心衰患者T细胞CD25水平及卡维地洛的体外影响

目的:观察慢性心衰患者外周血T细胞CD25表达水平,探讨卡维地洛(Car)体外对外周血T细胞CD25表达的作用。方法:分离外周血单个核细胞,用卟啉醇肉豆蔻酸乙脂(PMA)+A23187作为刺激剂,加或不加Car,分组体外培养。以流式细胞仪检测T淋巴细胞活化抗原CD25/CD3。结果:(1)与正常人相比,慢性心衰患者T细胞CD25 表达水平明显升高(P<0.001);(2)与对照组相比,10μMCar可以抑制T淋巴细胞CD25抗原的表达(P<0.05)。结论:慢性心衰患者外周血T细胞CD25表达水平明显升高;Car体外可以抑制其表达。     

免疫和结缔组织疾病

984161系统性红斑狼疮病人T和B淋巴细胞凋亡的观察/石成钢…//上海免疫学杂志一1998.18(2)一1 01~103 观察22例,发现患者淋巴细胞凋亡率在培养O、24、48小时均较正常组显著增高,并以CD才T细胞和CD九B细胞在活动期SLE病人中凋亡更突出。患者自身抗体产生愈多,其细胞凋亡率愈高,疾病活动度增高,凋亡率也较高。提示SLE病人的淋巴细胞凋亡在体外加速.与其自身抗体产生有关,在其发病机制中起一定的作用。同时此对指导治疗有重要意义。表3参8(李伟岚)984162抗神经元和抗心磷脂抗体对中枢型SLE诊断的愈义/张晓…//上海免疫学杂志一1998,18(…     

CD154在活动期系统性红斑狼疮B淋巴细胞中的表达

目的探讨CD154在系统性红斑狼疮(SLE)中的异常表达及其在狼疮发病中的作用机制。方法16例活动期SLE及14名正常健康人,分离外周血CD19阳性B细胞,以流式细胞仪荧光抗体标记检测其CD154的表达,并体外观察抗CD154抗体对外周血B细胞的增生及IgG分泌的影响。结果①活动期SLE外周血B淋巴细胞中CD154阳性率(36±17)%及表达强度(364±238)均显著高于正常健康人,后者分别为(10±8)%,124±97%(P均<0.001);②体外单独培养,活动期SLE外周血B淋巴细胞自身即可异常增生并分泌IgG,[3H]-TdR掺入及体外培养上清液IgG浓度与正常人相比,差异有显著性(P值分别<0.0001和0.001)。抗CD154抗体可显著抑制活动期SLE外周血B淋巴细胞的异常增生及IgG的异常分泌,[3H]TdR掺入及体外培养上清液IgG浓度与对照抗体组相比,差异有显著性(P值分别为0.027和0.034)。结论CD154在活动期SLE的B淋巴细胞中有异常表达,其异常调控可能是导致分泌自身抗体B细胞克隆增生的主要原因。     

系统性红斑狼疮患者CD1的表达与疾病活动性的相关性

目的研究系统性红斑狼疮(SLE)患者外周血CD1的表达与疾病活动性之间的关系。方法用流式细胞仪检测了47例SLE患者外周血单个核细胞CD1c及CD1d的表达及淋巴细胞亚群百分数。结果SLE活动组病人CD1c+及CD1d+细胞百分率显著增高(P<0.05),CD4+细胞百分率显著降低(P<0.01),CD3+、CD8+细胞百分率正常,CD20+细胞数增高(P<0.01)。稳定期病人CD1c+及CD1d+细胞百分率正常,CD4+、CD8+、CD20+细胞百分率均正常。SLE患者CD1c+、CD1d+细胞阳性率与患者SLEDAI的评分有显著的相关性(r=0.68与r=0.66,P<0.01),与抗dsDNA抗体的表达有显著相关性(r=0.36与r=0.41,P<0.05);SLE患者CD1c+细胞阳性率与抗磷脂抗体(ACA)的表达有显著的相关性(r=0.68,P<0.01),与血清C3水平有显著相关性(r=-0.35,P<0.05)。经治疗后CD1c及CD1d的表达明显下降。结论系统性红斑狼疮患者外周血CD1c与CD1d的表达与疾病的活动性明显相关,CD1c、CD1d可能在SLE脂类抗原及核酸类抗原的递呈及抗dsDNA抗体、抗磷脂抗体的产生中起重要作用。     

