Department of Ophthalmology and Visual Science, Kochi Medical School, Japan. k.fukuda@kochi−u.ac.jp
Allergol Int. 2011 Sep;60(3):355-63. doi: 10.2332/allergolint.10-OA-0247. Epub 2011 Apr 25.
Common features of allergic or atopic ocular and skin diseases are the participation of Th2 lymphocytes and eosinophils and colonization by Staphylococcus aureus. To examine the role of interaction between Th2 cells and bacterial infection in tissue eosinophilia, we determined the effects of Th2 cytokines and peptidoglycan derived from the cell wall of S. aureus on corneal fibroblasts.
Chemokine concentrations and the cell surface expression of adhesion molecules were determined by ELISAs, and chemokine and adhesion molecule mRNAs were quantitated by real-time PCR analysis. Signaling by the transcription factor NF-κB was evaluated by immunoblot and immunofluorescence analyses as well as by assay of DNA binding activity.
Among Th2 cytokines tested, only interleukin (IL)-4 and IL-13 induced a low level of eotaxin release by corneal fibroblasts, as did peptidoglycan. However, the combination of peptidoglycan and either IL-4 or IL-13 induced a marked synergistic increase both in eotaxin release (without affecting that of IL-8) and in the abundance of eotaxin mRNA. The combination of peptidoglycan and IL-4 or IL-13 also synergistically increased the surface expression of VCAM-1, but not that of ICAM-1. Peptidoglycan activated NF-κB in corneal fibroblasts, and inhibitors of NF-κB attenuated eotaxin release induced by peptidoglycan alone or in combination with IL-4 or IL-13.
Interaction of innate and adaptive immunity, as manifested by synergistic stimulation of eotaxin and VCAM-1 expression in corneal fibroblasts by peptidoglycan and Th2 cytokines, may play an important role in tissue eosinophilia associated with ocular allergy.
过敏性或特应性眼部和皮肤疾病的共同特征是 Th2 淋巴细胞和嗜酸性粒细胞的参与以及金黄色葡萄球菌的定植。为了研究 Th2 细胞与细菌感染在组织嗜酸性粒细胞中的相互作用的作用，我们测定了 Th2 细胞因子和来源于金黄色葡萄球菌细胞壁的肽聚糖对角膜成纤维细胞的影响。
通过 ELISA 测定趋化因子浓度和细胞表面黏附分子的表达，通过实时 PCR 分析定量趋化因子和黏附分子的 mRNA。通过免疫印迹和免疫荧光分析以及 DNA 结合活性测定评估转录因子 NF-κB 的信号转导。
在所测试的 Th2 细胞因子中，只有白细胞介素（IL）-4 和 IL-13 诱导角膜成纤维细胞释放低水平的嗜酸性粒细胞趋化因子，肽聚糖也是如此。然而，肽聚糖与 IL-4 或 IL-13 的组合诱导了明显的协同增加，无论是嗜酸性粒细胞趋化因子的释放（不影响 IL-8 的释放）还是嗜酸性粒细胞趋化因子 mRNA 的丰度。肽聚糖与 IL-4 或 IL-13 的组合也协同增加了 VCAM-1 的表面表达，但不增加 ICAM-1 的表面表达。肽聚糖激活了角膜成纤维细胞中的 NF-κB，NF-κB 的抑制剂减弱了肽聚糖单独或与 IL-4 或 IL-13 联合诱导的嗜酸性粒细胞趋化因子的释放。
固有免疫和适应性免疫的相互作用，表现为肽聚糖和 Th2 细胞因子协同刺激角膜成纤维细胞中嗜酸性粒细胞趋化因子和 VCAM-1 的表达，可能在与眼部过敏相关的组织嗜酸性粒细胞增多中发挥重要作用。
