BAFF and BAFF-R of peripheral blood and spleen mononuclear cells in idiopathic thrombocytopenic purpura

BAFF (B-cell activating factor belonging to the TNF family) is an essential component of B-cell homeostasis, and is required for the normal survival and development of B cells. To further explore the role of this cytokine in the pathogenesis of idiopathic thrombocytopenic purpura (ITP), BAFF/BAFF-R (one of receptors of BAFF) expression levels were determined and the correlation between the clinical parameters and the BAFF expression levels was analyzed. A total of 57 patients with ITP were enrolled and 25 age and sex-matched healthy volunteers served as controls. Serum was obtained from 41 patients with ITP and 22 healthy volunteers and was analyzed with a commercial human soluble BAFF (sBAFF) ELISA kit. BAFF and BAFF-R mRNA expression of peripheral blood (PB) (n = 42) and splenocytes (SP) (n = 8) mononuclear cells (MNC) were determined by real-time quantitative PCR. The SPMNC of normal controls came from three hereditary spherocytosis patients who underwent splenectomy. The untreated patients with ITP had higher serum BAFF levels (Median 1430 pg/ml, Range: 534–5787 pg/ml) than those of normal controls (Median 1120 pg/ml, Range: 640–2376 pg/ml, p = 0.006) and treated ITP group (Median 662 pg/ml, Range 267–1265 pg/ml, p = 0.000). On the other hand, serum BAFF levels of treated patients with ITP were lower than those of normal controls (p = 0.001). There was a weak correlation (the Pearson correlation coefficient is ? 0.242) between platelet count and BAFF (p = 0.064). However, BAFF levels did not correlate with platelet associated immunoglobulin or immunoglobulin levels. Moreover, the serum BAFF levels were not statistically different between acute and chronic ITP (p = 0.841). PBMNC of ITP had higher BAFF but not BAFF-R mRNA expression than that of normal controls. BAFF mRNA expression of SPMNC had a positive correlation with BAFF-R in ITP patients but not in PBMNC of normal controls and untreated ITP patients. The BAFF-R mRNA expression of SPMNC was shown to be 15.29 times higher than that of PBMNC in ITP. BAFF might contribute to autoimmunity and disease development in ITP. However, BAFF serum level must be carefully considered as a surrogate marker of disease activity in ITP.     

重症特发性血小板减少性紫癜的治疗体会

目的探讨常规剂量强的松并输注浓缩血小板、大剂量静脉注射丙种球蛋白(IVIG)或大剂量甲基强的松龙(甲强龙)、或二者联合治疗重症特发性血小板减少性紫癜(ITP)的效果。方法根据病情和病人经济能力将118例次病人分为常规剂量的强的松加输注浓缩血小板组(简称血小板组)37例;用大剂量IVIG/甲强龙组(简称药物组)39例;大剂量IVIG/甲强龙联合输注浓缩血小板组(简称联合组)42例。比较3组治疗前、后血小板计数。同时观察临床出血情况。结果治疗后外周血小板计数,血小板组升高(15.40±12.14)×109/L(p<0.001),药物组升高(17.98±14.01)×109/L(p<0.001),联合组升高(43.26±13.20)×109/L,(p<0.001)。联合组高于血小板组和药物组(p<0.001)。治疗3天后临床出血改善率,血小板组60%(22/37),药物组54%(21/39),联合组90%(38/42)。5例发热病人输注浓缩血小板后外周血小板计数未见升高。结论重症ITP病人,大剂量IVIG/甲强龙联合输注浓缩血小板较单纯输注浓缩血小板或大剂量IVIG/甲强龙疗效好,手工采和机单采浓缩血小板的输注无效性的发生率尚待进一步观察。     

