Changes of CD4＋ CD25＋ Regulatory T Cells in Peripheral Blood in Patients with Hepatocellular Carcinoma Before and after TACE

This study investigated the changes of CD4＋ CD25＋ regulatory T cells （Tregs） in periph-eral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoem-bolization （TACE）. The proportion of CD4＋ CD25＋ Tregs among CD4＋ T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups： 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4＋CD25＋ Tregs among CD4＋ T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant （P〈0.01） between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4＋ CD25＋ Tregs among CD4＋ T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE （P〈0.01）, whereas, that in group B was increased significantly 1 month after TACE （P〈0.01）. It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4＋CD25＋ Tregs in peripheral blood. TACE did not significantly affect the level of CD4＋ CD25＋ Tregs within short time （such as 1 week）. The proportion of CD4＋CD25＋ Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.     

Influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells of asthmatic rats

Objective： To investigate the influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells（CD4^＋CD25^＋ Tr） of asthmatic rats, and elucidate the possible mechanism of Danshen Injection in treatment of asthma. Methods： 30 Wister rats were randomly divided into control group, asthma group and Danshen Injection treated group. Bronchoalveolar lavage fluids （BALF） were collected, and cytology studies were conducted. Lung tissues were obtained and pathologic analyses were done with hematoxylin and eosin stain （HE）. Flow cytometry was used to detect the CD4^＋CD25^＋ Tr ratio in peripheral blood mononuclear cells （PBMCs）. Results： Total cell, the percentage of lymphocytes, neutrophils and eosinophils （Eos） in BALF of Danshen Injection-treated group were lower than that in asthma group （P〈0.05, P〈0.01）. Compared with asthma group, less infiltration of inflammatory cells in lung tissues was observed in Danshen Injection-treated group. CD4^＋CD25^＋ Tr of asthma group was lower than that of control and Danshen Injection treated group （P〈0.05）. Conclusion： Danshen Injection can suppress airway inflammation of asthmatic rats, probably by increasing the number of CD4^＋CD25^＋ Tr.     

Involvement of CD4^＋CD25^＋ regulatory T cells in the pathogenesis of polycythaemia vera

Background Regulatory T cells （mreg） have been shown to play an important role in the regulation of hematopoietic activity. However, there is no information about the effect of Treg cells in the pathogenesis of polycythaemia vera （PV）. Methods In this study, we investigated the percentage and function of Treg cells in the peripheral blood of 21 PV patients and 25 healthy donors, mreg cells were identified and characterized as CD4^＋CD25^＋FOXP3^＋ by flow cytometry. The suppressive activity of CD4^＋CD25^＋ Treg cells was assessed by the proliferation and cytokine secretion of the co-cultured CD4^＋CD25^- fractions. Results The results showed that the percentage of Treg cells in the peripheral blood of PV patients significantly increased compared to healthy controls （（10.93±4.02）% vs （5.86±1.99）%, P 〈0.05）. Moreover, the mRNA and protein expression of FOXP3 was higher in CD4^＋CD25^＋ Treg cells. Coordinately, when co-cultured with the activated CD4^＋CD25^- cells, the CD4^＋CD25^＋ Treg cells showed enhanced suppressive function in PV. Yet, the underlying mechanism for the increased frequency and function of CD4^＋CD25^＋ Treg cells is still to be clarified. Conclusion Treg cells expansion might account for the abnormal T cell immunity in PV patients and thus contribute to the pathogenesis of PV.     

