原发性干燥综合征外周血PBMC及唇腺组织EB病毒表达的研究
发布时间：2018-07-20 14:49
【摘要】：目的:原发性干燥综合征(Primary Sj?gren’s Syndrome,p SS)是以侵犯唾液腺、泪腺等外分泌腺体,以大量淋巴细胞浸润为特征的自身免疫性疾病。多项研究发现,p SS的发病与EB病毒(Epstein-Barr virus,EBV)的感染相关。EB病毒核抗原-1(Epstein-Barr Nuclear Antigen-1,EBNA-1)和EB病毒核抗原-2(Epstein-Barr Nuclear Antigen-2,EBNA-2)是EBV潜伏期表达的核心抗原。通过检测p SS患者外周血单个核细胞(Peripheral blood mononuclear cell,PBMC)EBV DNA拷贝数,以及测定p SS患者唇腺组织中EBNA-1和EBNA-2的表达情况,探讨EBV与p SS发病的关系。方法:应用Real-time荧光定量PCR定量检测32例p SS患者与28例正常对照PBMC中EBV DNA拷贝数。通过免疫组化的方法测定32例p SS患者及6例正常唇腺组织中EBNA-1和EBNA-2的表达情况(以细胞胞浆、胞核中出现棕黄色颗粒状物质为阳性)。结果:(1)p SS患者EB病毒DNA检测阳性率100%(32/32),对照组EB病毒DNA检测阳性率89.2%(25/28),经检验,差异无统计学意义(P0.05)。(2)32例p SS患者EB病毒DNA拷贝数均值为26.2±10.85 copies/μg,28例对照组EB病毒DNA拷贝数均值为8.6±5.21 copies/μg,经检验,患者组EB病毒DNA拷贝数均值明显高于对照组,差异具有统计学意义(P0.05)。(3)32例p SS患者临床表现与EB病毒DNA拷贝数比较显示,腮腺肿大组较非腮腺肿大组EB病毒DNA拷贝数差异有统计学意义(P0.05)。32例p SS患者实验室检查结果与EBV-DNA之间的相关性分析显示,Ig G与病毒DNA拷贝数呈正相关(p0.05)。(4)EBNA-1和EBNA-2在部分p SS患者唇腺组织浸润的淋巴细胞,唇腺导管上皮细胞中可见阳性表达,而对照组唇腺组织未见阳性表达。p SS唇腺组织中EBNA-1的阳性率为68.8%(22/32),EBNA-2的阳性表达率62.5%(20/32),较正常对照组比较均有显著统计学意义(P0.05)。(5)EBNA-1在p SS唇腺组织中阳性和阴性的患者在临床表现和实验室检查方面无明显差异性(P0.05)。EBNA-2在p SS唇腺组织中阳性和阴性的患者在临床表现和实验室检查方面无明显差异性(P0.05)。结论:(1)EBV复制可能参与p SS的发病。(2)p SS腮腺肿大的患者提示可能EBV复制活跃,加重发病。(3)高Ig G提示与EBV复制活跃相关。(4)EBNA-1、EBNA-2可能通过参与EBV感染而与p SS的发病相关。
[Abstract]:Objective: primary Sjgren's Syndromeg syndrome (SS) is an autoimmune disease characterized by invasion of exocrine glands such as salivary glands and lacrimal glands, and massive lymphocytic infiltration. Many studies have found that Epstein-Barr virus (EBV) infection and Epstein-Barr Nuclear Antigen-1EBNA-1 (EBNA-1) and Epstein-Barr Nuclear Antigen-2( EBNA-2) are the core antigens of EBV expression. The expression of EBNA-1 and EBNA-2 in labial gland of patients with PSS was determined by detecting the copy number of EBV DNA in peripheral blood mononuclear cells (PBMCs) of patients with PSS, and the relationship between EBV and PSS was studied. Methods: Real-time quantitative PCR was used to detect EBV DNA copy number in PBMC of 32 patients with PSS and 28 normal controls. The expression of EBNA-1 and EBNA-2 in 32 cases of PSS and 6 cases of normal labial gland were detected by immunohistochemical method. Results: (1) Epstein-Barr virus DNA positive rate was 100% (32 / 32) in patients with PSS and 89.2% (25 / 28) in controls. There was no significant difference (P0.05). (2). The average copy number of EBV DNA in 32 patients with PSS was 26.2 卤10.85 copies/ 渭 g. The average value of EBV DNA copy number in 28 cases of control group was 8.6 卤5.21 copies/ 渭 g, and the average value of EBV copy number in patients with PSS was significantly higher than that in control group. The difference was statistically significant (P0.05). (3). The clinical manifestations of 32 patients with PSS were compared with Epstein-Barr virus copy number (EBV). Epstein-Barr virus DNA copy number in parotid enlargement group was significantly different from that in non-parotid enlargement group (P0.05). The correlation analysis between EBV-DNA and EBV-DNA in 32 patients with PSS showed that Ig was positively correlated with viral DNA copy number (p0.05). (_ 4) EBNA-1 and EBNA-2 in the region. Lymphocytes infiltrated in labial gland of patients with PSS, Positive expression was found in epithelial cells of labial gland ducts. The positive rate of EBNA-1 was 62.8% (22 / 32) in the labial gland of the control group, which was significantly higher than that in the control group (P0.05). The positive rate of EBNA-1 in the labial gland tissue of PSS was 62.5% (20 / 32), which was significantly higher than that in the control group (P0.05). The positive rate of EBNA-1 in PSS labial gland was significantly higher than that in the control group (P0.05). There was no significant difference between clinical manifestation and laboratory examination (P0.05). There was no significant difference in clinical manifestation and laboratory examination between patients with EBNA-2 positive and negative in PSS labial gland (P0.05). Conclusion: (1) EBV replication may be involved in the pathogenesis of PSS. (2) the patients with PSS parotid gland enlargement may be active in EBV replication and aggravate the disease. (3) the high IgG hint may be related to EBV replication activity. (4) EBNA-1 EBNA-2 may be associated with the pathogenesis of PSS by participating in EBV infection.
