系统性红斑狼疮患者CD4~+CD25~+CD127~(low/-)调节性T细胞中Foxp3的表达及免疫抑制功能研究

目的 研究系统性红斑狼疮(SEE)外周血中调节性T细胞不同标志以及调节性T细胞在SLE发病中的作用;探讨CD127与Foxp3的相关性,明确CD127定义调节性T细胞的特异性;鉴定CD4~+CD25~+CD127~(low/-)T淋巴细胞免疫抑制功能.方法 ①采用四色直接荧光素标记法和多参数流式细胞术检测40例SLE患者(19例初发和21例缓解)及15名健康对照外周血CD4~+CD25~+T淋巴细胞、CD4~+CD25~+CD127~(low-)T淋巴细胞、CD4~+CD25~+Foxp3~+T淋巴细胞、CD4~+CD25~(high)T淋巴细胞、CD4~+CD25~(high)CD127~(low/-)T淋巴细胞、CD4~+CD25~(high)Foxp3~+T淋巴细胞以及CD4~+CD127~(low/-)Foxp3~+T淋巴细胞占CD4~+T淋巴细胞的比率,并且将7种调节性T细胞比率与外周血抗双链DNA(dsDNA)等抗体及SLE疾病活动指数(SLEDA1)评分等进行相关性分析.②以流式细胞分选术结合细胞培养技术,检测和分析3例SLE患者和4名健康人外周血中CD4~+CD25~+CD127~(low/-)调节性T细胞对CD4~+CD25~-效应性T细胞增殖的抑制作用.采用两样本均数的t检验,重复测量的方差分析,Pearson相关与Spearman相关分析进行统计学处理.结果 ①SLE患者组7种调节性T细胞比率分别为(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%,(0.97±0.28)%,(0.69±0.23)%和(0.71±0.35)%.与健康对照组比较:SLE患者组前6种调节性T细胞比率均低于健康对照组(P<0.05).②SLE患者组:CD4~+CD25~+Foxp3~+、CD4~+CD25~(high)Foxp3~+T淋巴细胞比率与IgA呈正相关;CD4~+CD25~(high)CD127~(low/-)T淋巴细胞比率与抗SSB抗体呈正相关.③SLE患者初发组和缓解组比较:SLE患者初发组7种调节性T细胞中除CD4~+CD127~(low/-)Foxp3~+T淋巴细胞比率外,其余均低于缓解组(P<0.05).④SLE患者初发组治疗前后比较:激素治疗前6种调节性T细胞比率均低于激素治疗后(P<0.05).⑤SLE患者初发组、缓解组和对照组中,CD4~+CD25~+T淋巴细胞及CD4~+CD25~(high)T淋巴细胞中Foxp3的表达与CD127低表达均呈正相关.⑥SLE患者、健康人CD4+CD25-效应性T细胞的体外增殖都可以被自身CD4~+CD25~+CD127~(low/-)调节性T细胞所抑制,但SLE患者的抑制率明显低于健康对照.结论 SLE的免疫异常可能与调节性T细胞的数量和功能缺陷有关;CD127可能代替Foxp3作为调节性T细胞特异性的表面标记物.     

细胞免疫参数对结直肠癌的评估价值

目的:评估结直肠癌患者抗肿瘤免疫状态.方法:应用流式细胞术检测100例未转移结直肠癌患者、100例伴有转移结直肠癌患者和100例健康志愿者外周血CD4+、CD8+、NK、B、CD4+CD25HighCD127lowTreg、Th/Treg值,采用单因素方差分析进行比较,分析差异.结果:转移性结直肠癌患者组分别与正常对照组和非转移性结直肠癌患者组相比,CD4+CD25HighCD127lowTreg细胞升高(7.72%±2.20%vs6.08%±1.47%,5.91%±1.55%,均P<0.05),CD4+T细胞降低(34.04%±8.71%vs37.83%±7.62%,37.68%±8.89%,均P<0.05),Th/Treg值降低(4.70±1.72vs6.47±2.54,6.81±4.09,均P<0.05).结直肠癌患者与正常对照组CD8+T细胞、NK细胞、B细胞三组两两相比,均无统计学意义.结论:转移性结直肠癌患者免疫功能紊乱,主要表现为CD4+T细胞、Th/Treg值降低,CD4+CD25HighCD127lowTreg细胞升高.     

