大剂量地塞米松治疗初诊免疫性血小板减少性紫癜的疗效观察

目的：观察大剂量地塞米松、常规剂量泼尼松两种方案治疗初诊免疫性血小板减少性紫癜（ITP）的临床疗效和不良反应.方法：45例初诊ITP分为两组,治疗组20例,地塞米松40 mg/d,静脉滴注,连用4d;对照组25例,泼尼松1 mg·kg^-1 ·d^-1,晨起顿服,3～4周后若血小板稳定,则开始减量,减量至10 mg/d时维持治疗4～6个月.结果：治疗后第4天治疗组与对照组血小板计数值为（35±18.2）×10^9/L和（16 ±6.4）×10^9/L,两组比较差异有统计学意义（P＜0.05）;治疗组和对照组总有效率分别为85％和76％,两组差异无统计学意义（P＞0.05）;长期反应率分别为52.6％和31.8％,两组比较差异有统计学意义（P＜0.05）;对照组所有病例皆出现不同程度类库欣氏表现;2例继发血糖升高;3例继发感染.治疗组不良反应较对照组明显减轻.结论：大剂量地塞米松治疗初诊ITP血小板上升速度快,长期反应率高,不良反应小,可作为临床一线方案.     

特发性血小板减少性紫癜外周血单个核细胞Toll样受体表达研究

目的了解特发性血小板减少性紫癜(ITP)外周血单个核细胞(PBMCs)Toll样受体(TLRs)的mRNA表达情况。方法采用RT-PCR方法(SYBR GreenⅠ荧光染料结合法),对30名ITP患者和30名健康者的PBMCs中所表达的TLR2、4、6、7、9 mRNA进行相对定量检测。结果ITP组TLR2和TLR7的mRNA相对表达量均明显高于对照组(P<0.05),而TLR4、TLR6、TLR9的mRNA相对表达量两组间无明显差异(P>0.05)。结论TLR2和TLR7在ITP患者中表达水平上调,可能在ITP发病机制中起重要作用。     

树突细胞及其Toll样受体4在特发性血小板减少性紫癜发病中的作用

目的 研究特发性血小板减少性紫癜(ITP)患者树突细胞(DC)免疫表型及其Toll样受体4(TLR4)的表达,探讨其在ITP发病机制中的作用.方法 取ITP完全缓解患者及未达完全缓解患者治疗前后及正常人外周血,分离单个核细胞,加入细胞因子rhGM-CSF及rhIL-4诱导DC,用流式细胞术检测DC表型;酶联免疫吸附法检测DC培养上清液中IL-12p70.应用RT-PCR检测DC的TLR4表达.结果 21例ITP完全缓解患者治疗前DC的CD80和CD86阳性表达率分别为(51.60±13.47)%和(61.50±15.93)%,15例未达完全缓解者CD80和CD86分别为(53.29±19.49)%和(62.91±18.43)%,均高于正常对照的(36.03±15.43)%和(40.28±11.49)%(P＜0.01).治疗后ITP缓解患者组CD80和CD86表达率分别降为(36.48±15.19)%和(44.05±17.70)%,与治疗前比较差异有统计学意义(P＜0.01),与对照组相比差异无统计学意义(P＞0.05);未完全缓解组CD80和CD86分别降为(52.30±20.98)%和(49.79±20.28)%,但与治疗前相比差异均无统计学意义(P＞0.05),CD80仍高于正常对照组(P＜0.05),而CD86与对照组比较差异亦无统计学意义(P＞0.05).地塞米松(DXM)治疗缓解组患者治疗前DC培养上清IL-12p70为(67.52±14.43)pg/ml,明显高于正常对照的(39.78±10.03)pg/ml(P＜0.01),治疗后IL-12 p70的表达降至(43.90±8.49)pg/ml,与治疗前比较差异有统计学意义(P＜0.01),与正常对照组相比无明显差异;而未缓解组患者DC培养上清IL-12p70则由治疗前的(65.35±12.52)pg/ml降至(48.45±9.68)pg/ml(P＜0.01),但仍高于正常对照(P＜0.05).治疗缓解组ITP患者DC的TLR4 mRNA相对表达水平为0.69±0.17,明显高于正常对照组的0.31±0.09(P＜0.01),治疗后ITP患者DC的TLR4 mRNA相对表达水平降为0.35±0.11(P＜0.01),与正常对照组相比无明显差别;未缓解组DC的TLR4 mRNA相对表达水平由治疗前的0.65±0.09降至治疗后的0.52±0.21(P＜0.01),但后者仍高于正常对照组(P＜0.01).结论 DC可能通过其Toll样受体及细胞因子的分泌在ITP发病中起重要作用.     

