目的研究B、T淋巴细胞衰减因子(B and T lymphocyte attenuator,BTLA)及其配体疱疹病毒侵入介体(herpes virus entry mediator,HVEM)在单纯疱疹性角膜基质炎(herpetic stromal keratitis,HSK)小鼠角膜组织中及外周血CD4+T细胞上的表达水...目的研究B、T淋巴细胞衰减因子(B and T lymphocyte attenuator,BTLA)及其配体疱疹病毒侵入介体(herpes virus entry mediator,HVEM)在单纯疱疹性角膜基质炎(herpetic stromal keratitis,HSK)小鼠角膜组织中及外周血CD4+T细胞上的表达水平,探讨共抑制信号BTLA-HVEM是否参与了CD4+T细胞介导的HSK免疫病理反应。方法将106PFU的单纯疱疹病毒I型(herpes simplex virus type 1,HSV-1)KOS毒株接种于BALB/c鼠的角膜上建立HSK动物模型,分别于角膜接种病毒前(0d),接种病毒后的第3、7、10、14、21天,用毛细管取小鼠的左眼眼眶静脉窦血1mL,分离淋巴细胞,行荧光抗体染色,用流式细胞仪检测CD3+CD4+BTLA+T细胞和CD3+CD4+HVEM+T细胞阳性率;在裂隙灯显微镜下观察小鼠角膜变化;免疫组织化学方法检测BTLA蛋白及HVEM蛋白在角膜组织中的表达。结果 BALB/c鼠的角膜接种HSV-1后的1～5d,角膜擦拭液中均检测出HSV-1复制,表明小鼠感染了单纯疱疹病毒。裂隙灯显微镜观察显示:角膜接种HSV-1后第3天,所有小鼠均患了急性上皮性角膜炎,并于感染后1周内痊愈,自病毒接种后第8天起,小鼠出现角膜基质炎的改变,表现为角膜基质呈灰白色混浊,角膜基质混浊于病毒接种后的第10天达到高峰,持续至第14天后逐渐减轻。流式细胞仪检测显示,小鼠外周血淋巴细胞中CD3+CD4+BTLA+T细胞和CD3+CD4+HVEM+T细胞的阳性率,在角膜接种病毒前(0d)分别为(3.15±0.60)%和(9.84±1.06)%,在角膜接种病毒后第10天(HSK疾病程度最严重时)分别增加到(20.47±3.15)%和(45.18±3.90)%(与0d相比差异均有显著统计学意义,均为P<0.01)。免疫组织化学方法检查HSK小鼠角膜组织中BTLA和HVEM的蛋白表达结果一致:HSK临床表现最严重时,即病毒接种后第10天时,BTLA蛋白和HVEM蛋白在角膜组织中表达最强,主要表达于角膜基质层内浸润的炎性细胞上,角膜上皮层和内皮层也有表达。结论在HSK小鼠模型中,BTLA及其配�展开更多
Purpose:To investigate the effect of NF-κB inhibitor PDTC on inflammation response in mice with herpetic stromal keratitis (HSK). Methods:A total of 120 female BALB/c mice,aged 4 -6 weeks, were treated by scuffing th...Purpose:To investigate the effect of NF-κB inhibitor PDTC on inflammation response in mice with herpetic stromal keratitis (HSK). Methods:A total of 120 female BALB/c mice,aged 4 -6 weeks, were treated by scuffing the epithelium of the right cornea with the tip of a 30G syringe needle, and approximately 1×106 PFU of HSV-1virus was seeded onto the corneal surface. The eye was then exposed to 170 mJ/cm2 of UV light 7 weeks later, in order to induce the relapse of HSK. PDTC eye drops were adjusted to various concentrations (0.1,1.0, and 10 mg/ml) with PBS; PBS only was used on control animals.All animals received PDTC or PBS eye drops 6 times/day,1 drop/time, for 7 consecutive days. Mice were sacrificed at 8h, and 1, 3, and 7d after administration. Corneal tissues were prepared for the detection of IL-1β, IL-4, IL-6, and IL-12 by ELISA and histological study. Results: Neutrophilic leukocytes and lymphocytes were found in the corneas of HSK mice. ELISA results showed fluctuating expressions of IL-1β, IL-4, IL-6, and IL-12. The IL-1β and IL-4 expressions were significantly lower in the high dose group than in the control group (P<0.05), while IL-6 and IL-12 expressions were significantly higher in the high dose group than in the control group (P<0.05). Conclusion:A high concentration of PDTC can suppress the expression of certain inflammatory factors in HSK mice while improving the expression of others.展开更多