异基因骨髓间充质干细胞对类风湿关节炎患者淋巴细胞的影响

目的 探讨在体外环境中,异基因骨髓间充质干细胞(BMSCs)对类风湿关节炎(RA)患者T、B淋巴细胞的增殖和功能成熟的影响.方法 采集健康供者的骨髓标本,经密度梯度离心分离、纯化获得其BMSCs,进行体外培养扩增.同时采集RA患者的外周血,分离单个核细胞.将来源于健康供者的BMSCs与来源于RA患者的单个核细胞在体外进行共培养,同时分别加入T淋巴细胞和B淋巴细胞刺激物.分别检测异基因BMSCs对RA患者T、B淋巴细胞增殖的影响;正常BMSCs对RA患者T淋巴细胞增殖周期和凋亡的影响;BMSCs对RA患者外周血T淋巴细胞CD3、CD4、CD8、CD25表达的影响;以及BMSCs对B细胞分泌IgG的影响.结果 正常骨髓来源的BMSCs对RA患者T、B淋巴细胞增殖均有抑制作用,并且这种抑制作用与BMSCs的剂量呈依赖性;与BMSCs共培养组的T细胞主要处于G0/G1期,而进入细胞增殖周期的细胞比例减少,同叶与BMSCs共培养组的凋亡比例(15.2±0.6)%明显低于单纯T细胞活化组(28.2±1.8)%;与BMSCs共培养后CD3+CD4+T细胞表达阳性率(34±6)较对照组(44±7)降低(P<0.05),CD25+的表达下降,但CD4+CD25+调节性T细胞数(4.9±2.3)增加(P<0.05).在SAC刺激下,健康人BMSCs与RA患者外周血淋巴细胞共培养后IgG分泌升高.结论 异基因BMSCs对RA患者T、B淋巴细胞的增殖和功能成熟均有影响,BMSCs可能在RA发病和病情进展中起一定的作用,同时证明利用MSCs来调节RA的免疫功能紊乱进行生物治疗是可行的.     

老年慢性阻塞性肺疾病患者淋巴细胞对肺炎链球菌荚膜多糖的反应能力

目的分析慢性阻塞性肺疾病(COPD)老年人体外培养的淋巴细胞在肺炎链球菌荚膜多糖干预环境下活化情况。方法研究对象分为老年COPD,健康老年,健康年轻3组,分离外周单个核细胞以PPV23干预培养,流式细胞技术检测CD3+细胞、CD19+细胞表面CD69表达。结果健康年轻组淋巴细胞对荚膜多糖抗原干预有显著反应,健康老年组T、B淋巴细胞及老年COPD组B淋巴细胞对多糖抗原干预无反应,老年COPD组T淋巴细胞对多糖抗原反应与健康年轻人组相反。结论 CD69为标志的各组人群T、B淋巴细胞对肺炎链球菌抗原刺激的反应能力不同,COPD老年人及健康老年人进行PPV23免疫后难以产生同健康年轻人相当的免疫效果。     

