Pityrosporum and Candida specific and non-specific humoral, cellular and cytokine responses in atopic dermatitis patients

BACKGROUND: Recent cytokine (RT-PCR, ELISA) analyses of inflammation in atopic dermatitis (AD) have suggested a role for IL-4, IL-5 and IFNgamma. Pityrosporum ovale and Candida albicans are important allergens in some patients with AD of the seborrhoic head, neck and shoulder region. In AD patients, the saprophytic yeasts induce IgE responses while they usually induce TH1 type responses. The cytokine responses induced by yeasts in AD are sparsely investigated. OBJECTIVE: To characterize the P. ovale- and C. albicans-specific and non-specific humoral, lymphoproliferative and cytokine (IL-2, 4, 5 and IFNgamma) responses in AD. METHODS: Fifteen AD patients and seven healthy controls (HC) were included. Ficoll-isolated PBMC were stimulated by PHA and laboratory-generated extracts of P. ovale and C. albicans. Lymphocyte proliferation was measured by 3H-thymidine incorporation and cytokine production by sandwich-ELISAs. The antigen-specific IgG and IgE antibodies were analysed by ELISA and nitrocellulose RAST. RESULTS: Pityrosporum ovale- and C. albicans-specific IgE (both P < 0.001) and P. ovale-induced PBMC proliferation (P < 0.02) were elevated in AD. In general, the IL-4/IFNgamma ratio induced by P. ovale was higher than that induced by C. albicans (P < 0.01). The PHA-induced IL-2 (P < 0.05) and IL-4 responses (P < 0.005), and the C. albicans-induced IL-5 response (P < 0.02) and IFNgamma response (P < 0.01), were elevated in AD. A network of correlations was seen between serum total and the yeast-specific IgE, P. ovale-specific lymphoproliferation, PHA-induced IL-2, IL-4 and IL-5, and C. albicans-induced IL-5. CONCLUSION: The cytokine profiles found in this study support the role of TH0 or TH1 cells by the side of TH2 cells in the pathogenesis of atopic dermatitis. Pityrosporum ovale appears to be associated more with IL-4 responses and C. albicans with IFNgamma responses.     

帕金森病人外周血单个核细胞分泌IL-2的实验研究

目的 探讨帕金森病病人中外周血单个核细胞(PBMC)分泌IL-2的能力.方法 采用酶联免疫吸附法(ELISA)分别检测体外培养的PBMC上清液、及PBMC接受刀豆蛋白(ConA)刺激后上清液中IL-2含量.结果 PD Ⅰ期患者中未受刺激的PBMC分泌的IL-2显著高于PDⅡ-Ⅲ期患者(P＜0.05),接受ConA刺激后,以PD Ⅱ-Ⅲ期患者的PBMC生成IL-2最低,显著低于正常对照组(P＜0.05),刺激后同刺激前相比各组IL-2都有升高,但仅健康对照组刺激前后有显著差异(P＜0.05).结论 帕金森病患者的外周血单个核细胞的分泌IL-2能力存在异常.     

Dietary nucleic acids promote a shift in Th1/Th2 balance toward Th1-dominant immunity

BACKGROUND: Dietary sources of nucleic acids and their relative components are known to affect host immune function; however, it has not yet been clarified whether such dietary nucleic acids influence the pathogenesis of allergic reaction. OBJECTIVE: The purpose of this study is to elucidate the effect of dietary nucleic acids on Th1/Th2 balance. METHODS: Both human flora-associated and specific pathogen-free BALB/c mice were maintained on either nucleic acid-free, or -supplemented diets. The effects of nucleic acids on both in vivo antibody levels and in vitro splenocyte cytokine production were compared using these mice. RESULTS: Supplementation of nucleic acids caused a reduction in the serum antibody levels of total IgM, IgG, IgG1, and IgE in the human flora-associated mice without affecting the composition of intestinal flora. In contrast, there was no significant difference of the serum IgG2a levels between nucleic acid-free and -supplemented mice. Such a phenomenon as that, the supplementation of dietary nucleic acids reduces the serum IgE or IgG1 levels, but not the IgG2a level, was also seen in the specific pathogen free mice. Moreover, when the mice were systematically challenged with ovalbumin, the supplementation of nucleic acids also suppressed the serum ovalbumin-specific IgE and IgG1 antibody levels as well as in vitro IL-4 and IL-10 secretion, while enhancing both the serum ovalbumin-specific IgG2a antibody levels and in vitro IFN gamma secretion. CONCLUSION: These results suggested that dietary nucleic acids may play an important role in promoting a shift in Th1/Th2 balance toward Th1-dominant immunity.     