免疫性血小板减少症患者树突状细胞的分布特点及其活化状态的研究

免疫性血小板减少症（ITP）患者树突状细胞（DC）存在异常。本研究旨在分析ITP患者DC的分布特点及其活化状态,以进一步明确ITP患者DC异常的机制。将ITP患者分为初诊纽、难治组、有效组（完全反应＋有效）,流式细胞术检测各组患者外周血、骨髓和／或脾脏中髓系树突状细胞（mDC）,浆细胞样树突状细胞（pDC）的分布比例,各部位mDC、pDC细胞分化程度、成熟mDC和pDC抗原CD80、CD86的表达。结果表明,各组患者各部位mDC比例均较pDC高,难治组各部位mDC、pDC比例较初诊组、有效组高,且难治组中脾脏mDC比例明显高于其他部位;各组pDC比例各部位差别不明显;各组患者各部位mDC、pDC的CD86的表达较CD80高,各组mDC、pDC的CD80的表达比例均较低,各部位差别不明显;难治组各部位mDC、pDC的CD86表达较初诊组、有效组高,且难治组中脾脏mDC的CD86表达明显高于其他部位。结论：ITP患者mDC、pDC的分布存在异常,脾脏mDC分布比例较高,分化较成熟,脾脏mDC可能与ITP发病较为密切。     

大剂量地塞米松对特发性血小板减少性紫癜患者B细胞活化因子及调节性T细胞的影响

目的 研究大剂量地塞米松治疗前后特发性血小板减少性紫瘕(ITP)患者B细胞活化因子(BAFF)和调节性T(Treg)细胞的变化.以及BAFF对Treg细胞的作用.方法 用ELISA方法 检测21例慢性活动性ITP患者治疗前后血浆BAFF水平;用流式细胞术检测患者治疗前后及外周血单个核细胞培养体系中Treg细胞的比例.结果 治疗前ITP患者的BAFF水平[(599.70±199.40)pg/ml]较正常对照[(454.50±132.50)pg/ml]明显升高(P<0.05),Treg细胞比例明显降低[(1.56±6.73)%](P<0.01);大剂量地塞米松治疗后,ITP患者的BAFF水平[(296.9±119.7)pg/ml]较治疗前明显下降(P<0.01),且低于正常对照组(P<0.01);Treg细胞比例明显升高[(5.94±2.22)%](P<0.01),且高于正常对照[(4.08±1.08)%](P<0.01).ITP患者BAFF水平和Treg比例与血小板计数无相关性(P>0.05).体外实验显示,rhBAFFO组和rhBAFF20组Treg数量无明显改变(P>0.05).结论 慢性ITP患者的BAFF水平增高,Treg细胞数量减少.大剂量地塞米松可降低患者的BAFF水平,增加Treg细胞的数量.BAFF对Treg细胞无明显影响.     

特发性血小板减少性紫癜患者调节性T细胞和Toll样受体4的分析

目的 探讨调节性T细胞和Toll样受体4(TLR4)在特发性血小板减少性紫癜(ITP)发病机制中的作用.方法 应用流式细胞术检测60例ITP患者和30例正常人外周血中CD<,4>+CD<,25>+T细胞及CD<,80>的表达水平.结果 与正常对照组相比,ITP患者外周血中CD<,4>+CD<,25>+T细胞及CD<,80>的表达水平差异有统计学意义(P<0.05).结论 CD<,4>+CD<,25>+T细胞及CD<,80>可能在ITP免疫发病机制中有重要作用.     

雷公藤内酯醇对系统性红斑狼疮患者树突状细胞Toll样受体-9表达的影响

目的:检测雷公藤内酯醇对体外培养的系统性红斑狼疮(SLE)患者树突状细胞(DC)Toll样受体-9(TLR9)表达的影响。方法:体外培养SLE患者DC,用终浓度分别为20、40、60μg/L的雷公藤内酯醇刺激细胞3、6、12h,混合淋巴细胞反应观察DC刺激T细胞后T细胞增殖的变化,RT-PCR检测TLR9 mRNA在DC中的表达情况。结果:与健康对照者相比,SLE患者DC表达CD11c+、CD123+的百分率明显下降(P<0.05);雷公藤内酯醇以剂量依赖的形式抑制SLE患者DC刺激同种T细胞增殖;以剂量和时间依赖的形式下调SLE患者DCTLR9 mRNA表达。结论:雷公藤内酯醇可抑制SLE患者DC刺激同种T细胞增殖,雷公藤内酯醇可能通过下调SLE患者DCTLR9的表达而发挥免疫抑制作用。     

