Yamagami Satoru, Yokoo Seiichi, Amano Shiro, Ebihara Nobuyuki
Department of Corneal Tissue Regeneration, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
Invest Ophthalmol Vis Sci. 2007 Oct;48(10):4476-81. doi: 10.1167/iovs.06-1543.
To characterize the main population of bone marrow-derived cells (BMCs) in human normal subconjunctiva and make a comparison with BMCs in the corneal stroma and epithelium.
Normal human donor corneas with attached conjunctiva were examined by fluorescence microscopy after single and double staining for multiple markers. CD68(+) cells were separated from the conjunctival tissues by using magnetic beads, and the expression of toll-like receptor (TLR) 2 and TLR4 was examined. Surface markers of CD68(+) cells were compared with those of BMCs from the corneal stroma and epithelium.
CD45(+) cells were detected in the substantia propria of the conjunctiva, and approximately 60% of these cells were CD68(+). All the CD68(+) cells expressed HLA-DR and CD14. CD68(+) cells isolated from conjunctival tissues expressed TLR2 and TLR4 on flow cytometry. BMCs in both the corneal stroma and the subconjunctiva expressed scavenger receptor CD163. Macrophage mannose receptor CD206 was expressed by BMCs in the substantia propria of the conjunctiva, but not by BMCs in the corneal stroma or epithelium.
These findings demonstrated that the main population of BMCs in the substantia propria of normal human conjunctiva is CD68(+)CD14(+)HLA-DR(+) cells. These BMCs express scavenger receptor, macrophage mannose receptor, TLR2, and TLR4 and may play a role in adaptive and innate immune responses in the human ocular surface. These cells are phenotypically different from the CD68(-)CD206(-) monocyte- lineage cells found in the corneal stroma and the CD11c(+)CD68(-)CD163(-)CD206(-) dendritic cells residing in the corneal epithelium.
鉴定人正常结膜中骨髓来源细胞(BMCs)的主要群体,并与角膜基质和上皮中的BMCs进行比较。
对带有结膜的正常人供体角膜进行多种标志物的单染和双染后,用荧光显微镜检查。使用磁珠从结膜组织中分离出CD68(+)细胞,并检测Toll样受体(TLR)2和TLR4的表达。将CD68(+)细胞的表面标志物与角膜基质和上皮中的BMCs的表面标志物进行比较。
在结膜固有层中检测到CD45(+)细胞,其中约60%的细胞为CD68(+)。所有CD68(+)细胞均表达HLA-DR和CD14。从结膜组织中分离出的CD68(+)细胞在流式细胞仪上表达TLR2和TLR4。角膜基质和结膜下的BMCs均表达清道夫受体CD163。巨噬细胞甘露糖受体CD206在结膜固有层的BMCs中表达,但在角膜基质或上皮的BMCs中不表达。
这些发现表明正常人结膜固有层中BMCs的主要群体是CD