蛋白芯片技术检测免疫性血小板减少性紫癜患者细胞因子的变化
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摘要
研究背景：免疫性血小板减少性紫癜(immune thrombocytopenic purpura,ITP)或称原发性血小板减少性紫癜(Primary thrombocytopenic purpura)或特发性血小板减少性紫癜(idiopathic thrombocytopenic purpura,ITP)是最常见的出血性疾病,发病率为1.6-10／10万。ITP虽为良性疾病,但容易复发,进展为难治性ITP。美国一项关于ITP患者健康相关的生活质量(Health-related quality of life, HRQOL)显示,ITP患者生活质量低于癌症患者,还低于常见慢性疾病高血压和关节炎,与糖尿病相当。尽管目前ITP有多种治疗方法,但仍无根治的治疗方案,并且其发病机制至今尚未完全阐明。其经典机制认为是由于自身抗体致敏的血小板被单核一巨噬细胞系统过度破坏所致。近年来研究发现,除体液免疫外,细胞免疫在ITP的发病中亦起着重要作用,对ITP自身免疫发病机制的深入研究令许多新的定向免疫干预措施进入临床研究阶段。蛋白芯片是一种高通量监测系统,通过靶分子和捕捉分子相互作用来监测蛋白分子之间的相互作用。蛋白芯片能够同时检测多种细胞因子,有助于阐明细胞因子在ITP发病中的作用,为靶向治疗提供理论依据。
目的：检测ITP患者外周血中507种细胞因子的变化,旨在利用蛋白芯片技术筛选出可能与ITP发病相关的细胞因子,并通过聚类分析发现相关细胞因子在信号通路中的变化。
材料和方法：
(1)病例与正常对照的选择：2009年8月至2010年4月在我院就诊的ITP患者共17例,其中初发12例,复发6例,缓解2例。均符合ITP的诊断标准。初发患者血小板计数1-82*10^9／L,中位血小板计数25*10^9／L。复发患者血小板计数6-41-10^9／L,中位血小板计数17-10^9／L。缓解患者血小板计数128-154*10^9／L。排除ITP患者合并有病毒性肝炎、糖尿病、高血压、心血管疾病、妊娠、活动性感染者。选取正常对照7例,均为健康志愿者,无感染、无病毒性肝炎、无免疫性疾病及免疫抑制剂应用史、抗HIV抗体阴性、肝肾功能正常,其血小板计数128-263*10^9／L。
(2)应用细胞因子抗体芯片盒RayBioTM人L系列抗体芯片I检测ITP患者的外周血中507种细胞因子变化。
结果：实验发现总计有400种目标蛋白在疾病不同时期出现了上调或下调,其中110种目标蛋白出现上调,175种目标蛋白出现下调,另有115种蛋白在疾病不同时期既出现过上调,也出现过下调。初发ITP患者相比正常对照组,35种目标蛋白出现上调,130种出现下调；复发ITP患者相比正常对照组,99种目标蛋白出现上调,189种出现下调；缓解患者相比正常对照组,94种目标蛋白出现上调,144种出现下调。通过通路分析,共发现6条感兴趣的通路：细胞因子间受体相互作用通路；JAK-STAT信号通路；TGF-beta信号通路；趋化因子信号通路；MAPK信号通路；TLR信号通路。
结论：蛋白芯片能以极小的样本量检测高通量的细胞因子,是筛选ITP相关的细胞因子的比较理想的技术方法。我们观察到细胞因子间受体相互作用通路、JAK-STAT信号通路、TGF-beta信号通路、趋化因子信号通路、MAPK信号通路以及TLR信号通路在ITP中有变化,可能与ITP发病有关。
Background and objective:Immune thrombocytopenic purpura is the most common hemorrhagic disease. The incidence rate is 1.6-10/100,000. Although ITP is a benign disease, it usually relapses and progress into refractory ITP. A research on health related quality of life(HRQOL) in USA shows that the quality of life of ITP patients is beneath not only the common people but patients of other chronic diseases such as hypertension and arthritis.Despite of multiple therapies,ITP has no radical method yet.The pathogenesis remains unknown yet.The classical mechanism is that platelets sensitized by autoantibody are over destroyed through mononuclear-macrophage system. Recent researches find out that cellular immunity is also important in the pathogenesis besides humoral immunity.The in-depth study in the autoimmunity of pathogenesis of ITP has lead many new oriented immunologic interventions into clinical research. Protein array is a kind of high throughput monitoring system. It monitors the protein-protein interaction through the target molecules and capturing molecules interaction.Protein array can detect varieties of cytokines, which does help to clarify the pathogenesis of ITP and provide a theoretical basis for the targeted therapy.
