Granulocyte colony-stimulating factor promotes the generation of regulatory DC through induction of IL-10 and IFN-alpha

We have recently demonstrated that G-CSF promotes the generation of human T regulatory (T(REG)) type 1 cells. In this study, we investigated whether the immunomodulatory effects of G-CSF might be mediated by DC. CD14(+) monocytes were cultured with serum collected after clinical administration of G-CSF (post-G), which contained high amounts of IL-10 and IFN-alpha. Similar to incompletely matured DC, monocytes nurtured with post-G serum acquired a DC-like morphology, expressed high levels of costimulatory molecules and HLA-DR, and exhibited diminished IL-12p70 release and poor allostimulatory capacity. Importantly, post-G DC-like cells were insensitive to maturation stimuli. As shown by neutralization studies, IFN-alpha and, even more pronounced, IL-10 contained in post-G serum inhibited IL-12p70 release by post-G DC-like cells. Furthermore, phenotypic and functional features of post-G DC-like cells were replicated by culturing post-G monocytes with exogenous IL-10 and IFN-alpha. Post-G DC-like cells promoted Ag-specific hyporesponsiveness in naive allogeneic CD4(+) T cells and orchestrated a T(REG) response that was dependent on secreted TGF-beta 1 and IL-10. Finally, neutralization of IL-10 and IFN-alpha contained in post-G serum translated into abrogation of the regulatory features of post-G DC-like cells. This novel mechanism of immune regulation effected by G-CSF might be therapeutically exploited for tolerance induction in autoimmune disorders.     

Trimodality of serum Dermatophagoides farinae-specific IgE RAST levels in atopic dermatitis patients.

Serum Dermatophagoides farinae-specific IgE RAST (DF IgE) levels were assayed in 122 atopic dermatitis patients at the first examination. From the statistical study, we found trimodality of the individual variability of serum DF IgE levels. We hypothesize that serum DF IgE levels may be controlled by a pair of allelic genes.     

哮喘患者IL—10水平与IgE含量的关系

本文用ELISA、细胞培养、RT-PCR方法研究哮喘患者的血浆和单个核细胞IL-10的水平及其与IgE的关系。结果显示32例哮喘患者血浆、细胞培养上清IL-10值和外周血单个核细胞IL-10转录水平均比20例正常人明显降低,血浆IL-10降低组不IgE明显高于血浆IL-10正常组。IL-10是一种抗炎因子,过敏性哮喘患者IL-10浓度降低说明其对过敏性炎症的抑制能力减弱,与哮喘的发病有关。     

儿童原发性单纯性肾病综合征患者白细胞介素4及IgE水平的相关性

本文报道应用抗人 IL-4McAb 及 PcAb 建立了双抗体夹心检测人 IL-4ELISA 法.对20例原发性单纯性肾病综合征患者、16例相应年龄组健康儿童及6例非肾病综合征患儿进行了 IL-2,IL-4 及 IgE水平的测定,并分析了 IL-4及 IgE 的相关性.结果表明,健康儿童 IgE 水平为50-100U/ml,IL-4水平为400-500pg/ml;肾病综合征患儿 IgE 水平为300->1000U/ml,IL-4水平为1080-4000Pg/ml.患儿 IgE 水平的升高与 IL-4水平的升高呈正相关,R 值为0.691.     

Passivein vitro sensitization of human basophils with dermatophagoides farinae specific IgE

Summary Basophils from allergic or non-atopic donors were depleted for their native membrane IgE by acid treatment and then passively sensitized by Dermatophagoïdes farinae specific IgE containing sera. Histamine release experiments were performed with a highly purified allergen (Df 11) on native cells, acid treated cells and passively sensitized cells. In the sensitization procedure, the quantity of the basophil-bound serum IgE is dependent on the concentration of the sensitizing serum IgE and the histamine release capacity is specifically acquired. It is shown that after passive sensitization basophil membrane IgE density as well as basophil sensitivity to Df 11 in histamine release experiments depend on the ratio [Df 11 specific IgE/total IgE] in the sensitizing serum.     

IL-4和IL-10调节嗜碱性粒细胞CXCR4表达及功能

目的 研究IL-4和IL-10对人嗜碱性粒细胞上CXC趋化性细胞因子受体-4（CXCR4）表达和配体SDF-1α（Chemokine stromal cell-derived factor-1 alpha)功能的调节。方法 嗜碱性粒细胞的纯化技术,流式细胞术,实时定量逆转录PCR（RT-PCR）,胞内游离Ca^2 的变化。趋 化性技术和组胺释放等方法进行测定与分析。结果 CXCR4大量表达在人外周血静息嗜碱性粒细胞上,IL-4可显著上调CXCR4蛋白和mRNA的表达,而IL-10则明显下调其表达,SDF-1α可通过CXCR4诱导嗜碱性粒细胞中游离Ca^2 增加,激活嗜碱性粒细胞使之产生趋化性游走并释放组胺,此活性可被抗CXCR4单抗所阻断。结论 IL-4和IL-10是CXCR4表达和功能重要的调节细胞因子,CXCR4-SDF-1α复合物相互作用对嗜碱性粒细胞的聚集和活化起着重要作用。     

