Cole N, Bao S, Willcox M, Husband A J
Cooperative Research Centre for Eye Research and Technology, School of Optometry, University of New South Wales, Sydney, New South Wales 2052, Australia.
Infect Immun. 1999 May;67(5):2497-502. doi: 10.1128/IAI.67.5.2497-2502.1999.
Strains of Pseudomonas aeruginosa causing keratitis can be either cytotoxic (6206) or invasive (6294), while a strain (Paer1) causing contact lens-induced acute red eye has been shown to be neither. In situ hybridization was used to examine the location and identity of cells expressing interleukin-6 (IL-6) mRNA in the murine cornea and changes in expression in response to infection with different strains of P. aeruginosa. The number of IL-6-positive cells was determined by image analysis. IL-6 protein levels were measured by an enzyme-linked immunosorbent assay. BALB/c mice were challenged by use of the wounded-cornea model with P. aeruginosa 6294, 6206, or Paer1 (2 x 10(6) CFU). At time intervals up to 24 h, postchallenge corneal tissue was probed for IL-6 mRNA. IL-6 mRNA expression was rapidly elevated in the epithelium in response to strains 6294 and 6206. At the conclusion of the experiments, infiltrating inflammatory cells also stained positively for IL-6 mRNA. In contrast, corneas challenged with strain Paer1 showed significant upregulation of IL-6 mRNA only at 4 h postchallenge. Three distinct patterns of IL-6 mRNA expression in the mouse cornea occur in response to these three ocular isolates of P. aeruginosa. The data obtained for mRNA expression in the cornea for all three strains of P. aeruginosa correlated well with IL-6 protein analysis of whole-eye homogenates. Differences in the cytokine responses to these strains correlate with differences in the pathology associated with each strain and may offer an opportunity to develop strategies for the improved management of ocular inflammation.
引起角膜炎的铜绿假单胞菌菌株可以是细胞毒性的(6206)或侵袭性的(6294),而引起隐形眼镜诱导的急性红眼的菌株(Paer1)已被证明两者都不是。原位杂交用于检查在小鼠角膜中表达白细胞介素-6(IL-6)mRNA的细胞的位置和特性,以及对不同铜绿假单胞菌菌株感染的表达变化。通过图像分析确定IL-6阳性细胞的数量。通过酶联免疫吸附测定法测量IL-6蛋白水平。使用角膜损伤模型用铜绿假单胞菌6294、6206或Paer1(2×10⁶CFU)攻击BALB/c小鼠。在长达24小时的时间间隔内,对攻击后角膜组织检测IL-6 mRNA。响应于菌株6294和6206,上皮中IL-6 mRNA表达迅速升高。在实验结束时,浸润的炎性细胞对IL-6 mRNA也呈阳性染色。相比之下,用菌株Paer1攻击的角膜仅在攻击后4小时显示IL-6 mRNA的显著上调。响应于这三种铜绿假单胞菌眼部分离株,小鼠角膜中出现三种不同的IL-6 mRNA表达模式。所有三种铜绿假单胞菌菌株在角膜中获得的mRNA表达数据与全眼球匀浆的IL-6蛋白分析良好相关。对这些菌株细胞因子反应的差异与每种菌株相关的病理学差异相关,并且可能提供开发改善眼部炎症管理策略的机会。