组织原位记忆T细胞在系统性红斑狼疮的表达及与皮损的相关性研究

发布时间：2018-03-16 23:26

本文选题：红斑狼疮　切入点：系统性　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨系统性红斑狼疮(systemic lupus erythematosus,SLE)组织原位记忆T细胞(tissue resident memory T cells,Trm cells)与SLE皮损的相关性,为研究SLE患者皮损的形成机制提供新思路,寻找狼疮皮损治疗的新靶点。方法:本研究连续性纳入2015年7月至2016年4月于安徽省立医院风湿免疫科门诊或住院部初诊的SLE患者20例(男性2例,女性18例),所有患者均符合1997年美国风湿病学会(American College of Rheumatology,ACR)制定的SLE分类标准。健康对照10例选自同期我院健康体检中心体检者(男性1例,女性9例),平均年龄(26.4±4.8)岁。皮肤对照组10例选自同期我院美容中心。所有研究对象均被告知并签署知情同意书。收集SLE患者及健康对照外周血,分离PBMC后流式细胞术检测SLE患者和健康对照外周血及T淋巴细胞亚群中CCR7-CD45RA-效应记忆T细胞(effector memory T cells,Tem cells)和CCR7-CD45RA+效应T细胞(effector T cells,Teff cells)亚群的比例变化;收集SLE患者皮损处皮肤组织及健康对照正常部位皮肤组织,免疫荧光法观察SLE患者及健康对照真皮和表皮间Ig A、Ig G、Ig M、C3免疫复合物沉积情况;免疫组织化学法检测SLE患者皮损处皮肤组织及健康对照正常部位皮肤组织中CD4、CD8以及Trm细胞的特征性表面标志CD103分子表达,比较CD103阳性细胞在SLE患者皮损组织和健康对照正常皮肤组织中的表达。结果:(1)与健康对照相比,SLE患者外周血中CD4+Tem、CD4+Teff、CD8+Tem、CD8+Teff细胞在外周血T淋巴细胞亚群中所占比例均明显升高,差异有统计学意义(12.56±3.40 vs 8.19±2.53,p=0.004;2.54±1.52 vs 1.34±0.82,p=0.029;15.31±3.62 vs 7.05±2.99,p=0.000;13.11±5.38 vs 3.73±1.33,p=0.000);(2)与健康对照相比,SLE患者CD4+Tem、CD4+Teff、CD8+Tem、CD8+Teff细胞在外周血中的比例均明显升高,差异有统计学意义(8.42±2.67 vs 5.81±1.97,p=0.011;1.55±0.96 vs 0.79±0.51,p=0.008;10.36±3.60 vs 5.27±2.39,p=0.000;8.16±4.10 vs 2.56±0.80,p=0.000);(3)皮肤组织免疫荧光染色荧光显微镜下观察到在SLE患者皮损处表皮和真皮交界处有大量Ig A、Ig M、补体C3呈线型沉积,而健康对照者皮肤组织中没有免疫复合物沉积;(4)皮肤组织病理与健康对照相比,SLE患者皮损处组织HE染色可见真皮层中浸润淋巴细胞明显增多。免疫组织化学染色显示浸润淋巴细胞中表达CD4、CD8、CD103分子的细胞数量均较健康对照有不同程度增多。结论:SLE患者体内存在Tem细胞和Teff细胞的异常活化,其表达较健康对照明显升高。Trm细胞局部浸润导致SLE患者皮损部位皮肤组织表皮和真皮下免疫复合物沉积。Trm细胞浸润可能与SLE患者皮损形成相关。
[Abstract]:Objective: To investigate the effects of systemic lupus erythematosus (systemic lupus, erythematosus, SLE) in situ memory T cells (tissue resident memory T cells, Trm cells) associated with SLE lesions, and provide new ideas for the study on the formation mechanism of the skin lesions of patients with SLE, to find a new target for the treatment of lupus lesions. Methods: This study included continuity from July 2015 to April 2016 in Anhui Provincial Hospital Department of rheumatology outpatient or inpatient department of newly diagnosed SLE patients 20 cases (2 cases, male 18 cases of female), all the patients were consistent with the 1997 American College of Rheumatology (American College of Rheumatology, ACR SLE) classification standard. 10 healthy subjects were selected from the same period in our hospital health examination center examination (male 1 cases, female 9 cases), average age (26.4 + 4.8) years old. The skin beauty center in our hospital 10 cases of control group were selected from the same period. All the subjects were informed and signed informed consent. Collect SL E patients and healthy control peripheral blood PBMC after separation by flow cytometry in patients with SLE and healthy control peripheral blood T lymphocyte subsets and CCR7-CD45RA- effect in memory T cells (effector memory T cells, Tem cells) and CCR7-CD45RA+ (effector T cells T cells, Teff cells) subsets proportion; collect SLE patients and healthy skin tissue and normal skin tissue site, immunofluorescence of SLE patients and healthy controls between the dermis and epidermis Ig A, Ig G, Ig M, C3 immune complex deposition; detection of skin tissue in normal position in CD4 lesions of patients with SLE and healthy skin tissue by immunohistochemical method CD8, Trm and cell surface markers characteristic of the expression of CD103, CD103 positive cells were expressed in normal skin tissues in the lesions of patients with SLE and healthy. Results: (1) and health controls, SLE patients The peripheral blood CD4+Tem, CD4+Teff, CD8+Tem, CD8+Teff cells were significantly increased in the proportion of peripheral blood T lymphocyte subsets in proportion, the difference was statistically significant (12.56 + 3.40 vs 8.19 + 2.53, p=0.004; 2.54 + 1.52 vs 1.34 + 0.82, p=0.029; 15.31 + 3.62 vs 7.05 + 2.99, P =0.000 13.11 + 5.38; 3.73 + 1.33 vs, p=0.000); (2) compared with healthy controls, CD4+Tem patients, SLE CD4+Teff, CD8+Tem, the proportion of CD8+Teff cells in peripheral blood were significantly increased, the difference was statistically significant (8.42 + 2.67 vs 5.81 + 1.97, p=0.011; 1.55 + 0.96 vs 0.79 + 0.51. P=0.008; 10.36 + 3.60 vs 5.27 + 2.39, p=0.000; 8.16 + 4.10 vs 2.56 + 0.80, p=0.000); (3) skin tissue immunofluorescence staining under fluorescence microscope observed a large number of Ig A, at the junction of the skin lesions of patients with SLE Ig M in the epidermis and dermis, complement C3 linear deposition, and healthy control skin tissue without immune complex Deposit; (4) compared with healthy skin tissue pathological lesions of patients with SLE, tissue HE staining in the dermis infiltrating lymphocytes increased significantly. Immunohistochemical staining showed that the expression of CD4, CD8 lymphocytes, cell number of CD103 molecules were compared with healthy controls with different degree increased. Conclusion: the aberrant activation of Tem cells and Teff cells of SLE patients compared with healthy controls, the expression of.Trm was significantly higher in patients with SLE cell infiltration in infiltrating lesions of skin tissue under the epidermis and dermal immune complex deposition of.Trm cells may be associated with lesions of patients with SLE.

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

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