ITGAM基因单核苷酸多态性与中国汉族系统性红斑狼疮的相关性分析

发布时间：2018-03-06 06:17

本文选题：ITGAM　切入点：CD11b　出处：《南京医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：背景:系统性红斑狼疮(systemic lupus erythematosus,SLE)是一种慢性自身免疫性疾病,常累及多系统多器官、病程迁延反复。患者最突出特征是免疫系统的紊乱和自身抗体的大量产生。系统性红斑狼疮有明显的遗传易感性。最近国际上多个研究小组通过病例对照研究,发现整合素ITGAM(又称为CD11b)外显子3中的单核苷酸位点(R77H)rs1143679与多种人群中的SLE易感性密切相关,该突变体使SLE病人临床表现更加严重,伴有更多的自身抗体。但迄今为止,缺乏中国汉族人群的ITGAM基因多态性与临床SLE病人表型相关性的研究报道,且机制未明。美国严俊小组采用条件型基因敲除CD11b技术,在动物和细胞水平证实了CD11b参与了对自身反应性B细胞的负向调节功能,该部分结果已发表在nature communications杂志上。为了在临床患者中得到验证,本研究小组和美国严俊小组合作申报了国家自然基金(编号81228018)。我们希望从临床方面通过分析大量SLE患者的CD11b基因多态性,综合临床资料,分析狼疮患者的CD11b基因型和临床表型间的关系,以及B细胞功能的改变。这将为阐明系统性红斑狼疮的发病机制,开发以CD11b为靶点的基因工程药物奠定坚实的基础。一中国汉族系统性红斑狼疮患者中ITGAM基因单核苷酸多态性分析目的分析中国汉族系统性红斑狼疮(SLE)患者中ITGAM基因rs1143679的单核苷酸多态性(single nucleotide polymorphism,SNP),并阐明该 SNP 与 SLE 临床表型的相关性。方法采用病例对照的研究方法,应用PCR-RFLP以及直接测序技术对中国汉族人群中984例系统性红斑狼疮和928例健康对照进行多态性检测,分析基因型和等位基因频率的分布差异,并与临床表型进行相关性分析。结果(1)SLE患者中ITGAMrs1143679A等位基因频率为1.17%,大大低于欧美国家的基因型频率,与日本及泰国地区接近。(2)ITGAMrs1143679AG基因型与SLE易感性、狼疮肾炎,抗心磷脂抗体有相关性(P0.05),而与皮肤红斑、光过敏、口腔溃疡、浆膜炎、关节炎表现、血液系统表现、神经系统表现以及SLEDA没有统计学差异(P0.05)。结论ITGAM rs1 143679 SNP与中国汉族人群系统性红斑狼疮易感性有关,并可能参与了狼疮肾炎的发生发展。二ITGAM突变SLE患者B细胞上CD11b的改变及其凋亡目的:探讨ITGAM突变对于SLE患者B细胞表达CD11b以及其凋亡的意义。方法:采用流式细胞术检测11例ITGAM突变SLE患者外周血B细胞CD11b表达和其凋亡率,并与11例非突变SLE患者、11例健康对照组进行比较。结果:(1)突变组活动期表达CD11b的B淋巴细胞百分率为(32.6±5.4)%,与未突变组(35.2±6.5)%差异无统计学意义(P=0.32),但与健康对照组(25.6±4.9)%比较,表达均上调(P=0.01);缓解期B淋巴细胞CD11b表达可回调至正常水平,与活动期比较差异有显著性意义(P0.01)(2)突变患者活动期B淋巴细胞凋亡率为(15.9±3.5)%,低于未突变组(26.8±5.8)%,差异均具统计学意义(P=0.001)。但较健康对照组差异均上调(P=0.001)。缓解期突变组与未突变组B淋巴细胞凋亡率均下降,但两者之间仍有差异,且与活动期比较有统计学差异。结论:CD11b突变不改变B细胞表面CD11b分子的表达水平,但凋亡减少。
[Abstract]:Background: systemic lupus erythematosus (systemic lupus, erythematosus, SLE) is a chronic autoimmune disease, often involving multiple organ systems, repeated course of disease. The most prominent feature is a large number of patients with immune system disorders and autoantibodies in systemic lupus erythematosus. There are obvious genetic susceptibility. The multiple the research team through a case-control study, found that integrin ITGAM (also called CD11b) in exon 3 of single nucleotide polymorphisms (R77H) is closely related to the susceptibility of SLE rs1143679 and other groups, the clinical manifestations of patients with SLE mutant is more serious, with more autoantibodies. But so far, the research reported lack of correlation between ITGAM gene Chinese polymorphism in Han population and clinical phenotype of SLE patients, and the mechanism is not clear. The United States team strict conditional knockout of CD11b technology in animal and cell level certificate The CD11b was involved in autoimmune B cells negative regulation function, the results have been published in the journal communications nature. In order to get the verification in clinical patients in the study group and the United States Yan Jun Group cooperation to declare the National Natural Science Foundation (No. 81228018). We hope that through the analysis of the CD11b gene from clinical aspects a large number of polymorphisms in patients with SLE, clinical data, analysis of the relationship