Depletion of CD4^+CD25^+ regulatory T cells can promote local immunity to suppress tumor growth in benzo[a]pyrene-induced forestomach carcinoma

AIM： To elucidate the distribution of CD4^＋CD25^＋ regulatory T cells （Tregs） in different lymphoid tissues and its local enhancement on tumor growth before and after depletion of CD4^＋CD25^＋ Tregs. METHODS： Female ICR mice were garaged with benzo[a]pyrene （BaP） to induce forestomach carcinoma. CD4^＋CD25^＋ Tregs were intraperitoneally depleted with monoclonal antibody PC61. These mice were divided into BaP-only, BaP ＋ IgG, BaP ＋ PC61, and control groups. The forestomach of mice was dissected for histological analysis, and tunnel test was performed for apoptosis of tumor cells. CD4^＋CD25^＋ Tregs were sorted from different lymphoid tissues and expression of Foxp3, IL-10, and chemokine receptors was analyzed by flow cytometry, semi-quantitative and veal-time polymerase chain reaction. RESULTS： The mice gavaged with only BaP showed increased forestomach papilloma and carcinoma at wk 16 and 32. The proportion of CD4^＋CD25^＋ Tregs was significantly higher in peri-stomach regional lymph nodes than in other lymphoid tissues. These CD4^＋CD25^＋ Tregs in regional lymph nodes expressed higher levels of Foxp3 and IL-10, enriched in the CD62L-subset, and CCR1 and CCR5 chemokine receptors. In mice gavaged with BaP ＋ PC61, the number of tumor nodules and tumor volume decreased significantly with massive infiltrating cells and apoptosis of tumor cells. In the draining regional lymph nodes, the number of CD4^＋CD25^＋ Tregs also decreased significantly. CONCLUSION： Inducible and activated CD4^＋CD25^＋ Tregs in the draining regional lymph nodes suppress host local immunity during tumor growth. Depletion of CD4^＋CD25^＋ Tregs can promote host local immunity to suppress tumor growth.

CD4^+CD25^+CD127^(low/-)调节性T细胞在寻常型银屑病发病机制中的作用

目的:探讨CD4^+CD25^+CD127^(low/-)调节性T细胞(Treg)在寻常型银屑病(PV)发病机制中的作用。方法:利用流式细胞仪检测外周血Treg细胞的数量;MTT法检测Treg细胞对自身CD4^+CD25-T细胞增殖的抑制作用;采用酶联免疫吸附试验法检测血清中白介素-10(IL-10)、转化生长因子-β1(TGF-β1)的含量。结果:PV患者外周血Treg细胞数量与正常对照组差异无统计学意义(P>0.05),但对自身CD4^+CD25-T细胞增殖的抑制百分率降低(P<0.01);PV患者血清IL-10含量较正常对照组低(P<0.01),血清TGF-β1的含量较正常对照组高(P<0.01)。结论:PV患者Treg细胞的功能缺陷及分泌IL-10的降低可能参与PV的发病机制。

半乳凝素-10在CD4^+ CD25^+ CD127^(low/-)调节性T细胞中的表达与意义

目的探讨半乳凝素-10(galectin-10)在CD4+CD25+CD127low/-调节性T细胞(regulatory T cell,Treg)中的表达及其与Foxp3的相关性。方法通过流式细胞术分选健康人外周血和胃癌患者癌旁淋巴结中Treg细胞与CD4+CD25-CD127+T细胞,经微量抽提RNA后,用RT-PCR检测galectin-10和Foxp3的mRNA表达水平。结果galectin-10和Foxp3的mRNA显著表达于Treg细胞,几乎不表达于CD4+CD25-CD127+T细胞。健康人外周血和胃癌患者癌旁淋巴结中的结果一致。结论ga-lectin-10基因高表达于Treg细胞中,可以作为Treg细胞一项新的标志物。

Analysis of CD4^+CD25^+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

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.