Upon prolonged allergen exposure IL-4 and IL-4Ralpha knockout mice produce specific IgE leading to anaphylaxis

BACKGROUND: IL-4 and IL-13 are key regulators in atopic disorders and both signal through the receptor chain IL-4Ralpha. IL-4 and IL-13 are also the only cytokines known to induce class switching to IgE. We sought to compare allergen-specific IgE responses and allergic reactivity of wild-type (wt) mice with IL-4-/- and IL-4Ralpha-/- mice, which lack both IL-4 and IL-13 functions. METHODS: BALB/c wt, IL-4-/- and IL-4Ralpha-/- mice were immunized with ovalbumin intranasally or intraperitoneally and specific antibody titers were measured by ELISA. Bronchoalveolar lavage fluids and lung tissue were analyzed cytologically and histologically. Allergic reactivity was determined by active cutaneous anaphylaxis and anaphylactic shock. RESULTS: wt mice immunized intranasally or intraperitoneally showed high titers of specific IgE 3 and 6 weeks after primary sensitization, resulting in cutaneous anaphylaxis and anaphylactic shock upon challenge. Intranasal sensitization resulted in airway eosinophilia and goblet cell metaplasia. In contrast, IL-4-/- and IL-4Ralpha-/- mice showed no specific IgE after 3 weeks, but produced high titers after 6 weeks. At this time cutaneous anaphylaxis and anaphylactic shock could be induced as in wt mice, but lung pathology was absent. CONCLUSIONS: We conclude that upon long-term allergen exposure, alternative switch mechanisms independent of IL-4 and IL-4Ralpha may induce IgE but not asthma-like lung pathology. This may be relevant for the development of allergic disease, since long-term allergen exposure is a frequent condition during allergic sensitization.     

IL-18 induction of IgE: dependence on CD4+ T cells, IL-4 and STAT6

Overproduction of immunoglobulin E (IgE) and T helper cell type 2 (TH2) cytokines, including interleukin 4 (IL-4), IL-5 and IL-13, can result in allergic disorders. Although it is known that IL-4 is critical to the polarization of na?ve CD4+ T cells to a TH2 phenotype, both in vitro and in many in vivo systems, other factors that regulate in vivo IL-4 production and TH2 commitment are poorly understood. IL-18, an IL-1-like cytokine that requires cleavage with caspase-1 to become active, was found to increase IgE production in a CD4+ T cells-, IL-4- and STAT6-dependent fashion. IL-18 and T cell receptor-mediated stimulation could induce na?ve CD4+ T cells to develop into IL-4-producing cells in vitro. Thus, caspase-1 and IL-18 may be critical in regulation of IgE production in vivo, providing a potential therapeutic target for allergic disorders.     

Elimination of IgE regulatory rat CD8+ T cells in vivo increases the co-ordinate expression of Th2 cytokines IL-4, IL-5 and IL-10.

Immunization of rats with soluble antigen (ovalbumin) and the castor bean toxin, ricin, eliminates a subpopulation of CD8+ T cells which suppress IgE responses in vivo. This treatment also reduces the ability of splenic T cells to produce interferon-gamma (IFN-gamma) and enhances their capacity to make interleukin-4 (IL-4). In this report we describe the effect of immunization with ricin and antigen on the expression of mRNA for other T-helper type 2 (Th2) cytokines--IL-5 and IL-10--and their relationship to serum IgE and IL-4 mRNA expression. Splenocytes were taken from rats at different times after immunization with antigen or ricin and antigen and activated in vitro with phorbol myristate acetate (PMA) and ionomycin for 6 hr and total RNA extracted and reverse transcribed. Cytokine gene expression was detected using a quantitative polymerase chain reaction (PCR). Expression of IL-4, IL-5, and IL-10 was increased 7-20-fold 11 days after immunization with ricin and antigen (from 0.107% to 0.769% beta-actin for IL-4, from 0.0167% to 0.381% beta-actin for IL-5, and from 0.0581% to 0.954% beta-actin for IL-10), and preceded maximum serum IgE levels by 4-5 days. There was no increase in IgE or mRNA for these cytokines in rats immunized with antigen alone. The level of IL-4, IL-5, and IL-10 expression declined rapidly after 12 days. Our results suggest that immunization with antigen and ricin preferentially induces a Th2 response, and that CD8+ T cells may play a part in down-regulating the development of Th2 T cells.     