Immune formulation-assisted conventional therapy on anti-infective effectness of multidrug-resistant Mycobacterium tuberculosis infection mice

Objective:To study the effect of immune formulation-assisted conventional therapy on antiinfective ability of multidrug-resistant Mycobacterium tuberculous infection mice.Methods:BALB/c mice were used as experimental animals,multidrug-resistant Mycobacterium tuberculosis infection models were built,randomly divided into model group,moxifloxacin group,thymopentin group and combined treatment group and given corresponding drug intervention,and then colony numbers in the spleen and lung,T lymphocyte subset contents and programmed death-1(PD-1) expression levels in peripheral blood were detected.Results:Colony numbers in lung and spleen of moxifloxacin group and thymopentin group were significantly lower than those of model group and colony numbers in lung and spleen of combined treatment group were significantly lower than those of moxifloxacin group and thymopentin group:contents of CD3~+CD4~+T cells,Thl and Thl7 in peripheral blood of moxifloxacin group and thymopentin group were higher than dtose of model group,and contents of CD3~+CD8~+T cells.Th2 and Treg were lower than those of model group;contents of CD3~+CD4~+T cells.Th 1 and Th 17 in peripheral blood of combined treatment group were higher than those of moxifloxacin group and thymopentin group,and contents of CD3~+CD8~+T cells.Th2 and Treg were lower than those of moxifloxacin group and thymopentin group:PD-I expression levels on T lymphocyte,B lymphocyte and monocyte surface in peripheral blood of moxifloxacin group and thymopentin group were lower than those of model group,and PD-I expression levels on T lymphocyte.B lymphocyte and monocyte surface in peripheral blood of combined treatment group were lower than those of moxifloxacin group and thymopentin group.Conclusions:Immune formulation thymopentin can enhance the anti-infective ability of multidrug-resistant Mycobacterium tuberculosis infection mice,decrease bacterial load in lung and spleen,and enhance immune function.     

脾栓塞对肝硬化患者外周血T淋巴细胞亚群的影响

目的 :研究部分脾栓塞治疗脾功能亢进对肝硬化患者T淋巴细胞亚群的影响。方法 :57例肝硬化并脾功能亢进患者 ,32例采用部分脾栓塞治疗 ,2 5例采用脾全切治疗 ,1 5例正常人作为对照 ,比较两组治疗前后外周血T淋巴细胞亚群的变化。结果 :肝硬化患者外周血T淋巴细胞 (CD3+ )、T辅助 /诱导淋巴细胞亚群 (CD4+ )、T辅助淋巴细胞 /T抑制淋巴细胞亚群 (CD4+ /CD8+ )明显低于正常人 (P <0 .0 1 ) ;脾栓塞组治疗前后CD3+ 、CD4+ 、CD8+ 、CD4+ /CD8+ 无明显差异 (P >0 .0 5) ;脾全切组治疗后CD3+ 、CD4+ 、CD4+ /CD8+ 均较治疗前及部分脾栓塞组治疗后明显降低 (P <0 .0 1 )。结论 :肝硬化患者外周血CD3+ 、CD4+ 、CD4+ /CD8+ 降低 ,部分脾栓塞治疗脾功能亢进 ,对肝硬化患者外周血T淋巴细胞亚群无明显影响 ,显著优于脾全切治疗     

Vasoactive intestinal peptide induces CD4+,CD25+ T regulatory cells with therapeutic effect in collagen-induced arthritis

OBJECTIVE: CD4+,CD25+ T regulatory cells (Treg) control the immune response to a variety of antigens, including self antigens, and may offer opportunities to intervene in the course of autoimmune diseases. Several models support the idea of the peripheral generation of CD4+,CD25+ Treg from CD4+,CD25- T cells, but little is known about the endogenous factors and mechanisms controlling the peripheral expansion of CD4+,CD25+ Treg. We undertook this study to investigate the capacity of the vasoactive intestinal peptide (VIP), an immunosuppressive antiarthritic neuropeptide, to induce functional Treg in vivo during the development of collagen-induced arthritis (CIA). METHODS: We measured the number of CD4+,CD25+ Treg following VIP administration to CIA mice, and we characterized their phenotype and their ability to suppress activation of autoreactive T cells. We determined the capacity of VIP to induce Treg in vitro as well as the use of Treg in the treatment of CIA, measuring the clinical evolution and the inflammatory and autoimmune components of the disease. RESULTS: The administration of VIP to arthritic mice resulted in the expansion of CD4+,CD25+,Foxp3+ Treg in the periphery and joints, which inhibited autoreactive T cell activation/expansion. VIP induced more efficient suppressors on a per-cell basis. The VIP-generated CD4+,CD25+ Treg transfer suppressed and significantly ameliorated the progression of the disease. CONCLUSION: These results demonstrate the involvement of the generation of Treg in the therapeutic effect of VIP on CIA. The generation of highly efficient Treg by VIP ex vivo could be used as an attractive therapeutic tool in the future, avoiding the administration of the peptide to patients with rheumatoid arthritis.     