Isolation and identification of CD4^+CD25^+ regulatory T cells in rat

Objective： To establish a stable and high efficient method for collection of CD4^＋CD25^＋ regulatory T cells from rats in vitro. Methods： CD4^＋CD25^＋ regulatory T cells were isolated from the rat splenic cells through two steps by magic cell sorting （MACS） system. The first step was negative selection of CD4^＋T cells by cocktail antibodies and anti-IgG magic microbeads, and the second step was positive selection of CD25^＋T cells by anti-CD25 PE and anti-PE magic microbeads. The purity and viability of separated cells were measured by flow cytometry （FACS） and Trypan blue staining. The suppressive ability of seperated cells on the proliferation of CD4^＋CD25^- T cells was assessed by cell proliferation assay. Results： The purity of negatively enriched CD4^＋ T cells was 79%-87% （83.6%±2.5% ） , and the purity of positively enriched CD4^＋CD25^＋ T cells was 86%- 93% （ 90.2±1.8% ） with the viability of 92%～95% （92.8% ± 3.4% ）. The enriched cells significantly suppressed the proliferation of CD4^＋CD25^- T cells in mixed lymphocyte culture （P 〈 0.05）. Conclusion： An effective method can be established for enrichment of CD4^＋CD25^＋ regulatory T cells in two steps by MACS, with satisfied cell purity, viability and function.

CD4^+CD25^(high) Regulatory Cells in Peripheral Blood of NSCLC Patients

The proportion and changes of CD4^＋CD25^high regulatory T cells （Trs） in peripheral blood of non-small cell lung cancer （NSCLC） patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets （CD3^＋, CD4^＋ and CD8^＋） and CD4^＋CD25^high Tr cells. The results showed that the proportion of CD4^＋CD25^high Tr cells in NSCLC group was significantly higher than in control group [（4.36 ±2.07） % vs （2.04±1.03） %, P〈0.01]. The proportion of CD4^＋CD25^ high Tr cells in late stage was higher than that in early stage [stages Ⅰ ＋Ⅱ （2.264±0.6） %; stage Ⅲ（3.284± 1.38） %; stage IV （6.06 4±4.08） %] （P〈0.05）. Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4^＋CD25^high Tr cells in peripheral blood was worse （P=0.0026）. In conclusion, the relative increase in CD4^＋CD25^high Tr cells in peripheral blood may be related to im- munosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4^＋CD25^high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4^＋CD25^high Tr cell therapy to treat NSCLC patients may be an effective strategy.

新风胶囊对类风湿关节炎活动期伴贫血患者外周血CD4^+CD25^+CD127^(lo)调节性T细胞的影响

目的:探讨类风湿关节炎(RA)活动期伴贫血患者外周血CD4+CD25+CD127lo调节性T细胞(Treg)的变化及新风胶囊对其的影响。方法:40例RA活动期伴贫血的患者按随机数字表法分为新风胶囊组(XFC)和正清风痛宁组各20例,均1月1疗程,1疗程后观察疗效;同时设健康志愿者正常对照组20例。采用流式细胞仪检测三组外周血中CD4+CD25+CD127loTreg的数量及其在CD4+T淋巴细胞中所占的比例,并研究其与临床症状积分、疾病活动度指标(DAS28分值)、实验室指标之间的关系。结果:①与正常对照组相比,RA活动期伴贫血患者外周血CD4+CD25+CD127loTreg数量明显下降(P<0.01),活动期伴贫血患者及正常对照组外周血CD4+CD25+CD127loTreg占CD4+T淋巴细胞的百分比分别为(2.75±1.01)%和(4.18±0.89)%,前者明显低于后者(P<0.01)。②RA活动期伴贫血患者CD4+CD25+CD127loTreg表达水平与临床症状积分、DAS28评分、血沉(ESR)、C反应蛋白(CRP)、血小板(PLT)呈负相关(P<0.05),与血红蛋白(HGB)、血清铁(SI)及补体C3、C4呈正相关(P<0.05),而与类风湿因子(RF)、红细胞(RBC)、血清铁蛋白(SF)、转铁蛋白(TF)无明显相关性(P>0.05)。③两治疗组相比,XFC组在总有效率及改善关节症状及部分实验室指标方面与正清风痛宁组相似(P>0.05),但在上调CD4+CD25+CD127loTreg表达水平、降低DAS28分值及升高HGB、降低PLT数量方面显著优于正清风痛宁组(P<0.05)。结论:RA活动期伴贫血患者外周血中CD4+CD25+CD127loTreg表达水平低,并且与疾病活动度及贫血相关指标密切相关,提示该细胞参与了RA的发病和疾病的活动,同时也可能为RA引起贫血的原因之一,XFC可上调CD4+CD25+CD127loTreg表达水平,升高HGB、降低PLT数量,改善RA患者临床症状、改善贫血,可能为XFC治疗机制的重要方面。

