RhoA、CD4^(+)CD25^(+)调节性T细胞、MYBL2在胃癌患者中的表达及对预后和生存时间的影响

目的:探讨RhoA、CD4^(+)CD25^(+)调节性T细胞、成髓细胞瘤转录因子第2亚型(MYBL2)在胃癌患者中的表达及对预后和生存时间的影响。方法:选取2017年1月至2019年1月于本院就诊的134例胃癌患者术后癌组织标本作为研究对象,另选取其中62例胃癌患者相应的癌旁组织标本及40例同期非胃癌患者正常胃黏膜组织标本进行对照,检测癌组织、癌旁组织、正常胃黏膜组织中RhoA、CD4^(+)CD25^(+)调节性T细胞、MYBL2的表达情况,分析其对胃癌预后及生存时间的影响。结果:RhoA,CD4^(+)CD25^(+)调节性T细胞、MYBL2在癌组织中的阳性率均明显高于癌旁组织和正常胃黏膜组织(P<0.05)。胃癌患者术后平均生存时间为(28.61±1.34)个月,其中RhoA、CD4^(+)CD25^(+)调节性T细胞、MYBL2阳性患者的生存时间均短于阴性患者(P<0.05)。RhoA(+)、CD4^(+)CD25^(+)调节性T细胞(+)、MYBL2(+)是胃癌患者预后的危险因素(P<0.05)。结论:检测RhoA,CD4^(+)CD25^(+)调节性T细胞、MYBL2的表达可作为胃癌病情严重程度、预后及生存时间评估的重要补充手段。

外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平对早期宫颈癌患者腹腔镜根治术后预后的预测价值

目的分析外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平对早期宫颈癌(CC)患者腹腔镜根治术后预后的预测价值。方法招募2018年9月至2020年9月于儋州市人民医院接受腹腔镜根治术治疗的早期CC患者204例,根据患者术后随访期间预后情况分为预后不良组(43例)和预后良好组(161例)。比较两组临床资料。采用Spearman秩相关分析外周血CD4^(+)CD25^(+)调节性T细胞水平与CD8^(+)CD28^(+)调节性T细胞水平的相关性。采用多因素logistic回归分析早期CC患者腹腔镜根治术后预后不良的影响因素。采用受试者工作特征(ROC)曲线评估外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平对早期CC患者腹腔镜根治术后预后不良的预测价值。结果预后不良组CD4^(+)CD25^(+)调节性T细胞、癌胚抗原(CEA)、糖类抗原125(CA125)水平,术后切缘阳性占比以及术中宫旁浸润占比高于预后良好组,CD8^(+)CD28^(+)调节性T细胞水平低于预后良好组,差异有统计学意义(P<0.05)。Spearman秩相关分析结果显示,早期CC患者外周血CD4^(+)CD25^(+)调节性T细胞水平与CD8^(+)CD28^(+)调节性T细胞水平呈负相关(r_(s)=-0.478,P<0.05)。多因素logistic回归分析结果显示,较高的CEA、CA125、CD4^(+)CD25^(+)调节性T细胞水平是促进早期CC患者腹腔镜根治术后预后不良发生的独立危险因素(P<0.05),较高的CD8^(+)CD28^(+)调节性T细胞水平是抑制早期CC患者腹腔镜根治术后预后不良发生的独立保护因素(P<0.05)。ROC曲线分析结果显示,外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平能有效预测早期CC患者腹腔镜根治术后预后不良(P<0.05),两项指标联合可进一步提高预测效能[AUC(95%CI)=0.939(0.898~0.979),P<0.001],灵敏度和特异度分别为86.00%、88.20%。结论外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平与早期CC患者腹腔镜根治术后预后不良有关,二者能有效预测早期CC患者腹腔镜根治术后预后不良。

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。

An Association between Immunosenescence and CD4^+CD25^+ Regulatory T Cells: A Systematic Review

Objective Age-related increment of the prevalence of CD4^＋CD25^＋ regulatory T （Treg） cells were described controversially, and whether such changes explain immune dysfunction in the elderly is still unclear. The aim of this systematic review is to evaluate the role of the Tregs in immunosenescence. Methods Medline and manual searches were performed to identify all published epidemiological and animal studies investigating the efficacy of the association between immunosenescence and Treg cells. Results It was founded that the frequency, phenotypic characteristics, and number/function of Tregs were altered significantly with aging. Medical conditions in individuals with advanced ageas well as apoptosis intensity of Treg cells had an impact on the accumulation of Tregs which in turn could deteriorate cytotoxic activity of CD8＋ T and NK cells and production of IL-2. The range of immune cells that could be suppressed by Treg cells was quite wide and covered CD4^＋CD25^＋ T cells, NK cells, dendritic cells and even monocytes. These changes were observed both in humans and experimental animals. Besides, it was believed that frequency of Tregs increased with age and was accompanied by intensified suppressive activity for Tregs in patients, for example, with Alzheimer disease （AD） and Parkinson disease （PD）. The impaired condition of CD4＋ T cells, so-called immunosenescence, rendered transplant recipients less responsive to an allogeneic kidney graft, an effect that was limited to transplant recipients who were aged over 60 years. Conclusions Treg cells are associated with immunosenescence. All these changes contribute to the aging-related decline of immune responses and lead to the higher risk of immune-mediated diseases, cancer or infections in aged individuals.

