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.

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.

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.

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.

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.

Changes of CD4^+CD25^+ Regulatory T Cells in Patients with Acute Coronary Syndrome and the Effects of Atorvastatin

The function of CD4＋CD25＋ regulatory T lymphocytes （Treg） in patients with acute coronary syndrome （ACS） and the effects of atorvastatin were investigated. Forty-eight patients with ACS were randomly divided into two groups： group C receiving conventional therapy （n=24）, and group C＋A receiving conventional therapy＋atorvastatin （10 mg/day, n=24）. T lymphocytes from ACS patients （before and 2 weeks after the treatment） or 18 healthy subjects were separated and the flow cytometry was used to measure the percentage of Treg. The inhibitory ability of Treg on effector T cells was determined by mixed lymphocyte reaction （MLR）. ELISA was used to measure the serum levels of cytokines （IL-10, TGF-β1 and IFN-γ） before and after treatment. The results showed that as compared with normal control group, Treg percentage was decreased significantly （P〈0.01）, the inhibitory ability of Treg on the T lymphocytes proliferation was reduced （P〈0.01）, IFN-γ levels were increased and IL-10 and TGF-β1 levels were lowered in ACS patients. After treatment with atorvastatin, Treg percentage and the inhibitory ability of Treg on T lymphocytes proliferation were significantly increased in ACS patients. Serum IFN-γ was decreased significantly, while IL-10 and TGF-β1 were elevated significantly as compared with the non-atorvastatin group. The number of Treg was positively correlated with serum TGF-β1, but negatively with serum IFN-γ and CRP. It was concluded that ACS was associated with decreased number and defected function of Treg, which may play an important role in initiating immune-inflammatory response in ACS. The inhibitory effects of atorvastatin on inflammation in ACS may be due to its beneficial effects on Treg and restoration of immune homeostasis.

葡萄膜炎患者外周血CD4^＋CD25^high调节性T细胞的表达

目的:通过检测葡萄膜炎患者外周血中CD4+CD25high调节性T细胞的表达水平,初步探讨其在葡萄膜炎发病中的作用。方法:取16例正常健康对照及30例不同类型活动期葡萄膜炎患者,采取抗凝外周血,进行三色免疫荧光染色、流式分析。结果:葡萄膜炎患者外周血中CD4+CD25highT细胞的百分率与正常健康对照(8.56%±4.22%vs5.72%±3.11%)相比,显著升高(P=0.022),而且与患者病情严重程度明显相关(P=0.007)。另外CD4+CD25highCTLA4+T细胞在葡萄膜炎患者外周血中比例较正常对照明显升高(P=0.012)。结论:葡萄膜炎患者外周血中CD4+CD25highTreg细胞的异常表达可能参与疾病的发生发展,并与疾病的活动密切相关。

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.

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.

肺癌患者CD4^+ CD25^(high) Foxp3^+调节性T细胞的格局变化及意义

目的:研究肺癌患者外周血(PBMC)及肿瘤浸润淋巴细胞(TIL)中CD4+CD25h ighFoxp3+调节性T细胞(Treg)的比例改变,探讨其在抗肿瘤免疫中的调节作用。方法:分离肺癌患者PBMC及TIL,FACS分析CD4+/CD8+T细胞的比值及CD4+CD25h ighT细胞占CD4+T细胞的比例。Real-tim e PCR检测Treg特异性转录因子Foxp3基因在PBMC及TIL中的表达。结果:肺癌患者PBMC及TIL中CD4+/CD8+比值降低;而CD4+CD25h ighT细胞在CD4+T细胞中所占比例升高;Foxp3基因仅在TIL中高表达,而在PBMC中低或不表达,表明肿瘤局部的CD4+CD25h ighT细胞主要是CD4+CD25h ighFoxp3+Treg。结合临床资料分析显示Treg在肺腺癌比例较高。结论:CD4+CD25h ighFoxp3+Treg在肺癌患者肿瘤浸润淋巴细胞中明显升高,可能与其通过细胞与细胞间接触抑制CD8+T细胞的杀伤效应,最终发挥免疫抑制效应相关。