前列腺素E1抑制慢性乙型重型肝炎树突状细胞成熟及CD4~+/CD8~+T细胞活性的观察

目的研究前列腺素E1对慢性乙型重型肝炎树突状细胞成熟及CD4+/CD8+T细胞活性的影响。方法抽取前列腺素E1治疗10 d前后的慢性乙型重型肝炎患者外周静脉血,密度梯度离心法分离淋巴细胞,贴壁培养获得PBMC,经重组人白细胞介素4(rhIL-4)、重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)、加或不加前列腺素E1刺激培养8 d获得树突细胞(DC)。经流式细胞仪检测DC表达的HLA-DR、CD83、CD86,ELISA测定分泌的IFNγ、IL-12p70。悬浮T细胞部分检测CD4+/CD8+T细胞比例及其细胞表面CD69、HLA-DR表达;部分经IL-2培养,用于检测DC体外诱导的T淋巴细胞毒活性。结果前列腺素E1治疗慢性乙型重型肝炎,对患者CD4+/CD8+T细胞比例无明显影响,可抑制CD4+/CD8+T细胞活化分子CD69、HLA-DR的表达;前列腺素E1体外培养的DC低表达HLA-DR、CD83、CD86,分泌IFNγ、IL-12p70均下降,体外诱导的自体淋巴细胞毒活性减弱,与未加用前列腺素E1比较差异有统计学意义(P<0.01)。结论前列腺素E1治疗慢性乙型重型肝炎,可抑制CD4+/CD8+T细胞活化;体外培养可抑制DC成熟,诱导减弱自体淋巴细胞毒活性。     

失代偿期乙型肝炎肝硬化患者骨髓间充质干细胞对自体外周血淋巴细胞及调节性T细胞亚群增殖的影响

目的研究乙型肝炎肝硬化失代偿期患者骨髓间充质干细胞(BMSCs)对其外周血中淋巴细胞、调节性T细胞亚群(Tregs)增殖的影响。方法 12例乙型肝炎肝硬化失代偿期患者骨髓分离培养出间充质干细胞,采用倒置显微镜观察培养后的贴壁细胞形态、采用流式细胞仪技术观察间充质细胞免疫表型;在植物血凝素(PHA)刺激下、自体血清作为培养基,实施BMSCs和患者自体外周血淋巴细胞共培养,分为A组(BMSCs+淋巴细胞+PHA接触培养)、B组(BMSCs+淋巴细胞+PHA非接触培养)、C组(淋巴细胞+PHA培养)、D组(淋巴细胞),均72 h后观察BMSCs对外周血淋巴细胞增殖及Tregs细胞表达的影响。结果流式细胞仪分析结果显示:培养3代后的间充质干细胞主要高表达T细胞亚群(Tregs)中的CD44(95.26%)、CD29(92.18%),低表达CD34(1.27%)、CD25(1.33%)。C组淋巴细胞增殖率显著高于A、B、D组(P0.05),A、B两组显著高于D组(P0.05),但A、B两组差异无统计学意义(P0.05)。A、B、C组Tregs细胞表达率显著高于D组(P0.05),A、B两组显著高于C组(P0.05),A、B两组差异无统计学意义(P0.05)。结论乙型肝炎肝硬化失代偿期患者BMSCs对其外周血中淋巴细胞增殖具有抑制作用、同时调上调T细胞亚群表达,BMSCs可能具有免疫抑制作用。     

系统性红斑狼疮患者外周血CD4+ CD8+和CD22+淋巴细胞活化分子CD69的表达

目的探讨系统性红斑狼疮(SLE)患者外周血淋巴细胞(PBL) T细胞(CD4+、CD8+)和B细胞(CD22+)活化分子CD69的表达.方法应用双染色流式细胞术检测CD4、CD8、和CD22细胞亚群CD69分子;在植物凝集素(PHA)刺激后20 h淋巴细胞亚群CD69分子的表达.结果①SLE患者PBMC的CD69分子活动期高于静止期(P＜0.001)和正常对照组(P＜0.01)的表达,SLE静止期患者与正常对照组CD69表达差异无显著性(P＞0.05).②进一步分析CD4+、CD8+和CD22+淋巴细胞亚群的CD69的表达,其中,SLE活动期患者CD4+细胞的CD69表达显著高于静止期(P＜0.001)和正常对照组(P＜0.01)的表达,SLE静止期患者与正常对照组CD69表达差异无显著性(P＞0.05);CD8+细胞活动期高于静止期患者(P＜0.05),其余组间差异无显著性(P＞0.05);CD22+B细胞各组间差异无显著性.③PHA刺激20 h后,CD4+、CD22+B细胞的CD69表达,活动期显著高于静止期患者和正常对照组(P＜0.01).结论 SLE患者外周血CD4+T细胞和CD22+B细胞存在着异常的活化,这种淋巴细胞的异常活化是SLE重要的发病机制之一.     