HBeAg(+)与HBeAg(-)乙型肝炎病毒携带者外周血CD4~+CD25~+调节性T淋巴细胞比例的变化及意义

目的探讨HBeAg(+)与HBeAg(-)HBV携带者外周血中CD4+CD25+调节性T淋巴细胞(Treg)的变化及其意义。方法 HBeAg(+)与HBeAg(-)HBV携带者两组各50例患者,应用流式细胞仪测定外周血CD3+T淋巴细胞所占比例、CD4+和CD8+T淋巴细胞所占比例及其比值、CD4+CD25+Treg所占比例;以同期门诊健康体检者20例为健康对照组。结果与健康对照组相比,HBeAg(+)与HBeAg(-)HBV携带者,在外周血CD3+T淋巴细胞、CD4+与CD8+T淋巴细胞所占比例差异无统计学意义。CD4+与CD8+T淋巴细胞比值,在HBeAg(+)HBV携带者有升高趋势,但与HBeAg(-)HBV携带者和对照组比较差异无统计学意义。HBeAg(+)HBV携带者,CD4+CD25+Treg所占比例较对照组明显升高,而HBeAg(-)HBV携带者,CD4+CD25+Treg所占比例较对照组明显降低,差异均有统计学意义。结论尽管肝功能在正常范围,HBeAg(+)与HBeAg(-)HBV携带者外周血CD4+CD25+Treg比例不同,HBeAg(+)HBV携带者高于健康对照组,而HBeAg(-)HBV携带者低于健康对照组,提示HBeAg(+)与HBeAg(-)HBV携带者处于不同的免疫状态。     

Low number of regulatory T cells in skin lesions of patients with cutaneous lupus erythematosus

OBJECTIVE: To define the phenotype and function of CD4+,CD25+ regulatory T cells (Treg) in patients with cutaneous lupus erythematosus (CLE), a heterogeneous autoimmune disease characterized primarily by inflammatory skin lesions. METHODS: The number of Treg in skin specimens obtained from patients with various subtypes of CLE was investigated by immunohistochemical analysis, using anti-Foxp3 and anti-CD4 monoclonal antibodies. Furthermore, characterization of peripheral blood CD4+,CD25+ Treg from normal healthy donors and patients with CLE was carried out by flow cytometry, analyzing the expression of Foxp3 and Treg subpopulations. We also purified CD4+,CD25(high) Treg obtained from patients with CLE and tested the sensitivity of these cells to CD95L-mediated apoptosis. RESULTS: Quantitative analysis of CD4+ T cells in skin lesions from patients with CLE revealed that the number was similar to that in lesions from patients with other chronic inflammatory diseases, but the number of Foxp3+ Treg in CLE was significantly reduced. There was no correlation between disease subtype and the frequency of Foxp3+ Treg in the skin of patients with CLE. In peripheral blood, no significant differences were observed in the number and phenotype of CD4+,CD25+ Treg or in the sensitivity to apoptosis of CD4+,CD25(high) Treg derived from patients with CLE and those derived from normal healthy donors. CONCLUSION: These data suggest that an organ-specific abnormality of Treg in the skin underscores the importance of analyzing Treg in the affected tissue. Such a local process might give insight into the pathogenic mechanisms of CLE and differs from a global peripheral dysfunction as reported for patients with a systemic manifestation of the disease.     