Effects of estrogen on CD4^+ CD25^+ regulatory T cell in peripheral blood during pregnancy

Objective To investigate the effects of estrogen(E_2)level on regulatory T cells(Treg)in peripheral blood during pregnancy.Methods:A total of 30 healthy non-pregnant women were selected as control group,90 pregnant women of early,middle and late pregnancy and 30 postpartum women at 1 month after parturition were selected as experimental groups including early pregnancy group,middle pregnancy group and late pregnancy group;the proportions of CD4^+CD25^+Treg and CD4^+CD25^+CD127^-Treg among CD4 T cells were detected by flow cytometry;the serum estrogen content in peripheral blood was detected by electrochemical immune luminescence method.Results:E_2 level was coincident with the change of Tregs number during pregnancy.The estrogen content in peripheral blood increased gradually from early pregnancy to late pregnancy,then decreased significantly after parturition,and the level at 1 month after parturition down to the level in non-pregnancy group(P>0.05);the level of E_2 in pregnancy groups were significantly higher than those in non-pregnancy group(P<0.01);and there were significant differences among early pregnancy group,middle pregnancy group and late pregnancy group(P<0.05).The proportions of CD4^+CD25^+Treg and CD4^+CD25^+CD127^-Treg in pregnancy groups were significantly higher than those in non-pregnancy group(P<0.05),but decreased significantly after parturition,and there was no significant difference between non-pregnancy group and postpartum women group(P>0.05):the proportions in middle and late pregnancy groups were significantly higher than those in early pregnancy group(P<0.05).but decreased slightly in late pregnancy group,there was no significant difference between late pregnancy group and middle pregnancy group(P>0.05).There was correlation between Tregs number with estrogen level during pregnancy.The proportion of CD4^+CD25^+Treg and CD4^+CD25^+CD 127^-Treg were positively correlated with estrogen level.Conclusions:High proportion of CD4^+CD25^+Trcg and CD4^+CD25^+CD127^-Treg is closely related to the high level of E,during pregnancy.It suggested that high level of estrogen may induce an increase of CD4^+CD25^+Treg in peripheral blood.and then influence the immune function of pregnant women.The results of this experiment might play an important role of estrogen in immune-modulation during pregnancy.

大肠癌患者外周血CD_4^+CD_(25)^+CD_(127)^(lo/-)调节性T细胞的含量及临床意义

目的探讨大肠癌患者外周血CD4+、CD25+、CD127lo/-调节性T细胞(Treg)的变化及意义。方法流式细胞术检测36例大肠癌患者及30例健康人外周血中CD4+、CD25+、CD127lo/-Treg的变化;ELISA检测同一标本血清中转化生长因子β(TGF-β)的表达水平。结果36例大肠癌患者外周血中CD4+、CD25+、CD127lo/-Treg占CD4+、淋巴细胞的比例为(6.92±1.31)%,与正常对照组(4.91±1.24)%相比差异有显著性(P<0.05)。20例腺癌、7例黏液癌、9例未分化癌患者外周血中CD4+、CD25+、CD127lo/-Treg比例分别为(6.81±1.52)%、(6.99±1.21)%、(7.08±1.17)%,各组间比较差异无显著性(P>0.05),均高于对照组的(4.91±1.24)%,差异有显著性(P<0.05)。Ⅳ期大肠癌患者外周血CD4+、CD25+、CD127lo/-Treg高于Ⅱ期大肠癌患者(P<0.05)。大肠癌组外周血清中TGF-β水平显著高于正常对照组,随着TGF-β水平增加,CD4+、CD25+、CD127lo/-Treg比例逐渐增高,两者呈正相关。结论大肠癌患者外周血中CD4+、CD25+、CD127lo/-Treg表达增高,与临床分期相关,但与组织学分类无关,导致机体免疫抑制,可能参与大肠癌发生。