Objective:To detect the changes of 507 kinds of cytokines in ITP patients, screen out the cytokines possibly related to the pathogenesis of ITP, and then observe their changes in the signaling pathways by cluster analysis.
Method:
(1)Patients and controls:During August 2009 to April 2010,17 ITP patients were enrolled by the diagnostic criteria for ITP, including 12 primary,2 CR,3 relapsed patients.The primary patients'platelet counts range from 1-82*10-9/L,the mean count is 25*10-9/L. The relapsed patients'platelet counts range from 6-41*10-9/L,the mean count is 17*109/L. The relapsed patients'platelet counts range from 128-154*109/L. Patients complicated with viral hepatitis,diabetes,hypertension,cardiovascular diseases,pregnancy, active infection,or connective tissue diseases were excluded. The control group consisted of 7 adult healthy volunteers.Their platelet counts range from 128-263*10-9/L.
(2) Detection of changes of 507 kinds of cytokines in ITP patients by RayBio(?) Biotin Label-based Human Antibody Array I.
Results:It was found that 400 kinds of cytokines have changed, of which 110 elevated,175 decreased, while 115 elevated or decreased during the different stages of the disease. Compared with the healthy volunteers, the primary ITP patients showed 35 elevated and 130 decreased cytokines, the relapsed patients showed 99 elevated and 189 decreased cytokines,the CR patients showed 94 elevated and 144 decreased cytokines.Through the pathway analysis,we discovered 6 interested signaling pathways:cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, TGF-beta signaling pathway,chemokine signaling pathway,MAPK signaling pathway, TLR signaling pathway.
Conclusion:Protein array,is an ideal method to screen the cytokines in ITP with little sample volume. The change in the cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, TGF-beta signaling pathway,chemokine signaling pathway,MAPK signaling pathway and TLR signaling pathway we observed maybe related to the pathogenesis in ITP.
目的：检测ITP患者外周血中507种细胞因子的变化,旨在利用蛋白芯片技术筛选出可能与ITP发病相关的细胞因子,并通过聚类分析发现相关细胞因子在信号通路中的变化。
材料和方法：
(1)病例与正常对照的选择：2009年8月至2010年4月在我院就诊的ITP患者共17例,其中初发12例,复发6例,缓解2例。均符合ITP的诊断标准。初发患者血小板计数1-82*10^9／L,中位血小板计数25*10^9／L。复发患者血小板计数6-41-10^9／L,中位血小板计数17-10^9／L。缓解患者血小板计数128-154*10^9／L。排除ITP患者合并有病毒性肝炎、糖尿病、高血压、心血管疾病、妊娠、活动性感染者。选取正常对照7例,均为健康志愿者,无感染、无病毒性肝炎、无免疫性疾病及免疫抑制剂应用史、抗HIV抗体阴性、肝肾功能正常,其血小板计数128-263*10^9／L。
(2)应用细胞因子抗体芯片盒RayBioTM人L系列抗体芯片I检测ITP患者的外周血中507种细胞因子变化。
结果：实验发现总计有400种目标蛋白在疾病不同时期出现了上调或下调,其中110种目标蛋白出现上调,175种目标蛋白出现下调,另有115种蛋白在疾病不同时期既出现过上调,也出现过下调。初发ITP患者相比正常对照组,35种目标蛋白出现上调,130种出现下调；复发ITP患者相比正常对照组,99种目标蛋白出现上调,189种出现下调；缓解患者相比正常对照组,94种目标蛋白出现上调,144种出现下调。通过通路分析,共发现6条感兴趣的通路：细胞因子间受体相互作用通路；JAK-STAT信号通路；TGF-beta信号通路；趋化因子信号通路；MAPK信号通路；TLR信号通路。
结论：蛋白芯片能以极小的样本量检测高通量的细胞因子,是筛选ITP相关的细胞因子的比较理想的技术方法。我们观察到细胞因子间受体相互作用通路、JAK-STAT信号通路、TGF-beta信号通路、趋化因子信号通路、MAPK信号通路以及TLR信号通路在ITP中有变化,可能与ITP发病有关。
Background and objective:Immune thrombocytopenic purpura is the most common hemorrhagic disease. The incidence rate is 1.6-10/100,000. Although ITP is a benign disease, it usually relapses and progress into refractory ITP. A research on health related quality of life(HRQOL) in USA shows that the quality of life of ITP patients is beneath not only the common people but patients of other chronic diseases such as hypertension and arthritis.Despite of multiple therapies,ITP has no radical method yet.The pathogenesis remains unknown yet.The classical mechanism is that platelets sensitized by autoantibody are over destroyed through mononuclear-macrophage system. Recent researches find out that cellular immunity is also important in the pathogenesis besides humoral immunity.The in-depth study in the autoimmunity of pathogenesis of ITP has lead many new oriented immunologic interventions into clinical research. Protein array is a kind of high throughput monitoring system. It monitors the protein-protein interaction through the target molecules and capturing molecules interaction.Protein array can detect varieties of cytokines, which does help to clarify the pathogenesis of ITP and provide a theoretical basis for the targeted therapy.
