Zhan Hong, Towler Hamish M A T, Calder Virginia L
Department of Clinical Ophthalmology, Institute of Ophthalmology, University College London, London, United Kingdom.
Invest Ophthalmol Vis Sci. 2003 Sep;44(9):3906-10. doi: 10.1167/iovs.02-0665.
To characterize phenotypically the primary epithelial cells obtained from normal bulbar conjunctival biopsy specimens and a human conjunctival epithelial cell line (Wong-Kilbourne derivative of Chang conjunctiva; ChWK) with regard to their immunostimulatory function.
The effects on expression of HLA-DR and costimulatory molecules (CD80, CD86, ICAM-1, and CD40) on normal conjunctival epithelial cells and ChWK treated with various cytokines (IFN-gamma, TNF-alpha, IL-4, and IL-13) were determined using flow cytometry and confocal microscopy.
Epithelial cells were successfully grown from conjunctival explants and cultures passaged three times, while retaining their cell surface markers. At least 97% of primary epithelial cells (n = 10) and more than 96% of ChWK cells (n = 10) were cytokeratin positive by flow cytometry and immunocytochemistry and demonstrated epithelial cell morphology. Both primary conjunctival epithelial cells and ChWK had a low basal expression of HLA-DR and ICAM-1, and both were upregulated by IFN-gamma. For ChWK cells, CD80 and CD86 were constitutively expressed at low levels. CD80 was significantly upregulated after IFN-gamma treatment (P = 0.043), whereas IL-4 induced a significant upregulation of CD86 (P = 0.039). Treatment with IL-13 and TNF-alpha did not induce significant effects.
The ability of conjunctival epithelial cells to express costimulatory molecules suggests a proinflammatory role for conjunctival epithelial cells.
对从正常球结膜活检标本和人结膜上皮细胞系(张氏结膜的黄 - 基尔伯恩衍生物;ChWK)获得的原代表皮细胞的免疫刺激功能进行表型特征分析。
使用流式细胞术和共聚焦显微镜测定各种细胞因子(γ干扰素、肿瘤坏死因子 -α、白细胞介素 -4和白细胞介素 -13)处理后对正常结膜上皮细胞和ChWK上HLA - DR和共刺激分子(CD80、CD86、细胞间黏附分子 -1和CD40)表达的影响。
结膜外植体成功培养出上皮细胞,传代三次,同时保留其细胞表面标志物。通过流式细胞术和免疫细胞化学检测,至少97%的原代表皮细胞(n = 10)和超过96%的ChWK细胞(n = 10)细胞角蛋白呈阳性,并表现出上皮细胞形态。原代结膜上皮细胞和ChWK细胞的HLA - DR和细胞间黏附分子 -1基础表达水平均较低,并均被γ干扰素上调。对于ChWK细胞,CD80和CD86以低水平组成性表达。γ干扰素处理后CD80显著上调(P = 0.043),而白细胞介素 -4诱导CD86显著上调(P = 0.039)。白细胞介素 -13和肿瘤坏死因子 -α处理未诱导显著效应。
结膜上皮细胞表达共刺激分子表明结膜上皮细胞具有促炎作用。