特发性血小板减少性紫癜患者T细胞及B细胞亚群变化的研究

目的 探讨特发性血小板减少性紫癜(idiopathic thrombocytopenic purpura,ITP)患者T、B淋巴细胞亚群的变化.方法 从大理州人民医院收集ITP患者和健康者的外周血,用Sysmex血液分析仪及流式细胞术分析T细胞及B细胞亚群.结果 与健康人群比较,ITP患者CD3+T细胞百分率无明显变化,CD4+T细胞百分率降低(42.39％±12.12％,P＜0.05),CD8+T细胞百分率升高(46.93％±11.99％,P＜0.05);CD19+B细胞百分率升高(14.11％±10.28％,P＜0.05),T2B细胞百分率(34.51％±9,70％,P＜0.05)、成熟B细胞百分率(30.91％±7.12％,P＜0.05)及记忆性B细胞百分率(23.31％±8.23％,P＜0.05)增多,差异均有统计学意义.ITP患者T1 B细胞、Kappa+B细胞、Lambda+B细胞百分率与健康人差异无统计学意义.结论 ITP患者外周血T细胞亚群和B细胞亚群分布发生了变化,这种变化在ITP发病中的作用尚需进一步研究.     

Dose-response relationship in the treatment of idiopathic thrombocytopenic purpura with intravenous immunoglobulin

Summary The dose-response relationship in the intravenous immunoglobulin treatment of idiopathic thrombocytopenic purpura was studied in 20 adult patients in a multicenter prospective crossover trial. The rate of response increases from 3 out of 11 (27%) to 6 out of 10 treatment periods (60%) by raising the 7S-IgG dose given on 5 consecutive days from 164.50±24.55 to 359.65±58.62 mg/kg body weight. The onset and duration of response as well as the peak platelet count were found to be independent of the doses. A long-term benefit induced by intravenous immunoglobulin treatment could be achieved in 2 out of 14 patients with chronic idiopathic thrombocytopenic purpura.Abbreviations IgG Immunoglobulin G - ITP Idiopathic thrombocytopenic purpura     

特发性血小板减少性紫癜共刺激分子表达的研究进展

特发性血小板减少性紫癜（ITP）是一种自身免疫性疾病,其发病机制尚未完全明确,目前已证实,ITP患者体内存在T、B淋巴细胞的异常活化和B淋巴细胞依赖Th细胞辅助而产生自身抗血小板抗体。在T、B淋巴细胞相互作用和产生自身抗体的病理过程中,CD80、CD86与其配体CD28、CTLA4结合及CD40与其配体CD40L相互作用提供的共刺激信号起了重要的作用。研究表明共刺激分子过度表达有可能激活自身反应性T淋巴细胞,导致自身免疫病的发生。     

Perisinusoidal fibrosis of the liver in patients with thrombocytopenic purpura

Summary 10 patients with thrombocytopenic purpura (TP) underwent splenectomy. Eight of these patients had idiopathic TP (certain or probable). All had normal liver function tests. Liver histology of the surgical biopsy was normal with the exception of a non specific mild portal infiltration in 6 cases. On Sirius red staining the perisinusoidal network was normal in 3 cases, mildly or moderately increased in 5 cases and often associated with perivenular fibrosis. Collagen types I, III, IV, laminin and fibronectin were increased in the 8 biopsies tested. On semi-thin sections, numerous Kupffer cells were observed. Under the electron microscope, sinusoidal abnormalities were very similar in all 7 patients studied: numerous Kupffer cells containing abundant lysosomes, numerous collagen bundles in the Disse space, active endothelial cells, transformation of some perisinusoidal cells into cells with some of the characteristics of fibroblasts (increased RER) and myofibroblasts (peripheral condensations of the filamentous network), increased fragments of basement membrane-like material. In two cases there was an increase in the number of perisinusoidal cells loaded with lipids. The similarity of the lesions and the absence of other fibrogenic causes (except in 2 cases) suggest that TP may represent another group of diseases with perisinusoidal fibrosis. The aetiology of fibrosis remains unknown but platelet derived growth factor and activated macrophages may play a major role.     