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.

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.

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.

Increase of CD4^+CD25^+ T cells in Smad3^(-/-) mice

AIM： To investigate the changes of lymphocyte subpopulations, especially CD4^＋CD25^ T regulatory cells in Smad3^-/- mice. METHODS： Hematological changes and changes of lymphocyte subpopulations were detected in Smad3＂/- mice using cell counter and flow cytometry, respectively, and compared to their littermate controls. RESULTS： The numbers of neutrophils and lymphocytes in peripheral blood were significantly increased in Smad3^-/- mice compared to littermate controls. CD19^＋ expressing cells in blood and spleen, and CD8^＋ T cells in thymus were all markedly decreased in Smad3^-/- mice. More important, Smad3^-/- mice had an increased population of CD4^＋CD25^＋ T cells in peripheral lymphoid tissues, including thymus, spleen, and lymph nodes. CONCLUSION： These observations suggest that the changes of lymphocyte subpopulations might play a role in susceptibility to inflammation of Smad3^-/- mice.

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.

Effect of CD4^+CD25^+ regulatory T cells in the development of anterior chamber-associated immune deviation

AIM: To investigate whether CD4(+)CD25(+) regulatory T (Treg) cells play a role in the development of anterior chamber-associated immune deviation (ACAID). METHODS: The dynamic changes in the frequency of CD4(+)D25(+) T cells, CD4(+)D25(+) FoxP3(+) T cells and CD4(+)CD25(+) PD-1(+) T cells from spleens of mice with ACAID were analyzed by flow cytometry. Foxp3 mRNA expression in purified CD4(+)CD25(+) T cells was analyzed using real-time PCR. The suppressive effect of purified CD4(+)CD25(+) T cells on the proliferation of CD4(+)CD25(-) T cells was evaluated by [H-3] thymidine incorporation. A blocking experiment was performed to further address the role of CD4(+)CD25(+) T cells in ACAID. The expression of IL-10 in purified CD4(+)CD25(+) T cells was evaluated by ELISA. RESULTS: Increased frequencies of CD4(+)CD25(+) T cells, CD4(+)CD25(+) Foxp3(+) T cells and CD4(+)CD25(+) PD-1(+) T cells were observed in ACAID. The CD4(+)CD25(+) T cells from mice with ACAID showed enhanced suppressive effect on the proliferation of CD4(+)CD25(-) T cells. Treatment of BALB/c mice with anti-CD25 antibody after injection of OVA into the anterior chamber significantly inhibited the induction of ACAID. Furthermore, purified CD4(+)CD25(+) T cells from ACAID mice secreted IL-10. CONCLUSION: Our results demonstrate that Treg cells are induced in the mice undergoing ACAID. These Treg cells may play a role in the development of ACAID.

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.

Immunotherapy of rat glioma without accumulation of CD4^+CD25^+FOXP3^+ regulatory T cells

Immunotherapy may be used for the treatment of glioblastoma multiforme; however, the induced immune response is inadequate when either T cells or dendritic cells are used alone. In this study we established a novel vaccine procedure in rats, using dendritic cells pulsed with C6 tumor cell lysates in combination with adoptive transfer of T lymphocytes from syngenic donors. On day 21 after tumor inoculation, all the rats were sacrificed, the brains were harvested for calculation of glioma volume, cytolytic T lymphocyte responses were measured by cytotoxic assay, and the frequency of regulatory T lymphocytes （CD4＋CD25~FOXP3~） in the peripheral blood was investigated by flow cytometric analysis. The survival rate of rats bearing C6 glioma was observed. Results showed that the co-immunization strategy had significant anti-tumor potential against the pre-established C6 glioma, and induced a strong cytolytic T lymphocyte response in rats. The frequency of peripheral blood CD4＊CD25＊FOXP3＊ regulatory T lymphocytes was significantly decreased following the combination therapy, and the rats survived for a longer period. Experimental findings indicate that the combined immunotherapy of glioma cell lysate-pulsed dendritic cell vaccination following adoptive transfer of T cells can effectively inhibit the growth of gliomas in rats, boost anti-tumor immunity and produce a sustained immune response while avoiding the accumulation of CD4＋CD25＋FOXP3＋ regulatory r lymphocytes.

