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.

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.

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.

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.

Downregulation of CD4+CD25+ regulatory T cells may underlie enhanced Th1 immunity caused by immunization with activated autologous T cells

Regulatory T cells （Treg） play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti-tumor immunity. We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti-tumor immunity and upregulated Thl responses in vivo. However, the underlying molecular mechanisms are not well understood. Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated autologous T cells. We found that Foxp3 expression decreased in CD4＋CD25＋ T cells from the immunized mice. Moreover, CD4＋CD25＋Foxp3＋ Treg obtained from immunized mice exhibited diminished immunosuppression ability compared to those from naive mice. Further analysis showed that the serum of immunized mice contains a high level ofanti-CD25 antibody （about 30 ng/ml, p〈0.01 vs controls）. Consistent with a role ofanti-CD25 response in the downregulation of Treg, adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice. The triggering of anti-CD25 response in immunized mice can be explained by the fact that CD25 was induced to a high level in the ConA activated autologous T cells used for immunization. Our results demonstrate for the first time that immunization with attenuated activated autologous T cells evokes anti-CD25 antibody production, which leads to impeded CD4＋CD25＋Foxp3＋ Treg expansion and function in vivo. We suggest that dampened Treg function likely contributes to enhanced Thl response in immunized mice and is at least part of the mechanism underlying the boosted anti-tumor immunity.

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.

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.

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.

慢性丙型肝炎患者外周血CD4^+ CD25^(high)调节性T细胞与抗病毒疗效的关系

目的探讨慢性丙型肝炎患者外周血CD4+CD25high调节性T细胞(Treg)与抗病毒疗效的关系。方法选取83例完成抗病毒治疗和随访的慢性丙型肝炎患者作为研究对象(慢性丙型肝炎组)。分别在治疗前和治疗12、48周及随访24周时,采用流式细胞术检测外周血中CD4+CD25highTreg百分比,并进行血清丙型肝炎病毒核糖核酸(HCV-RNA)定量和肝功能指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)等指标的检测。设立正常对照组(n=32)。结果与正常对照组比较,治疗前慢性丙型肝炎组外周血中的CD4+CD25highTreg百分比和血清ALT、AST均显著升高(P<0.05)。在治疗12周时,32例患者获得早期病毒学应答(EVR),外周血CD4+CD25highTreg百分比和HCV-RNA定量结果与治疗前和未获得EVR患者比较,差异均有统计学意义(P<0.01),但血清ALT无明显变化。在治疗48周时,慢性丙型肝炎病毒学应答组(HCV-RNA定量结果低于最低检测限)(n=51)患者外周血中CD4+CD25highTreg百分比、HCV-RNA定量结果和血清ALT均显著低于治疗前和无病毒学应答组(n=32)(P<0.01),慢性丙型肝炎无病毒学应答组患者各项检测指标治疗前后比较差异无统计学意义(P>0.05)。治疗后随访24周时,病毒学应答组中有39例患者获得了持续病毒学应答(SVR),外周血中的CD4+CD25highTreg百分比、HCV-RNA定量及血清ALT均显著低于治疗前和未获得SVR患者(P<0.01);而未获得SVR患者的各项检测指标均较治疗结束时(48周末)明显升高(P<0.05)。结论慢性丙型肝炎患者抗病毒治疗及随访过程中外周血中CD4+CD25highTreg百分比的变化对预测疗效和复发有一定的价值。

