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Mitsuhiro Iwata, Koichi Soya, Mitsuru Sawa, Takashi Sakimoto, David G. Hwang; CD40 Expression in Normal Human Cornea and Regulation of CD40 in Cultured Human Corneal Epithelial and Stromal Cells. Invest. Ophthalmol. Vis. Sci. 2002;43(2):348-357.
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purpose. To determine whether CD40–CD40 ligand (CD40L) interaction
plays a role in corneal inflammatory responses, the expression of CD40
and CD40L on normal human cornea was investigated. In addition, using
cultured human corneal epithelial (HCE) and human corneal stromal (HCS)
cells, the regulation of CD40 expression in human corneal cells
investigated, including that induced by proinflammatory cytokines such
as interferon (IFN)-γ and tumor necrosis factor (TNF)-α.
methods. Frozen optimal cutting temperature (OCT) compound–embedded sections of
corneal tissues obtained from 18 normal human corneas were examined by
an immunoperoxidase staining technique with anti-CD40 and anti-CD40L
monoclonal antibodies (mAbs). Also, cultured HCE and HCS cells, with
IFN-γ (250–1000 U/mL) or TNF-α (500–4000 U/mL) treatment for 1 to
4 days and with no treatment, were stained by the immunofluorescence
technique with mAbs and analyzed by flow cytometry.
results. The area of positive staining for CD40 showed a topographical
difference. The limbal epithelial cells were predominantly positive for
CD40. Positive staining was also found to a lesser extent on the cells
in the basal layer of peripheral corneal epithelium. Epithelial cells
of the central cornea showed no immunoreactivity for CD40. Corneal
stromal cells were negative for CD40 in most of the donor tissues
(positive: 5 of the 18 corneas). Endothelial cells were distinctly
negative for CD40. Cultured HCE cells were also positive but decreased
in positive cell number with lengthening culture period. None or less
than 5% of the cultured HCS cells were CD40 positive. IFN-γ enhanced
CD40 expression on both cell types. In contrast, TNF-α enhanced CD40
on HCE but not on HCS cells. No component cells of normal human cornea
or cultured HCE and HCS cells showed immunoreactivity for CD40L.
conclusions. In the human cornea, CD40 is expressed predominantly on limbal
epithelial cells and also on cultured HCE cells with high proliferative
potential. In addition, the expression of CD40 is induced on cultured
HCE and HCS cells differentially by proinflammatory cytokines, such as
IFN-γ and TNF-α.
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