负载HBcAg对慢性HBV感染患者树突状细胞的活化作用及功能影响

目的:探讨免疫耐受期慢性乙型肝炎(CHB)患者外周血树突状细胞(DC)负载HBcAg后细胞表型及免疫功能的改变。方法:从CHB免疫耐受期患者外周血分离培养DC,在DC成熟前,加入重组HBcAg表位肽,诱导HBV特异性DC分化成熟,流式细胞仪检测DC表面共刺激分子CD1a、CD80、CD83的表达水平,应用淋巴细胞增殖试验评估DC功能。结果:负载不同剂量的HBcAg后,DC细胞活化增强,DC细胞共刺激分子标志物CD1a、CD80以及CD83表达率均明显上升,且随着抗原负载剂量的增加,表达率进一步增加,各组之间差异具有统计学意义(均P<0.001);未负载HBcAg的DC细胞激活的淋巴细胞反应较弱,负载HBcAg的DC细胞刺激同种异体健康成人T淋巴细胞增殖能力增强,且随着负载抗原剂量的加大,T淋巴细胞的增殖能力进一步提高,与未负载抗原的对照组相比,差异具有统计学意义(F=428.14,P=0.000)。结论:体外负载HBcAg刺激CHB患者DC细胞可增强其有效抗原提呈能力,促进T淋巴细胞增殖。     

Rituximab-induced serum sickness in refractory immune thrombocytopenic purpura

Serum sickness may occur in patients treated with chimeric monoclonal antibody. Rituximab, an anti-CD20 chimeric monoclonal antibody, is used with increasing frequency in chronic immune thrombocytopenic purpura (ITP). Rituximab is relatively safe; however, serum sickness is reported in 1-20% of patients, more commonly among those with autoimmune conditions. We describe a case of serum sickness in a patient with ITP and review the literature of rituximab-induced serum sickness.     

Sustained remission of platelet counts following monoclonal anti-CD20 antibody therapy in two cases of idiopathic autoimmune thrombocytopenia and neutropenia

Idiopathic autoimmune thrombocytopenia and neutropenia (ITN) is a primary haemocytopenic disorder clinically characterised by recurrent mucocutaneous bleeding episodes and infections. Unlike in simple idiopathic thrombocytopenic purpura, the platelet deficiency of ITN tends to be chronic and difficult to treat. We describe two patients with ITN who obtained sustained remission of their platelet counts after therapy with the chimeric monoclonal anti-CD20 antibody Rituximab. In one of two cases, Rituximab also induced prolonged normalisation of the neutrophil count and disappearance of auto-antibodies. Our observations indicate that disturbed B-cell function plays a central role in the pathogenesis of ITN. Anti-CD20 antibody therapy seems to constitute a safe and efficient alternative to corticosteroids for the management of ITN patients with chronic thrombocytopenia.     

B-cell depletion with rituximab as treatment for immune hemolytic anemia and chronic thrombocytopenia

BACKGROUND AND OBJECTIVES: Rituximab reacts specifically with the CD20 antigen and induces B-cell depletion. This could interfere with the production of autoantibodies in some immune diseases. The objective of this study was to assess the effects of rituximab in autoimmune hemolytic anemia and thrombocytopenia. DESIGN AND METHODS: Seven patients (one with cold agglutinin disease, two with warm antibody autoimmune hemolytic anemia, four with chronic idiopathic thrombocytopenic purpura) previously refractory to conventional treatments were treated with weekly infusions of rituximab, 375 mg/m2, for 4 weeks. Only treatment with steroids, if strictly necessary, was allowed during the period of rituximab administration, but only patients who reached steroid suspension were considered responders. The pharmacokinetics of rituximab were quantified during therapy and the follow-up period. RESULTS: All patients had marked, even if temporary, B-cell depletion. Three patients, 1 with cold agglutinin disease (CAD) and 2 with chronic idiopathic thrombocytopenic purpura (ITP), had a complete hematologic response. In the patient with cold agglutinin disease a decrease in the agglutinin titer was observed. The hematologic improvement was prompt, appearing by the second or third infusion of rituximab. The response duration was CAD 96+, ITP 17+ and 13+ weeks in these 3 patients. Treatment tolerance was satisfactory and no infections or other late events were registered. Serum rituximab concentrations appeared to be similar to those calculated in a historical control group of patients with follicular non-Hodgkin's lymphoma who received rituximab as consolidation of response after first-line CHOP chemotherapy. INTERPRETATION AND CONCLUSIONS: Rituximab appeared to be active and safe in some patients with refractory autoimmune hemolytic anemia and thrombocytopenia. These results, along with data from literature, suggest that this agent may have a therapeutic role in autoimmune diseases.     