胃癌患者外周血CD4^+CD25^+CD127^(lo/-)调节性T细胞的表达及意义

目的:探讨胃癌患者外周血中CD4+CD25+CD127lo/-调节性T细胞(regulatory T cells,Treg)的表达及意义。方法:采用流式细胞术检测26例胃癌患者、30例胃溃疡患者和20例健康志愿者外周血中CD4+CD25+CD127lo/-Treg、细胞毒性T细胞(cytotoxic T lympho-cytes,CTL)和NK细胞,采用ELISA法检测血清中γ-干扰素(interferon γ,IFN-γ)的表达水平。结果:胃癌患者外周血中CD4+CD25+CD127lo/-Treg占CD4+T淋巴细胞的百分比为(6.12±2.08)%,高于胃溃疡患者和健康志愿者(P均<0.05)。胃癌患者外周血中CD4+CD25+CD127lo/-Treg水平与CTL、NK细胞及IFN-γ呈负相关。结论:胃癌患者外周血中具有免疫抑制活性的CD4+CD25+CD127lo/-Treg水平较高,对胃癌患者具有肿瘤免疫抑制作用。

规律血液透析患者外周血CD4^(+)CD25^(+)调节性T细胞的检测与思考

目的探讨CD4^(+)CD25^(+)CD127^(low)调节性T细胞在终末期肾脏病血液透析患者中的变化及临床意义。方法选取2019年1月—2020年12月在包头医学院第二附属医院肾内科初诊的血液透析患者20名(除外各种免疫疾病、感染及肿瘤相关的终末期肾脏病)为观察组,选择同期20名性别、年龄等匹配的健康志愿者为对照组。观察组首次透析前抽取空腹静脉血,并于规律透析治疗6个月抽取透析前的空腹静脉血,对照组在开始试验时、试验6个月分别抽取空腹静脉血,采用流式细胞术测定外周血CD4^(+)CD25^(+)CD127^(low)Treg细胞的百分比,采用酶联免疫吸附法检测IL-10水平,比较观察指标的变化。结果观察组与对照组比较,CD4^(+)CD25^(+)CD127^(low)Treg细胞比例水平及IL-10水平降低,差异有统计学意义(P<0.05)。观察组的血液透析患者透析6个月后,CD4^(+)CD25^(+)CD127^(low)Treg细胞比例水平及IL-10水平较透析前升高,差异有统计学意义(P<0.05);与对照组相比CD4^(+)CD25^(+)CD127^(low)Treg细胞比例水平及IL-10水平降低,差异有统计学意义(P<0.05)。对照组CD4^(+)CD25^(+)CD127^(low)Treg细胞比例2次结果差异无统计学意义(P>0.05)。结论终末期肾脏病患者存在CD4^(+)CD25^(+)CD127^(low)Treg细胞及IL-10比例减少,说明功能紊乱,血液透析有助于改善机体的免疫功能。

血清IL-18、IL-22、CD4^(+)CD25^(+)CD127^(low)Treg水平联合检测对狼疮性肾炎患者活动期的诊断效能

目的:分析血清白细胞介素-18(IL-18)、白细胞介素-22(IL-22)、CD4^(+)CD25^(+)CD127^(low)调节性T细胞(Treg)水平联合检测对狼疮性肾炎患者活动期的诊断效能。方法:回顾性分析2022年1月至2023年9月该院收治的64例狼疮性肾炎患者的临床资料,设为研究组,并根据病情严重程度将其分为活动期、稳定期,另选取同期64名健康体检者设为对照组。比较两组及不同病情严重程度患者血清IL-18、IL-22、CD4^(+)CD25^(+)CD127^(low)Treg水平,并分析各指标水平与狼疮性肾炎患者病情严重程度的相关性,采用受试者工作特征(ROC)曲线分析各指标水平单项及联合检测对狼疮性肾炎患者活动期的诊断效能。结果:研究组血清IL-18、IL-22水平均高于对照组,CD4^(+)CD25^(+)CD127^(low)Treg水平低于对照组,差异有统计学意义(P<0.05);活动期患者血清IL-18、IL-22水平均高于稳定期患者,CD4^(+)CD25^(+)CD127^(low)Treg水平低于稳定期患者,差异有统计学意义(P<0.05);Pearson相关性分析结果显示,血清IL-18、IL-22水平与狼疮性肾炎患者病情严重程度均呈正相关(r>0,P<0.05),CD4^(+)CD25^(+)CD127^(low)Treg水平与狼疮性肾炎患者病情严重程度呈负相关(r<0,P<0.05);ROC曲线分析结果显示,血清IL-18、IL-22、CD4^(+)CD25^(+)CD127^(low)Treg水平单项及联合检测诊断狼疮性肾炎患者活动期的曲线下面积分别为0.828、0.817、0.788、0.921,联合检测诊断效能高于三者单项检测。结论:IL-18、IL-22、CD4^(+)CD25^(+)CD127^(low)Treg水平联合检测诊断狼疮性肾炎患者活动期的效能高于三者单项检测。