Objective:To detect the changes of 507 kinds of cytokines in ITP patients, screen out the cytokines possibly related to the pathogenesis of ITP, and then observe their changes in the signaling pathways by cluster analysis.
Method:
(1)Patients and controls:During August 2009 to April 2010,17 ITP patients were enrolled by the diagnostic criteria for ITP, including 12 primary,2 CR,3 relapsed patients.The primary patients'platelet counts range from 1-82*10-9/L,the mean count is 25*10-9/L. The relapsed patients'platelet counts range from 6-41*10-9/L,the mean count is 17*109/L. The relapsed patients'platelet counts range from 128-154*109/L. Patients complicated with viral hepatitis,diabetes,hypertension,cardiovascular diseases,pregnancy, active infection,or connective tissue diseases were excluded. The control group consisted of 7 adult healthy volunteers.Their platelet counts range from 128-263*10-9/L.
(2) Detection of changes of 507 kinds of cytokines in ITP patients by RayBio(?) Biotin Label-based Human Antibody Array I.
Results:It was found that 400 kinds of cytokines have changed, of which 110 elevated,175 decreased, while 115 elevated or decreased during the different stages of the disease. Compared with the healthy volunteers, the primary ITP patients showed 35 elevated and 130 decreased cytokines, the relapsed patients showed 99 elevated and 189 decreased cytokines,the CR patients showed 94 elevated and 144 decreased cytokines.Through the pathway analysis,we discovered 6 interested signaling pathways:cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, TGF-beta signaling pathway,chemokine signaling pathway,MAPK signaling pathway, TLR signaling pathway.
Conclusion:Protein array,is an ideal method to screen the cytokines in ITP with little sample volume. The change in the cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, TGF-beta signaling pathway,chemokine signaling pathway,MAPK signaling pathway and TLR signaling pathway we observed maybe related to the pathogenesis in ITP.
引文
1.McMillan R, Bussel JB, George JN, et al.Self-reported health-related quality of life in adults with chronic immune thrombocytopenic purpura. Am J Hematol,2008,83(2):150-154.
2. Rodeghiero F, Stasi R, Gernsheimer T, et al.Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children:report from an international working group.Blood,2009,113(11):2386-2393.
3.Harrington WJ, Sprague CC, Minnich V, et al.Immunologic mechanisms in idiopathic and neonatal thrombocytopenic purpura. Ann Intern Med,1953,38(3):433-469.
4. Shulman NR, Marder VJ, Weinrach RS. Similarities between known antiplatelet antibodies and the factor responsible for thrombocytopenia in idiopathic purpura. Physiologic,serologic and isotopic studies.Ann N Y Acad Sci,1965,124(2):499-542.
5.Mylvaganam R, Garcia R0, et al.Depressed functional and phenotypic properties of T but not B lymphocytes in idiopathic thrombocytopenic purpura. Blood,1988,71(5):1455-1460.
6. Sayeh E, Sterling K, Speck E, et al.IgG antiplatelet immunity is dependent on an early innate natural killer cell-derived interferon-gamma response that is regulated by CD8+ T cells.Blood, 2004,103(7):2705-2709.