Dysfunctional CD4+,CD25+ regulatory T cells in untreated active systemic lupus erythematosus secondary to interferon-alpha-producing antigen-presenting cells

OBJECTIVE: To explore whether there are extrinsic factors that impair the suppressive function of CD4+,CD25+ regulatory T cells in patients with untreated active systemic lupus erythematosus (SLE). METHODS: We studied 15 patients with untreated active SLE, 10 patients with SLE in remission, and 15 healthy control subjects. Percentages of CD4+,CD25+,FoxP3+ Treg cells and levels of forkhead box P3 (FoxP3) protein were analyzed by flow cytometry. Expression of messenger RNA (mRNA) for FoxP3 in purified Treg cell populations was assessed by real-time polymerase chain reaction analysis. Experiments examining Treg cell function in SLE were designed to distinguish primary from secondary T cell dysfunction. Levels of interferon-alpha (IFNalpha) in supernatants from the function assays were determined with an IFN-stimulated response element-luciferase reporter assay. RESULTS: The percentage of CD4+,CD25+, FoxP3+ cells in peripheral blood was significantly increased in SLE patients as compared with controls (mean +/- SEM 9.11 +/- 0.73% versus 4.78 +/- 0.43%; P < 0.0001). We found no difference in FoxP3 expression at either the mRNA or protein level in any CD4+,CD25+ T cell subset from SLE patients as compared with controls. Antigen-presenting cells (APCs) from SLE patients were responsible for decreased Treg cell activity and could also render dysfunctional Treg cells from healthy control subjects. CD4+,CD25+ Treg cells from SLE patients exhibited normal suppressive activity when cultured with APCs from healthy controls. A partial Treg cell blockade effect was induced by the high levels of IFNalpha derived from SLE patient APCs. CONCLUSION: We suggest that blockade of Treg cell-mediated suppression by IFNalpha-producing APCs in SLE patients may contribute to a pathogenic loss of peripheral tolerance in this disease.     

急性冠状动脉综合征患者血清调节性T细胞的变化及其临床意义

目的通过检测急性冠状动脉综合征患者(ACS)外周血中调节性T细胞(Treg细胞)的数量变化,了解Treg细胞与ACS发生发展的关系及意义。方法入选40例急性心肌梗死患者(心梗组)、40例不稳定型心绞痛患者(心绞痛组)和40名健康人(对照组)。所有入选受试对象外周血血液样本均采用流式细胞术检测CD4+CD25+CD127-的结果来定义各组外周血中Treg细胞表达的百分比,比较各组的Treg细胞表达百分比,同时比较心梗后12、48、72h Treg细胞表达百分比变化。常规检测血常规、血脂。结果外周血中Treg细胞表达的百分比[心绞痛组(3.20%±0.92%)比对照组(3.76%±1.18%)]有所下降,差异有统计学意义(P<0.05),心梗后12h(2.12%±0.89%)患者外周血中Treg细胞表达的百分比也明显下降(P<0.001);心梗后12 h(2.12%±0.89%)、48 h(3.0%±0.89%)、72 h(3.57%±0.92%)的Treg细胞百分比逐渐递增。心梗后各时间点两两比较,差异均存在统计学意义(P<0.05)。心梗后72h Treg细胞百分比与对照组相比差异无统计学意义。心梗组常规检测血常规中白细胞[(10.34±2.48)×109/L]、中性粒细胞(0.80±0.06)较正常值显著异常升高,且中性粒细胞与Treg细胞百分比呈负性相关(相关系数r为-0.517,P<0.05)。结论急性冠脉综合征患者Treg细胞数量减少,且与炎症反应呈负性相关,故可推测Treg细胞是ACS患者免疫稳态失衡和炎症反应的标志物之一。     

Suppressor activity among CD4+,CD25++ T cells is discriminated by membrane-bound tumor necrosis factor alpha