Th1 (IL-2, interferon-gamma (IFN-γ)) and Th2 (IL-10, IL-4) cytokine production by peripheral blood mononuclear cells (PBMC) from patients with systemic lupus erythematosus (SLE)

We investigated the production of IL-2, IFN-γ, IL-10 and IL-4 by PBMC from 24 patients with SLE and 10 healthy individuals. Basal and mitogen-stimulated (lipopolysaccharide and phytohaemagglutinin (LPS + PHA)) cytokine production was determined in a whole blood assay (WBA). Supernatants were collected and assayed with specific ELISAs. Although the IL-2 and IFN-γ contents did not differ significantly between patients and controls under both conditions, statistically significant correlations were found between each cytokine and disease activity (SLAM index) after stimulation (respectively, r= 0.501, P = 0.01 and r = 0.631, P = 0.001). PBMC IL-10 production was significantly higher for patients than controls (P = 0.05), but no correlation between IL-10 levels and the SLAM index was obtained. IL-4 production was not statistically different between SLE patients and controls. For stimulated WBAs, the IL-10/IL-2 and IL-10/IFN-γ ratios were significantly correlated with disease severity (P = 0.02; P = 0.001, respectively). Overall, our data suggest that SLE is characterized by an elevated production of IL-10, reflecting the basal state of activation of the immune system. During exacerbation of SLE, IL-2 and IFN-γ are synthesized in larger amounts and may cause the tissue damage observed.     

The profiles of interleukin (IL)-2, IL-6, and interferon-gamma production by peripheral blood mononuclear cells from house-dust-mite-allergic patients: a role for IL-6 in allergic disease

We have developed a model to measure cytokine production by peripheral blood mononuclear cells (PBMC) in vitro. In this report, we examine the production of interleukin-2 (IL-2), IL-6, and interferon-gamma (IFN-γ) by PBMC of house-dust-mite ( Dermatophagoides pteronyssinus )-allergic subjects. When stimulated with specific allergen ( D. pteronyssinus ), PBMC of patients produced significant levels of IL-2 and high levels of IL-6, but little or no IFN-γ. Nonatopic control PBMC also produced IL-6, although at lower levels, but no IL-2 or IFN-γ. A ubiquitous antigen, streptokinase/streptodornase (SKSD), induced high levels of IL-2 in patients, but only low levels of IFN-γ and IL-6. Nonatopic controls produced similar levels of IL-2 and IL-6, but high levels of IFN-γ to SKSD. IL-2 and IFN-γ levels induced by the T-cell mitogen phytohaemagglutinin (PHA) were similar in patient and control groups, but IL-6 levels were significantly lower in the patients. IgE synthesis in vitro was shown only in atopic PBMC cultures stimulated with specific allergen. The major points can be summarized as 1) IL-2 production by atopic patients in response to allergen; 2) IL-6 production to allergen by both atopic and nonatopic patients, but significantly increased in atopic patients; and 3) defective IFN-γ production by atopic patients to both allergen and antigen. These findings suggest that IL-6 may be important in the immune response to inhalent allergens such as D. pteronyssinus , possibly by creating a cytokine environment favourable to a T^H2 response, and that atopic patients exhibit a generalized defect of IFN-γ production, not related to the response to allergen.     

