类风湿关节炎患者外周血Th17细胞、血清IL-17和IL-6检测水平与疾病活动性的研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
第一部分:Th1 7细胞亚群的测定在类风湿关节炎患者不同活动期变化的研究
     目的:
     1.应用流式细胞仪检测类风湿关节炎(reuhmatoid arthritis, RA)患者外周血Thl7细胞水平。
     2.探讨RA患者外周血Th17细胞水平的检测,在判断疾病活动程度中的可能作用。
     方法:
     1.采用流式细胞仪检测63例次诊断明确的RA患者、15例健康对照者外周血Th17细胞水平。
     2.将63例次RA患者按照DAS28评分分为高、中、低活动组,并初步探讨Th17细胞水平的测定在RA患者不同病情活动程度中意义。
     结果:
     1.RA患者与健康对照者相比,外周血Th17水平前者明显高于后者。
     2.按照DAS28评分分组,高、中、低活动组的RA患者与健康对照者外周血Th17细胞水平存在组间差异,高活动组外周血Th17细胞水平明显高于健康对照组,余组间比较,无明显差异。
     3.以双手X线分级分组,同样与健康对照组进行多变量组间比较,各组间Th17细胞水平无明显差异。
     结论:
     运用流式细胞仪,通过对患者外周血进行Th17细胞检测,可监测患者病情,对患者病情活动严重程度进行评估。
     第二部分:CBA方法测定类风湿关节炎患者血清I L-17、I L-6表达水平
     目的:
     通过检测RA患者血清中白介素17(interleukin 17)与白介素6(interleukin 6)表达水平,分析其在RA不同活动期变化的意义。
     方法:
     1.流式细胞仪微球捕获芯片技术CBA (Cytometric Bead Array)测定63例次(IL-6为62例次)RA患者和18例健康对照者血清IL-17、IL-6表达水平。
     2.按照DAS28评分分组,高、中、低活动组的RA患者与健康对照者血清IL-17、IL-6水平进行多组变量组间比较。
     结果:
     1.RA患者与健康对照者相比,血清IL-17、IL-6水平前者明显高于后者。
     2.按照DAS28评分分组,高、中、低活动组的RA患者与健康对照者外周血IL-17水平存在组间差异,高活动组外周血Th17细胞水平明显高于健康对照组,余组间比较,无明显差异;IL-6水平在各组间无明显差异。
     3.以双手X线分级分组,同样与健康对照组进行多变量组间比较,各组间IL-17水平无明显差异。
     结论:
     RA患者血清中的IL-17和IL-6与健康人存在明显差异,联合Th17细胞的检测可作为预测RA疾病进展及病情活动的潜在生物学标志物。
PartⅠ:Determination of Th17 cell subsets in patients of rheumatoid arthritis at different Period of activity
     Objictive:
     Detected by flow cytometry with rheumatoid arthritis peripheral the level of blood Th17cells.
     Methods:
     1.63 patients' which were diagnosed as RA and 15 healthy controls'Th17 cells were detected by flow cytometry.
     2.63 patients of RA were divided into high, medium and low activity group in accordance with DAS28 scores, which did multiple comparison with healthy control group. Preliminary study the level of Th17 cells in the determination of disease activity in RA patients with different levels of significance.
     Results:
     1.The level of Th17 cells of the patients which were diagnosed as RA is higher than healthy controls.
     2. The level of Th17 cells is higher in high activity group than in healthy control group. There is no significant difference between other groups.
     3. There is no significant difference between these groups which were divided by X-ray film of both hands.
     Conclusion:
     We can monitor patient's condition by testing Th17 cells in peripheral blood from patients with flow cytometry, and evaluate the severity of disease activity of patients.
     PartⅡ:The expression of IL-17 and IL-6 in serum of patients with rheumatoid arthritis by CBA
     Objective:
     Analyze the significance of IL-17 and IL-6 in various activities period of patients with RA by detecting their expression in serum of patients.
     Methods:
     1. Determine IL-17 and IL-6 in serum of 63 RA patients and 18 healthy controls by Cytometric Bead Array(CBA).
     2.63 patients of RA were divided into high, medium and low activity group in accordance with DAS28 scores, which did multiple comparison of their expression of IL-17 and IL-6 with healthy control group.