OBJECTIVE: Previous studies have shown that the suppressive capacity of CD4+,CD25++ T cells is compromised in patients with rheumatoid arthritis (RA) and restored by anti-tumor necrosis factor alpha (anti-TNFalpha) therapy. Given the lack of specific cell surface markers for human Treg cells, this study aimed to define surface markers for identifying and enriching Treg cells with enhanced regulatory ability within the CD4+,CD25++ T cell compartment and to provide additional understanding of the effects of anti-TNFalpha antibodies in humans. METHODS: The expression of membrane-bound TNFalpha in human peripheral blood CD4+ T cells was analyzed by flow cytometry in healthy individuals and RA patients before and after anti-TNFalpha treatment. Membrane-bound TNFalpha-positive and TNFalpha-negative CD4+,CD25++ T cells were purified by fluorescence-activated cell sorting, and their suppressive capacity was assessed in vitro by a standard suppression assay. RESULTS: A substantial number of CD4+,CD25++ T cells expressed membrane-bound TNFalpha. Membrane-bound TNFalpha-positive CD4+,CD25++ T cells displayed reduced antiinflammatory cytokine production and less potent suppressor capacity, since 4 times more cells were required to achieve 50% inhibition compared with their membrane-bound TNFalpha-negative counterparts. Treatment of RA patients with TNFalpha-specific antibodies led to a reduction in the number of membrane-bound TNFalpha-positive CD4+,CD25++ T cells from peripheral blood. CONCLUSION: Our data indicate that the absence of membrane-bound TNFalpha on CD4+,CD25++ T cells can be used to characterize and enrich for Treg cells with maximal suppressor potency. Enrichment of membrane-bound TNFalpha-negative CD4+,CD25+ cells in the CD4+,CD25++ T cell compartment may contribute to restoring the compromised suppressive ability of CD4+,CD25++ T cell populations in RA patients after anti-TNFalpha treatment.     

Treg cells suppress osteoclast formation: a new link between the immune system and bone

OBJECTIVE: To investigate whether Treg cells can suppress osteoclast differentiation, and to define a new potential link between the immune system and the skeleton. METHODS: Regulatory CD4+,CD25+,Foxp3+ T cells were isolated and purified from the spleen and cocultured with CD11b+ osteoclast precursor cells isolated from bone marrow. Osteoclastogenesis and bone erosion were assessed by tartrate-resistant acid phosphatase staining and pit resorption assay, respectively. In addition, Transwell experiments and cytokine-blocking experiments were performed to define the mechanisms of interaction between Treg cells and osteoclasts. RESULTS: CD4+,CD25+,Foxp3+ T cells, but not CD4+,CD25- T cells, dose dependently inhibited macrophage colony-stimulating factor- and RANKL-dependent osteoclast formation. Pit formation was inhibited by up to 80% when Treg cells were added. The blockade of osteoclast formation was not based on the alteration of RANKL/osteoprotegerin balance but was essentially dependent on direct cell-cell contact via CTLA-4. Treg cell-mediated expression of transforming growth factor beta, interleukin-4 (IL-4), and IL-10 contributed but was not essential to the inhibitory effect on osteoclastogenesis. CONCLUSION: These data show that CD4+,CD25+,Foxp3+ Treg cells suppress osteoclast formation, provide a new link between the immune system and bone, and extend our knowledge on regulation of bone homeostasis by the immune system.     

Vasoactive intestinal peptide induces CD4+,CD25+ T regulatory cells with therapeutic effect in collagen‐induced arthritis

Objective

CD4+,CD25+ T regulatory cells (Treg) control the immune response to a variety of antigens, including self antigens, and may offer opportunities to intervene in the course of autoimmune diseases. Several models support the idea of the peripheral generation of CD4+,CD25+ Treg from CD4+,CD25− T cells, but little is known about the endogenous factors and mechanisms controlling the peripheral expansion of CD4+,CD25+ Treg. We undertook this study to investigate the capacity of the vasoactive intestinal peptide (VIP), an immunosuppressive antiarthritic neuropeptide, to induce functional Treg in vivo during the development of collagen‐induced arthritis (CIA).

Methods

We measured the number of CD4+,CD25+ Treg following VIP administration to CIA mice, and we characterized their phenotype and their ability to suppress activation of autoreactive T cells. We determined the capacity of VIP to induce Treg in vitro as well as the use of Treg in the treatment of CIA, measuring the clinical evolution and the inflammatory and autoimmune components of the disease.

Results

The administration of VIP to arthritic mice resulted in the expansion of CD4+,CD25+,Foxp3+ Treg in the periphery and joints, which inhibited autoreactive T cell activation/expansion. VIP induced more efficient suppressors on a per‐cell basis. The VIP‐generated CD4+,CD25+ Treg transfer suppressed and significantly ameliorated the progression of the disease.

Conclusion

These results demonstrate the involvement of the generation of Treg in the therapeutic effect of VIP on CIA. The generation of highly efficient Treg by VIP ex vivo could be used as an attractive therapeutic tool in the future, avoiding the administration of the peptide to patients with rheumatoid arthritis.