大剂量地塞米松冲击疗法治疗小儿重症血小板减少性紫癜

采用大剂量地塞米松冲击疗法治疗小儿重症特发性血小板减少性紫癜(ITP) 18例 ,报告如下。1　病例与方法1.1　病例 :ITP患者诊断符合文献〔1〕标准 ,并经外周血及骨髓等检查确诊。根据临床情况 (表 1)分为 2组。1.2　方法 :确诊后治疗组立即给予地塞米松 2 mg/kg静注 ,每日 1次 ,共 3～ 5日 ,同时给予利血生、氨肽素、止血敏等常规治疗 ,并每日查血小板计数 (BPC)。对照组予泼尼松 2 mg· kg- 1· d- 1口服 ,4～ 8周 ,其它治疗同治疗组。表 1　 2组患儿临床情况组别 例数(例 )年龄(x± s,岁 )病程(x± s,d)出血情况 (例 )皮肤口腔鼻衄血…     

大剂量丙种球蛋白联合地塞米松治疗血小板减少性紫癜患儿的疗效观察

目的探讨大剂量丙种球蛋白联合地塞米松治疗血小板减少性紫癜患儿的临床疗效。方法选取2016年8月～2018年11月我院收治的血小板减少性紫癜患儿116例,依照治疗方案不同分为观察组与对照组各58例。对照组予以地塞米松治疗,观察组予以大剂量丙种球蛋白+地米塞松治疗。对比两组疗效、临床恢复情况及治疗前后T淋巴细胞亚群水平。结果观察组总有效率93.10%(54/58)高于对照组79.31%(46/58)(P<0.05);观察组出血控制、住院、血小板升高及恢复正常时间短于对照组(P<0.05);治疗后,观察组CD3+、CD4+水平较对照组高(P<0.05)。结论大剂量丙种球蛋白联合地塞米松治疗血小板减少性紫癜患儿效果显著,可改善患儿免疫功能,缓解出血症状,促进血小板恢复,缩短患儿恢复时间。     

Management of patients with refractory immune thrombocytopenic purpura

In immune thrombocytopenic purpura (ITP), thrombocytopenia is a result of both increased platelet destruction and insufficient platelet production. In adults, the course is commonly chronic, but most patients never experience serious bleeding even with severe thrombocytopenia. In case series of consecutive adult patients identified at the time of diagnosis, the frequency of death from bleeding is low, < 1%. The goal of treatment is only to prevent bleeding, not to correct the platelet count to normal. All current treatments are designed to diminish the increased platelet destruction, either by immunosuppression or splenectomy. The frequency of death from complications of treatment is similar to the frequency of death from bleeding. Perhaps because of increasing recognition of both the infrequent occurrence of serious bleeding and the risks of immunosuppressive treatment and splenectomy, data from case series across the past 30 years suggest a trend toward less therapy and fewer splenectomies among patients with ITP. However treatment is necessary for patients with severe and symptomatic thrombocytopenia. Splenectomy remains the most effective treatment for ITP, with two-thirds of patients achieving durable complete remissions. Immunosuppressive agents, including rituximab and combinations of agents, may be less effective than splenectomy in achieving complete remissions and the remissions may also be less durable. New agents for patients with ITP are currently in development that enhance platelet production, rather than diminish platelet destruction. In preliminary reports, these agents have been effective in maintaining safe platelet counts in patients with chronic ITP that was refractory to splenectomy and other treatments.     

Decreased plasma cytokines are associated with low platelet counts in aplastic anemia and immune thrombocytopenic purpura