TF3-H4对CD4^+CD25^+调节性T细胞和CD4^+CD25^-效应性T细胞早期活化的影响

目的观察1',2',3',7'-四氢茶黄素-3,3'-双没食子酸酯(TF3-H4)对CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞早期活化的影响,分析TF3-H4对两群功能相反的CD4+T细胞的作用。方法免疫磁珠分离BALB/c小鼠脾脏CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞,加入ConA或PDB,和TF3-H4共同孵育12 h后收集细胞,流式细胞仪分析细胞表面早期活化标志CD69的表达。结果在ConA和PDB的作用下,两群细胞早期活化标志CD69的表达均升高。TF3-H4(20 mg.L-1)不能抑制PKC激动剂PDB激活的CD4+CD25-效应性T细胞CD69的表达,却能抑制ConA激活的CD4+CD25-效应性T细胞CD69表达;同时,TF3-H4也能明显抑制ConA激活的CD4+CD25+调节性T细胞的CD69表达。结论茶黄素衍生物TF3-H4可能经由TCR活化途径的上游抑制CD4+CD25-效应性T细胞活化;TF3-H4对CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞早期活化的抑制作用,可能是该类化合物发挥抗炎、抗肿瘤作用的机制之一。

人CD4^+CD25^+调节性T细胞系的建立与功能分析

目的 :建立可在体外长期培养的人CD4+CD2 5+调节性T细胞系并研究其免疫生物学特性。方法 :用流式分选的方法从健康人外周血淋巴细胞中得到CD4+CD2 5+T细胞 ,并对其进行体外长期培养、扩增 ;淋巴细胞转化实验分析其免疫抑制功能 ,流式细胞法分析其表型。结果 :经对人CD4+CD2 5+T细胞的长期培养扩增 ,获得具有免疫抑制功能的调节性T细胞系。该T细胞系对经TCR的刺激不敏感 ,且能抑制同一来源或同种异型CD4+CD2 5-T细胞的活化 ,大剂量IL 2可以逆转其抑制功能。长期培养的CD4+CD2 5+T细胞 ,膜表面CD2 5和CTLA 4分子持续高表达 ,而CD4+CD2 5-T细胞CD2 5和CTLA 4分子的表达呈周期性变化。结论 :将CD4+CD2 5+T细胞体外扩增培养成系 ,其功能和表型与同一来源的CD4+CD2

肺癌和乳腺癌患者外周血中CD4^+CD25^+T细胞及foxp3 mRNA的变化特点及临床意义

目的:探讨肺癌、乳腺癌患者外周血中CD4+CD25+调节性T细胞水平的变化及临床意义。方法:应用流式细胞术检测9例肺癌、9例乳腺癌患者及10例非肿瘤患者外周血单个核细胞(PBMC)中CD4+CD25+T细胞占CD4+T细胞的比例,应用RT-PCR技术检测人外周血PBMCfoxp3基因的表达。结果:流式细胞术检测显示,肺癌及乳腺癌患者外周血PBMC中CD4+CD25+T细胞占CD4+T细胞的比例[(21·30±11·26)%]高于非肿瘤患者[(14·19±8·56)%](P<0·05),肺癌及乳腺癌患者外周血PBMC中foxp3mRNA的表达水平高于非肿瘤患者表达水平(P<0·05),肺癌和乳腺癌患者外周血CD4+CD25+调节性T细胞比例增加。结论:CD4+CD25+调节性T细胞在维持自身免疫稳定的同时对肿瘤免疫具有抑制作用,该细胞群比例的增加可能参与肿瘤的发生。