雌激素刺激活动性狼疮肾炎外周血单个核细胞的IL-10表达

探讨性激素对活动性狼疮肾炎外周血单个核细胞 (PBMC)Th2细胞活化的影响。采用ELISA、RT PCR等方法检测了雌激素对 19例活动性狼疮肾炎和 16例健康人外周血单个核细胞 (PBMC)IL 10表达的调节作用。活动期狼疮肾炎PBMCIL 10mRNA和蛋白含量明显高于正常人 (P <0 0 5 ) ;与正常对照相比 ,雌激素明显提高了活动性狼疮肾炎PBMCIL 10蛋白和mRNA水平 (P <0 0 5 ) ,雄激素的加入明显抑制了雌激素诱导的活动性狼疮肾炎PBMCIL 10蛋白和mRNA水平 (P <0 0 5 )。雌激素、雄激素比例失调可能参与了活动性狼疮肾炎IL 10的高效表达     

Interleukin-4 and interferon-gamma production by peripheral blood mononuclear cells from food-allergic patients

The aim of this study was to investigate whether patients with food allergy had a cytokine imbalance of interleukin (IL)-4 and interferon-gamma (IFN-γ) production. Diagnostic procedures including skin prick tests, determination of food-specific serum IgEs, and positive double-blind, placebo-controlled, food challenges identified 15 adult patients. They were compared with 15 age- and sex-matched healthy subjects. Peripheral blood mononuclear cells (PBMC) were incubated for 24, 48, and 72 h in the presence of phytohemagglutinin plus phorbol myristate acetate. After mitogen stimulation, culture supernatants from patients with food allergy contained significantly less IFN-γ but increased IL-4 when compared with healthy controls. Secretion of IL-4 was maximal at 24 h and IFN-γ secretion was maximal at 72 h. There was no correlation between cytokine secretion in vitro and serum IgE level. These findings demonstrated that an imbalance of IL-4 and IFN-γ production is present in food allergy, as documented in other allergic diseases, but other mechanisms are probably also involved.     

IFN-γ production in response to IL-18 or IL-12 stimulation by peripheral blood mononuclear cells of atopic patients

BACKGROUND: Several studies have shown that interleukin (IL)-4 and interferon-gamma (IFN-gamma) are important for the regulation of immunoglobulin E (IgE) production and that IL-18 and IL-12 induce IFN-gamma. OBJECTIVE: IFN-gamma production in response to IL-18 or IL-12 stimulation was investigated in peripheral blood mononuclear cells (PBMCs) of atopic patients with various levels of serum IgE. METHODS: Cytokine production from PBMCs was measured following stimulation with a non-specific stimulator (phytohemagglutinin: PHA), IL-18 or IL-12 in 12 healthy controls and 26 atopic patients with various serum IgE levels. RESULTS: IFN-gamma production by IL-18-stimulated PBMCs was positively correlated with IFN-gamma production by IL-12-stimulated PBMCs (P < 0.05). However some atopic patients showed discrepancy between the levels of IFN-gamma production stimulated by IL-12 and by IL-18. CONCLUSIONS: The results shown here suggest the presence of abnormalities in the IL-12 and/or IL-18 signalling pathways, such as genetic defects in the atopic patients.     

Chlamydia pneumoniae enhances the Th2 profile of stimulated peripheral blood mononuclear cells from asthmatic patients

Chlamydia pneumoniae is a cause of respiratory infection in adults and children. There is evidence for an association between atypical bacterial respiratory pathogens and the pathogenesis of asthma. We compared T helper (Th) responses in C. pneumoniae – infected peripheral blood mononuclear cells (PBMC) in patients with or without asthma. PBMC (1 × 10⁶/mL) from asthmatic patients (N = 11) and non-asthmatic controls (N = 12) were infected or mock-infected for 1 h +/− C. pneumoniae TW-183 at a multiplicity of infection (MOI) = 1 and MOI = 0.1, or cultured for 24 h +/− Lactobacillus rhamnosus GG (LGG). Interleukin (IL)-4, IL-10, IL-12, Interferon (IFN)-gamma and total IgE levels were measured in supernatants (ELISA). C. pneumoniae infection led to an increase (>50%) of IgE levels in PBMC from asthmatics, compared with mock-infected on day 10; IgE wasn’t detected in non-asthmatics. C. pneumoniae – infected PBMC from asthmatics increased levels of IL-4 and IFN-gamma after 24 h, compared with PBMC alone; levels of IL-10 and IL-12 were low. When uninfected-PBMC from asthmatics were LGG-stimulated, after 24 h, IL-4 was undetectable, but IL-10, IL-12, and IFN-gamma increased, compared with PBMC alone. Thus, C. pneumoniae infection has the ability to induce allergic responses in PBMC of asthmatics, as evidenced by production of Th2 responses and IgE.     