     

Function of CD4+,CD25+ Treg cells in MRL/lpr mice is compromised by intrinsic defects in antigen-presenting cells and effector T cells

OBJECTIVE: Naturally occurring CD4+,CD25+ Treg cells are central in the maintenance of peripheral tolerance. Impaired activity and/or a lower frequency of these cells is involved in the emergence of autoimmunity. We undertook this study to analyze relative proportions and functional alterations of Treg cells in MRL/lpr mice. METHODS: The frequency of CD4+,CD25+ T cells in the peripheral blood of healthy and autoimmune mice was compared by flow cytometry. The capacity of CD4+,CD25+ T cells to inhibit the proliferation and cytokine secretion of CD4+,CD25- T cells was assessed after polyclonal activation. RESULTS: MRL/lpr mice exhibited a normal percentage of CD4+,CD25 high T cells, and forkhead box P3 messenger RNA and protein expression in Treg cells was not altered. However, MRL/lpr Treg cells displayed a reduced capacity to suppress proliferation and to inhibit interferon-gamma secretion by syngeneic effector CD4+,CD25- T cells, as compared with syngeneic cocultures of CBA/J T cells. Moreover, effector MRL/lpr CD4+,CD25- T cells were substantially less susceptible to suppression even when cultured with CBA/J or MRL/lpr Treg cells. Crossover experiments led us to conclude that in MRL/lpr mice, each partner engaged in T cell regulation displays altered functions. Molecules involved in suppressive mechanisms (CTLA-4 and CD80/CD86) are underexpressed, and antigen-presenting cells (APCs) produce raised levels of interleukin-6, which is known to abrogate suppression. CONCLUSION: Our results suggest that although the frequency and phenotype of Treg cells in MRL/lpr mice are similar to those in normal mice, Treg cells in MRL/lpr mice are not properly stimulated by APCs and are unable to suppress proinflammatory cytokine secretion from effector T cells.     

经导管肝动脉化疗栓塞术对原发性肝癌患者细胞免疫及调节T细胞的影响

目的 探讨原发性肝癌(PLC)患者经导管肝动脉化疗栓塞术(TACE)前后细胞免疫及调节T细胞(Treg)的变化及其临床价值.方法 采用流式细胞术检测健康志愿者及PLC患者介入治疗术前1 d与术后1个月外周血CD4+、CD8+T细胞及调节T细胞的比例变化;ELISA法检测外周血IL-35的含量.结果 PLC患者外周血中C...     

Proinflammatory mediator-induced reversal of CD4+,CD25+ regulatory T cell-mediated suppression in rheumatoid arthritis

OBJECTIVE: We previously demonstrated that CD4+,CD25+ regulatory T (Treg) cells are present in increased numbers in the synovial fluid (SF) of rheumatoid arthritis (RA) patients and display enhanced suppressive activity as compared with their peripheral blood (PB) counterparts. Despite the presence of these immunoregulatory cells in RA, chronic inflammation persists. The purpose of the present study was to investigate whether particular proinflammatory mediators that are associated with RA could abrogate CD4+,CD25+ Treg-mediated suppression. METHODS: Monocyte phenotype was determined by flow cytometry and cytokine levels by enzyme-linked immunosorbent assay. Magnetically sorted CD4+,CD25- and CD4+,CD25+ T cells derived from the PB and SF obtained from RA patients were stimulated alone or in coculture with anti-CD3 monoclonal antibody (mAb) and autologous antigen-presenting cells, in the absence or presence of anti-CD28 mAb or the proinflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), or IL-7. RESULTS: Monocytes from the SF of RA patients displayed increased expression of HLA class II molecules, CD80, CD86, and CD40 as compared with PB-derived monocytes, indicating their activated status. Mimicking this increased costimulatory potential, addition of anti-CD28 mAb to cocultures of CD4+,CD25- and CD4+,CD25+ T cells resulted in reduced CD4+,CD25+ Treg-mediated suppression in both PB and SF. Furthermore, IL-7 and, to a limited extent, TNFalpha, both of which are produced by activated monocytes and were detected in SF, abrogated the CD4+,CD25+ Treg-mediated suppression. In contrast, IL-6 did not influence Treg-mediated suppression. CONCLUSION: Our findings suggest that the interaction of CD4+,CD25+ Treg cells with activated monocytes in the joint might lead to diminished suppressive activity of CD4+,CD25+ Treg cells in vivo, thus contributing to the chronic inflammation in RA.     