     Results:
     1. The level of IL-17 and IL-6 of the patients which were diagnosed as RA is higher than healthy controls.
     2. The level of IL-17 is higher in high activity group than in healthy control group. There is no significant difference between other groups. There is no significant difference in IL-6 between all groups.
     3. There is no significant difference between these groups which were divided by X-ray film of both hands.
     Conclusion:
     There is significant difference in serum of IL-17 and IL-6 between RA patients and healthy people. Join in detection of Th17 cells in RA may predict disease progression and determine disease activity by these biomarkers.
引文
[1]叶任高,陆再英.内科学第6版[M],北京:人民卫生出版社,2004,885.
    [2]曲世晶,李小峰,王彩虹,等.甲氨蝶呤联合环磷酰胺对胶原诱导性关节炎大鼠滑膜细胞凋亡作用的研究[J],中国药物与临床,2008,8(6):443-445.
    [3]牛红青,李小峰,张莉芸,等.周期联合甲氨蝶呤环磷酰胺对类风湿关节炎患者外周血淋巴细胞周期及凋亡的影响[J],山西医药杂志,2008,37(4):306-308.
    [4]Haus E, Lakatua DJ, Swoyer J, et al. Chronobiology in hematology and immunology. Am J Anat, 1983,168:467-517.
    [5]Genestier L,Paillot R,Fournel S, et al. Immunosuppressive proper-ties of methotrexate: apoptosis and clonal deletion of activated pe-ripheral T cells. J Clin Invest,1998,102:322-328
    [6]Marie-Christophe Boissier, Eric Assier, Ge'raldine Falgarone, et, al. Shifting the imbalance from Thl/Th2 to Th17/treg:The changing rheumatoid arthritis paradigm[J]. Editorial/Joint Bone Spine 2008(75):373-375.
    [7]韩根成,沈倍奋.Th17细胞分化、调节及效应研究进展[J].生物化学与生物物理进展,2008,35(2):117-123.
    [8]Jinfang Zhu, William E. Paul. CD4T cells:fates, functions, and faults [J]. Blood.2008, (112): 1557-1569.
    [9]王丹丹,孙凌云.CD4+CD25+调节性T细胞在自身免疫性肝病中的研究进展[J].中华风湿病学杂志.2008,12(10):713-715.
    [10]. Cbabaud M, Lubberts E, Joosten L, et al. Arthritis Res,2001; 3(3):168-177.
    [11]. Cai L, Yin JP, Starovasnik MA, et al. Cytokine,2001; 16(1):10-21.
    [12]. Koshy PJ, Henderson N, Logan C, et al. Ann Rheum Ds,2002; 61(8):704-713.
    [13]. Prevoo ML, vant'Hof MA, Kuper HH, et al. Modified disease activity scores that include twenty eight joint counts. Development and validation in a prospective longitudinal study of patients with rheumatoid arthritis (J).Arthritis Rheum,1995; 38(1):31524.
    [14]Astbury C, Platt R, Dixon JS et al. Optimizing the assessment of disease activity during treatment with anti-rheumatoid drugs. British Journal of Rheumatology,1993,32:467-473.
    [15]. Suarez-Almazor ME, Belseck E, Shea B, et al. Cyclophosphamide for treating rheumatoid arthritis. Cochrane Database Syst Rev,2000, (4):CD001157.
    [16]Notley CA, Inglis JJ, Alzabin S, McCann FE, McNamee KE, Williams RO. Blockade of tumor necrosis factor in collageninduced arthritis reveals a novel immunoregulatory pathway for Thl and Th17 cells. J Exp Med 2008; 205:2491-7.
    [17]Young DA, Hegen M, Ma HL et al. Blockade of the interleukin-21/interleukin-21 receptor pathway ameliorates disease in animal models of rheumatoid arthritis. Arthritis Rheum 2007;56:1152-63.
    [18]. van Bezooijen RL, van der Wee2Pals L, Papapoulos SE, et al. Interleukin 17 synergises wit htumour necrosis factor alpha to induce cartilage dest ruction in vit ro [J]. Ann Rheum Dis,2002, 61 (10):870-876.