Altered production of IgE and IgA induced by IL-4 in peripheral blood mononuclear cells from patients with IgA nephropathy.

In order to elucidate the factors responsible for altered immunoglobulin production in patients with IgA nephropathy (IgAN), the in vitro effects of IL-4 and interferon-gamma (IFN-gamma) on the synthesis of IgE and IgA by peripheral blood mononuclear cells (PBMC) were studied. Spontaneous IgE and IgA synthesis by PBMC was significantly increased in patients with IgA nephropathy compared with controls. The maximum amounts of IgA and IgE synthesis by PBMC after stimulation with IL-4 were almost the same both in patients with IgAN and in controls. The enhancement rate of IL-4-induced IgE and IgA synthesis was significantly lower in IgAN than in the controls, suggesting in vivo preactivation of PBMC in IgAN patients. IFN-gamma suppressed IgA and IgE synthesis by PBMC from IgAN patients as well as controls. However, the suppressive effect on IgE synthesis was less prominent in patients with IgAN. These results suggested that altered IL-4 action might be involved in the development of IgA nephropathy.     

Effects of ONO-1078 (pranlukast) on cytokine production in peripheral blood mononuclear cells of patients with bronchial asthma

BACKGROUND: ONO-1078 (pranlukast) is a leukotriene receptor antagonist developed in Japan. This drug has been shown to be useful in oral treatment of bronchial asthma. The present study was undertaken to assess the effects of this drug on the production of cytokines in the peripheral blood mononuclear cells of patients with asthma under stimulation with specific antigens. METHODS: Peripheral blood mononuclear cells from mite antigen-positive asthmatic patients (immunoglobulin E RAST score > 3) were incubated for 72 h in the presence of mite antigen (10 microg/mL). The supernatant of the culture was subjected to enzyme-linked immunosorbent assay (ELISA) to quantify interleukin (IL) -4, IL-3, IL-5, and granulocyte macrophage-colony stimulating factor (GM-CSF). Other peripheral blood mononuclear cells from the same patients were incubated for 72 h in the presence of both mite antigen (10 microg/mL) and ONO-1078 (0.5, 1, or 10 microg/mL), followed by ELISA of the supernatant to quantify the cytokines. RESULTS: Production of IL-4, IL-5, and GM-CSF by mononuclear cells under stimulation with mite antigen was markedly suppressed when they were exposed to ONO-1078 at a concentration of 10 microg/mL. CONCLUSION: The results suggest that ONO-1078 acts directly on peripheral blood mononuclear cells and that blockade of leukotriene receptors on blood mononuclear cells by the cysteinyl-leukotriene receptor antagonist (LTRA) pranlukast (ONO-1078) can dose-dependently inhibit release of immunoreactive TH2-type cytokines (IL-3, IL-4, GM-CSF, and possibly IL-5), but not of the TH1-type cytokine IL-2, when stimulated by mite allergen in vitro. The data may provide clues to the mechanism by which a number of LTRA including zafirulukast and montelukast can reduce airway, sputum and blood eosinophil counts in clinical asthma. It supports animal studies showing that anti-IL-5 antibodies partially block cys-LT-induced airway eosinophilia, suggesting that cys-LTs may cause secondary release of IL-5 from an unknown cell-type. These findings indicate that ONO-1078 suppresses the production of IL-4 (a cytokine that affects IgG antibody production), IL-5, and GM-CSF (cytokines that affect eosinophil activation) by peripheral blood mononuclear cells under stimulation with specific antigens in patients with bronchial asthma. Because of its anti-inflammatory effects, ONO-1078 should be useful in the treatment of bronchial asthma.     