妊娠期特发性血小板减少性紫癜的诊断及治疗方式选择

目的 探讨妊娠期特发性血小板减少性紫癜(GITP)的临床及实验室特点,以提高这种疾病的诊断和治疗水平.方法 回顾性采集了44例GITP患者的诊断治疗经过及相关临床和实验室资料,以同期住院的148例妊娠期血小板减少症(GT),18例妊娠高血压病相关的血小板减少(GHT)以及同一年龄段的42例女性非妊娠ITP患者为对照,进行对比分析和统计学处理.结果 与GT组患者相比,GITP患者发病较早,临床出血倾向较重,初发血小板和病程中最低血小板计数均明显低于前者.GITP组患者血小板相关免疫球蛋白(PAIgG)阳性率高于GT和GHT组.GITP组剖宫产率高于GT组,与GHT组相当.3组患者术中和产后出血的发生率及新生儿Apgar评分异常的比例无显著性差异.GITP组新生儿体重与其他两组比较,差异不显著,而GHT组新生儿体重明显低于GT组.GITP组新生儿血小板减少的发生率高于其它两组,但差别无统计学意叉.GT和GHT孕妇分娩后,血小板数目多可恢复正常.GITP组分娩后也有改善,但血小板变化不如其他两组显著,常需要进一步治疗.和单纯ITP患者相比,GITP患者病情较轻,对糖皮质激素的反应率差别不大.结论 GITP患者病情较GT为重,但处理适当,对母体及新生儿影响不大.因此应早期诊断和密切监测.妊娠本身不会加重ITP 病情,但分娩也不会明显减轻症状,GITP患者分娩后多需要进一步治疗.     

近年血小板减少性紫癜文献分析

本文通过对1995-2003年有关血小板减少性紫癜文献年代分布、期刊分布、著者、文献主题词分布等方面进行统计分析,寻找其研究热点和发展趋势,为临床研究及治疗提供借鉴。     

Polarization and apoptosis of T cell subsets in idiopathic thrombocytopenic purpura

It is well-known that idiopathic thrombocytopenic purpura （ITP） is an acquired organ-specific autoimmune hemorrhagic disease and dysfunctional cellular immunity is considered important in the pathophysiology of ITP. However, polarization patterns and apoptosis profiles of T lymphocytes remain unclear. In this study, we investigated the polarization of T cell subsets, the expressions of apoptotic proteins Fas/FasL on the subsets and the level of anti-apoptotic gene bcl-2 and bax mRNA. It was demonstrated that the ratios of Thl/Th2 and Tcl/Tc2 in ITP children were increased obviously and that the average percentages were increased clearly for Thl and Th2, but not for Tcl and Tc2. In ITP children, the enhancing expressions were detected for FasL on Thl and Tcl and for Fas on Th2 and Tc2. With increasing level of bcl-2 mRNA and decreasing expression of bax mRNA in ITP children, the ratio of bcl-2/bax mRNA was improved obviously, which was positive correlated with the ratio of Thl/Th2. Taken together, our findings indicate that ITP is a Thl predominant disease. This polarization pattern of T cell subsets might be related to the high ratio of bcl-2/bax mRNA and the abnormal expressions of Fas and FasL on T cell subsets.     

