Expansion of blood IgG4+ B,TH2, and regulatory T cells in patients with IgG4-related disease

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Circulating CD4+CD8+ T lymphocytes in patients with Kawasaki disease

We serially monitored cell surface antigen expression on mononuclear cells in peripheral blood isolated from patients with Kawasaki disease (KD), and found, for the first time, that a markedly increased number of CD4⁺CD8⁺ T lymphocytes was present in some of the patients (11 of the 24 cases). The cases of five of these 11 patients were complicated with coronary artery lesion (CAL); the 13 patients with normal numbers of CD4⁺CD8⁺ T lymphocytes did not have CAL. The patients' age, sex and grade of systemic inflammation evaluated by peripheral leucocyte count and serum C-reactive protein levels were not correlated to the number of CD4⁺CD8⁺ T lymphocytes. Other cell surface antigen characteristics of the CD4⁺CD8⁺ T lymphocytes included CD3⁺, CD45RA⁺, CD45RO⁺, CD16^?, and HLA-DR⁺. These results indicate that the surface antigen characteristics of the KD peripheral blood examined were the same as those of Epstein–Barr virus infection without CD45RA⁺. These findings provide useful information for the analysis of the pathogenesis of KD.     

CD4+CD25+免疫调节性T细胞对CD4+T细胞介导的移植物抗宿主病预防作用的研究

目的研究CD4+CD25+免疫调节性T(Treg)细胞在小鼠骨髓移植后,对移植物抗宿主病的预防作用及其作用机制.方法用C3H(H-2k)小鼠骨髓作为供体,提取C3H(H-2k)小鼠CD4+T及CD4+CD25+T细胞,C3H×B6(H-2k/b)F1小鼠为骨髓移植的受者.在受者接受致死量全身放射后,输注供者去除T细胞的骨髓(ATBM),使其造血功能重建(ATBM组).于不同的实验组给予CD4+(CD4组)T细胞,CD4+CD25+T(CD25组)或二者同时输注(CD4/CD25组).观察各组小鼠移植物抗宿主病(GVHD)的发生率.结果所有10只ATBM组小鼠至骨髓移植后60天仍全部存活,无GVHD发生;所有10只CD4组小鼠在骨髓移植10天内全部死于GVHD(P＜0.01);所有5只CD25组小鼠于骨髓移植后60天仍全部存活,无明显GVHD发生(P＞0.05);同样,所有6只CD4/CD25组小鼠至骨髓移植后仍全部存活,无明显GVHD发生(P＞0.05).结论在同种异基因小鼠的骨髓移植模型中,CD4+CD25+T不诱导GVHD的发生,并有预防CD4+T细胞介导的GVHD发生的作用.     

自身免疫性肝炎CD4+CD25+high调节性T细胞的研究

目的探讨CD4^＋CD25^＋high调节性T细胞（Tr）在自身免疫性肝炎（AIH）发病中的作用。方法用流式细胞仪技术比较分析AIH患者16例、慢性乙型肝炎（CHB）22例及键康正常人20例外周血中的CD4^＋CD25^＋high细胞，同时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组外周血中CD4^＋CD25^＋high／CD4^＋百分比显著低于正常组（P〈0、05）和CHB组（P〈O．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 Tr细胞下降可能是自身免疫性肝炎发病的原因之一。     

Maintenance of alveolitis in patients with chronic beryllium disease by beryllium-specific helper T cells