Expression profiling of IL-10-regulated genes in human monocytes and peripheral blood mononuclear cells from psoriatic patients during IL-10 therapy

Interleukin-10 (IL-10), originally identified as an inhibitor of pro-inflammatory cytokine production, exerts multiple immunomodulatory functions. Its ability to inhibit a Th1 response has been used in clinical trials for the treatment of inflammatory diseases including psoriasis. However, little is known about the molecular mechanisms of IL-10 functions. We aimed at identifying possible mediators of in vitro IL-10 treatment in monocytes by gene chip technology using Hu95a Affymetrix mRNA arrays with 12,000 genes. To prove relevance of the identified genes for the clinical situation we compared these in vitro results with genes being regulated by IL-10 in peripheral blood mononuclear cells from psoriatic patients undergoing IL-10 therapy. A high proportion of the 1,600 genes up-regulated and 1,300 genes down-regulated in vitro was found to be similarly regulated in vivo. Some genes, which were previously unknown to be regulated by IL-10, can be assigned to known IL-10 functions like e.g. the increase of pathogen clearance. Other new potentially immunomodulating genes have been identified to be regulated by IL-10, but their impact needs to be experimentally evaluated. We could confirm a recently reported up-regulation of heme oxygenase-1 (HO-1). However, we demonstrate that the anti-inflammatory mechanisms of IL-10 remain functional even when HO-1 is irreversibly inhibited.     

Effects of interleukin-4 and interferon-γ on the secretion of IgG4 from human peripheral blood mononuclear cells

Whereas IgE antibodies are linked with allergy, IgG4 antibodies may reflect the state of immunity and protection against a particular antigen. It has been shown that interleukin (IL)-4 is required for induction of IgE synthesis. In order to elucidate the role of IL-4 in the production of IgG4 and to compare IgG4 and IgE regulatory processes, we quantified these immunoglobulin isotypes after in vitro culture of peripheral blood mononuclear cells (PBMC) in the presence of IL-4. The production of IgG4 was increased by IL-4 under the same conditions which are optimal for IgE production but not among PBMC from all donors, depending on the magnitude of spontaneous IgG4 secretion: IL-4 was effective only when the spontaneous secretion of IgG4 was < 7% of the total IgG secretion; it had no effect when spontaneous IgG4 production was >7% of total IgG. The IL-4-induced IgE response was consistently obtained when IgG4 was < 7% of total IgG but was markedly diminished or absent when IgG4 was > 7% of total IgG. If Staphylococcus aureus strain Cowan 1 (SAC) was present during the 48-h preincubation step, spontaneous IgG4 production was increased, but the stimulatory effect of this mitogen on immunoglobulin production, including IgG4, was markedly blocked by the addition of IL-4. In contrast, IL-4-induced IgE synthesis was strongly blocked by the presence of SAC. Finally, secretion of IgG4 (spontaneous and IL-4-induced) was suppressed among cells from most donors by interferon-γ (IFN-γ). These results suggest that IL-4 has opposite effects on in vitro IgG4 production and that the in vitro synthesis of both IgG4 and IgE appears to be regulated similarly by IL-4 and IFN-γ, whereas additional signals promote the production of one isotype in preference to the other. It is possible that activated B cells respond to IL-4 less well than do nonactivated cells.     

系统性红斑狼疮患者外周血单个核细胞中IL—4和IL—13 mRNA的表达

目的探讨Th2型细胞因子在系统性红斑狼疮(SLE)发病机制中的作用及其意义.方法应用逆转录-聚合酶链反应(RT-PCR)检测了34例活动期SLE患者和30例正常人外周血单个核细胞(PBMC)中IL-4和IL-13mRNA的表达水平.结果活动期SLE患者IL-4和IL-13的阳性表达率与正常人对照组相比均无明显差异(P>0.05);活动期SLE患者PBMC中IL-4和IL-13mRNA的平均表达水平(0.938 6±0.168 9,0.898 3±0.115 3)均明显高于正常人对照组(0.5494±0.151 0,0.608 5±0.090 3),差异非常显著(P<0.001).结论Th2型细胞因子IL-4和IL-13在SLE患者中呈高水平表达,这可能与SLE患者外周血T细胞高度活化、功能异常有关.     