间充质干细胞体外对ITP患者T淋巴细胞分泌功能的影响

目的： 体外观察间充质干细胞（MSCs）对特发性血小板减少性紫癜（ITP）患者T淋巴细胞分泌细胞因子功能的影响。方法: 采用Ficoll分离和体外贴壁、传代培养,扩增出骨髓MSCs;通过Ficoll分离法和尼龙棉柱法获取ITP患者外周血T淋巴细胞。以经丝裂霉素（MMC）处理后不同数量（2×103、1×104、5×104 cells/well）的MSCs作为基底层细胞,接种体外分离纯化的异体ITP患者T淋巴细胞,分别于2 d、4 d、6 d后各自收集培养上清,用酶联免疫吸附试验(ELISA)法动态测定T淋巴细胞分泌白细胞介素2(IL-2)、干扰素-γ(IFN-γ)、白细胞介素4(IL-4)、白细胞介素10(IL-10)水平的变化。结果: ITP患者T淋巴细胞分泌细胞因子IL-2、IFN-γ较正常人高(P<0.05),IL-4、IL-10较正常人低(P<0.05)。MSCs可显著抑制ITP患者或正常对照组T淋巴细胞分泌IL-2、IFN-γ(P<0.05),且随MSCs数量的增加,抑制增强(P<0.05),共培养4 d、6 d时作用明显强于2 d时(P<0.05);MSCs可促进ITP患者T淋巴细胞分泌IL-4、IL-10 (P<0.05),且随MSCs量的增加,促进作用增强(P<0.05),对IL-10的作用随时间延长而增强(P<0.05),但对IL-4的作用在培养第2 d、4 d、6 d时无显著差异(P>0.05);在正常对照组,当MSCs数量>1×104可以促进T淋巴细胞分泌IL-4 与IL-10 (P<0.05),且随MSCs数量的增加,作用增强(P<0.05),共培养4 d、6 d时作用明显强于2 d时(P<0.05)。结论: MSCs能够在体外调节ITP患者辅助性T细胞1 (Thl)和辅助性T细胞2 (Th2)反应平衡,可使ITP患者Th1极化状态部分改善。     

特发性血小板减少性紫癜淋巴细胞凋亡及凋亡蛋白表达的研究

目的 :探讨淋巴细胞凋亡及凋亡蛋白表达的变化与特发性血小板减少性紫癜 (ITP)的关系。方法 :采用流式细胞仪检测 2 5例ITP患儿治疗前后外周血淋巴细胞凋亡率和Fas、FasL蛋白表达 ;采用酶联免疫夹心法 (ELISA)测定ITP患儿血清sFas、sFasL的水平。结果 :治疗后血小板恢复正常的患儿淋巴细胞凋亡率 (3 35± 1 4 3)、Fas(32 17± 6 5 4 )和FasL(41 14±6 76 )蛋白表达增加 ,与正常对照组 (0 85± 0 15、2 2 0 4± 4 5 4、2 0 83± 3 75 )和治疗前 (0 92± 0 17、2 3 0 3± 5 95、18 88± 4 5 7)相比差异有显著性意义 (P <0 0 0 1) ;治疗前ITP患者血清sFas含量为 15 5 4± 5 2 6 ,sFasL含量为 0 6 8± 0 2 3,均显著高于正常对照 (5 92± 1 78、0 33± 0 11,P <0 0 0 1)和治疗后组 (6 3± 1 92、0 36± 0 12 ,P <0 0 0 1)。结论 :ITP患者治疗后淋巴细胞凋亡增加可能与其治疗转归有关 ,sFas、sFasL的异常可能参与了ITP的免疫病理过程。     

Platelet factor 4 and the response to plasma exchange in the treatment of thrombotic thrombocytopenic purpura

Summary We report two patients with thrombotic thrombocytopenic purpura who were subjected to plasma exchange. In one case, the plasma levels of platelet factor 4, measured shortly after plasma exchange, increased significantely during plasma exchange. This was followed, however, by a failure to respond to therapy. Repeated plasmapheresis over 3 weeks gave no therapeutic benefit and reversible deep coma occurred. This patient recovered completely after treatment with vincristine. In the second patient, a decline in platelet factor 4 was observed after plasma exchange. This was accompanied by improvement of the patient's condition and a slow rise in platelet count. Plasma exchange was again carried out in this patient because of a recurrence of thrombotic thrombocytopenic purpura 3 years later; again decreased platelet factor 4 plasma levels were observed after plasma exchange and again a therapeutic response followed. Platelet factor 4, therefore, seems to be an effective and early index for the therapeutic benefit of plasma exchange in thrombotic thrombocytopenic purpura.Abbreviations PF4 platelet factor 4 - TG beta-thromboglobulin - PE plasma exchange - TTP thrombotic thrombocytopenic purpura - 5% HSA 5% human albumin     