Analysis of Epstein-Barr viral DNA load,EBV-LMP2 specific cytotoxic T-lymphocytes and levels of CD4^＋CD25^＋ T ceils in patients with nasopharyngeal carcinomas positive for IgA antibody to EBV viral capsid antigen

Background Epstein-Barr virus （EBV） is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma （NPC）, which is a common cancer in Southeastern Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 （LMP2）-specific cytotoxic T-lymphocytes （CTLs） in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4^＋CD25^＋T cells in such patients.
Methods From February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carriers positive for EBV viral capsid antigen （EBV-IgA-VCA） and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay, real-time polymerase chain reaction （PCR） and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4^＋CD25^＋T cells, respectively.
Results The EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carriers and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carrier samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages Ⅲ or Ⅳ had significantly higher viral loads compared with those at stages Ⅰ or Ⅱ. A significantly higher percentage of CD4^＋CD25^＋ T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC （Ⅲ and Ⅳ） had significantly higher percentages than the patients with early stages （Ⅰ and Ⅱ）. Conclusions Patients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor cells that express immunodominant LMP2 proteins.
Controlling the activity of CD4^＋CD25^＋T cells and elevating CD8^＋ cells specific for LMP2 epitopes could be an effective immunotherapy for patients with NPC.     

Changes of Regulatory T Cells in Graves＇ Disease

The immune mechanism of Graves＇ diseases （GD） and the roles of regulator T cells were investigated. In 32 patients with GD （GD group） and 20 healthy volunteers （control group）, flow cytometry was used to detect the proportion of CD4^＋CD25^＋ cells, MACS to isolate CD4^＋ CD25^＋ cells, RT-PCR to assay the expression of FOXP3, and ELISA to test the leyel of IL-10, respectively. It was found that there was no significant change in the proportion of CD4^＋CD25^＋ T cells between GD group and control group （P〉0.05）, while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group （P〈0.01 and P〈0.05, respectively）. In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.     

The expression of CD40 and CD40L on the surface of peripheral blood mononuclear cells in asthmatic rats and the effect of anti-CD40L McAb on Th1 and Th2 cytokines

Objective： To investigate the expression of CD40 and CD40 ligand （CD4OL） on the surface of peripheral blood mononuclear cells（PBMCs） in asthmatic rats and the effect of anti-CD40L McAb on cytokines of PBMCs. Methods： Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs in asthmatic rats. After the PBMCs was treated with anti-CD40L McAb, ELISA was used to detect the IL-4 and IFN-γ levels of culture supernatants. Results： Compared with the normal control group, the expression of CD40 and CD40L of PBMCs in asthmatic rats increased （P 〈 0.05）. Compared with the untreated group, the level of IL-γ and the ratio of IL-4/IFN-γ decreased after the PBMCs was treated with anti-CD40L McAb（P 〈 0.05）. Conclusion： The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats was up-regulated. Anti-CD40L McAb can rectify the imbalance of Th1 and Th2 cytokines.     

外周血调节性T细胞在儿童与成人重症肌无力发病机制中的作用

目的 初步探讨外周血CD4+CD25+调节性T细胞在重症肌无力(MG)发病中的作用.方法 用流式细胞术分别检测儿童和成人MG外周血CD4+CD25+调节性T细胞的数最及其表达Foxp3和CTLA-4的情况.结果 儿童MG外周血CD4+CD25+调节性T细胞的数量明显高于正常儿童对照组,但其表达Foxp3水平反而大大低于对照组,成人MG比较差异无统计学意义.所有MG患者外周血CD4+CIY25+调节性T细胞表达的表面分子CTLA-4与正常对照比较,差异无统计学意义.结论 儿童MG患者外周血CD4+CD25+调节性T细胞存在着数量改变和功能缺失.提示儿童MG发病可能与外周血调节性T细胞免疫耐受功能受损有关.     