Chronic beryllium disease is characterized by the accumulation of helper/inducer T cells, macrophages, and granulomas in the lungs. To evaluate the hypothesis that the proliferation of CD4+ (helper/inducer) T cells in the lungs of patients with this disorder is maintained by local activation of beryllium-specific T-cell clones, we studied T cells obtained from peripheral blood and by bronchoalveolar lavage in eight patients and five healthy controls. The proliferation of T cells in response to beryllium in vitro was confined to the CD4+ T cells from the patients and was dependent on the presentation of antigen in the presence of both major histocompatibility complex class II antigens and functional interleukin-2 receptors. T cells from the patients' lungs had a significantly greater response to beryllium than did T cells from their peripheral blood (stimulation index, 103 vs. 5; P less than 0.01). Lines and clones of cells developed from T cells from the patients' lungs showed dose-dependent proliferation in response to beryllium but did not respond to recall antigens or to other metals. Although all beryllium-specific T-cell clones were CD4+ and none were CD8+ (suppressor/cytotoxic), all beryllium-specific clones studied had different rearrangements of T-cell antigen receptors, suggesting that the response to beryllium involved T cells with diverse specificities for beryllium. We conclude that in patients with chronic beryllium disease, beryllium acts as a class II-restricted antigen, stimulating local proliferation and accumulation in the lung of beryllium-specific CD4+ (helper/inducer) T cells. Hence, chronic beryllium disease is a hypersensitivity disease in which beryllium is the specific antigen.     

Neuropilin-1+T细胞与CD4+CD25+调节性T细胞的比较

目的:分析Neuropilin-1 T细胞(Nrp-1 T细胞)与经典CD4 CD25 调节性T细胞(Treg)的关系并比较二者的免疫调节作用。方法:流式细胞术分析BALB/c小鼠脾脏T细胞上Nrp-1与CD4、CD25的表达关系并分选Nrp-1 T细胞及CD4 CD25 Treg,通过B16-F10-luc-G5黑色素肿瘤细胞体外培养实验并利用萤光成像系统,观察比较两种细胞对NK细胞杀伤B16-F10-luc-G5黑色素瘤细胞的影响。结果:CD4 CD25 Treg中表达Nrp-1的比例为(27.28±1.17)%,明显高于CD4 CD25-T细胞的(1.63±0.08)%(P<0.01);在体外实验中,Nrp-1 T细胞与CD4 CD25 Treg均能抑制NK细胞杀伤B16-F10-luc-G5黑色素瘤细胞,Nrp-1 T细胞组的肿瘤细胞数目在6、24、48、72h分别为984±15、1015±14、1261±21、1323±38,高于CD4 CD25 Treg组的931±4、983±8、1201±18、1256±18,两组肿瘤细胞数目在各时间点均有统计学意义(P<0.01)。结论:经典CD4 CD25 Treg中表达Nrp-1的细胞比例较高,Nrp-1 T细胞有负性免疫调节作用,抑制功能比CD4 CD25 Treg更强,可以作为一类新的Treg亚群。     

Human CD4+CD25+ T cells derived from the majority of atopic donors are able to suppress TH1 and TH2 cytokine production

BACKGROUND: Recently, it has been established that CD4(+)CD25(+) T cells with regulatory capacity are present in human peripheral blood, inhibiting allogeneic proliferation and cytokine production of preactivated CD4(+)CD25(-) respond-er T cells. OBJECTIVE: The aim of this study was to analyze in an allergen-specific setting whether such regulatory CD4(+)CD25(+) T cells also exist and function normally in atopic individuals, especially concerning the inhibition of T(H)2 cytokines. METHODS: For this purpose, CD4(+)CD25(-) or CD4(+)CD25(+) T cells from donors allergic to grass or birch pollen (mainly with rhinitis) or from healthy nonatopic donors were stimulated in the presence of autologous, mature, monocyte-derived, allergen-pulsed dendritic cells, and the preactivated CD4(+)CD25(+) T cells were added to CD4(+)CD25(-) T cells during restimulation. RESULTS: CD4(+)CD25(+) T cells from the nonatopic donors and from the majority of the patients investigated proliferated poorly, produced fewer cytokines, and inhibited the proliferation and T(H)1 (IFN-gamma) and T(H)2 (IL-4 and IL-5) cytokine production of CD4(+)CD25(-) T cells but not IL-10 production. The suppression of CD4(+)CD25(-) T cells by CD4(+)CD25(+) T cells was at least partially antigen unspecific and not reversible with anti-IL-10, anti-transforming growth factor beta, or anti-cytotoxic T lymphocyte-associated antigen 4 mAb but was reversible with IL-2. In some atopic patients preactivated CD4(+)CD25(+) T cells reproducibly showed strong proliferative responses, produced higher amounts of IL-4 and IL-10 than CD4(+)CD25(-) T cells, and suppressed only the IFN-gamma production of CD4(+)CD25(-) T cells. CONCLUSION: These data indicate that regulatory CD4(+)CD25(+) T cells are present and functional in most atopic patients with allergic rhinitis and are able to inhibit T(H)1, as well as T(H)2, cytokine production.     