银屑病患者体外系统血液固有免疫反应中CD4^+ CD25^+ T细胞及CD4^+ CD25^(high) T细胞的变化与意义

目的用体外系统血液固有免疫反应体系研究银屑病患者外周血CD4+ CD25+T细胞及CD4+ CD25highT细胞的变化,并探讨其在银屑病发病机制中的作用。方法将灭活大肠杆菌悬液(3×108/mL)0.2mL作为免疫原激活剂加入枸橼酸抗凝的全血细胞悬液0.2mL和血浆0.3mL中,37℃水浴1h,用流式细胞仪测定CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例。结果在加入大肠杆菌的银屑病全血细胞组(银屑病实验组)CD4+ CD25highT细胞比例(1.88%)明显高于银屑病对照组(1.41%)(P﹤0.01),CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例(16.86%,1.88%)明显低于加入大肠杆菌的健康实验组(24.26%,2.81%)(P﹤0.01);银屑病对照组CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例(15.97%,1.41%)较健康对照组明显降低(21.75%,2.17%)(P﹤0.01);健康实验组CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例群(24.26%,2.81%)均明显高于健康对照组(21.75%,2.17%),(P﹤0.05)。结论银屑病患者CD4+ CD25+T细胞及CD4+ CD25highT细胞比例降低,外来抗原刺激后比例升高,但与健康正常人存在差异。这可能与其复杂的系统免疫学发病机制相关。

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.

慢性乙型肝炎患者外周血CD4^+CD25^(+high)调节性T细胞对免疫状态的影响

目的探讨CD4+ CD25+high调节性T(Tr)细胞在慢性乙型肝炎(CHB)患者外周血的变化及其对免疫状态的影响。方法用流式细胞仪技术分析比较42例CHB患者及20例正常健康人外周血中CD4+CD25+highTr细胞,同时检测CD4+T细胞和CD8+T细胞。结果与健康对照组相比,CHB组CD4+CD25+highTr细胞百分率显著升高(P<0.05),而CD4+T细胞显著下降(P<0.05);CD8+T细胞虽有增加趋势,但无差异(P>0.05)。CHB组CD4+CD25+highTr细胞百分率与HBV DNA载量有一定的关系,但与血清ALT水平无相关(P>0.05)。结论CHB患者外周血CD4+CD25+highTr细胞升高可能是导致机体免疫功能下降的原因之一。

CD4^+CD25^(high) Tr细胞在非小细胞肺癌中的表达及意义

目的:比较非小细胞肺癌(NSCLC)患者外周血和肿瘤浸润淋巴细胞(TIL)中CD4+CD25high Tr细胞比例的变化与肺癌分期的关系,探讨患者局部肿瘤微环境和全身免疫状态与CD4+CD25+Tr细胞的关系。方法:分离NSCLC患者和健康人外周血单个核细胞(PBMC),取NSCLC患者新鲜肺癌组织抽提TIL。用流式细胞仪检测CD4+T细胞和CD4+CD25high Tr细胞。结果:NSCLC患者PBMC中CD4+CD25high Tr/CD4+T细胞比例较健康人显著升高(P<0.01)。Ⅱ～ⅢA期NSCLC患者PBMC和TIL中CD4+CD25high Tr/CD4+T细胞比例较Ⅰ期患者显著升高。结论:CD4+CD25high Tr细胞可能参与肿瘤产生的宿主免疫抑制反应及其发展。

