摘要
背景:干燥综合征是以口干、眼干为主要临床表现的自身免疫性疾病,病理上以腺体尤其是外分泌腺炎症细胞浸润及次级生发中心形成为主要特点。目前关于该病的发病机制尚不清楚,对该病无针对性有效治疗手段。Th17细胞和Treg细胞是近年来发现的辅助性T细胞的新亚群,对免疫性疾病的发生有关键调节作用。目前关于Th17、Treg在该病中的研究较少。另外B细胞在该病的发病过程中起关键作用,而B细胞的激活需要B细胞激活因子(BAFF)的作用,BAFF与该疾病的发生发展的具体关系目前尚不清楚。
目的:评价干燥综合征患者Th17、Treg、记忆性B细胞的外周表达及其与临床相关性,评价BAFF的外周表达及其与临床的相关性。
方法:干燥综合征国际合作联盟(SICCA)与北京协和医院合作确诊的原发性干燥综合征(pSS)患者41例,患者家属7例及正常健康献血者27例为样本,采集外周血用流式细胞仪分析Th17、Treg、记忆性B细胞,ELISA法检测血浆中的B细胞激活因子(BAFF)浓度,并且结合患者临床资料进行分析。
结果:患者Th17细胞数目较正常人显著升高(5.02±1.95%vs3.8%±1.0%,p=0.004):记忆性B细胞数目较正常人显著升高(11.1%±5.1%vs9.24%±6.99%,p=0.0002):BAFF浓度较正常人有显著性升高(1.18±0.72ng/ml vs0.43±0.19ng/ml p=0.0005):Treg数目较正常人无明显差异(3.14±1.63%vs2.8%±1.2%,p=0.098)。患者及家属检测结果显示Th17数目无显著性差异(5.26%±1.35%vs5.2%±2.1%,p=0.94);Treg数目无显著性差异(3.17±1.62%vs3.08%±1.08%p=0.90);记忆性B细胞数目无显著性差异(16.26%±8.3%vs8.75%±3.82%,p=0.088);患者BAFF水平较其家属有显著性升高(1.29±0.46ng/ml vs0.56±0.11ng/ml p=0.015)。同时结合临床资料分析得出Thl7与IgG具有正相关关系(r=0.457,p=0.005),但对评估疾病严重程度的临床症状、体征及血清学IgM.IgA、补体C3、补体C4、口腔科、眼科检查并无相关关系(p>0.05)。Treg与唇腺病理Focus评分成正相关关系(r=0.480,p=0.005)。CD27+B细胞与5min未刺激唾液量成正相关(r=0.345,p=0.021),与眼科角结膜评分成负相关关系(r=-0.321p=0.041), BAFF与临床症状、体征及血清学IgM、IgA、IgG、补体C3、补体C4、口腔科、眼科检查并无相关关系(p>0.05)。
结论:1Th17在患者及一级亲属中较正常人高表达,与IgG正相关,说明Th17可能与干燥综合征发病有关。2Treg表达较正常人无差异,说明Treg与疾病的关系目前尚不清楚。3记忆B细胞在患者及一级亲属高表达,与临床指标负相关,说明记忆B细胞在疾病发生发展过程中有重要作用。4患者BAFF水平明显高于正常人及一级亲属,与临床指标不相关,说明BAFF可能参与疾病的发生,但与疾病的严重程度不平行。
Background. Primary Sjogren's syndrome is an autoimmune disease with both organ-specific and systemic manifestations. pSS affects the salivary and lacrimal glands preferentially but may frequently also involve other exocrine glands. It is characterized by specific pathological features:the formation of ectopic lymphoid tissue. The mechanism of pSS is unclear and there is not any specific therapies either.Thl7and Treg are newly defined subgroups of helper T cell and are considered critical to the development of autoimmune diseases. B cell plays important role in the occurrence of pSS. However there is still fewer researches on the expression of Th17, Treg and memory B cell. B-lymphocyte activating factor (BAFF) is a key survival factor for B-cells and ensures their existence through reducing apoptotic clearance.
Objective. To investigate the expression of CD4+IL-17+Th17cell, CD4+CD25+regulatory T cells (Tregs) and memory B cell, in the peripheral blood of patients with primary Sjogren's syndrome (pSS) and healthy people. Then to examine BAFF level in pSS and in healthy controls. To analyze the correlations of the cells with clinical symptoms and lab tests.