文摘目的研究B、T淋巴细胞衰减因子(B and T lymphocyte attenuator,BTLA)及其配体疱疹病毒侵入介体(herpes virus entry mediator,HVEM)在单纯疱疹性角膜基质炎(herpetic stromal keratitis,HSK)小鼠角膜组织中及外周血CD4+T细胞上的表达水平,探讨共抑制信号BTLA-HVEM是否参与了CD4+T细胞介导的HSK免疫病理反应。方法将106PFU的单纯疱疹病毒I型(herpes simplex virus type 1,HSV-1)KOS毒株接种于BALB/c鼠的角膜上建立HSK动物模型,分别于角膜接种病毒前(0d),接种病毒后的第3、7、10、14、21天,用毛细管取小鼠的左眼眼眶静脉窦血1mL,分离淋巴细胞,行荧光抗体染色,用流式细胞仪检测CD3+CD4+BTLA+T细胞和CD3+CD4+HVEM+T细胞阳性率;在裂隙灯显微镜下观察小鼠角膜变化;免疫组织化学方法检测BTLA蛋白及HVEM蛋白在角膜组织中的表达。结果 BALB/c鼠的角膜接种HSV-1后的1～5d,角膜擦拭液中均检测出HSV-1复制,表明小鼠感染了单纯疱疹病毒。裂隙灯显微镜观察显示:角膜接种HSV-1后第3天,所有小鼠均患了急性上皮性角膜炎,并于感染后1周内痊愈,自病毒接种后第8天起,小鼠出现角膜基质炎的改变,表现为角膜基质呈灰白色混浊,角膜基质混浊于病毒接种后的第10天达到高峰,持续至第14天后逐渐减轻。流式细胞仪检测显示,小鼠外周血淋巴细胞中CD3+CD4+BTLA+T细胞和CD3+CD4+HVEM+T细胞的阳性率,在角膜接种病毒前(0d)分别为(3.15±0.60)%和(9.84±1.06)%,在角膜接种病毒后第10天(HSK疾病程度最严重时)分别增加到(20.47±3.15)%和(45.18±3.90)%(与0d相比差异均有显著统计学意义,均为P<0.01)。免疫组织化学方法检查HSK小鼠角膜组织中BTLA和HVEM的蛋白表达结果一致:HSK临床表现最严重时,即病毒接种后第10天时,BTLA蛋白和HVEM蛋白在角膜组织中表达最强,主要表达于角膜基质层内浸润的炎性细胞上,角膜上皮层和内皮层也有表达。结论在HSK小鼠模型中,BTLA及其配�
基金supported by the Fund for Medical Sciences of Guangdong Province (A2009463)the Internal Fund of Shantou International Eye Center (10-002)
文摘Purpose:To investigate the effect of NF-κB inhibitor PDTC on inflammation response in mice with herpetic stromal keratitis (HSK). Methods:A total of 120 female BALB/c mice,aged 4 -6 weeks, were treated by scuffing the epithelium of the right cornea with the tip of a 30G syringe needle, and approximately 1×106 PFU of HSV-1virus was seeded onto the corneal surface. The eye was then exposed to 170 mJ/cm2 of UV light 7 weeks later, in order to induce the relapse of HSK. PDTC eye drops were adjusted to various concentrations (0.1,1.0, and 10 mg/ml) with PBS; PBS only was used on control animals.All animals received PDTC or PBS eye drops 6 times/day,1 drop/time, for 7 consecutive days. Mice were sacrificed at 8h, and 1, 3, and 7d after administration. Corneal tissues were prepared for the detection of IL-1β, IL-4, IL-6, and IL-12 by ELISA and histological study. Results: Neutrophilic leukocytes and lymphocytes were found in the corneas of HSK mice. ELISA results showed fluctuating expressions of IL-1β, IL-4, IL-6, and IL-12. The IL-1β and IL-4 expressions were significantly lower in the high dose group than in the control group (P<0.05), while IL-6 and IL-12 expressions were significantly higher in the high dose group than in the control group (P<0.05). Conclusion:A high concentration of PDTC can suppress the expression of certain inflammatory factors in HSK mice while improving the expression of others.