Rituximab in the treatment of adult acquired hemophilia A: a systematic review

Rituximab is a monoclonal chimeric antibody to the CD20 antigen, which has proven to be effective in the treatment of non-Hodgkin lymphomas. Recently, rituximab has also been employed in many non-malignant autoimmune disorders (i.e., idiopathic thrombocytopenic purpura, thrombotic thrombocytopenic purpura, connective tissue disorders and autoimmune hemolytic anemia) in which it has been used with the aim of interfering with the production of pathologic antibodies. Moreover, this agent has also shown to be effective in the treatment of acquired antibodies against factor VIII. Through a careful literature search, the current knowledge on rituximab therapy in adult acquired hemophilia A is presented in this review. Although mostly based on uncontrolled studies, the literature data suggest that this drug can be useful in the treatment of disorders of acquired inhibitors to factor VIII. However, large, prospective, randomized trials are needed to confirm these positive preliminary results.     

Should rituximab be used before or after splenectomy in patients with immune thrombocytopenic purpura?

PURPOSE OF REVIEW: The anti-CD20 monoclonal antibody rituximab has been used to treat patients with chronic immune thrombocytopenic purpura. This review discusses whether the optimal timing for this therapy is before splenectomy, or after failure of splenectomy. RECENT FINDINGS: No study has directly compared rituximab to splenectomy in patients with chronic immune thrombocytopenic purpura. Rituximab produces an initial response in approximately 60% of cases, with no significant difference between splenectomized and nonsplenectomized patients. Long-term complete responses are observed in 15-20% of cases. Adverse events related to the drug were usually mild or moderate, with a low incidence of infections. Long-term safety data, however, are still lacking. Deaths have been reported for 2.9% of immune thrombocytopenic purpura cases treated with rituximab, but they could not be attributed to the study drug. SUMMARY: Both the response rate and the response duration appear lower following rituximab than following splenectomy. Although the side effects may be fewer, there is insufficient evidence to support the replacement of splenectomy with rituximab as a second-line treatment of chronic immune thrombocytopenic purpura outside a clinical trial. At the present time, the use of immunotherapy before splenectomy can be recommended only in patients at high risk for splenectomy and in those not willing to undergo surgery.     

Drug insight: the mechanism of action of rituximab in autoimmune disease--the immune complex decoy hypothesis

Inflammatory responses to cell-associated or tissue-associated immune complexes are key elements in the pathogenesis of several autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus and immune thrombocytopenic purpura. Effector cells, such as monocytes, macrophages and neutrophils, bind immune complexes in a process mediated by Fcgamma receptors, and these cells then initiate inflammatory reactions that lead to tissue destruction. Rituximab is an anti-CD20 monoclonal antibody that suppresses inflammation effectively in autoimmune diseases. It was initially approved by the FDA for the treatment of B-cell lymphomas and later for rheumatoid arthritis refractory to anti-tumor necrosis factor therapies. Rituximab is hypothesized to suppress disease injury in autoimmune diseases by promoting rapid and long-term elimination of circulating and possibly lymphoid-tissue-associated B cells. We suggest, however, that a different mechanism may underlie much of the therapeutic action of rituximab in autoimmune diseases: binding of tens of thousands of rituximab-IgG molecules to B cells generates decoy sacrificial cellular immune complexes that efficiently attract and bind Fcgamma receptor-expressing effector cells, which diminishes recruitment of these effector cells at sites of immune complex deposition and, therefore, reduces inflammation and tissue damage.     