原发性肝癌和乙型肝炎肝硬化患者外周血淋巴细胞凋亡率、T细胞亚群和NK细胞的水平变化和临床意义

目的通过检测乙型肝炎肝硬化和合并乙型肝炎病毒感染的原发性肝细胞癌患者的外周血的粒细胞、单核细胞、NK细胞、T细胞及其亚群和淋巴细胞及其凋亡率,探讨两者在细胞免疫方面的差异.方法用Ficoll Hypaque离心法分离外周血单个核细胞(PBMNC),流式细胞仪检测外周血T细胞及其亚群、NK细胞和淋巴细胞、单核细胞和粒细胞,AnnexinV/FITCKit检测凋亡细胞.结果乙型肝炎肝硬化患者的外周血单核细胞、粒细胞、淋巴细胞、CD3-CD16 CD56 NK细胞、CD3 T细胞、CD3 CD4 T细胞、CD3 CD4 T细胞和CD4 CD8 T细胞比值,均与正常对照组无显著性差异(P＞0.05);但淋巴细胞凋亡率明显低于对照组(P＜0.01).原发性肝癌外周血CD3-CD16 CD56 NK细胞、单核细胞和CD4 /CD8 T细胞比值与肝硬化组和正常对照组比较,均无显著性差异(P＞0.05),而粒细胞明显升高(P＜0.05);CD3 T细胞、CD3 CD4 T细胞和CD3 CD8 T细胞均较另两组明显减少(P＜0.05),淋巴细胞及其凋亡率均明显低于另两组(P＜0.01).结论乙型肝炎肝硬化患者的外周血细胞免疫只发生不显著的变化,但淋巴细胞的凋亡率明显降低.原发性肝癌外周血的细胞免疫和淋巴细胞凋亡率均明显低下.     

M受体拮抗剂对提高慢性阻塞性肺疾病患者调节性T细胞表达的影响

目的 观察COPD稳定期患者体内是否存在CD_8~+CD_(25)~+Foxp_3~+调节性T细胞(Treg细胞),以及M受体拈抗剂噻托溴铵对其表达的影响.方法 2007年10月至2008年3月,选择23例COPD稳定期患者,每日1次吸入噻托溴铵18 μg,连续治疗3个月.分别在治疗前后榆查患者的肺功能和检测外周血中T细胞亚群的数量.两组间均数比较采用配对t检验,相关分析采用直线相关分析.结果 COPD稳定期患者在吸入噻托溴铵治疗后,外周血巾CD_4~+为(36±46)%,CD_8~+CD_(25)~+Treg 细胞为(21±21)%,明显高于治疗前的(28±10)%和(8±8)%;CD_4~+CD_(25)~+为(4±3)%,明显低于治疗前的(10±7)%,差异均有统计学意义(t值为2.72～3.78,P<0.01和P<0.05).CD_8~+、CD_8~+CD_(25)~+和CD_4~+CD_(25)~+Treg细胞均有表达,但治疗前后无明显变化.治疗后肺通气功能得到明显改善,FEV_1、FEV_1占预计值%和FEV_1/FVC分别由(1.0±0.3)L、(35±10)%和(41±8)%提高到(1.1±0.3)L、(40±11)%和(45±11)%,差异均有统计学意义(t值为2.37～2.65,均P相似文献   