7. Da Wei Huang, Brad T Sherman, Richard A Lempicki.Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.Nature Protocols,2009,4(1):44-57.
8.张鲁勤,葛晋源,等.特发性血小板减少性紫癜患者血浆白细胞介素18及其有关细胞因子.中国实验血液学杂志,2003,11(6)：662-664.
9. Shan NN, Zhu XJ, et al.Interleukin 18 and interleukin 18 binding protein in patients with idiopathic thrombocytopenic purpura. Br J Haematol,2009,144(5):755-761.
10. Aderem A, Ulevitch RJ. Toll-like receptors in the induction of the innate immune response.Nature,2000,406(6797):782-787.
1 Francesco Rodeghiero, Roberto Stasi,et al.Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children:report from an international working group.Blood,2009,113(11):2386-2393.
2 Mylvaganam R, Garcia RO, Depressed functional and phenotypic properties of T but not B lymphocytes in idiopathic thrombocytopenic purpura. Blood,1988,71(5):1455-60.
3 Harrington WJ, Sprague CC, Minnich V, et al.Immunologic mechanisms in idiopathic and neonatal thrombocytopenic purpura. Ann Intern Med,1953,38(3):433-69.
4 Shulman NR, Marder VJ, Weinrach RS. Similarities between known antiplatelet antibodies and the factor responsible for thrombocytopenia in idiopathic purpura. Physiologic, serologic and isotopic studies.Ann N Y Acad Sci,1965,124(2):499-542.
5 McMillan R. Antiplatelet antibodies in chronic adult immune thrombocytopenic purpura:assays and epitopes.J Pediatr Hematol Oncol,2003,25(Suppl 1):S57-61.
6 Sayeh E, Sterling K, Speck E, et al.IgG antiplatelet immunity is dependent on an early innate natural killer cell-derived interferon-gamma response that is regulated by CD8+ T cells.Blood, 2004,103(7):2705-2709.
7 Semple JW, Fre edmanJ. Increased antiplatelet lymphocyte reactivity in patients with autoimmune thrombocytopenia(AITP). Blood,1991,78(10):2619-2625.
8 Ware RE,Howard TA. Elevated numbers of gamma-delta(gamma delta+) T lymphocytes in children with immune thrombocytopenic purpura. J Clin Immunol.1994,14(4):234-237.
9 Shimomura T, Fujimura K, Takafuta T, et al.Oligoclonal accumulation of T cells in peripheral blood from patients with idiopathic thrombocytopenic purpura. Br J Haematol.1996,95(4):732-737.
10 Kuwana M, Kawakami Y, Ikeda Y. Suppression of autoreactive T-cell response to glycoprotein Ⅱb/Ⅲa by blockade of CD40/CD154 interaction:implication for treatment of immune thrombocytopenic purpura[J].Blood,2003,101(2):621-623.
11 Ogawara H, Handa H, et al.High Th1/Th2 ratio in patients with chronic idiopathic thrombocytopenic purpura. Eur J Haematol,2003,71(4):283-288.
12 Olsson B, Andersson P0, JernasM, et al.T-cell mediated cytotoxicity toward platelets in chronic idiopathic thrombocytopenic purpura. NatMed,2003,9(9):1123-1124.
13 Panitsas FP, Theodoropoulou M, Kouraklis A, et al.Adult chronic idiopathic thrombocytopenic purpura(ITP) is the manifestation of a type-1 polarized immune response.Blood,2004,103(7):2645-2647.
14 Stasi R, Del Poeta G, Stipa E, et al.Response to B-cell depleting therapy with rituximab reverts the abnormalities of T cell subsets in patients with idiopathic thrombocytopenic purpura. Blood,2007,110(8):2924-2930.
15 Tag LM, Ezz-Eldeen AM, et al.Serum IL-2 and platelet-associated immunoglobulins are good prognostic markers in immune thrombocytopenic purpura. Egypt J Immunol,2004,11(2):121-132.
16张鲁勤,葛晋源,等.特发性血小板减少性紫癜患者血浆白细胞介素18及 其有关细胞因子.中国实验血液学杂志,2003,11(6)：662-664.
17 Shan NN, Zhu XJ, et al.Interleukin 18 and interleukin 18 binding protein in patients with idiopathic thrombocytopenic purpura. Br J Haematol,2009,144(5):755-761.