Rituximab in cold agglutinin disease

PURPOSE: Cold agglutinin disease is a chronic auto-immune hemolytic anemia related to a lymphoproliferative disorder with a degenerative potential and no codified treatment. This rare affection is related to the production of anti-erythrocytes immunoglobulins M. They are responsible of hemolytic crises sometimes severe and vascular acrosyndrom when submitted to cold temperature. Before rituximab, a monoclonal antibody targeted against the B lymphocyte CD20 antigen, no treatment was really efficient. METHODS: We present 5 patients who have been treated with 4 weekly rituximab perfusions, and then we proceed to a review of the literature concerning the other 23 similar cases. RESULTS: With a good tolerance, the treatment allowed a remission in all the cases (4 partial, 1 complete). Among the 23 observations published, the rate of answer was 21/23 (of which 14 gave completes). CONCLUSION: Rituximab is an alternative treatment of cold agglutinin disease.     

Response to B-cell depleting therapy with rituximab reverts the abnormalities of T-cell subsets in patients with idiopathic thrombocytopenic purpura

Rituximab, an anti-CD20 monoclonal antibody, has been used to treat autoimmune disorders such as idiopathic thrombocytopenic purpura (ITP). However, its mechanisms of action as well as the effects on cellular immunity remain poorly defined. We investigated the changes of different peripheral blood T-cell subsets, the apoptosis profile, as well as the changes of T-cell receptor (TCR) beta-variable (VB) region gene usage of CD4+ and CD8+ T-cell subpopulations following rituximab therapy. The study involved 30 patients with chronic ITP who received rituximab, of whom 14 achieved a durable (> 6 months) response. Compared with the control group, pretreatment abnormalities of T cells in ITP patients included an increase of the Th1/Th2 ratio and of the Tc1/Tc2 ratios (P < .001), increased expression of Fas ligand on Th1 and Th2 cells (P < .001), increased expression of Bcl-2 mRNA (P = .003) and decreased expression of bax mRNA (P = .025) in Th cells, and expansion of oligoclonal T cells with no preferential use of any TCR VB subfamily. These abnormalities were reverted in responders at 3 and 6 months after treatment, whereas they remained unchanged in nonresponders. Our findings indicate that in patients with ITP, response to B-cell depletion induced by rituximab is associated with significant changes of the T-cell compartment.     

Successful treatment of autoimmune hepatitis and idiopathic thrombocytopenic purpura with the monoclonal antibody, rituximab: case report and review of literature.

Rituximab, a chimeric monoclonal anti-CD20 antibody, has shown activity in several autoimmune disorders. We describe a case of a 52 years old female who was diagnosed with idiopathic thrombocytopenic purpura and concomitant autoimmune hepatitis (AIH), both non-responsive to steroids. She was subsequently treated with rituximab, which resulted in a rapid increase in her platelet count and an unexpected normalization of her hepatic biochemical tests. Both her platelet count and her hepatic biochemical tests remained normal for over 5 months. In this case, rituximab showed an impressive clinical response for the treatment of AIH, and it may be considered as an alternative treatment in patients who do not respond to corticosteroid therapy. Prospective randomized studies in AIH are needed to validate this observation.     

Successful treatment of severe thrombotic thrombocytopenic purpura with the monoclonal antibody rituximab

The only established treatment for patients with thrombotic thrombocytopenic purpura (TTP) is plasma exchange against fresh frozen plasma. For cases refractory to plasma exchange, no generally treatment schedule exists. One option is immunosuppressive therapy with corticosteroids and vincristine. Rituximab is a chimeric monoclonal antibody directed against the CD20 antigen, and it has been successfully used in B-cell malignancies and is being investigated in autoimmune diseases. Its efficacy in TTP has not yet been determined. We report two female patients with severe TTP refractory to multiple courses of plasmapheresis, high-dose steroid treatment, and vincristine who responded after adding rituximab while continuing plasmapheresis.