Alterations of peripheral CD4^(+)CD25^(+)Foxp3^(+)T regulatory cells in mice with STZ-induced diabetes

Complications arising from abnormal immune responses are the major causes of mortality and morbidity in diabetic patients.CD4^(+)CD25^(+)T regulatory cells(Tregs)play pivotal roles in controlling immune homeostasis,immunity and tolerance.The effect of hyperglycemia on CD4^(+)CD25^(+)Tregs has not yet been addressed.Here we used streptozotocin(STZ)-induced diabetic mice to study the effects of long-term hyperglycemia on CD4^(+)CD25^(+)Tregs in vivo.Four months after the onset of diabetes,the frequency of CD4^(+)CD25^(+)Foxp3^(+)T regulatory cells was significantly elevated in the spleen,peripheral blood lymphocytes(PBLs),peripheral lymph nodes(pLNs)and mesenteric LNs(mLNs).CD4^(+)CD25^(+)Tregs obtained from mice with diabetes displayed defective immunosuppressive functions and an activated/memory phenotype.Insulin administration rescued these changes in the CD4^(+)CD25^(+)Tregs of diabetic mice.The percentage of thymic CD4^(+)CD25^(+)naturally occurring Tregs(nTregs)and peripheral CD41Helios1Foxp31 nTregs were markedly enhanced in diabetic mice,indicating that thymic output contributed to the increased frequency of peripheral CD4^(+)CD25^(+)Tregs in diabetic mice.In an in vitro assay in which Tregs were induced fromCD4^(+)CD25^(+)T cells by transforming growth factor(TGF)-b,high glucose enhanced the efficiency of CD4^(+)CD25^(+)Foxp3^(+)T inducible Tregs(iTregs)induction.In addition,CD4^(+)CD25^(+)T cells from diabetic mice were more susceptible to CD4^(+)CD25^(+)Foxp3^(+)TiTreg differentiation than those cells from control mice.These data,together with the enhanced frequency of CD4^(+)CD25^(+)Foxp3^(+)T iTregs in the periphery of mice with diabetes,indicate that enhanced CD4^(+)CD25^(+)Foxp3^(+)T iTreg induction also contributes to a peripheral increase in CD4^(+)CD25^(+)Tregs in diabetic mice.Our data show that hyperglycemia may alter the frequency of CD4^(+)CD25^(+)Foxp3^(+)T Tregs in mice,which may result in late-state immune dysfunction in patients with diabetes.