18 Webber NP, Mascarenhas JO, et al.Functional properties of lymphocytes in idiopathic thrombocytopenic purpura. Hum Immunol,2001,62(12):1346-1355.
19 Andersson PO, Stockelberg D, Jacobsson S,Wadenvik H. A transforming growth factor-betal-mediated bystander immune suppression could be associated with remission of chronic idiopathic thrombocytopenic purpura. Ann Hematol,2000,79(9):507-513.
20 Langrish CL, Chen Y, Blumenschein WM, et al.IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. J Exp Med,2005,201(2):233-240.
21 Weaver CT, Harrington LE, Mangan PR, et al.Thl7:an effector CD4 T cell lineage with regulatory T cell ties. Immunity,2006,24(6):677-688.
22 Ma D, Zhu X, et al.Profile of Th17 cytokines(IL-17, TGF-p, IL-6) and Thl cytokine(IFN-γ)in patients with immune thrombocytopenic purpura. Ann Hematol,2008,87(11):899-904.
23 Guo ZX, Chen ZP, et al.The role of Th17 cells in adult patients with chronic idiopathic thrombocytopenic purpura. Eur J Haematol,2009,82(6):488-489.
24 Zhu X, Ma D, Zhang J et al.Elevated Interleukin-21 Correlated to Th17 and Thl Cells in Patients with Immune Thrombocytopenia. J Clin Immunol. 2009,10 [Epub ahead of print].
25孙雨梅,张彦明,等.CD4+CD25+调节性T细胞在特发性血小板减少性紫癜患者中的变化及意义.临床血液学杂志,2008,21(5)：469-473.
26 Zhang F, Chu X. Cell-mediated lysis of autologous platelets in chronic idiopathic thrombocytopenic purpura. Eur J Haematol,2006,76(5):427-431.
27 Yoshimura C, Nomura S,Nagahama M, et al.Plasma-soluble Fas(AP021, CD95) and soluble Fas ligand in immune thrombocytopenic purpura. Eur J Haematol,2000,64(4):219-224.
28 B Zhou, et al.Multi-dysfunctional pathophysiology in ITP. Critical Reviews in Oncology/Hematology,2005,54(2):107-116.
29 Shenoy S, Mohanakumar T, Chatila T, et al.Defective apop tosis in lymphocytes and the role of IL-2 in autoimmune hematologic cytopenias. Clin Immunol,2001,99(2):266-275.
30钟永根,封蔚莹,等.特发性血小板减少性紫癜患者外周血T细胞Fas-FasL和caspase-3凋亡相关蛋白的表达及其意义.中华血液学杂志,2008,29(5)：329-332.
2. Rodeghiero F, Stasi R, Gernsheimer T, et al.Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children:report from an international working group.Blood,2009,113(11):2386-2393.
3.Harrington WJ, Sprague CC, Minnich V, et al.Immunologic mechanisms in idiopathic and neonatal thrombocytopenic purpura. Ann Intern Med,1953,38(3):433-469.
4. Shulman NR, Marder VJ, Weinrach RS. Similarities between known antiplatelet antibodies and the factor responsible for thrombocytopenia in idiopathic purpura. Physiologic,serologic and isotopic studies.Ann N Y Acad Sci,1965,124(2):499-542.
5.Mylvaganam R, Garcia R0, et al.Depressed functional and phenotypic properties of T but not B lymphocytes in idiopathic thrombocytopenic purpura. Blood,1988,71(5):1455-1460.
6. Sayeh E, Sterling K, Speck E, et al.IgG antiplatelet immunity is dependent on an early innate natural killer cell-derived interferon-gamma response that is regulated by CD8+ T cells.Blood, 2004,103(7):2705-2709.
7. Da Wei Huang, Brad T Sherman, Richard A Lempicki.Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.Nature Protocols,2009,4(1):44-57.
8.张鲁勤,葛晋源,等.特发性血小板减少性紫癜患者血浆白细胞介素18及其有关细胞因子.中国实验血液学杂志,2003,11(6)：662-664.
9. Shan NN, Zhu XJ, et al.Interleukin 18 and interleukin 18 binding protein in patients with idiopathic thrombocytopenic purpura. Br J Haematol,2009,144(5):755-761.