支气管哮喘患者血清IL-10与IgE水平变化的研究

分别用ELISA、IRMA法对50例支气管哮喘急性发作患者和48名对照者血清IL-10、IgE进行测定.结果是: 支气管哮喘患者IL-10含量明显低于对照组(P＜0.01), 而IgE含量则明显高于对照组(P＜0.01), 两者之间呈明显负相关(r=-0.18,P＜0.01).结论是: 支气管哮喘患者IL-10不足、IgE水平增高可能成为其发病的重要原因之一.     

Counter regulation of the high affinity IgE receptor, FcεRI, on human airway dendritic cells by IL-4 and IL-10

Background:  Immunoglobulin E is a signalling molecule within the environment of the respiratory tract, the high affinity receptor for which, FcεRI, is expressed by dendritic cells (DC). Little is known, however, of the expression and function of FcεRI on DC in the human respiratory tract.
Methods:  CD1c⁺ DC were purified from surgically resected nasal turbinates of 11 atopic and 12 nonatopic patients with chronic rhinosinusitis. Expression of FcεRI was determined by flow cytometry. Cytokine production by DC was determined by cytometric bead array.
Results:  Expression of FcεRI was significantly elevated on respiratory tract dendritic cells (RTDC) from atopic as compared to nonatopic patients. Activation of RTDC through FcεRI induced production of the pro-inflammatory cytokines IL-6 and TNF-α, and the anti-inflammatory cytokine IL-10. The production of IL-6 and TNF-α was elevated in atopic compared to nonatopic patients studied. Conversely IL-10 production was elevated in nonatopic patients. Concomitant activation of FcεRI and stimulation of RTDC with IL-4 inhibited production of IL-10 by RTDC. Neutralization experiments with anti-IL-10 Ab enhanced whereas addition of exogenous IL-10 to RTDC inhibited FcεRI-mediated inflammatory cytokine production.
Conclusion:  The function of FcεRI on RTDC from patients with rhinosinusitis is susceptible to counter regulation by IL-4 and IL-10.     

CD4+ Foxp3+ regulatory T cells mediate Toxoplasma gondii-induced T-cell suppression through an IL-2-related mechanism but independently of IL-10

Acute Toxoplasma gondii infection comprises an immunosuppression stage, characterized by a reduction in T-cell proliferation in vitro. Treg cells maintain the homeostasis of the immune system, but their role in T. gondii-induced suppression has not been addressed. We show herein that immunosuppression, affecting both CD4(+) and CD8(+) T-cell proliferation, concurs with a reduction in Treg-cell number. The residual Treg cells, however, are activated and display an increased suppressive capacity. We show that selective elimination of Treg cells using Foxp3(EGFP) mice leads to a full recovery of CD4(+) and CD8(+) T-cell proliferation. After Treg-cell removal, a reduced production of IL-10 was observed, but IL-2 levels were unchanged. The numbers of IL-10-producing Treg cells also increased during infection, although the in vitro neutralization of this cytokine did not modify T-cell proliferation, suggesting that IL-10 does not mediate the Treg-mediated suppression. However, addition of rIL-2 in vitro fully restored T-cell proliferation from infected animals. Thus, we show that Treg cells mediate the T-cell suppression observed during acute T. gondii infection through an IL-2-dependent mechanism. Our results provide novel insights into the regulation of the immune response against T. gondii.     

Anti-inflammatory effects of IL-4 and IL-10 on human polymorphonuclear leukocytes

Inflammatory responses are strictly regulated by coordination of pro-inflammatory and anti-inflammatory mediators. Interleukin-4 (IL-4) and interleukin-10 (IL-10) have typically the biologic anti-inflammatory effects on monocytes, but uncertain effects on polymorphonuclear leukocytes (PMNs). The PMNs are the first line of cellular response for host defense during acute inflammation. To modify hyper-inflammatory reaction with biologic anti-inflammatory mediators, we have determined the biologic anti-inflammatory activities of IL-4 and IL-10 on human PMNs. Human PMNs were pretreated with IL-4 or IL-10 and then stimulated with formyl methionyl leucyl phenylalanine (fMLP) for times indicated. The level of H2O2, interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) were determined in the each cell free supernatants. fMLP plays the role of a typical pro-inflammatory agent and, at least in determined conditions, down-regulated TNF release. IL-4 acts as an anti-inflammatory mediator but IL-10 did not show its anti-inflammatory activities on fMLP-stimulated human PMNs. IL-4 and IL-10 have different anti-inflammatory mechanisms. Perhaps, IL-10 needs co-factors to act as an anti-inflammatory mediator.     