INVESTIGATION OF REGULATORY T CELLS IN PATIENTS WITH NON-SMALL CELL LUNG CANCER

Objective To evaluate the prevalence of CD4 ^＋ CD25^high regulatory T cells （ Treg cells） in the peripheral blood mononuclear cells （PBMC） and tumor-infiltrating lymphocytes （TIL） of patients with non-small cell lung cancer （NSCLC） and to investigate immunosuppression to the progression of cancer. Methods Peripheral blood and tumor tissues were collected from 20 patients with NSCLC at the time of surgery. None of the patients received surgery, radiotherapy, chemotherapy, or other medical interventions before this study. Cancer stages of the patients were Ⅰ-Ⅲ A. Venous blood samples were obtained from 20 health donors. PBMC were isolated from blood samples by differential centrifugation over Ficoll-Hypaque. TILs were isolated from tumors by differential centrifugation over Ficoll-Hypaque and Percoll. Percentage of CD4^＋ CD25^highTr/CD4＋T in PBMC and TIL was assessed by the flow cytometry. Results The percentage of CD4^ ＋ CD25high Tr/ CD4 ^＋T in PBMC [ （4. 87 ± 1.22 ） % ] of NSCLC patients was significantly higher than that in healthy donors [ （ 2.36 ± 0. 72 ） % ] （ P 〈 0.01 ）. The percentage of CD4^＋ CD25^highTr/ CD4^＋ T in PBMC [ （5.40 ± 1.20） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （3. 87 ± 0. 22 ） % ] （ P 〈 0. 01 ）. The percentage of CD4 ＋ CD25hiShTr/ CD4 ＋ T in TIL[ （ 8. 66 ±0. 76） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （ 7. 04 ± 0. 80） % ] （ P 〈 0. 01 ）. Conclusion The prevalence of CD4 ^＋ CD25^highTreg cells in PBMC and TIL of NSCLC patients was significantly higher than that in healthy donors. These Treg cells may be preventing appropriate antitumor immune responses. The population of CD4^ ＋ CD25^highTreg cells in PBMC and TILs of NSCLC patients with Ⅱ-Ⅲ A stage was significantly higher than that of NSCLC patients with Ⅰ stage. These Treg cells may facilitate development of tumors.

CD4^＋ CD25^ ＋CD127^low/-调节性T细胞在乳腺癌患者外周血的表达及临床意义

目的探讨CD4^＋ CD25^ ＋CD127^low/-调节性T细胞（regulatory Tcells，Treg）、T细胞亚群在乳腺癌患者外周血的表达及临床意义，并进行手术前、手术后、TNM分期的对比分析。方法应用流式细胞术检测57例乳腺癌患者、60例正常妇女外周血CD4^＋ CD25^ ＋CD127^low/-调节性T细胞和淋巴细胞亚群表达百分率。其中乳腺癌患者于手术前、手术后两个时间点分别采血。结果乳腺癌患者、健康志愿者外周血CD4’CD25’CDl27““。调节性T细胞占CD4^＋淋巴细胞的比例分别为（6．92±1．51）％、（4．38±1．31）％。乳腺癌患者CD4^＋ CD25^ ＋CD127^low/-调节性T细胞表达高于健康对照组，差异有统计学意义（P〈0．05），CD3^＋CD4^＋细胞低于健康对照组，差异有统计学意义（P〈0．05），NK细胞（CD3-／CD16＋56＋）明显降低，差异有统计学意义（P〈0．05）。Treg细胞与CD3^＋CIM^＋细胞、NK（CD3-/CD16^＋56^＋）细胞呈明显负相关。乳腺癌患者手术前外周血CD4^＋ CD25^ ＋CD127^low/-调节性T细胞水平升高，手术后外周血Treg细胞水平降低，CD3^＋CD4^＋细胞比例升高，NK细胞比例增高。不同TNM分期乳腺癌患者外周血Treg表达比例不同，Ⅳ期CD4^＋ CD25^ ＋CD127^low/-Treg表达比例明显高于Ⅰ、Ⅱ期，疾病越严重，CD4^＋ CD25^ ＋CD127^low/-Treg表达水平越高。结论CD4^＋ CD25^ ＋CD127^low/-调节性T细胞对乳腺癌患者具有肿瘤免疫抑制作用，可作为评估乳腺癌患者疾病严重程度的一项指标。

肺癌患者外周血CD4^＋CD25^HiCD127^Lo调节性T细胞检测的临床意义

目的探讨肺癌患者外周血中CD4^＋CD25^HiCD127^Lo调节性T细胞（Treg）占CD4^＋淋巴细胞比例的变化及其意义。方法采用流式细胞术检测肺癌患者30例（鳞癌20例、腺癌10例）及健康人（对照组）20名外周血中Treg的变化；ELISA法检测同一标本血清中自细胞介素10（IL-10），转化生长因子-β（TGF—β）的表达水平。结果鳞癌、腺癌患者外周血中Treg占CD4^＋T淋巴细胞的比例分别为（6．81±1．52）％、（7．08±1．17）％，两组间比较差异无统计学意义（P〉0．05），二者均高于对照组的（4．91±1．24）％，差异有统计学意义（P〈0．05）；肺癌组血清中IL—10水平为（24．36±4．02）ng／ml，高于健康对照组的（21．53±4．30）ng／ml；肺癌组血清中TGF—β水平为（218．49±35．23）ng／ml，高于健康对照组的（124．31±20．32）ng／ml。结论肺癌患者外周血中Treg比例明显升高，可能是导致免疫抑制的原因之一，并且可能通过分泌IL-10和TGF—β,发挥免疫抑制的效应。