Summary. Background: We previously found plasma levels of CD40 ligand (CD40L), chemokine (C‐X‐C motif) ligand 5 (CXCL5), chemokine (C‐C motif) ligand 5 (CCL5) and epidermal growth factor (EGF) to be low in aplastic anemia (AA) patients and to be correlated with platelet count. Objectives: To study the association of CD40L, CXCL5, CCL5 and EGF with platelets. Methods: We measured cytokines in the plasma of immune thrombocytopenic purpura (ITP) and AA patients using the Luminex assay and confirmed the results in a mouse model and in vitro experiments. Results: Both ITP and AA showed similarly low levels of CD40L, CXCL5, CCL5 and EGF, compared with healthy controls. In ITP, levels of these proteins were significantly greater in patients with higher platelet counts than in those with lower platelet counts. In a murine thrombocytopenia model, levels of CD40L, CXCL5, CCL5 and EGF decreased with platelet count after immune‐mediated destruction, while the cytokine levels increased when the platelet count recovered. In vitro, concentrations of these cytokines in the supernatants of platelet suspensions were proportional to platelet numbers, and levels in sera prepared by simple blood coagulation were equivalent to those in platelet‐rich plasma‐converted sera. mRNA expression for CXCL5, CCL5 and EGF was higher in platelets than in megakaryocytes, peripheral blood mononuclear cells, granulocytes and non‐megakaryocytic bone marrow cells. Conclusions: Plasma CD40L, CXCL5, CCL5 and EGF are mainly platelet‐derived, suggesting a role of platelets in immune responses and inflammation. Measurement of CD40L, CXCL5, CCL5 and EGF in human blood allowed testable inferences concerning physiology and pathophysiology in quantitative platelet disorders.     

Combination of high-dose dexamethasone and antiretroviral therapy rapidly improved and induced long-term remission of HIV-related thrombocytopenic purpura

We present a case of HIV-related thrombocytopenic purpura (HIV-ITP) successfully treated with high-dose dexamethasone and antiretroviral therapy (ART). Although high-dose dexamethasone is regarded as the first-line therapy in adult patients with non-HIV ITP, there is limited information on treatment of HIV-ITP and long-term prednisone therapy is considered the standard therapy. High-dose dexamethasone is preferable to conventional long-term prednisone therapy, because of fewer side effects mainly due to shorter steroid use. The ART helps achieve long-term remission for HIV-ITP, although this therapy lacks an immediate effect. In our patient, administration of high-dose dexamethasone resulted in rapid rise in platelet count and ART maintained long-term remission of HIV-ITP. The combination therapy is potentially suitable strategy for the treatment of patients with HIV-ITP and severe thrombocytopenia or bleeding.     

Splenic macrophages maintain the anti-platelet autoimmune response via uptake of opsonized platelets in patients with immune thrombocytopenic purpura

Summary.  Background:  Immune thrombocytopenic purpura (ITP) is an autoimmune disease primarily caused by IgG anti-platelet autoantibodies. Activation of autoreactive CD4⁺ T cells upon recognition of cryptic GPIIb/IIIa peptides presented by antigen-presenting cells (APCs) is a critical step for triggering and maintaining the pathogenic anti-platelet autoantibody response. Objectives:  We investigated which APCs carry the cryptic peptides of GPIIb/IIIa that activate autoreactive CD4⁺ T cells in ITP patients. Methods:  GPIIb/IIIa-reactive T-cell lines generated from ITP patients were cultured with autologous freshly isolated splenic macrophages, B cells or dendritic cells. To further investigate how the macrophages presented the antigenic GPIIb/IIIa peptides, we prepared macrophages from the peripheral blood monocytes of the same patients during remission. Results:  Macrophages induced the proliferation of GPIIb/IIIa-reactive T-cell lines without an exogenous antigen, but B cells and dendritic cells required GPIIb/IIIa peptides to stimulate the T cells. Macrophages derived from peripheral blood during remission required an exogenous antigen to induce the GPIIb/IIIa-reactive T-cell line response, but could elicit a response without added antigen if they were preincubated with platelets from ITP patients with platelet-associated anti-GPIIb/IIIa antibodies or healthy platelets pretreated with ITP platelet eluates. The T-cell response was inhibited by anti-FcγRI antibody. Finally, cultured macrophages that captured opsonized platelets promoted anti-GPIIb/IIIa antibody production in mixed cultures of autologous GPIIb/IIIa-reactive T-cell lines and B cells. Conclusions:  Splenic macrophages that take up opsonized platelets via FcγRI are major APCs for cryptic GPIIb/IIIa peptides, and are central to the maintenance of anti-platelet autoantibody production in ITP patients.     

慢性特发性血小板减少性紫癜患者在氟美松或静脉注射人免疫球蛋白治疗后体内细胞因子谱的变化

目的研究慢性特发性血小板减少性紫癜（CITP）患者经氟美松或静脉注射用人免疫球蛋白治疗后辅助性T细胞（Th）细胞因子谱的变化。方法对CITP、系统性红斑狼疮伴有血小板减少（SLETP）、再生障碍性贫血（AA）患者及正常对照分别用ELISA及RIA方法测定细胞因子谱。用RT—PCR检测辅助性T细胞相关的转录因子c-Mar及T-bet。结果经氟美松和／或静脉注射人免疫球蛋白治疗的CITP患者均取得了良好的反应,体内IL-2水平显著低于正常对照。但是Th2细胞因子IL-4水平显著高于正常对照。用PHA对CITP患者外周血单个核细胞（PBMC）刺激后,IL-13的分泌量明显增高,而正常对照PBMC受刺激后IL-13水平很低。结论CITP患者经氟美松／静脉注射人免疫球蛋白治疗后倾向于Th2细胞因子分泌,这可能是患者获得病情缓解的机制之一。     