过敏性哮喘患者血浆IL-4与IgE及嗜碱性细胞关系的研究

对30例过敏性哮喘患者和24例正常人分别检测了血浆IL-4、IgE、全血嗜碱性细胞值和以尘螨作为刺激剂的嗜碱性细胞释放介质能力,可观察到哮喘组IL-4平均值为35.2ng/L,虽高于正常组的20.33ng/L,但无显著差异,10％哮喘患者的IL-4值>150ng/L,而正常人全部低于此值。哮喘组血浆IgE平均值为486.80IU相似文献   

Regulation of surface FcepsilonRI expression on human eosinophils by IL-4 and IgE

BACKGROUND: Recent studies have demonstrated that eosinophils from allergic patients express low levels of FcepsilonRI on their surface, but the regulatory mechanisms of eosinophil surface FcepsilonRI expression are not fully understood. We investigated whether IL-4 and IgE, which are reported to regulate surface FcepsilonRI expression on human mast cells, are able to affect surface FcepsilonRI expression in normal human eosinophils. METHODS: Eosinophils purified from peripheral blood were cultured with IL-5 and with or without IL-4 and/or IgE, and surface FcepsilonRI expression was analyzed by flow cytometry using an anti-FcepsilonRI mAb, CRA-1. RESULTS: Apparent FcepsilonRI expression (approximately 1% of mast cell FcepsilonRI levels) was observed in eosinophils cultured with both IL-4 and IgE. A combination of IL-4 (>or=1 ng/ml) and IgE (>or= 0.5 microg/ml) was necessary for the maximal induction of surface FcepsilonRI expression. In the presence of IL-4 and IgE, eosinophils cultured for 2 days demonstrated low but statistically significant levels of surface FcepsilonRI, which reached a plateau after 7 days of culture. However, cross-linkage of surface FcepsilonRI molecules by CRA-1 or anti-IgE did not induce any eosinophil activation. CONCLUSIONS: IL-4 and IgE can affect the levels of surface FcepsilonRI on normal human eosinophils. FcepsilonRI expression on eosinophils may be regulated by a mechanism similar to that in mast cells.     

IL-4 and IL-10 modulate autoimmune haemolytic anaemia in NZB mice

New Zealand Black (NZB) mice spontaneously develop autoimmune haemolytic anaemia (AIHA). Here the effect of injecting NZB mice with plasmids encoding IL-4 (pIL-4) or IL-10 (pIL-10) on NZB disease was tested. Both constructs delayed the development of anaemia as judged by increased haematocrit values as compared with controls, but neither altered the IgG1 to IgG2 red blood cell (RBC) bound autoantibody levels. The increased haematocrit value was associated temporally with increased RBC bound IgG in NZB mice treated with pIL-10, but not pIL-4. By contrast, up-regulation of splenic macrophage FcgammaRIIb2 mRNA was associated temporally with increased haematocrit values in NZB mice given pIL-4. However, no such increase occurred in NZB mice that inhaled a peptide containing a dominant T-cell epitope, although this treatment is known to bias the autoimmune response towards Th2 and to reduce the severity of anaemia. It is considered that IL-4 treatment, in part, ameliorates NZB anaemia by increasing the expression of the inhibitory FcgammaRIIb2 and thereby reducing the capacity of splenic macrophages to phagocytose autoantibody coated RBC, but that this mechanism does not explain the beneficial effects of the inhaled peptide.     

Regulation of anaphylactic IgG1 antibody production by IL-4 and IL-10

BACKGROUND: Different cytokines have been implicated in the regulation of isotype expression in primary and secondary antibody responses. The aim of this study was to assess the regulation of anaphylactic IgG1 and IgE antibodies by IL-4, IL-10 and IFN-gamma at different time points of the antibody response against PI, an immunosuppressive fraction of Ascaris suum extract, and ovalbumin (OVA). METHODS: Wild-type or cytokine-deficient C57BL/6 or BALB/c mice were immunized with PI or OVA in different adjuvants. Twenty days later, they were boosted with the respective antigen. IgG1 and IgE antibodies produced during primary and secondary responses were measured by passive cutaneous anaphylaxis. RESULTS: PI induced low levels of anaphylactic IgG1 antibodies in the primary response and moderate levels after the antigenic booster, which were IL-4-dependent. In the absence of IL-10 and IFN-gamma, PI-specific IgG1 and IgE enhanced significantly, indicating that these cytokines downregulated antibody production in primary and secondary responses. The IgG1 response to OVA in aluminium hydroxide or complete Freund's adjuvant was IL-4-dependent in the beginning of the primary response. Later on, it became only partially regulated by IL-4 in C57BL/6 mice and IL-4-independent in Th2-prone BALB/c mice. In contrast, IgE antibodies depended exclusively upon IL-4 during the entire time course. CONCLUSIONS: These results indicate, first, that the IL-4 dependency of anaphylactic IgG1 antibody production, mainly in the secondary response, varies among mouse strains, and, second, that the nature of the antigen determines whether IL-10 and IFN-gamma limit the potential to make large amounts of anaphylactic IgG1 and IgE.