【学位授予单位】：青岛大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R593.2
[Abstract]:Objective: primary Sjgren's Syndromeg syndrome (SS) is an autoimmune disease characterized by invasion of exocrine glands such as salivary glands and lacrimal glands, and massive lymphocytic infiltration. Many studies have found that Epstein-Barr virus (EBV) infection and Epstein-Barr Nuclear Antigen-1EBNA-1 (EBNA-1) and Epstein-Barr Nuclear Antigen-2( EBNA-2) are the core antigens of EBV expression. The expression of EBNA-1 and EBNA-2 in labial gland of patients with PSS was determined by detecting the copy number of EBV DNA in peripheral blood mononuclear cells (PBMCs) of patients with PSS, and the relationship between EBV and PSS was studied. Methods: Real-time quantitative PCR was used to detect EBV DNA copy number in PBMC of 32 patients with PSS and 28 normal controls. The expression of EBNA-1 and EBNA-2 in 32 cases of PSS and 6 cases of normal labial gland were detected by immunohistochemical method. Results: (1) Epstein-Barr virus DNA positive rate was 100% (32 / 32) in patients with PSS and 89.2% (25 / 28) in controls. There was no significant difference (P0.05). (2). The average copy number of EBV DNA in 32 patients with PSS was 26.2 卤10.85 copies/ 渭 g. The average value of EBV DNA copy number in 28 cases of control group was 8.6 卤5.21 copies/ 渭 g, and the average value of EBV copy number in patients with PSS was significantly higher than that in control group. The difference was statistically significant (P0.05). (3). The clinical manifestations of 32 patients with PSS were compared with Epstein-Barr virus copy number (EBV). Epstein-Barr virus DNA copy number in parotid enlargement group was significantly different from that in non-parotid enlargement group (P0.05). The correlation analysis between EBV-DNA and EBV-DNA in 32 patients with PSS showed that Ig was positively correlated with viral DNA copy number (p0.05). (_ 4) EBNA-1 and EBNA-2 in the region. Lymphocytes infiltrated in labial gland of patients with PSS, Positive expression was found in epithelial cells of labial gland ducts. The positive rate of EBNA-1 was 62.8% (22 / 32) in the labial gland of the control group, which was significantly higher than that in the control group (P0.05). The positive rate of EBNA-1 in the labial gland tissue of PSS was 62.5% (20 / 32), which was significantly higher than that in the control group (P0.05). The positive rate of EBNA-1 in PSS labial gland was significantly higher than that in the control group (P0.05). There was no significant difference between clinical manifestation and laboratory examination (P0.05). There was no significant difference in clinical manifestation and laboratory examination between patients with EBNA-2 positive and negative in PSS labial gland (P0.05). Conclusion: (1) EBV replication may be involved in the pathogenesis of PSS. (2) the patients with PSS parotid gland enlargement may be active in EBV replication and aggravate the disease. (3) the high IgG hint may be related to EBV replication activity. (4) EBNA-1 EBNA-2 may be associated with the pathogenesis of PSS by participating in EBV infection.
【学位授予单位】：青岛大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R593.2
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