系统性红斑狼疮患者外周血Th2型细胞因子受体水平的研究

目的探讨Th2型细胞因子受体(IL-4R、IL-6R、IL-10R)在系统性红斑狼疮(SLE)发病机制中的作用及其临床意义.方法应用逆转录酶-聚合酶链反应(RT-PCR)检测89例SLE患者和30例正常人外周血单核细胞(PBMC)中IL-4R、IL-6R和IL-10R mRNA的表达水平;用ELISA法检测血清中IL-4、IL-6和IL-10水平.结果①活动期SLE患者和非活动期SLE患者及正常人Th2型细胞因子受体阳性表达率为100%.②PBMC中,IL-4R mRNA表达水平活动期SLE患者(1组)为0.604±0.147,非活动期SLE患者(2组)为0.40±0.13,正常人(3组)为0.37±0.07;IL-6R mRNA表达水平1组为0.90±0.27,2组为0.52±0.11,3组为0.57±0.24;IL-10R mRNA表达水平1组为0.87±0.29,2组为0.72±0.21,3组为0.68±0.14.1组和2组间比较无显著性差异(P＜0.05),1组和3组间比较有显著性差异(P=0.00),2组和3组间比较无显著性差异(P＞0.05).③血清中IL-4、IL-6和IL-10水平1组显著高于2组和3组(P＜0.05),2组显著高于3组(P＜0.05).活动期SLE患者和非活动期SLE患者血清中的IL-4水平与PBMC上的IL-4R的表达水平呈正相关(r=0.622和r=0.859,P＜0.05).活动期SLE患者和非活动期SLE患者血清中的IL-6水平与PBMC上的IL-6R的表达水平呈正相关(r=0.887和r=0.615,P＜0.05).活动期SLE患者和非活动期SLE患者血清中的IL-10水平与PBMC上的IL-10R的表达水平呈正相关(r=0.888和r=0.787,P＜0.05).结论Th2型细胞因子及其受体的异常表达可能在SLE疾病活动和发展过程中起重要作用.检测SLE患者PBMC中Th2型细胞因子受体的表达水平可作为疾病的活动性指标之一.     

系统性红斑狼疮患者外周血单个核细胞中白细胞介素-10mRNA表达的研究

目的 探讨白细胞介素(IL-10)在系统性红斑狼疮(SLE)中的作用。方法 采用逆转录多聚酶链反应(RT-PCR)及酶联免疫吸附法(ELISA)测定40例SLE患者和20例正常对照组外周血单核细胞(PBMC)IL-10mRNA表达及IL-10自发分泌水平。结果 SLE患者PBMC自发分泌IL-10水平及其IL-10mRNA表达水平均显著高于正常对照组(P<0.01),其中SLE活动期明显高于非活动期(P<0.01),而非活动期又明显高于正常对照组(P<0.01)。结论 IL-10在SLE发病中起重要作用,PBMC分泌IL-10水平对SLE诊断和病情活动性监测有重要临床意义,拮抗SLE患者体内IL-10水平,将为SLE治疗开辟一条新途径。     

Apoptotic cells selectively suppress the Th1 cytokine interferon gamma in stimulated human peripheral blood mononuclear cells and shift the Th1/Th2 balance towards Th2

Apoptotic cells are readily recognized and engulfed by phagocytes and usually do not induce inflammation or tissue damage. Furthermore, they can actively suppress a pro-inflammatory response in phagocytes: In the presence of apoptotic cells, activated monocytes/macrophages produce more of the anti-inflammatory and immunoregulatory cytokines IL-10 and TGF-beta, but less of the pro-inflammatory cytokines TNFalpha, IL-1beta and IL-12. This immunoregulatory effect is most likely mediated by several receptors on monocytes/macrophages including the thrombospondin receptor (CD36). In addition to the modulation of cytokine secretion, apoptotic cell material inhibited the expression of MHC class II molecules on the surface of monocytes/macrophages. Decreased MHC II expression appeared to be mediated predominantly by increased IL-10 secretion in a para-/autocrine manner. Here, we show that the functional modulation of antigen-presenting monocytes/macrophages by apoptotic cells also influences T cell activation and function. When human peripheral blood mononuclear cells were stimulated with recall antigens in the presence of apoptotic cells, interferon gamma (IFN gamma) secretion was markedly suppressed, whereas secretion of the Th2 cytokine IL-4 was not significantly altered. Hence, apoptotic cells shift the T cell cytokine secretion pattern towards a Th2-like response. This Th2 shift can largely be prevented by neutralizing IL-10, indicating an important role of this cytokine for modulating T cell cytokine secretion patterns.