【作者】 郭洁；

【导师】 钱江； 袁一飞； 张锐；

【作者基本信息】 复旦大学 ， 眼科学， 2009， 硕士

【摘要】 目的:观察T辅助细胞因子IL-4、IFN-γ及IL-17在特发性泪腺炎及MRL/lpr小鼠泪腺炎中的表达,对T辅助细胞在特发性泪腺炎发病中的作用进行初步探讨。方法:1.收集2005年2月-2007年3月在复旦大学附属眼耳鼻喉科医院行手术治疗的特发性泪腺炎患者组织标本19例,免疫组化法行IL-4、IFN-γ及IL-17染色,观察三种因子阳性表达细胞的形态及分布特征,计算每高倍视野阳性细胞数量,并对结果进行统计学分析。2.观察3月龄MRL/lpr小鼠泪腺炎症病变并以BALB/c小鼠为正常对照;分别对两组小鼠的泪腺及颌下淋巴结行IL-4、IL-17、IFN-γ免疫组化染色,计算阳性细胞表达量并对结果进行统计学分析;同时对MRL/lpr小鼠颌下淋巴结行IL-4、IFN-γ、IL-17与CD4荧光双标染色。用ELISA法检测两组小鼠血清及24小时脾细胞培养上清液中IL-17的含量,并对结果进行统计学分析。结果:1.特发性泪腺炎以泪腺不同程度的淋巴细胞浸润及纤维增殖为特征。IL-4阳性细胞计数为40±29/HP,多分布于淋巴滤泡中心,IFN-γ阳性细胞计数为15±12/HP,多见于淋巴滤泡间区;IL-17在部分淋巴型泪腺炎中为中性粒细胞阳性着色(66.7%),在部分纤维型泪腺炎中为淋巴浆细胞阳性着色(42.9%),平均阳性细胞计数为5±4.4/HP,IL-4、IFN-γ、IL-17三种细胞因子表达量间差异存在统计学意义(P<0.01)。2.3月龄MRL/lpr小鼠泪腺中大量淋巴细胞浸润,其中IL-4及IFN-γ阳性细胞占总单核细胞百分比分别为30±14.5%及4±2.6%,IL-17阳性细胞数占总单核细胞百分比为4.5±3.4%,三种细胞因子表达量间差异存在统计学意义(P<0.01)。MRL/lpr小鼠颌下淋巴结中每高倍视野IL-4、IL-17、IFN-γ阳性细胞数分别为54±6.5,43±3.6,20±5,均存在与CD4+细胞共表达,其中IL-4、IL-17阳性细胞数量显著高于IFN-γ(P<0.01),且三种细胞因子阳性细胞数均较BALB/c小鼠显著增高(P<0.01)。MRL/lpr小鼠血清中IL-17含量为247±85pg/ml,较BALB/c小鼠血清IL-17含量104±31pg/ml显著增高(P<0.01),但24小时脾细胞培养上清液中IL-17含量在两组小鼠差异中无统计学意义。结论:1.特发性泪腺炎患者泪腺中IL-4阳性细胞数量较IFN-γ、IL-17明显增高,表明Th2型免疫反应可能在该病的免疫中起主导作用,IL-17参与该病免疫进程,可由中性粒细胞或淋巴浆细胞产生。2.IL-4是MRL/lpr小鼠泪腺病灶及颌下淋巴结中的主要表达因子,Th2型免疫可能在MRL/lpr小鼠泪腺炎中起到主导作用。IL-17是该病的重要参与因因子,在MRL/lpr小鼠泪腺病灶、颌下淋巴结及血清中表达较BALB/c小鼠明显升高,但其随病程的变化及与Th1、Th2的具体作用尚待进一步研究。更多还原

【Abstract】 Objective:To investigate the expression of T-helper(Th) cytokines interleukin(IL)-4, interferon(IFN)-γand IL-17 in lacrimal glands of idiopathic dacryoadenitis patients.To determine the distribution of IL-4,IFN-γand IL-17 in lacrimal gland lesions,submaxillary lymph nodes of MRL/lpr mice——a model for idiopathic dacryoadenitis,and investigate the level of IL-17 in serum and splenocyte culture supernatant of MRL/1pr mice.Methods:1.Nineteen surgery specimens of idiopathic dacryoadenitis patients were collected from Eye and ENT hospital Fudan Unversity.Sections of lacrimal glands were stained with antibodies to IL-4,IFN-γand IL-17 using immunohistochemical technique,and counted the number of cells positive stained under high power field. 2.3-month-old MRL/lpr mice and control BALB/c mice were prepared and lacrimal gland inflammation were evaluated.Frozen sections of lacrimal glands and submandibular lymph nodes from both mice were stained with antibodies to the cytokines IL-17 as well as IL-4 and IFN-γ.Stains for CD4 both with IL-17,IL-4,IFN-γwere taken in submandibular lymph nodes of MRL/lpr mice.The concentrations of IL-17 in serums and 24-hour splenocyte culture supernatants were detected by enzyme-linked immunosorbent assay(ELISA).Results:1.The number of cells staining for IL-4 was 40±29/HP,and cells positive stained were found almost in the center of lymph follicle.The number of cells staining for IFN-γwas 15±12/HP,and cells positive stained were found almost between lymph follicles,the difference between IL-4 and IFN-γwas significantly.For 12 lymphocytictype idiopathic dacryoadenitis patients,the neutrophilic granulocytes positive staining for IL-17 were found in eight patients(66.7%).And for seven sclerosing dacryoadenitis patients,lymphoplasmacytes positive staining for IL-17 were found in three patients(42.9%),the average of IL-17-producing cells was 5±4.4/HP, the difference between IL-17 and IL-4,IL-17 and IFN-γwas also significantly.2.Marked lacrimal gland inflammation was observed in all 3-month-old MRL/lpr mice. The proportion of cells staining positively for IL-4 was 30±14.5%in lacrimal glands of MRL/lpr mice,for IL-17 was 4.5±3.4%and for IFN-γwas 4±2.6%.The expression of IL-4 increased significantly than IFN-γand IL-17.In submandibular lymph nodes of MRL/lpr mice,the number of positive cells for IL-17,IL-4 and IFN-γwere 43±3.6,54±6.5,20±5 per HPF respectively,significantly increased than control BALB/c mice,and the expression of IL-4 and IL-17 was higher than IFN-γin MRL/lpr mice.Each cytokine was expressed by CD4+T cells in lymph nodes.The concentration of IL-17 in serum of 3-month-old MRL/lpr mice was 247±85 pg/ml which significantly raised than BALB/c mice with 104±31 pg/ml,but there was no difference for the concentration of IL-17 in splenocyte culture supernatant of two group mice.Conclusions:1.IL-4 is the Th-cytokine highest expressed in idiopathic dacryoadenitis, this result indicates Th2 immunity play a crucial role in the occurrence of idiopathic dacryoadenitis,IL-17-producing cells also participate the pathophysiology of idiopathic dacryoadenitis.2.Th2 immunity plays a crucial role in lacrimal gland inflammation of MRL/lpr mice. IL-17 expression is elevated in 3-month-old MRL/lpr mice,the result suggests IL-17 may play an important role in the pathophysiology of autoimmunity dacryoadenitis of MRL/lpr mice.更多还原