Biological features of intrahepatic CD4^(+)CD25^(+)T cells in the naturally tolerance of rat liver transplantation

The biological features of intrahepatic CD4^(+)CD25^(+)T regulatory cells in the naturally tolerance of rat liver transplantation were explored.Orthotopic liver transplan-tation was performed in two allogeneic rat strain combina-tions,one with fatal immunosuppression despite a complete major histocompatibility complex mismatch.The subjects were divided into three groups according to different donors and recipients[Tolerance group:LEW-to-DA;Rejection group:DA-to-LEW;Syngegnic group(control group):DA-to-DA].The proportion of intrahepatic CD4^(+)CD25^(+)T cells from three groups was determined by flow cytometry(FCM)in different time.The intrahepaitc CD4^(+)CD25^(+)T cells were isolated by magnetic activated cell sorting(MACS)method and identified by FCM.The Foxp3 mRNA was detected by reverse transcriptase polymerase chain reaction(RT-PCR).And their suppression on the proliferation of CD4^(+)CD25^(-)T effector cells was analyzed by cell proliferation assay in vitro.Beginning immediately after transplantation,the proportion of Treg cells increased over time in both allogeneic groups but was significantly greater in the Rejection group.The pro-portion of Treg cells declined after day 5,and such reduction was more dramatic in the Rejection group than in the Tole-rance group.Animals in the Tolerance group showed a second increase in the proportion after day 14.Intrahepatic CD4^(+)CD25^(+)T cells isolated from spontaneous tolerance models inhibited the proliferation of mixed lymphocyte reaction.The purity of CD4^(+)CD25^(+)Tcells sorted by MACS was 86%–93%.The CD4^(+)CD25^(+)T cells could specifically express the Foxp3 gene compared with CD4^(+)CD25^(-)T cells.In vitro,the spleen cells from LEW rats can irritate the proliferation of CD4^(+)CD25^(+)T cells more obviously than the syngegnic spleen cells.CD4^(+)CD25^(+)Tr cells could suppress the proliferation of CD4^(+)CD25^(-)T cells,but the inhibition was reversed by exo-genous IL-2(200 U/mL).The CD4^(+)CD25^(+)T regulatory cells specifically express the Foxp3 gene,which may play animpor-tant role in the induction of liver transplantation tolerance by suppressing the reaction of effective T cells.

CD4^＋CD25^＋ regulatory T lymphocytes in tuberculous pleural effusion

Background Active suppression by CD4^＋CD25^＋ regulatory T lymphocytes plays an important role in the down-regulation of T cell responses to foreign and self-antigens. This study was conducted to analyze whether the CD4^＋CD25^＋ regulatory T cells exist and function normally in tuberculous pleural effusion. Methods The percentages of CD4^＋CD25^＋ T cells in pleural effusion and peripheral blood from patients with tuberculous pleurisy and peripheral blood from healthy control subjects were determined by flow cytometry. The expression of forkhead transcription factor Foxp3 was also examined. CD4^＋CD25^＋ and CD4^＋CD25^-T cells from pleural effusion and blood were isolated, and were cultured to observe the effects of CD4^＋CD25^＋ T cells on proliferation response of CD4^＋CD25^- T cells in vitro. Results There were increased numbers of CD4^＋CD25^＋ T cells in tuberculous pleural effusion compared with peripheral blood from both patients with tuberculous pleurisy and normal subjects, and these cells demonstrated a constitutive high-level expression of Foxp3. Moreover, CD4^＋CD25^＋ T cells mediated potent inhibition of proliferation response of CD4^＋CD25^- T cells. Conclusion The increased CD4^＋CD25^＋ T cells in tuberculous pleural effusion express a high level of Foxp3 transcription factor, while potently suppressing the proliferation of CD4^＋CD25^- T cells.