CD4+ CD25+ regulatory T cells suppress CD4+ T-cell function and inhibit the development of Plasmodium berghei-specific TH1 responses involved in cerebral malaria pathogenesis

The infection of mice with Plasmodium berghei ANKA constitutes the best available mouse model for human Plasmodium falciparum-mediated cerebral malaria, a devastating neurological syndrome that kills nearly 2.5 million people every year. Experimental data suggest that cerebral disease results from the sequestration of parasitized erythrocytes within brain blood vessels, which is exacerbated by host proinflammatory responses mediated by cytokines and effector cells including T lymphocytes. Here, T cell responses to P. berghei ANKA were analyzed in cerebral malaria-resistant and -susceptible mouse strains. CD4+ T-cell proliferation and interleukin-2 (IL-2) production in response to parasite-specific and polyclonal stimuli were strongly inhibited in cerebral malaria-resistant mice. In vitro and in vivo depletion of CD4+ CD25+ regulatory T (T(reg)) cells significantly reversed the inhibition of CD4+ T-cell proliferation and IL-2 production, indicating that this cell population contributes to the suppression of T-cell function during malaria. Moreover, in vivo depletion of T(reg) cells prevented the development of parasite-specific TH1 cells involved in the induction of cerebral malaria during a secondary parasitic challenge, demonstrating a regulatory role for this cell population in the control of pathogenic responses leading to fatal disease.     

Activated CD4+ CD25+ T cells suppress antigen-specific CD4+ and CD8+ T cells but induce a suppressive phenotype only in CD4+ T cells

CD4(+) CD25(+) regulatory T cells are increasingly recognized as central players in the regulation of immune responses. In vitro studies have mostly employed allogeneic or polyclonal responses to monitor suppression. Little is known about the ability of CD4(+) CD25(+) regulatory T cells to suppress antigen-specific immune responses in humans. It has been previously shown that CD4(+) CD25(+) regulatory T cells anergize CD4(+) T cells and turn them into suppressor T cells. In the present study we demonstrate for the first time in humans that CD4(+) CD25(+) T cells are able to inhibit the proliferation and cytokine production of antigen specific CD4(+) and CD8(+) T cells. This suppression only occurs when CD4(+) CD25(+) T cells are preactivated. Furthermore, we could demonstrate that CD4(+) T-cell clones stop secreting interferon-gamma (IFN-gamma), start to produce interleukin-10 and transforming growth factor-beta after coculture with preactivated CD4(+) CD25(+) T cells and become suppressive themselves. Surprisingly preactivated CD4(+) CD25(+) T cells affect CD8(+) T cells differently, leading to reduced proliferation and reduced production of IFN-gamma. This effect is sustained and cannot be reverted by exogenous interleukin-2. Yet CD8(+) T cells, unlike CD4(+) T cells do not start to produce immunoregulatory cytokines and do not become suppressive after coculture with CD4(+) CD25(+) T cells.     

CD26 expression on CD4+T cells in patients with cutaneous leishmaniasis

Surrogate marker(s) of protection in human leishmaniasis is not well defined. In this study, T helper 1 (Th1) and Th2 cytokine profiles and CD26 expression on CD4(+) T cells in peripheral blood mononuclear cells of patients with healing or non-healing forms of cutaneous leishmaniasis (CL) stimulated with Leishmania antigens were assessed. The level of interferon (IFN)-gamma production was significantly higher in patients with healing or non-healing forms of CL than in healthy controls, but it was not significantly different between the two patient groups. The level of interleukin-5 production was significantly higher in patients with the non-healing form of CL than in the two other groups. There was a significant increase in the level of CD26 expression on CD4(+) T cells in patients with healing (P < 0.001) or non-healing (P = 0.025) forms of CL compared with the control group, but no significant difference was seen between the two patient groups. A weak positive correlation was seen between IFN-gamma production and CD26 expression on CD4(+) T cells of patients with the healing form of lesion (r = 0.54, P = 0.008), but this correlation was not observed in patients with the non-healing form of CL (r = 0.53, P = 0.078). Surface CD26 is not correlated with the clinical manifestation of CL or IFN-gamma production. Therefore, CD26 is not a surrogate marker for IFN-gamma production in CL.     