人类白细胞抗原DRB1 等位基因与儿童特发性血小板减少性紫癜的相关性研究

目的研究人类白细胞抗原(human leukocyte antigen, HLA)DRB1等位基因与儿童特发性血小板减少性紫癜(idiopathic thrombocytopenic purpura, ITP)的关系.方法用聚合酶链反应-序列特异的寡核苷酸探针杂交技术对42例ITP患儿进行了HLA-DRB1等位基因分型,同时用改良的血小板抗原单抗特异性固相化法检测了其中36例ITP患儿血清中的抗GPⅡb/Ⅲa和抗GPIb/Ⅰx自身抗体.结果与健康对照相比,ITP患儿HLA-DRB1*17基因频率显著升高(P<0.05,RR＝2.76,EF=0.1064),而HLA-DRB1*1202基因频率显著降低(P<0.025,RR＝0.20,PF=0.7616);慢性难治性ITP患儿与非难治性患儿相比,HLA-DRB1*11基因频率显著升高(χ2＝6.091,P<0.025),且具有DRB1*11的患儿主要(5/6)为女性年长患儿;抗GPⅡb/Ⅲa及抗GPIb/Ⅰx自身抗体的阳性率都与HLA-DRB1*02(15/16)基因显著相关(P分别为0.02和0.01),但难治性和非难治性ITP患儿间抗体阳性率差异无显著性(P>0.1).结论 DRB1*17可能与儿童ITP的易感性有关,而DRB1*1202则可能对儿童ITP的发病具有保护作用;具有DRB1*11基因的患儿易发展为慢性难治性ITP;血小板自身抗体与抗原表位的反应可能受DRB1*02限制,但自身抗体阳性与否并不能提示ITP患儿的预后.     

Lymphocyte subpopulations and immunoglobulins in idiopathic thrombocytopenic purpura (ITP)

Data gained from observations of the humoral and cellular immunity of 22 patients with ITP (11 in remission and 11 splenectomised) are presented. The amount of T-lymphocytes according to E-rosetting decreased significantly. In tests performed with monoclonal sera the amount of OKT-4 cells was significantly lower then normal, and the ratio of OKT-4/OKT-8 was also reduced. The C-4 complement fraction showed a significant reduction, and a rise was observed in the three Ig classes. No differences could be found, however, between patients in remission and in relapse, and between the splenectomised cases, those treated with prednisolone, and those who had no such treatment. Apart from the primary role of antiplatelet antibodies that cause thrombopenia, a complex disturbance in immune regulation also occurred in ITP, and this manifested with changes in the T-lymphocyte subpopulations. Its pathological progress still remains to be justified, but the plasma of healthy individuals and gammaglobulin have a favorable effect by blocking their receptors and they result in the cessation of the symptom and thrombocytopenia.     

Diffuse histiocytosis of the spleen and idiopathic thrombocytopenic purpura (ITP): histochemical and ultrastructural studies

A case of ITP where the spleen was infiltrated diffusely and extensively by foamy macrophages is described. Among these macrophages were also clusters of cells with the morphologic and tinctorial characteristics of ceroid or "sea-blue" histiocytes. The ultrastructural characteristics of the foamy and ceroid histiocytes are described. Transition of ceroid cells into foamy macrophages is demonstrated, and intermediate forms were observed. It is postulated that the ceroid or sea-blue histiocytes are macrophages that contain platelets at an early stage of digestion, and when the phagocytosis of these platelets is completed, the macrophages become foamy. Therefore, it is suggested that the foamy cells and ceroid or sea-blue histiocytes seen in the spleen in cases of ITP are macrophages that have ingested and metabolized the abnormal platelets.     