糖皮质激素对哮喘小鼠CD4^+CD25^+调节性T细胞的作用

目的建立哮喘小鼠动物模型并给予吸入糖皮质激素治疗,通过分析CD4+CD25+调节性T细胞及相关细胞因子在哮喘发病中的变化情况,以及观察糖皮质激素治疗对该细胞的影响和干预作用,探讨哮喘发作的新的免疫机制。方法分别制备哮喘组(OVA),哮喘吸入激素治疗组(OVA/GC)及对照组(NS)动物模型。所有小鼠于末次激发后24 h,取右肺中叶标本作病理切片,观察炎症改变;同时收集外周抗凝血及肺组织细胞悬液,通过流式细胞仪及酶联免疫吸附试验(ELISA)测定CD4+CD25+调节性T细胞占CD4+T细胞的百分比和血浆中IL-10和TGF-β1水平。结果①哮喘小鼠外周血及肺组织细胞悬液中CD4+CD25+调节性T细胞百分率明显降低(P<0.01),吸入糖皮质激素治疗后CD4+CD25+调节性T细胞百分率明显升高(P<0.01)。②IL-10在哮喘组较对照组明显降低(P<0.01),糖皮质激素治疗组IL-10明显升高(P<0.01)。③TGF-β1在哮喘小鼠较对照组明显降低(P<0.01),吸入糖皮质激素治疗后TGF-β1有所升高,但差异无显著性。结论①哮喘小鼠CD4+CD25+调节性T细胞和IL-10,TGF-β1数量和分泌水平降低,尤其是肺部CD4+CD25+调节性T细胞不能有效聚集,导致全身和局部对炎症反应的抑制作用减弱,可能是导致哮喘发生的原因之一。②糖皮质激素可通过增加CD4+CD25+调节性T细胞及IL-10数量和分泌水平,增强其免疫抑制功能而发挥治疗作用。③外周血和肺部CD4+CD25+调节性T细胞百分率变化具有一致性,监测外周血CD4+CD25+调节性T细胞变化情况可以了解肺部变化趋势。

鼻咽癌组织与外周血CD4^+ CD25^+调节性T细胞检测及临床意义

目的通过检测鼻咽癌患者肿瘤组织及外周血中CD4+T、CD8+T、CD4+CD25-T、CD4+CD25+T细胞的频数,寻找客观、全面评价鼻咽癌患者免疫状态的临床指标。方法采用流式细胞术检测40例初诊鼻咽癌患者及10例正常对照鼻咽部组织和外周血CD4+T、CD8+T、CD4+CD25-T、CD4+CD25+T细胞比例。结果鼻咽癌患者CD4+T细胞比例及CD4+/CD8+T比值均低于对照组(P<0.05),而CD8+T细胞两组间差异无统计学意义(P>0.05),但是CD4+/CD8+T比值在鼻咽癌组织与外周血间差异无统计学意义(P>0.05)。鼻咽癌组织及外周血中CD4+CD25+T细胞比例都高于对照组(P<0.05),同时癌组织中该细胞比例远远高于外周血(P<0.05)。在鼻咽癌组织中CD4+CD25+T细胞与CD8+T细胞、CD4+CD25-T细胞呈负相关(r分别为-0.70、-0.675,P<0.05),而在外周血中没有相关关系(P>0.05)。在不同T(原发肿瘤大小)组间,T4组的鼻咽癌组织中CD4+CD25+T细胞分别高于T1、T2、T3各组(P<0.05),而在T1、T2、T3各组间差异无统计学意义(P>0.05);鼻咽癌中CD4+CD25+T细胞比例与患者有无淋巴结转移并无关系(P>0.05);鼻咽癌组织中Ⅲ+Ⅳ期组CD4+CD25+T细胞比例高于Ⅰ+Ⅱ期组(P<0.05),而在外周血中两组间差异无统计学意义(P>0.05)。结论CD4+CD25+T细胞与鼻咽癌病程进展无相关性,但是联合检测患者肿瘤组织及外周血中CD4+CD25+T细胞的频数并结合既往CD4+/CD8+T比值会全面反应患者免疫状态,为临床治疗提供依据。

中国健康人外周血中具有CD4^+CD25^(nt/hi)CD127^(lo)特征的调节性T细胞频率

目的:初步确定健康人外周血中具有CD4+CD25nt/hiCD127lo特征的调节性T细胞(Treg)频率,为临床相关疾病的研究及Treg的分选提供参考。方法:采集312名8~60岁(5个年龄组)、不同性别健康人的静脉血,经三重免疫荧光染色,用流式细胞术分析CD4+CD25nt/hiCD127loTreg细胞频率,并观察细胞内Foxp3转录因子的表达。结果:健康人CD4+CD25nt/hiCD127loTreg细胞在外周血中约占CD4+T细胞的(6.55±0.11)%,各年龄组之间有差异(P=0.015),组内性别之间也存在统计学意义(P<0.05);CD25nt/hiCD127lo细胞特异性地表达Foxp3转录因子。结论:初步确定了中国健康人外周血中具有CD4+CD25nt/hiCD127lo表达特征的细胞频率,为Treg细胞的临床研究奠定了基础;CD25nt/hiCD127lo作为CD4+CD25+Treg细胞表面的特征性标志,可在分选时排除其他细胞干扰,获得较完整的Treg细胞。