CD4+CD25+ T cells as immunoregulatory T cells in vitro

We have further characterized the in vitro phenotype and function of anergic and suppressive CD4(+)25(+) T cells. Following TCR ligation, DO.11.10 CD4(+)25(+) T cells suppress the activation of OT-1 CD8(+)25(-) T cells in an antigen nonspecific manner. Although suppression was seen when using a mixture of APC from both parental strains, it was very much more marked when using F1 APC. APC pretreated with, and then separated from CD4(+)25(+) T cells did not have diminished T cell costimulatory function, suggesting that APC are not the direct targets of CD4(+)25(+) T cell regulation. CTLA-4 blockade failed to abrogate suppression by CD4(+)25(+) T cells in mixing experiments. Although CD4(+)25(+) T cells failed to respond following cross-linking of TCR, they could be induced to proliferate following the addition of exogenous IL-2, allowing the generation of a T cell line from CD4(+)25(+) T cells. After the first in vitro restimulation, CD4(+)25(+) T cells were still anergic and suppressive following TCR engagement. However, after three rounds of restimulation, their anergic and suppressive status was abrogated.     

成人隐匿性自身免疫性糖尿病患者CD4+CD25+T细胞的变化研究

目的：通过观察成人隐匿性自身免疫性糖尿病（LADA）患者CD4^＋CD25^＋T细胞的变化并与速发性1型糖尿病（T1 DM）比较，了解成人隐匿性自身免疫性糖尿病患者T细胞免疫功能的变化及与1型糖尿病的异同点。方法：LADA组24例，速发性T1DM18例，对照组20例，应用流式细胞技术测定3组人选者T细胞表面分子CD4、CD8、CD25、CD4^＋CD25^＋、CD8^＋CD25^＋、CD4^＋CD25^＋CD62L^＋，以百分比表示各表面分子阳性T细胞占外周血淋巴细胞的比例。结果：LADA与T1DM组CD4^＋T细胞、CD4／CD8比值明显高于健康对照组（P〈0．01），LADA与T1DM对比无差异。LADA组CD25^＋、CD4^＋CD25^＋、CD4^＋CD25^＋CD62L^＋T细胞高于健康对照组（P〈0．05），明显高于T1 DM组（P〈0．01）。T1 DM组CD25^＋、CD4^＋CD25^＋、CD4^＋CD25^＋CD62L^＋T细胞略低于健康对照组，但无统计学意义（P〉0．05）。结论：LADA患者外周血中诱导免疫耐受的CD4^＋CD25^＋、CD4^＋CD25^＋CD62L^＋T细胞较对照组升高并明显高于T1 DM患者。LADA患者胰岛B细胞功能下降较速发性T1 DM患者相对缓慢可能与CD4^＋CD25^＋T细胞的免疫保护有关。     

类风湿性关节炎患者CD4+CD25+Foxp3+调节性T细胞检测及意义

目的:研究CD4 CD25 FoxP3 调节性T细胞在类风湿关节炎患者(RA)外周血中的比例改变,并探讨其在疾病进程中的意义.方法:选取活动期及稳定期RA患者,采用细胞内染色的流式细胞术及定量PCR的方法,分别在蛋白质和mRNA水平检测FoxP3表达,并与正常人进行比较.结果:RA患者CD4和CD25双阳性细胞所占比例与对照组没有明显差异,而活动期患者外周血CD4 CD25high和CD4 CD25 FoxP3 细胞明显低于稳定期和对照组(P＜0.05).FoxP3 mRNA表达水平与蛋白质表达水平变化相一致.结论:类风湿性关节炎活动期时CD4 CD25 FoxP3 调节性T细胞明显减少,这群调节性T细胞可能参与了类风湿性关节炎的病理进程.     