非霍奇金淋巴瘤患者外周血CD4^+CD25^(high)调节性T细胞研究

本研究分析B细胞非霍奇金淋巴瘤(B-NHL)患者外周血T细胞亚群及CD4+CD25high调节T(Tr)细胞比例及其变化规律,初步探讨B-NHL免疫抑制机制,以及化疗对此免疫抑制功能的影响。采用流式细胞术检测42例初治B-NHL患者、36例化疗后达CR/PR患者及15例正常健康者(对照组)外周血CD3+、CD4+、CD8+和CD4+CD25highT细胞的比例。结果显示:B-NHL患者CD3+及CD4+T细胞比例、CD4/CD8比值均低于正常对照组,它们依次为(68.33±15.27)%,对照(72.06±9.26)%;(34.47±12.84)%,对照(42.45±9.2)%;1.36±0.26,对照1.92±0.20,但CD4+CD25highTr细胞比例明显高于正常对照组,为(4.10±1.21)%,对照(2.04±1.03)%,(P<0.001)。化疗后组CD4/CD8比值低于化疗前组(P<0.05);而外周血CD4+CD25highTr细胞比例,显著高于化疗前组和对照组(P<0.01)。结论:B-NHL初治化疗前及化疗后患者外周血CD4+CD25highTr细胞比例都显著升高,提示CD4+CD25highTr细胞可能是B-NHL患者免疫抑制的重要原因之一。

淋巴结核患者外周血CD4^+CD25^(high)FoxP3^+调节性T淋巴细胞以及血浆IFN-γ和IL-10水平及其临床意义

目的研究淋巴结核患者外周血CD4+CD25highFoxP3+调节性T淋巴细胞(Treg)和干扰素γ(IFN-γ)、白细胞介素10(IL-10)水平的变化及临床意义。方法采用流式细胞仪对100例淋巴结核患者及30名正常人外周血CD4+CD25highFoxP3+Treg和IFN-γ、IL-10水平进行检测。结果淋巴结核患者组与正常对照组比较,外周血CD4+CD25highFoxP3+Treg和IL-10水平均升高(P<0.05),而IFN-γ水平则降低(P<0.05)。在干酪样型、增殖型和混合型3种类型的淋巴结核病之间各指标差异均无统计学意义(P>0.05)。结论淋巴结核患者细胞免疫功能明显异常。CD4+CD25highFoxP3+Treg和IFN-γ、IL-10在淋巴结核的发病过程中发挥了重要作用。

不同病程寻常型银屑病患者外周血中CD4^+CD25^(high)调节性T细胞的比较与分析

目的:检测和分析不同病程寻常型银屑病患者外周血CD4+CD25high调节性T细胞(CD4+CD25highTreg)水平特点和意义。方法:选取进行期(70例)、稳定期(64例)和消退期(38例)寻常型银屑病患者,知情同意后,取外周抗凝全血,分离单一核细胞,采用流式细胞术检测不同病程寻常型银屑病患者外周血CD4+CD25highTreg的水平,并统计分析其特点与意义。结果:进行期、稳定期和消退期寻常型银屑病患者组外周血CD4+CD25highTreg与CD4+淋巴细胞的百分比分别为(2.86±0.9)%、(3.95±0.6)%和(4.77±0.1)%。稳定期和消退期寻常型银屑病患者组明显高于进行期组(t’=8.312,P<0.01;t=10.126,P=0.000),而消退期组明显高于稳定期组(t’=4.588,P<0.01)。值得注意的是,消退期寻常型银屑病患者组与正常对照组比较,差异显著,消退期组仍低于正常对照组(t=2.281,P=0.0264)。结论:不同病程寻常型银屑病患者外周血CD4+CD25highTreg水平存在显著差异,提示CD4+CD25highTreg与寻常型银屑病发生和病程发展关系密切。本研究为深入研究寻常型银屑病免疫学发病机制做出了初步的尝试与探索。

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.