    [19].Kirkham BW, Lassere MN, Edmonds J P, et al. Synovial membrane cytokine expression is predictive of joint damage progression in rheumatoid art hritis:A two2year prospective study (t he DAMAGE study cohort) [J]. A rthritis Rheum,2006,54 (4):1122-1131.
    [20]. Shiva Shahraral, QiQuan Huangl, Arthur M Mandelin.et, al.。TH-17 cells in rheumatoid arthritis. Arthritis Research & Therapy 2008,10:R93.
    [21]. le Grand A, Fermor B, Fmk C, et al. Arthritis Rheum,2001; 44(9):2078-2083.
    [22]Chabaud M, Durand JM, Buchs N et al. Human interleukin-17:a T cell-derived proinflammatory cytokine produced by the rheumatoid synovium. Arthritis Rheum 1999; 42:963-70.
    [23]Lubberts E, Joosten LA, Oppers B et al. IL-1-independent role of IL-17 in synovial inflammation and joint destruction during collagen-induced arthritis. J Immunol 2001; 167:1004-13.
    [24]Ziolkowska M, Koc A, Luszczykiewicz G et al. High levels of IL-17 in rheumatoid arthritis patients:IL-15 triggers in vitro IL-17 production via cyclosporin A-sensitive mechanism. J Immunol 2000; 164:2832-8.
    [25]Koenders MI, Devesa I, Marijnissen RJ et al. Interleukin-1 drives pathogenic Th17 cells during spontaneous arthritis in interleukin-1 receptor antagonist-deficient mice. Arthritis Rheum 2008; 58:3461-70.
    [26]Shahrara S, Huang Q, Mandelin AM II, Pope RM. TH-17 cells in rheumatoid arthritis. Arthritis Res Ther 2008; 10:R93.
    [27]Kirkham BW, Lassere MN, Edmonds JP et al. Synovial membrane cytokine expression is predictive of joint damage progression in rheumatoid arthritis:a two-year prospective study (the DAMAGE study cohort). Arthritis Rheum 2006; 54:1122-31.
    [28]Nakae S, Nambu A, Sudo K, Iwakura Y. Suppression of immune induction of collagen-induced arthritis in IL-17-deficient mice. J Immunol 2003; 171:6173-7.
    [29]Bush KA, Farmer KM, Walker JS, Kirkham BW. Reduction of joint inflammation and bone erosion in rat adjuvant arthritis by treatment with interleukin-17 receptor IgGl Fc fusion protein. Arthritis Rheum 2002; 46:802-5.
    [30]Lubberts E, Koenders MI, Oppers-Walgreen B et al. Treatment with a neutralizing anti-murine interleukin-17 antibody after the onset of collagen-induced arthritis reduces joint inflammation, cartilage destruction, and bone erosion. Arthritis Rheum 2004; 50:650-9.
    [31]Fujimoto M, Serada S, Mihara M et al. Interleukin-6 blockade suppresses autoimmune arthritis in mice by the inhibition of inflammatory Th17 responses. Arthritis Rheum 2008; 58:3710-9.
    1. Harrington L E, Hatton R D.Mangan P R, et al. Interleukin 17-p roducing CD4+effector T cells develop via a lineage distinct from the Thelper type 1 and 2 lineages[J]. Nat Immunol,2005,6 (11):1123-1132.
    2. Ivanov Ⅱ, McKenzie BS, Zhou L, et al. The orphan nuclear recep tor RORyt directs the differentiation p rogram of p ro-inflammatory IL-17+T helper cells[J]. Cell,2006,126 (6):1121-1133.
    3. VeldhoenM, Hocking RJ.Atkins CJ, et al. TGF beta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-p roducing T cells[J]. Immunity,2006,24 (9):179-189
    4. Bettelli E, Carrier Y, Gao W, et al. Recip rocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells. Nature,2006,441:235-238.
    5. Wahl SM. Transforming growth factor2beta:innately bipolar.CurrOp in Immunol,2007,19:55~62.
    6. Acosta-Rodriguez EV, Napolitani G, Lanzavecchia A, et al.Interleukins lbeta and 6 but not transforming growth factor beta are essential for the differentiation of interleukin 17-p roducing human T helper cells. Nat Immunol,2007, 8:942-949.
