300 cells/mm(2)). Dual-color immunostaining demonstrated that 100% of the CD45(+) cells coexpressed CD11b and 50% coexpressed F4/80. Approximately 30% of the total cells and 50% of the F4/80(+) cells coexpressed major histocompatibility complex (MHC) class II antigens. Very small to negligible numbers of cells expressed markers of dendritic cells (CD11c) or granulocytes (Ly6G). Markers for T-cells and NK cells were absent from the corneal stroma, indicating that all the cells identified in the stroma were of the myeloid lineage. CONCLUSIONS: The normal murinecorneal stroma contains a significant number of CD45(+) leukocytes. Most these cells express the CD11b marker, but not other dendrite, granulocyte, T-cell, or NK markers, placing them in the monocyte/macrophage lineage.
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