脐带血、成人外周血中CD4^+CD25^(high)调节性T细胞分布的比较

目的比较成人外周血及新生儿脐带血中CD4+CD25+调节性T细胞(Tr细胞)的数量。方法采集成年人外周血和新生儿的脐带血,以荧光素分别标记的抗CD4/抗CD25/抗CD3和抗CD4/抗CD8/抗CD3mAb染色后,流式细胞术进行分析。结果成人外周血中CD4+CD25highTr细胞的数量为(9.09±3.05)个/μL,占CD3+CD4+T细胞的(1.40±0.37)%。新生儿脐带血中CD4+CD25-T细胞和CD4+CD25highTr细胞的绝对数及其占CD3+CD4+T细胞的百分率,均明显高于成人外周血(P<0.01);而CD4+CD25intT细胞的绝对数及其占CD3+CD4+T细胞的百分率,则明显低于成人外周血(P<0.01)。结论新生儿脐带血中CD4+CD25highTr细胞、CD4+CD25intT细胞及CD4+CD25-T细胞的数量明显有异于成人外周血。

非小细胞肺癌外周血CD4^+CD25^(high)调节T细胞分析

目的:分析非小细胞肺癌(NSCLC)患者外周血CD4+CD25high调节T细胞(Tr)比例及变化规律,初步探讨其临床意义。方法:采用流式细胞仪检测61例非小细胞肺癌患者和15例正常健康者(对照组)外周血中CD3+、CD4+、CD8+及CD4+CD25highTr细胞比例,并进行分层分析。结果:NSCLC患者CD4+CD25highTr细胞比例明显高于正常对照组,分别为(4.36±2.07)%、(2.04±1.03)%,(P<0.01)。而且,随疾病进展外周血CD4+CD25highTr细胞比例逐渐升高,分别为Ⅰ+Ⅱ期(2.26±0.6)%;Ⅲ期(3.28±1.38)%,(P<0.05);Ⅳ期(6.06±4.08)%,(P<0.05)。Kaplan-Meier生存分析表明,外周血CD4+CD25highTr细胞比例较高者预后较差(P=0.0026)。结论:NSCLC患者外周血CD4+CD25highTr细胞比例升高,提示与肺癌免疫逃逸有关。去除这群细胞可有效诱导肿瘤免疫,为NSCLC治疗提供一种新的方法。

抗CD25单克隆抗体对CD^4+CD25^(high)调节性T细胞的影响

目的评价抗CD25单克隆抗体对活体肾移植受者外周血CD4+CD25high调节性T细胞(Treg)的影响。方法观察14例活体肾移植受者外周血CD4+CD25highT细胞百分比和FoxP3mRNA表达水平的变化,应用流式细胞仪FACSCalibur和CellQuest软件行双色流式细胞计数和结果分析,半定量逆转录-聚合酶链反应(RT-PCR)对肾移植受者外周血单个核细胞(MNC)的FoxP3基因水平进行检测。结果在移植术后3d、13d,CD4+CD25highT细胞占CD4+比例显著升高;在移植后1个月内,CD4+CD25highT细胞比例升高约至原来的3倍。移植术后13d、17d、27d时外周血FoxP3mRNA表达明显高于移植术后3d。在肾移植术后的不同时间抗CD25单克隆抗体使用组受者的CD25+T细胞百分比明显低于对照组(未使用抗CD25单克隆抗体)。抗CD25单克隆抗体使用组与对照组两组间比较,术后3d、13d,抗CD25单克隆抗体组CD4+CD25highT细胞占CD4+的比例明显降低,术后3d对照组、抗CD25单克隆抗体使用组分别为1.24%±0.15%、2.25%±0.24%,P=0.00;术后13d为3.75%±0.38%、7.28%±0.51%,P=0.00。然而,术后17d、27d后,抗CD25单克隆抗体使用组的CD4+CD25highT恢复到对照组水平,两组间CD4+CD25highT细胞占CD4+的比例无统计学差异,即第17d为3.72%、0.26%、4.43%、0.44%,P=0.17,27d为9.00%±0.30%、8.31%±0.33%,P=0.16。在肾移植术后不同时间,受者外周血FoxP3mRNA表达水平在两组之间相似,无统计学差异,说明FoxP3mRNA表达水平并没有受到体内注射抗CD25单克隆抗体影响。结论抗CD25单克隆抗体的使用对活体肾移植受者外周血CD4+CD25high调节性T细胞在一定时间具有暂时性的影响,而FoxP3mRNA表达水平没有受到影响。