卵巢癌患者CD8+ T细胞中调节性T细胞相关分子标志物的表达率及临床意义

目的 :检测卵巢癌患者组织及外周血CD8~+T细胞中调节性T细胞(regulatory T cells,Treg)相关分子标志物的表达率,探讨其在卵巢癌发生发展中的临床意义。方法:用流式细胞术检测31例卵巢癌患者、31例卵巢良性肿瘤患者和31例健康对照者的外周血,以及14例卵巢癌组织和12例卵巢良性肿瘤组织CD8~+T细胞中Foxp3、CD25、CD28、CTLA-4及GITR的表达率。结果:Foxp3和CTLA-4在卵巢癌组织CD8~+T细胞中的表达率显著高于卵巢良性肿瘤组织(P<0.05);CD28在卵巢癌组织CD8~+T细胞中的表达率显著低于卵巢良性肿瘤组织(P<0.05)。卵巢癌患者外周血CD8+T细胞中Foxp3、CD25、CTLA-4的表达率显著高于卵巢良性肿瘤患者和健康对照者(P<0.05),CD28的表达率显著低于卵巢良性肿瘤患者和健康对照者(P<0.05)。结论:卵巢癌组织及外周血CD8~+Treg所占的比例显著高于卵巢良性肿瘤组及健康对照组,且CD8~+T细胞中Foxp3的表达率可能对了解卵巢癌进展状况有一定帮助。     

CD4+CD25+调节性T细胞在儿童哮喘发病机制中的作用初探

目的探讨哮喘急性发作患儿CD4+CD25+调节性T(Tr)细胞数量变化及影响其发育成熟的因素.方法观察20例哮喘患儿及相同数量同龄对照组.用流式细胞术检测外周血单个核细胞(PBMC)CD4、CD25表面标志及CD4+CD25+细胞内白细胞介素(IL-10)和转化生长因子受体(TGF-β)表达.用逆转录-聚合酶链反应(RT-PCR) 和荧光定量聚合酶链反应(Real-time PCR)检测PBMC Foxp3及SOCS1 mRNA表达.结果急性发作期哮喘患儿CD4+CD25+Tr细胞百分率(6.5%±1.9%)明显低于同龄对照组(12.0%±2.3%),P<0.01 ;CD4+CD25++IL-10及CD4+CD25++TGF-β分泌细胞亦明显低于对照组.Foxp3及SOCS1mRNA表达也显著降低( Foxp30.12±0.05 vs 1.71±0.58,P<0.001;SOCS10.38±0.19 vs 2.14±0.41,P<0.001).结论哮喘患儿CD4+CD25+Tr细胞明显降低可能参与哮喘发病,Foxp3及SOCS1表达降低可能是导致CD4+CD25+Tr细胞发育障碍的重要因素.     

Improvement of quality of life in patients with concomitant allergic asthma and atopic dermatitis: one year follow-up of omalizumab therapy

Objective

Anti IgE treatment with omalizumab is efficacious in the treatment of patients suffering from allergic asthma, improving asthma control and improving quality of life. Furthermore, this approach could be beneficial for patients with concomitant atopic dermatitis. We assessed quality of life and asthma control in atopic patients with allergic asthma and concomitant atopic dermatitis versus those with asthma and without atopic dermatitis treated with omalizumab.

Methods

A total of 22 patients with severe allergic asthma were treated with omalizumab for 12 months. 13 patients with allergic asthma without concomitant atopic dermatitis (IgE 212 ± 224 IU/ml) and 9 patients with concomitant allergic asthma and atopic dermatitis (IgE 3,528 ± 2,723 IU/ml) were included. Asthma-related quality of life (AQLQ), atopic dermatitis related quality of life (DLQI), and asthma-related treatment were compared between both groups at baseline and after initiating omalizumab treatment.

Results

DLQI was significantly in favor of omalizumab after 2 months in the atopic dermatitis/asthma group (P = 0.01); AQLQ was improved after 6 months in the asthma group (P = 0.01), while no change was seen in AQLQ in the atopic dermatitis/asthma group (P = 0.12). Omalizumab controlled oral corticosteroid use more effective (P < 0.01) in patients with asthma and atopic dermatitis (in 9/9 cases) compared to patients with asthma alone (9/13). Baseline IgE as well as other factors do not predict response to omalizumab.

Conclusions

Omalizumab is effective in improving atopic dermatitis-related quality of life scores and modulates oral corticosteroid use in patients with concomitant asthma and atopic dermatitis in a positive fashion.     