Frequency and phenotypic analysis of CD4+CD25+ regulatory T cells in children with juvenile idiopathic arthritis.

BACKGROUND AND PURPOSE: The aim of this study was to investigate the frequency of CD4(+)CD25(+) regulatory T cells and their phenotypic expression in peripheral blood of children with active and non-active juvenile idiopathic arthritis (JIA) and healthy controls, to determine if their frequency or phenotypic expression is involved in the immunoregulation of this disease. METHODS: From October 2004 to October 2005, 55 JIA patients and 55 age- and gender-matched healthy controls were enrolled in the study at National Taiwan University Hospital. Flow cytometry was used to determine the frequency of CD4(+)CD25(+) and CD4(+)CD25(hi) in CD4(+) T cells and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) expression on CD4(+)CD25(+) by tricolor staining. Basic profiles, medication history, clinical symptoms and laboratory data were obtained by chart review and outpatient department interviews. RESULTS: There was no significant difference in expression of CD4(+)CD25(+) T cells between patients with inactive JIA and normal controls (13.74+/-3.25% vs 12.85+/-3.68%, p>0.05). The expression of CD4(+)CD25(hi) T cells was significantly lower in inactive JIA patients than in normal controls (1.89+/-1.01% vs 2.76+/-1.28%, p<0.01). The expression of CTLA-4 on CD4(+)CD25(+) T cells was also significantly lower in inactive JIA patients compared with controls (4.37+/-2.02% vs 6.33+/-2.57%, p<0.001). CONCLUSION: We speculate that a decreased frequency of CD4(+)CD25(hi) regulatory T cells and lower level of CTLA-4 expression on CD4(+)CD25(+) regulatory T cells might play a role in the immunoregulation of JIA.     

Decreased CD4+CD25+ T cells in peripheral blood of patients with systemic lupus erythematosus

Recent animal studies have shown that CD4+CD25+ T cells play a crucial role in the suppression of the immune response and that depletion of this subset of T cells might lead to development of autoimmune diseases. The aim of the present study was to investigate the levels of CD4+CD25+ T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE). Ninety-four SLE patients, 52 patients with rheumatoid arthritis (RA) and 50 age- and gender-matched healthy individuals were enrolled in the study. A flowcytometric method was applied in the measurement of CD4+CD25+ T cells. The results showed that patients with SLE had statistically lower levels of CD4+CD25+ T cells than did normal controls, when expressed as either percentages of peripheral blood mononuclear cells (PBMCs) (mean +/- SD, 8.49 +/- 6.36 versus 11.11 +/- 4.58%, P < 0.05) or absolute cell numbers (98.77 +/- 97.52 versus 213.93 +/- 104.52 cells/mm3, P < 0.05). In terms of CD25brightCD4+ T cells, defined as having a fluorescence intensity of CD25 expression exceeding 100, SLE patients still had significantly lower levels than did normal controls expressed as percentages of PBMCs (1.76 +/- 1.32 versus 3.73 +/- 1.30%, P < 0.05). No significant differences could be found between RA patients and normal controls. The overwhelming majority of CD4+CD25+ T cells belonged to CD45RO+ cells and most did not express the CD69 molecule. Although decreased CD4+CD25+ T cells were found in SLE patients, we failed to find a significant correlation between the levels of CD4+CD25+ T cells and disease activities of SLE. To the best of our knowledge, this is the first study to demonstrate that patients with SLE had decreased CD4+CD25+ T cells. However, the exact role of the decreased CD4+CD25+ T cells in the pathogenesis of SLE remains to be elucidated.     