抗CD25单克隆抗体对肾移植受者CD4^+CD25^(high)调节性T细胞的影响

目的评价抗CD25单克隆抗体对肾移植受者术后早期CD4+CD25high调节性T细胞(CD4+CD25 high Treg)的影响。方法2007年2-9月接受初次亲属活体供肾移植的受者41例,根据是否使用抗CD25单克隆抗体(商品名daclizumab)分为抗体组(21例)和对照组(20例)。其中抗体组在肾移植术前2h及术后第14天分别给予抗CD25单抗各50mg。在移植前及移植后第13、17、60天分别留取肝素抗凝外周血15ml。应用流式细胞仪测定两组受者外周血CD4+T细胞和CD4+CD25 high Treg比例的变化,半定量RT-PCR检测CD25 mRNA的表达变化。结果肾移植术后13、17、60d抗体组的CD25+T细胞占CD4+T细胞的比例(20%±8%、13%±7%、24%±9%)低于对照组(45%±6%、41%±5%、40%±6%),差异有统计学意义(P<0.05)。抗体组术后第17天CD4+CD25 high Treg占CD4+T细胞的比例为4.40%±0.26%,明显低于对照组(8.56%±0.36%,P<0.01);而术后13、60d抗体组CD4+CD25 high Treg所占比例分别为7.00%±0.47%、3.75%±0.19%,与对照组(分别为8.04%±0.32%、3.66%±0.31%)比较差异无统计学意义(P>0.05)。抗体组CD25 mRNA相对表达水平在给予第二次抗体前(术后第13天)为1.65±0.22,术后第17天为1.84±0.27,两者间差异无统计学意义(P>0.05)。对照组术后第17天CD25 mRNA相对表达水平为1.70±0.23,与抗体组比较差异无统计学意义(P>0.05)。结论两剂共100mg抗CD25单抗仅一过性地降低CD4+CD25 high Treg,不会影响其活化扩增,无损于术后早期的免疫耐受诱导及维持。

雷公藤多苷对IgA肾病CD_4～+CD_(25)～(high)调节性T细胞及Th1、Th2影响

目的:探讨CD4+CD25high调节性T细胞(Treg)及辅助性T细胞亚群(Th1、Th2)比例失衡在IgA肾病(IgAN)免疫发病机制中的作用,以及使用雷公藤多苷(tripterygium wilfordii glycoside,TW)治疗后对IgAN的Treg、Th1、Th2及Th1/Th2的比例影响。方法:用流式细胞分析仪检测30例IgAN患者,使用雷公藤多苷治疗前后的外周血Treg、Th1、Th2及Th1/Th2的比例。并与健康对照组进行比较,两组间比较采用t检验。结果:IgA N患者外周血Treg的比例明显高于健康人(P<0.01);IgAN患者外周血中,Th2细胞比例显著高于健康对照组(P<0.01);IgAN患者Th1/Th2比值显著低于健康对照组(P<0.01)。使用TW(1 mg·kg-1·d-1)治疗IgAN 3个月后,可显著降低IgAN患者外周血Treg(P<0.01),但治疗后Treg仍高于健康对照组(P<0.05);显著降低Th2细胞比例(P<0.05),显著升高Th1/Th2比值(P<0.01);且治疗后IgAN的尿蛋白明显降低(P<0.01),血清白蛋白升高(P<0.05)。结论:TW可改善IgAN患者体内存在T细胞亚群表达紊乱,对IgAN治疗具有一定作用。

自身免疫性肝病相关抗体与CD_4^+ CD_(25)^(+high)调节性T细胞的临床意义探讨

目的探讨自身免疫性肝炎(AIH)患者外周血CD4+CD25+high调节性T细胞的作用。方法抗核抗体酶联免疫分析法检测(ANA)。自身免疫性肝病谱线性免疫分析法(LIVER-LIA)测定抗肝肾微粒体1(LKM-1)、抗肝细胞溶质抗原1(LC-1)、抗线粒体-2(AMA-M2)、抗可溶性肝抗原/抗肝胰抗原(SLA/LP)。用流式细胞仪技术比较分析AIH患者51例、慢性乙型肝炎(CHB)30例及键康正常人20例外周血中的CD4+CD25+high调节性T细胞,用免疫组织化学法检测AIH和CHB患者肝组织Foxp3的表达情况。结果AIH占51%(51/100),AIH组外周血中CD4+CD25+high/CD4+百分率显著低于正常组(P<0.05)和CHB组(P<0.01),并且CHB组显著高于正常组(P<0.05);同时AIH组外周血中CD4+T细胞也显著高于CHB组(P<0.01);肝组织Foxp3+细胞主要分布于肝小叶内窦周隙、汇管区,AIH组肝组织Foxp3+表达显著低于CHB组(P<0.01)。结论CD4+CD25+high调节性T细胞下降可能是自身免疫性肝炎发病的原因之一。