    7. Mangan P R, Harrington L E, O!Quinn D B, et al. Transforming growth factor-beta induces development of the T (H)17 lineage. Nature,2006,441:231~234
    8.杨静,贾磊,李丽.抗原特异性Th17细胞的分化与调节[M].细胞与分子免疫学杂志.2008,24(3):213-216.
    9. Laurence A, Tato CM, Davidson TS, et al. Interleukin-2 signaling via STAT5 constrains T helper 17 cell generation. Immunity,2007,26: 371~381.
    10. Weaver CT, Hatton RD, Mangan PR, et al. IL-17 family cytokines and the expanding diversity of effector T cell lineages. Annu Rev Immunol,2007,25:821~852.
    11. Langrish C L, Chen Y, Blumenschein W M, et al. IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. J Exp Med,2005,201:233~240
    12. Wilson NJ, Boniface K, Chan JR, et al. Development,cytokine p rofile and function of human interleukin 17-producing helper T cells. Nat Immunol,2007,8:950-957.
    13. Zhou L, Ivanov II, Spolski R, et al. IL-6 p rograms T(H)-17 cell differentiation by p romoting sequential engagement of the IL-21 and IL-23 pathways. Nat Immunol,2007,8:967~974.
    14. Brustle A, Heink S, HuberM, et al. The development of inflammatory T (H)-17 cells requires interferon regulatory factor 4. Nat Immunol,2007,8:958-966.
    15. Laurence A, O'Shea JJ. T (H)-17 differentiation:of mice and men. Nat Immunol,2007,8:903~905.
    16. Park H, Li Z, Yang XO, et al. A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat Immunol,2005,6:1133~1141.
    17. le Grand A, Fermor B, Fmk C, et al. Arthritis Rheum,2001; 44(9):2078-2083.
    18. Aggareal S, Gurney AL. IL-17:prototype member of an emerging cytokine family.[J]. J Leukoc Boil,2002,71 (1):1-8.
    19. Spriggs MK. Interleukin-17 and its receptor[J]. Clin Immunol,1997,17 (6):366~369.
    20. Kolls J K,Linden A. Interleukin-17 Familymembers and inflammation[J]. immunity,2004,21 (4):467~476.
    21. HonoratiMC,Meliconi R, Pulsatelli L, et al. High in vivo expression of interleukin-17 recep tor in synovial endothelial cells and chondrocytes fromarthritis patients [J]. Rheumatology (Oxford),2001,40(5):522~527.
    22. Subramaniam SV, Cooper RS,Adunyah SE. Evidence for the involvement of JAK/STAT pathway in the signaling mechanism of interleukin 17 [J]. BiochemBiophy Res Commun,1999,262 (6):14~19.
    23. Jovanovic DV, Di Battista JA,Martel2Pelletier J, et al. IL17 stimulates the p roduction and exp ression of p roinflammatory cytokines, IL-1beta and TNF-alpha, by human macrophages[J]. J Immunol,1998,160 (7):3513-3521.
    24. Schwardner R, Yamaguchi K, Cao Z. Requirement of tumor necrosis factor recep tor associated factor (TRAF) 6 in interleukin 17 signal transduction[J]. J Exp Med,2000,191 (7):1233~1240.
    25. Ouyang W, Kolls JK, Zheng Y. The biological functions of T helper 17 cell effector cytokines in inflammation[J]. Immunity,2008,28 (4):454-467.
    26.唐碧霞,张烜,唐福林.IL-17与自身免疫性疾病关系的研究进展.基础医学与临床,2008,28(1):94-97.
    27.Hsu HC, Yang P,Wang J, et al. Interleukin17-producing T helper cells and interleukin 17 orchestrate autoreactive germinal center development in autoimmune BXD2 mice. Nat Immunol,2008,9:166-175.
    28. Cbabaud M, Lubberts E, Joosten L, et al. Arthritis Res,2001; 3(3):168-177.
    29. Cai L, Yin JP, Starovasnik MA, et al. Cytokine,2001; 16(1):10-21.
    30. Koshy PJ, Henderson N, Logan C, et al. Ann Rheum Ds,2002; 61(8):704-713.
    31. CaiL, Yin JP, StarovasnikMA, et al. Pathways bywhich interleukin-17 induces articular cartilage breakdown in vitro and in vivo [J].Cytokine,2001,16 (1):10~21.