哮喘儿童外周血淋巴细胞CD21、CD23及血清IgE测定

目的:研究哮喘儿童外周血淋巴细胞CD21、CD23表达与哮喘发病的关系。方法:采用流式细胞分析技术及免疫化学发光法检测48例哮喘发作期儿童和29例哮喘缓解期儿童外周血CD21+、CD23+、CD21+CD23+淋巴细胞百分率和血清IgE水平,并以30例健康儿童作对照。结果:哮喘发作组CD21+、CD23+、CD21+CD23+淋巴细胞百分率及血清IgE水平均高于哮喘缓解组和正常对照组(P<0.05)。哮喘缓解组CD23+淋巴细胞百分率和IgE水平明显高于正常对照组(P<0.05);哮喘发作组CD21+、CD23+、CD21+CD23+淋巴细胞百分率与血清IgE水平呈正相关(r=0.726,0.739,0.714,P均<0.05)。哮喘缓解组和正常对照组上述指标与血清IgE水平不相关。结论:CD21、CD23的表达在哮喘发病过程中发挥重要作用。     

哮喘患者CD4+T细胞对单核/巨噬细胞抗原递呈功能的影响

目的:探讨支气管哮喘患者CD4⁺T细胞对单核/巨噬细胞抗原递呈功能的影响。 方法：哮喘患者20例,健康对照22例,分别取静脉血5 mL,并 分离单核/巨噬细胞、CD4⁺T细胞和B细胞。每份血样分成4组：MΦ递呈抗原组、CD4⁺T细胞参与MΦ递呈抗原组、CD4⁺T细胞体外活化后对MΦ递呈抗原影响组及自然状态下CD4⁺T细胞对MΦ细胞递呈抗原影响组。各组用CTLL2P抗原刺激18 h后,洗去刺激原,与自体B细胞共同孵育10 d,吸取上清液,测定特异性IgM、IgE、IgG含量。 结果：哮喘患者CD4⁺T细胞与单核/巨噬细胞共同培养递呈抗原时,产生特异性IgM(A₄₉₀ nm)（0.033±0.031）低于健康人（0.098±0.062）（P<0.05）;产生特异性IgG(A₄₉₀ nm)（0.080±0.035）明显高于健康人（0.055±0.024）(P<0.05);产生特异性IgE(A₄₉₀ nm)（0.154±0.119）的调节作用与健康人（0.186±0.101）差异无显著性（P>0.05）;在无抗原刺激的自然状态下,哮喘患者CD4⁺T细胞与递呈抗原单核/巨噬细胞共同培养,也能产生高水平的特异性IgG（哮喘组：0.075±0.018,对照组：0.057±0.018;P<0.05）。 结论：哮喘患者CD4⁺T细胞对单核/巨噬细胞递呈抗原产生的各型免疫球蛋白均有不同程度的调节作用。     

哮喘患儿外周血淋巴细胞CD23、CD40L及IgE检测

目的:研究哮喘儿童白细胞表面分化抗原CD23及CD40L的表达与血清IgE的相关性。方法:哮喘发作儿童 40例,缓解期 25例;肺炎患儿 16例,正常儿童 28例,利用流式细胞分析技术检测外周血CD23、CD40L阳性淋巴细胞的百分率,同时用免疫化学发光法检测血清IgE水平。结果: 哮喘发作组CD23、CD40L的百分率及IgE水平均明显高于哮喘缓解组、肺炎组和正常组(P均<0. 01);哮喘缓解组 3项指标明显高于肺炎组和正常组 (P均 <0. 01);哮喘发作组CD23、CD40L与IgE呈正相关(r分别为 0. 74、0. 81,P均<0. 01)。结论:CD23、CD40L的表达在哮喘发作中发挥重要作用。     

CD4+CD25+调节性T细胞在过敏性哮喘和无症状的过敏者中表达的差异

目的:比较过敏性哮喘患者、无症状的过敏者和不过敏的健康人外周血CD4+CD25+调节性T细胞(Treg)数量的差异及对特异性过敏原刺激的反应.方法:选取急性发作期的尘螨过敏性哮喘患者20例,无症状的尘螨过敏者24例,不过敏的健康人22例,分离外周血单个核细胞(PBMC)并进行CFSE染色,分别在不刺激和1mg/L尘螨抗...