慢性HBV感染患者肝组织内CD4+CD25+调节性T淋巴细胞的表达

目的 观察慢性HBV感染患者免疫耐受期与免疫清除期肝组织中CD4+ CD25+调节性T细胞的表达及分布情况.方法 应用免疫组织化学法检测19例免疫耐受期及12例免疫清除期慢性乙型肝炎患者肝组织中FoxP3的表达,6例正常肝组织为对照.结果 FoxP3阳性信号位于淋巴细胞胞核内,阳性细胞主要聚集在汇管区,肝窦内亦可见散在单个淋巴细胞呈阳性.在免疫耐受期及免疫清除期患者肝组织中FoxP3较正常肝组织明显增加(P<0.01),免疫清除期患者肝组织内Fox P3明显高于免疫耐受期患者(P<0.01).免疫清除期患者肝组织中FoxP3阳性标记指数与ALT、HBeAg及HBV-DNA水平无明显相关性.免疫耐受期与免疫清除期两组相比,在年龄、ALT、TBIL、PTA、HBeAg及HBV-DNA水平方面差异均有统计学意义.结论 肝组织中CD4+ CD25+调节性T细胞在慢性HBV感染慢性化和抑制免疫,控制肝脏炎症反应方面可能起了重要作用.     

CD4+CD25+Treg在急性淋巴细胞白血病患者外周血中的表达及其体外实验研究

目的初步探讨CD4 CD25 调节性T细胞(CD4 CD25 regulatory T cells,CD4 CD25 Treg)在急性淋巴细胞白血病(acute lymphocytic leukemia,ALL)患者化疗前及化疗缓解后外周血中的表达水平,并研究患者血清能否诱导外周血CD4 CD25-T细胞转化为CD4 CD25 Treg。方法①采用流式细胞术分别检测ALL初诊组、化疗完全缓解或部分缓解组及正常对照组外周血中CD4 CD25 T细胞所占比例,然后通过荧光定量RT-PCR检测各组外周血中转录因子Foxp3mRNA的表达水平,并逐层分析比较。②采集正常人外周血单个核细胞后,对照组用正常人血清,实验组用患者血清并分别设浓度梯度进行培养,72h后采用流式细胞术、荧光定量RT-PCR分别检测CD4 CD25 T细胞和Foxp3mRNA表达。结果ALL化疗缓解组CD4 CD25 T细胞及Foxp3mRNA表达水平均明显高于ALL初诊组和正常对照组(P<0.05),后两者之间CD4 CD25 T细胞水平无统计学差异(P>0.05),但ALL初诊组Foxp3mRNA含量较正常对照组明显升高(P<0.01),差异具有统计学意义;并且血清培养对照组CD4 CD25 T细胞水平及Foxp3mRNA含量均明显低于实验组(P<0.05),且其表达并不随血清浓度的增加而升高。结论CD4 CD25 Foxp3 Treg在ALL初诊组及化疗缓解组患者外周血中比例明显升高,且初步表明患者血清中的可溶性物质可诱导外周血CD4 CD25 T细胞转化为CD4 CD25 Treg,提示CD4 CD25 Treg可能是ALL免疫抑制的一个重要原因。     

The role of CD4+CD25+ regulatory T cells in patients with Rheumatoid Arthritis]

CD4(+)CD25(+) regulatory T cells play an important role in preventing autoimmunity. We investigated the presence of CD4(+)CD25(+) regulatory T cells in the peripheral blood of patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and systemic sclerosis (SSc), using flow cytometry. The percentage of CD4(+)CD25(+) regulatory T cells was significantly decreased in RA, especially in patients with high serum levels of either CRP or MMP-3. In SSc and SLE, the percentage of CD4(+)CD25(+) regulatory T cells was higher in patients than in controls, but not significant. We also investigated the serum levels of IL-10, which influences the function of CD4(+)CD25(+) regulatory T cells and other regulatory T cells. In RA, on contrast to CD4(+)CD25(+) regulatory T cells, the serum levels of IL-10 increased in patients with higher serum levels of CRP, or MMP-3. In SLE and SSc, the serum level of IL-10 increased significantly in patients than in controls. These data thus indicated that CD4(+)CD25(+) regulatory T cells contributes to occurrence and progression of RA, and other regulatory T cells or cytokines contribute to occurrence and progression of SSc and SLE.