Coexistence of thrombotic thrombocytopenic purpura and immune thrombocytopenic purpura in an Asian woman: a case report

A 33-year-old Chinese woman with a history of immune thrombocytopenic purpura presented with heavy menstrual bleeding. She was found to have thrombocytopenia, plasma ADAMTS13 activity of 0%, and positivity for the plasma ADAMTS13 inhibitor. She was diagnosed with the coexistence of thrombotic thrombocytopenic purpura and immune thrombocytopenic purpura. The patient was treated by plasmapheresis, a glucocorticoid, and rituximab. Her platelet level returned to normal, and she was discharged 28 days after admission. The number of plasmapheresis sessions and the timing of rituximab administration may be the key aspects of management of patients with thrombotic thrombocytopenic purpura who have underlying immune dysfunction caused by diseases such as immune thrombocytopenic purpura.     

Modulation of immune response with cytotoxic T-lymphocyte-associated antigen 4 immunoglobulin-induced anergic T cells in chronic idiopathic thrombocytopenic purpura.

BACKGROUND: Platelet glycoprotein (GP)-reactive CD4+ T cells are essential for the stimulation and maintenance of antiplatelet autoantibody production in chronic idiopathic thrombocytopenic purpura (ITP). Blocking costimulatory signals could result in platelet-specific T-cell anergy. METHODS: GP-specific CD4+ T cells from patients with ITP were made anergic using cytotoxic T-lymphocyte-associated antigen 4 immunoglobulin (CTLA4-Ig). The CTLA4-Ig-induced GP-specific anergic T cells were investigated for their inhibitory function on GP-reactive T-cell proliferation and antibody production with in vitro culture systems. To further analyze their tolerizing mechanisms, we cocultured GP-anergic T cells with dendritic cells (DCs) from patients with ITP. RESULTS: Our studies demonstrated that the anergized GP-specific T cells have profound effects on both GP-specific T-cell proliferation and antibody production. These anergic T cells exerted their suppressive effects mainly in a cell contact-dependent manner, and they were not constitutively suppressive but required specific antigen stimulation to make DCs tolerogenic. The anergic T-cell-modulated DCs could induce the autoreactive T cells to be tolerant, and this effect was not restricted to T cells of the same specificity. CONCLUSION: Our studies demonstrate the efficacy of CTLA4-Ig in suppressing the pathologic autoimmune responses in ITP. These findings provide new insights into the underlying mechanisms of anergy induction in chronic ITP.     

特发性血小板减少性紫癜患者免疫状态分析

目的 探讨特发性血小板减少性紫癜(ITP)患者免疫状态的异常改变.方法 以52例ITP确诊患者为观察组,其中慢性ITP(CITP)组27例、急性ITP(AITP)组25例;以52例体检健康者为对照组.检测各组受试对象外周血T淋巴细胞亚群、血清免疫球蛋白及补体、红细胞免疫功能,并对结果 进行统计学分析.结果 观察组CD3+、CD4+、CD4+/CD8+、红细胞C3b受体花环检出率均低于对照组,CD8+、IgA、IgG、IgM、C3、C4、免疫复合物花环检出率高于对照组(P＜0.05);CITP组变化幅度大于AITP组(P＜0.05).结论 ITP患者免疫状态呈异常状态,且与疾病严重程度等有较大的相关性.     

Plateletpheresis for postsplenectomy rebound thrombocytosis in a patient with chronic immune thrombocytopenic purpura on romiplostim

Immune thrombocytopenic purpura (ITP) is an autoimmune disease in which IgG‐coated platelets are removed from circulation by the spleen, and platelet production is impaired due to increased thrombopoietin (TPO) clearance. Romiplostim, a novel TPO‐mimetic agent, is approved for patients with ITP that are unresponsive to traditional treatments. However, there is little experience when using this drug before splenectomy. We describe herein the case of a young female with chronic ITP who was treated with romiplostim, underwent splenectomy shortly thereafter, and required plateletpheresis for postoperative rebound thrombocytosis with concomitant neurologic symptoms. J. Clin. Apheresis 28:321–324, 2013. © 2013 Wiley Periodicals, Inc.