CD4^+ CD25^(high)调节性T细胞在异基因造血干细胞移植后的重建及其与aGVHD的相关性

目的:研究CD4+CD25high调节性T细胞在异基因造血干细胞移植后的重建情况及其与aGVHD相关性。方法:应用流式细胞技术及实时定量PCR技术检测22例异基因造血干细胞移植物中及移植后不同时期外周血中CD4+CD25+T细胞和CD4+CD25highT细胞占CD4+T细胞的比例(CD4+CD25+/CD4+、CD4+CD25high/CD4+)、CD4+CD25highT细胞与CD4+CD25+T细胞之比(CD4+CD25high/CD4+CD25+)以及FOXP3基因的表达。结果:①aGVHD组移植物中CD4+CD25high/CD4+及CD4+CD25high/CD4+CD25+比例均显著低于无aGVHD组(P<0.05)。②移植后早期,两组外周血中的CD4+CD25+/CD4+比例均较移植物中者显著升高,在aGVHD组CD4+CD25high/CD4+及CD4+CD25high/CD4+CD25+比例亦较移植物中者显著升高,而在无aGVHD组两者无显著升高,且低于aGVHD组。③移植后两组的FOXP3的表达均逐渐升高,但至移植后6个月仍低于健康对照组(P<0.05)。aGVHD组移植后外周血FOXP3的表达低于无aGVHD组(P<0.05)。结论:①移植后早期外周血CD4+CD25high调节性T细胞发生活化和增殖,有利于维持免疫稳态和控制aGVHD;②移植物中CD4+CD25high调节性T细胞的比例及其与活化的CD4+效应性T细胞之比可能影响aGVHD的发生;③aGVHD可影响CD4+CD25high调节性T细胞的数量或FOXP3的表达。

恶性肿瘤患者CD4^+CD25^+/CD4^+CD25^(high)调节性T细胞的研究

目的:探讨恶性肿瘤患者外周血CD4+CD25+/CD4+CD25high调节性T细胞(Regulatory T cell,Treg)水平的特点及其临床意义。方法:采用流式细胞术检测Treg水平,并进行分层分析。结果:62例恶性肿瘤患者外周血中CD4+CD25+Treg、CD4+CD25high Treg占T细胞的百分比分别为19.61%±8.17%和4.20%±1.90%,高于正常对照组(分别为P<0.05和P<0.001),它们与NK细胞呈负相关(r分别为-0.2361和-0.306)。随疾病进展,CD4+ CD25+Treg水平升高,肿瘤进展期(IV期)与前3期比较有极显著差异(P<0.001)。CD4+CD25high Treg占CD4+T细胞的百分比率逐渐升高,中期(Ⅲ期)患者与早期患者、正常对照组比较有极显著差异(P<0.001);晚期患者与中期患者比较有极显著差异(P<0.001)。结论:恶性肿瘤患者外周血CD4+CD25+/CD4+CD25high Treg水平的升高,与恶性肿瘤免疫功能低下及肿瘤的发生发展密切相关。