10. Aderem A, Ulevitch RJ. Toll-like receptors in the induction of the innate immune response.Nature,2000,406(6797):782-787.
1 Francesco Rodeghiero, Roberto Stasi,et al.Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children:report from an international working group.Blood,2009,113(11):2386-2393.
2 Mylvaganam R, Garcia RO, Depressed functional and phenotypic properties of T but not B lymphocytes in idiopathic thrombocytopenic purpura. Blood,1988,71(5):1455-60.
3 Harrington WJ, Sprague CC, Minnich V, et al.Immunologic mechanisms in idiopathic and neonatal thrombocytopenic purpura. Ann Intern Med,1953,38(3):433-69.
4 Shulman NR, Marder VJ, Weinrach RS. Similarities between known antiplatelet antibodies and the factor responsible for thrombocytopenia in idiopathic purpura. Physiologic, serologic and isotopic studies.Ann N Y Acad Sci,1965,124(2):499-542.
5 McMillan R. Antiplatelet antibodies in chronic adult immune thrombocytopenic purpura:assays and epitopes.J Pediatr Hematol Oncol,2003,25(Suppl 1):S57-61.
6 Sayeh E, Sterling K, Speck E, et al.IgG antiplatelet immunity is dependent on an early innate natural killer cell-derived interferon-gamma response that is regulated by CD8+ T cells.Blood, 2004,103(7):2705-2709.
7 Semple JW, Fre edmanJ. Increased antiplatelet lymphocyte reactivity in patients with autoimmune thrombocytopenia(AITP). Blood,1991,78(10):2619-2625.
8 Ware RE,Howard TA. Elevated numbers of gamma-delta(gamma delta+) T lymphocytes in children with immune thrombocytopenic purpura. J Clin Immunol.1994,14(4):234-237.
9 Shimomura T, Fujimura K, Takafuta T, et al.Oligoclonal accumulation of T cells in peripheral blood from patients with idiopathic thrombocytopenic purpura. Br J Haematol.1996,95(4):732-737.
10 Kuwana M, Kawakami Y, Ikeda Y. Suppression of autoreactive T-cell response to glycoprotein Ⅱb/Ⅲa by blockade of CD40/CD154 interaction:implication for treatment of immune thrombocytopenic purpura[J].Blood,2003,101(2):621-623.
11 Ogawara H, Handa H, et al.High Th1/Th2 ratio in patients with chronic idiopathic thrombocytopenic purpura. Eur J Haematol,2003,71(4):283-288.
12 Olsson B, Andersson P0, JernasM, et al.T-cell mediated cytotoxicity toward platelets in chronic idiopathic thrombocytopenic purpura. NatMed,2003,9(9):1123-1124.
13 Panitsas FP, Theodoropoulou M, Kouraklis A, et al.Adult chronic idiopathic thrombocytopenic purpura(ITP) is the manifestation of a type-1 polarized immune response.Blood,2004,103(7):2645-2647.
14 Stasi R, Del Poeta G, Stipa E, et al.Response to B-cell depleting therapy with rituximab reverts the abnormalities of T cell subsets in patients with idiopathic thrombocytopenic purpura. Blood,2007,110(8):2924-2930.
15 Tag LM, Ezz-Eldeen AM, et al.Serum IL-2 and platelet-associated immunoglobulins are good prognostic markers in immune thrombocytopenic purpura. Egypt J Immunol,2004,11(2):121-132.
16张鲁勤,葛晋源,等.特发性血小板减少性紫癜患者血浆白细胞介素18及 其有关细胞因子.中国实验血液学杂志,2003,11(6)：662-664.
17 Shan NN, Zhu XJ, et al.Interleukin 18 and interleukin 18 binding protein in patients with idiopathic thrombocytopenic purpura. Br J Haematol,2009,144(5):755-761.
18 Webber NP, Mascarenhas JO, et al.Functional properties of lymphocytes in idiopathic thrombocytopenic purpura. Hum Immunol,2001,62(12):1346-1355.
19 Andersson PO, Stockelberg D, Jacobsson S,Wadenvik H. A transforming growth factor-betal-mediated bystander immune suppression could be associated with remission of chronic idiopathic thrombocytopenic purpura. Ann Hematol,2000,79(9):507-513.
20 Langrish CL, Chen Y, Blumenschein WM, et al.IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. J Exp Med,2005,201(2):233-240.
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