Methods. Samples of peripheral venous blood were collected from41newly diagnosed pSS patients from SICCA and PUMCH, all females and27healthy controls and7family members of the patients. The levels of CD4+IL-17+Th17cell, CD4+CD25+regulatory T cells (Tregs) and memory B cell in the peripheral blood were measured by flow-cytometric assay. The level of BAFF in serum is tested with ELISA. The clinical indexes including IgG, IgA, IgM, C3, C4, Focus score,5min salivary rate, ocular score, schirmer test were collected to analyze the correlations.
Results. The level of CD4+IL-17+Th17in the blood of pSS patients was5.02%±1.95%, significantly higher than that of the healthy controls3.8%±1.0%(p=0.004). The level of CD4+CD25+Tregs in the blood of pSS patients was3.14%±1.63%, higher than that of the healthy controls2.8%±1.2%, but without statistical difference(p=0.098). The level of memory B cell in the blood of pSS patients was11.1%±5.1%, significantly higher than that of the healthy controls9.24%±6.99%(p=0.0002). There were no statistical differences between Th17, Treg, memory B cell in patients and in family members.(p>0.05). BAFF in pSS was1.18±0.72ng/ml, significantly higher than that of the healthy controls0.43±0.19ng/ml (p=0.0005) and also significantly higher than that of the family members0.56±0.11ng/ml (p=0.015). Th17level were significantly positively correlated with IgG(r=0.457,p=0.005). Treg level were significantly positively correlated with Focus score (r=0.480,p=0.005).CD27+B cell were significantly positively correlated with5min salivary rate (r=0.345,p=0.005) and significantly negatively correlated with ocular score (r=-0.321,p=0.041). BAFF was not correlated with IgG, IgA, IgM, C3, C4, Focus score,5min salivary rate, ocular score, schirmer test.(p>0.05)
Conclusion. Th17is overexpressed in pSS patients and relationships than in healthy people. It is significantly positively correlated with IgG. Th17may involve in pSS mechanism. There is no statistic difference between pSS and controls. Its role in pSS is less obvious. Similarly the memory B cell is overexpression in pSS patients and relationships than in healthy people. It is negatively correlated with clinical indexes. It plays important roles in the developing of the disease. BAFF level is statistically elevated in pSS compared with relationships and controls. It is proved to be not associated with clinical disease activity in pSS. It is not parallel with exocrine dysfunction. These finding suggest that BAFF may contribute to pSS by other mechanisms.
引文
1 Carsten B. Schmidt-Weber, PhD, Mu" beccel Akdis, MD, PhD, and Cezmi A. Akdis, MD Davos, Switzerland. TH17 cells in the big picture of immunology J Allergy Clin Immunol 2007; 120:247-54.
2 B. Afzali, G. Lombardi, R. I. Lechler and G. M. Lord Department of Nephrology and Transplantation, King's College London, Guy's and St Thomas'Hospital, UK.The role of T helper 17 (Th17) and regulatory T cells (Treg) in human organ transplantation and autoimmune disease
3 Cuong Q. Nguyen, Min H. Hu, Yi Li, Carol Stewart, and Ammon B. Peck Salivary Gland Tissue Expression of Interleukin-23 and Interleukin-17 in Sjo"gren's Syndrome Findings in Humans and Mice ARTHRITIS & RHEUMATISM Vol.58, No.3, March 2008, pp 734-743 DOI 10.1002/art.23214.2008, American College of Rheumatology
4 Arne Hansen,(?)l Peter E Lipsky,2 and Thomas Dorner11Charite Centers (CC) 12 and 14, Departments of Medicine and Transfusion Medicine, Charite-Universitatsmedizin Berlin, Charite-Platz 01,10098 Berlin, Germany 2Autoimmunity Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Building 10, Bethesda, MD 20892, USA.B cells in Sjogren's syndrome:indications for disturbed selection and differentiation in ectopic lymphoid tissue Arthritis Res Ther. 2007; 9(4): 218. Published online 2007 August 6:10.1186/ar2210.
5蔡丽敏 陈志强B细胞活化因子与干燥综合征国际皮肤性病学杂志2007年3月第33卷第2期
6 Jaime R. Darce, Bonnie K. Arendt, Sook Kyung Chang and Diane F. Jelinek, Department of Immunology, Mayo Clinic College of Medicine, Mayo Graduate School, Rochester, MN 55905 Divergent Effects of BAFF on Human Memory B Cell Differentiation into Ig-Secreting Cells The Journal of Immunology,2007,178 (9) P.5612-5622
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