between CD11b genotypes and clinical phenotypes in patients with systemic lupus erythematosus, and B cell function change. This will help to elucidate the pathogenesis of systemic lupus erythematosus, development and lay a solid foundation for CD11b gene engineering drug target. Objective to analyze China Han systemic lupus erythematosus ITGAM gene single nucleotide polymorphism analysis China Han patients with systemic lupus erythematosus (SLE) polymorphism of ITGAM gene in patients with rs1143679 (single Nucleotide polymorphism, SNP), and to clarify the relationship between SNP and clinical phenotype of SLE. Methods by using the case-control study, using PCR-RFLP and direct sequencing of Chinese in Han population of 984 cases of systemic lupus erythematosus and 928 healthy controls were tested for polymorphism detection, analysis of differences in genotype distribution and allele frequency. And the correlation with clinical phenotype. Results (1) in patients with SLE ITGAMrs1143679A allele frequency was 1.17%, significantly lower than the genotype frequency of Europe and the United States, Japan and Thailand and close to the area. (2) ITGAMrs1143679AG genotypes and susceptibility to SLE, lupus nephritis, correlation of anticardiolipin antibody (P0.05), and skin erythema light, allergy, oral ulcer, serositis, arthritis, hematological manifestations, no statistically significant differences between the nervous system and SLEDA (P0.05) ITGAM SNP and RS1 143679. Conclusion The susceptibility of systemic lupus erythematosus Chinese in Han population, and may be involved in the occurrence and development of lupus nephritis. Two ITGAM patients with SLE mutation in B cells CD11b change and apoptosis Objective: To investigate the expression of SLE ITGAM mutations on B cells in patients with CD11b and its significance of apoptosis. Methods: flow cytometry was used to detect 11 cases of B cell CD11b SLE mutation in patients with peripheral blood ITGAM expression and apoptosis rate, and mutation and 11 cases of non SLE patients, 11 healthy controls were compared. Results: (1) mutation group activity expression of B lymphocyte percentage for CD11b (32.6 + 5.4)%, and no mutation group (35.2 + 6.5) no significant the significance of differences, but% (P=0.32) and healthy controls (25.6 + 4.9)%, were up-regulated (P=0.01); remission of B lymphocyte CD11b expression can be back to normal levels, there were significant differences compared with the active phase (P0.01) (2) mutations in patients with active B lymph The apoptosis rate was (15.9 + 3.5)%, lower than non mutation group (26.8 + 5.8)%, the difference was statistically significant (P=0.001). But compared with the healthy control group were increased (P=0.001). The remission group and no mutation mutation group B lymphocyte apoptosis rate decreased, but there are still differences between them., and active period is statistically significant. Conclusion: the expression level of CD11b mutation does not alter the B cell surface CD11b molecules, but reduced apoptosis.

【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.241

【相似文献】