肿瘤患者化疗前后外周血NK/T及Treg细胞比例分析

目的探讨肿瘤患者化疗前后,外周血免疫激活细胞及免疫抑制细胞比例变化。 方法本研究通过分析38例健康人外周血NK/T细胞及Treg细胞流式分析结果,并跟踪本中心140例常见肿瘤患者(结直肠癌31例、胃癌21例、乳腺癌27例、肺癌61例),发病前后、化疗疗程中外周血中的NK/T细胞及Treg细胞比例。 结果恶性肿瘤患者外周血中Treg细胞比例显著高于健康对照组分别为(6.95%±8.96%)、(4.25%±2.59%),P<0.05, NK细胞比例也显著高于健康人分别为(18.56%±9.19%)、(12.37%±6.24%),P<0.01。因统计病例数目有限,对69例肿瘤患者化疗的后续随访中发现,随着化疗进程,NK细胞以及Treg的比例变化无统计学意义,但在结直肠癌及胃癌中,Treg细胞比例可见先降低后升高的趋势。 结论肿瘤患者外周血Treg细胞比例高于健康人,在化疗过程中NK细胞比例以及Treg细胞比例均高于化疗前水平。随着本研究的继续深入,可能进一步证实Treg细胞在肿瘤中富集,形成免疫抑制微环境,是通过抑制CD4^＋、CD8^＋T淋巴细胞及NK细胞的肿瘤杀伤效应,介导肿瘤免疫逃逸的可能。     

姜黄素对NOD糖尿病小鼠Th1细胞和CD4^+Treg细胞的作用

目的研究姜黄素对Th1细胞和CD4^+Treg细胞的作用,探讨姜黄素对非肥胖糖尿病(NOD)小鼠的治疗作用。方法将10周龄雌性NOD小鼠分为对照组和姜黄素组,分别用溶剂或姜黄素灌胃。第14周时测血糖。第20周时,流式细胞术分析外周血Th1细胞和CD4^+Treg细胞,分析胸腺CD4^+ CD8^-T细胞、CD8+CD4^-T细胞及CD4^+Treg细胞;分选脾CD4^+Treg细胞用于检测其抑制功能。结果姜黄素组血糖正常比例高于对照组。姜黄素组Th1细胞比例显著低于对照组(P<0.05),CD4^+Treg细胞比例与对照组差异无统计学意义(P>0.05),CD4^+Treg/Th1显著高于对照组(P<0.05)。姜黄素组胸腺CD4^+CD8^-T细胞、CD8^+CD4^-T细胞及CD4^+Treg细胞比例与对照组差异无统计学意义(P>0.05),胸腺总细胞数显著高于对照组(P<0.05)。姜黄素组CD4^+Treg细胞的抑制功能明显高于对照组(P<0.05)。结论姜黄素降低NOD小鼠外周Th1细胞、增加CD4^+Treg细胞抑制功能、缓解胸腺退化,延缓了NOD小鼠的高血糖症状。     

Proinflammatory mediator–induced reversal of CD4+,CD25+ regulatory T cell–mediated suppression in rheumatoid arthritis

Objective

We previously demonstrated that CD4+,CD25+ regulatory T (Treg) cells are present in increased numbers in the synovial fluid (SF) of rheumatoid arthritis (RA) patients and display enhanced suppressive activity as compared with their peripheral blood (PB) counterparts. Despite the presence of these immunoregulatory cells in RA, chronic inflammation persists. The purpose of the present study was to investigate whether particular proinflammatory mediators that are associated with RA could abrogate CD4+,CD25+ Treg–mediated suppression.

Methods

Monocyte phenotype was determined by flow cytometry and cytokine levels by enzyme‐linked immunosorbent assay. Magnetically sorted CD4+,CD25– and CD4+,CD25+ T cells derived from the PB and SF obtained from RA patients were stimulated alone or in coculture with anti‐CD3 monoclonal antibody (mAb) and autologous antigen‐presenting cells, in the absence or presence of anti‐CD28 mAb or the proinflammatory cytokines interleukin‐6 (IL‐6), tumor necrosis factor α (TNFα), or IL‐7.

Results

Monocytes from the SF of RA patients displayed increased expression of HLA class II molecules, CD80, CD86, and CD40 as compared with PB‐derived monocytes, indicating their activated status. Mimicking this increased costimulatory potential, addition of anti‐CD28 mAb to cocultures of CD4+,CD25– and CD4+,CD25+ T cells resulted in reduced CD4+,CD25+ Treg–mediated suppression in both PB and SF. Furthermore, IL‐7 and, to a limited extent, TNFα, both of which are produced by activated monocytes and were detected in SF, abrogated the CD4+,CD25+ Treg–mediated suppression. In contrast, IL‐6 did not influence Treg‐mediated suppression.

Conclusion

Our findings suggest that the interaction of CD4+,CD25+ Treg cells with activated monocytes in the joint might lead to diminished suppressive activity of CD4+,CD25+ Treg cells in vivo, thus contributing to the chronic inflammation in RA.