    32. Dudler J, Renggli ZulligerN,Busso N, et al. Effect of interleukin 17 on p roteoglycan degradationin murine knee joints[J]. Ann RheumDis,2000,597:529-532.
    33.Lubberts E, Joosten LA,Van deloo FA, et al. Reduction of interleukin-17-induced inhibition ofchonodrocyte p roteoglycan synthesis in intact murine articular cartilage by interleukin24 [J]. Arthritis Rheum,2000,43 (6): 1300-1306.
    34. Le Grand A, Fermor B, Fink C, et al. Interleukin-1, tumor necrosis factor alpha, and interleukin-17 synergistically upregulate nitricoxide and prostaglandin E prouction in exp lants of human os-teoar-thritic knee menisci [J]. Arthritis Rheum,2001,44(9):2078-2083.
    35. NumasakiM, Fukushi J, Ono M, et al. Interleukin-17 promotes angiogenesis and tumor growth[J]. Blood,2003, 101 (7):735-738.
    36. Kotake N, Udagawa N, Takahashi N, et al. IL217 in synovial fluids form paientswith rheumatoid arthritis is a potent stimulator of osteoclastogenesis[J]. Clin Invest,1999,103 (3) L1345~1352.
    37. Murphy CA,Langrish CL, Chen Y, et al. Divergent p roand anti-inflammatory roles for IL-23 and IL-12 in joint autoimmune inflammation [J]. J Exp Med,2003,198(12):1951-1957.
    38. Cho ML, Yoon CH, Hwang SY, et al. Effector function of type Ⅱ collagen-stimulated T cells from rheumatoid arthritis patients:Crosstalk between T cells and synovial fibroblasts[J]. Arthritis Rheum,2004,50 (3):776-784.
    39. Hwang SY, Kim HY. Exp ression of IL-17 homologs and their recep tors in the synovial cells of rheumatoid arthritis patients[J]. Mol Cell,2004,19 (2):180-184.
    40. Lubberts E, Joosten LA, van de Loo FA, et al. Reduction of interleukin-17-induced inhibition of chondrocyte p roteoglycan synthesis in intact murine articular cartilage by interleukin-4 [J]. Arthritis Rheum,2000,43(6):1300-1306.
    41. Hwang SY, Kim JY, ParkMK, et al. IL-17 induces p roduction of IL-6 and IL-8 in rheumatoid arthritis synovial Fibro-blasts via NF-kappaB-and P I3-kinase/Akt-depend-nt path ways [J]. Arthritis Res Ther,2004,6 (2):R120-R128.
    42. Kirkham BW, LassereMN, Edmonds JP, et al. Synovial membrane cytokine exp ression is p redictive of joint damage p rogression in rheumatoid arthritis:A two yearp rospective study(the DAMAGE study cohort) [J]. Arthritis Rheum,2006,54 (4):1122-1131.
    43. Yamada H,Nakashima Y, Okazaki K, et al. Th1 but not Th17 cells p redominate in the joints of patientswith rheumatoid arthritis [J]. Ann Rheum Dis,2008,67(9):1299-1304.
    44. Brentano F,Ospelt C, Stanczyk J, et al. Abundant exp ression of the IL-23 subunit p19, but low levels of bioactive IL-23 in the rheumatoid synovium [J]. Ann Rheum Dis,2008,14. [Epub ahead of print]
    45. Lubberts E. IL-17/Th17 targeting:On the road to p revent chronic destructive arthritis [J] Cytokine,2008, 41 (2):84-91.
    46. Bush KA, Walker JS, Lee CS, et al. Cytokine exp ression and synovial pathology in the initiation and spontaneous resolution phases of adjavan arthritis:interleukin17 exp ression is up regulated in early disease [J]. Clin Exp Immunol,2001,123 (3):487~495.
    47. Lubberts E, Koenders MI,OppersWalgreen B, et al. Treatment with a neutralizing antimurine interleukin-17 antibody after the onset of collagen induced arthritis reduces joint inflammation cartilage destruction and bone erosion[J]. Arthritis Rheum,2004,50 (2):650~659.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700