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Involvement of C-C Chemokine Ligand 2-CCR2 Interaction in Monocyte-Lineage Cell Recruitment of Normal Human Corneal Stroma¹

Nobuyuki Ebihara^*, Satoru Yamagami^2,, Seiichi Yokoo, Shiro Amano and Akira Murakami^*

^* Department of Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan; and Department of Corneal Tissue Regeneration, University of Tokyo Graduate School of Medicine, Tokyo, Japan

Bone marrow-derived cells (BMCs) reside in the anterior stroma of the central and paracentral cornea, as well as all stromal layers of the peripheral cornea, in normal human eyes. We investigated the factors regulating the constitutive distribution of BMCs in normal human corneal stroma. Cultured human corneal keratocytes expressed several chemokines (growth-related oncogene/CXCL1–3, IL-8/CXCL8, and MCP-1/CCL2) in the Ab array study. CCR2 and CCR7 mRNAs were detected in BMCs by multiplex RT-PCR. Keratocytes/corneal epithelial cells and BMCs selected from normal human donor corneas by using magnetic beads expressed MCP-1/CCL2 and CCR2 protein, respectively. BMCs isolated from human corneal stroma showed a chemotactic response to MCP-1/CCL2 in the Boyden chamber assay. The chemotactic effect of keratocyte supernatant was inhibited by blockade of MCP-1/CCL2. This is the first work on constitutive expression of CCR2 by BMCs from the corneal stroma and MCP-1/CCL2 by keratocytes/epithelial cells. Our findings suggest that the interaction between MCP-1/CCL2 and CCR2 determines the distribution of constitutive BMCs in normal human corneal stroma.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, and Science.

² Address correspondence and reprint requests to Dr. Satoru Yamagami, Department of Corneal Tissue Regeneration, Tokyo University Graduate School of Medicine, Hongo 7-3-1, Bunkyo-ku, Tokyo, Japan 113-8655. E-mail address: syamagami-tky{at}umin.ac.jp

³ Abbreviations used in this paper: BMC, bone marrow-derived cell; ELC, EBI 1-ligand chemokine; GRO, growth-related oncogene; SLC, secondary lymphoid-tissue chemokine; TIMP, tissue inhibitor of metalloproteinase.