Anti-endothelial cell antibodies from patients with thrombotic thrombocytopenic purpura specifically activate small vessel endothelial cells

Thrombotic thrombocytopenic purpura (TTP) is an uncommon disease of an unknown etiology, characterized by consumptive thrombocytopenia, microangiopathic hemolytic anemia, fever and acute thrombotic complications, especially within the cerebral circulation. Although anti-endothelial cell antibodies (AECA) have occasionally been shown to be present in TTP, their role in the pathogenesis of the disease has never been ascertained. In the current study we demonstrated the pathogenic activity of affinity-purified anti-endothelial cell F(ab)2 antibodies (AECA/TTP) from four consecutive patients with active TTP. These AECA/TTP bound to and activated only microvascular endothelial cells (EC) and not large vessel EC. The specificity of AECA/TTP binding to microvascular EC was confirmed by competition assay employing membranes derived from small and large vessels EC. Activation included enhanced IL-6 and von Willebrand factor release from the EC followed by increased expression of adhesion molecules P-selectin, E-selectin and vascular cell adhesion molecule-1 on the EC, as evaluated by ELISA. Increased expression of adhesion molecules was followed by an increase in monocyte adhesion to EC. The level of soluble thrombomodulin (TM) also increased in the culture medium of activated microvascular EC upon exposure to AECA/TTP antibodies and was directly correlated to a decrease in cell-associated TM. Our data suggest that AECA/TTP directed against microvascular EC could play a pathogenic role in the development of endothelial injury in TTP that leads to thrombosis.     

免疫性血小板减少性紫癜患者脾脏T淋巴细胞亚群的变化

目的 探讨免疫性血小板减少性紫癜(ITP)患者脾脏T淋巴细胞亚群的变化及其临床意义.方法 收集2005年12月至2008年5月本科室收治的ITP患者47例,分为激素依赖(SD)组29例和激素抵抗(SR)组18例,以急诊外伤性脾破裂行脾切除术者12例作为对照组.免疫组化法检测脾脏组织标本CD3~+、CD4~+和CD8~+细胞的表达情况,观察脾脏组织动脉周围淋巴鞘(PALS)中染色阳性细胞的百分数,并计算CD4~+与CD8~+细胞的比值.分析3组CD3~+、CD4~+,CD8~+细胞百分数和CD4~+/CD8~+比值的差异. 结果3组间PALS中CD3~+、CD4~+细胞百分数差异均无统计学意义(均P>0.05).对照组PALS中CD8~+T淋巴细胞百分数低于SD组和SR组(28.70±22.19比43.80±20.77,49.27±14.10,均P<0.05),而CD4~+/CD8~+比值高于SD组和SR组(6.27±4.64比0.95±0.93,0.89±0.51,均P<0.05).SD组和SR组PALS中CD8~+细胞百分数及CD4~+/CD8~+比值的差异均无统计学意义(均P>0.05).结论 ITP患者脾脏细胞免疫存在异常.术前激素治疗反应性与脾脏T淋巴细胞亚群百分数的改变无明显关联.     

慢性ITP患者血小板膜表面GPⅣ、GPⅤ特异性自身抗体的测定

目的：检测慢性ITP患者血小板膜表面GPⅣ、GPⅤ特异性自身抗体。方法：应用改良直接MAIPA(单克隆抗体俘获血小板抗原技术)检测血小板膜糖蛋白(GPⅣ、GPⅤ)特异性自身抗体。结果：慢性ITP患者血小板洗脱液中GPⅣ特异性和GPⅤ特异性自身抗体的阳性率分别为6．3％和4．8％。结论：血小板膜GPⅣ、GPⅤ分子是慢性ITP自身抗体的靶抗原，但多与GPⅡb／Ⅲa或GPⅠb自身抗原共存。