IL-5 release of CD4+ non-effector lymphocytes is increased in COPD--modulating effects of moxifloxacin and dexamethasone

T-lymphocytes are crucial in chronic obstructive pulmonary disease (COPD) pathogenesis. Especially T(H)1-lymphocytes are involved in local and systemic inflammation in COPD, yet the role of T(H)2-mediated immune-responses in COPD pathogenesis is poorly understood. The objective of this study was to examine IL-5 expression in T(H)2-lymphocytes in smokers with and without COPD ex vivo compared with non-smokers and to evaluate the effects of bacterial endotoxin (lipopolysaccharide, LPS) as well as two drugs often used for treatment of COPD exacerbation, corticosteroids and moxifloxacin. CD4(+) lymphocytes were isolated from the peripheral blood of non-smokers (NS; n=11), current smokers without airflow limitation (S; n=11) and smokers with COPD (n=11). Baseline IL-5 release of CD4(+) T-lymphocytes was significantly increased in COPD compared to S and NS. After T-cell activation and differentiation into T(H)2-lymphocytes, IL-5 release increased without differences between the cohorts. LPS reduced IL-5 release of ex vivo generated T(H)2-lymphocytes without differences in all cohorts. Moxifloxacin and dexamethasone significantly reduced IL-5 release in T(H)2-lymphocytes in the absence and presence of LPS without differences between groups. In summary, our data indicate that IL-5 might contribute to systemic inflammation in smokers with COPD and that T(H)2-based immune responses might be suppressed in response to gram-negative bacterial infections independent from smoking and disease status. Dexamethasone and moxifloxacin both have T(H)2-immunmodulating effects.     

Resveratrol impairs the release of steroid-resistant cytokines from bacterial endotoxin-exposed alveolar macrophages in chronic obstructive pulmonary disease

Airway inflammation in chronic obstructive pulmonary disease (COPD) is believed to be insensitive to corticosteroids. However, corticosteroids are recommended in COPD (GOLD stages III, IV) with frequent exacerbations. Resveratrol has anti-inflammatory properties and could be an alternative to corticosteroids in COPD therapy. We investigated the effect of dexamethasone versus resveratrol on the release of COPD-related inflammatory mediators (IL-6, IL-8, GM-CSF and MCP-1) and matrix-metalloprotease-9 (MMP-9) from alveolar macrophages exposed to gram-negative bacterial endotoxin (lipopolysaccharide, LPS). We compared never-smokers, current smokers without airway obstruction and current smokers with COPD. The cytokines and MMP-9 were measured in cell culture supernatants with ELISA. The release of IL-8 and MMP-9 from LPS-exposed alveolar macrophages was increased in COPD, the release of GM-CSF and IL-6 was decreased in COPD and the release of MCP-1 was without differences between the cohorts. Dexamethasone impaired the release of all cytokines and MMP-9 from LPS-exposed alveolar macrophages of all cohorts, but for IL-8 and GM-CSF this effect was reduced in COPD. In alveolar macrophages of COPD, there was an almost complete reduction in IL-6 release but only a partial reduction in IL-8, GM-CSF, MCP-1 and MMP-9 release demonstrating a partial corticosteroid-insensitivity. In contrast, resveratrol almost completely reduced the release of all cytokines and MMP-9 without significant differences between the cohorts. Our data provide evidence for a corticosteroid resistance of alveolar macrophage-dependent inflammatory responses induced by gram-negative bacteria in COPD and thus question the utility of corticosteroids in COPD therapy. Instead, resveratrol may prove an alternative.     

Lipoteichoic acid enhances IL-6 production in human synovial fibroblasts via TLR2 receptor, PKCδ and c-Src dependent pathways

Patients with rheumatoid arthritis (RA) are at increased risk of developing infections and appear to be particularly susceptible to septic arthritis. Lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria is an amphiphilic, negatively charged glycolipid. However, the effects of LTA on human synovial fibroblasts are largely unknown. We investigated the signaling pathway involved in IL-6 production stimulated by LTA in rheumatoid arthritis synovial fibroblasts (RASF). LTA caused concentration- and time-dependent increases in IL-6 production. LTA-mediated IL-6 production was attenuated by Toll-like receptor 2 (TLR2) monoclonal antibody or siRNA. Pretreatment with PKCδ inhibitor (rottlerin), c-Src inhibitor (PP2), AP-1 inhibitor (tanshinone IIA) and NF-κB inhibitor (PDTC and TPCK) also inhibited the potentiating action of LTA. However, focal adhesion kinase (FAK) mutant and siRNA did not affect LTA-mediated IL-6 production. Stimulation of cells with LTA increased the PKCδ and c-Src phosphorylation and kinase activity. LTA increased the accumulation of p-c-Jun and p-p65 in the nucleus, as well as AP-1 and NF-κB luciferase activity. LTA-mediated increase of AP-1 and NF-κB luciferase activity was inhibited by rottlerin and PP2 or TLR2 and PKCδ siRNA or c-Src mutant. Our results suggest that LTA-increased IL-6 production in human synovial fibroblasts via the TLR2 receptor, PKCδ, c-Src, AP-1 and NF-κB signaling pathways.     

Resveratrol Attenuates the Release of Inflammatory Cytokines from Human Bronchial Smooth Muscle Cells Exposed to Lipoteichoic Acid in Chronic Obstructive Pulmonary Disease

During bacterial infections, pathogen‐associated molecular patterns (PAMPs) induce cytokine/chemokine release in immunoactive cells. This increases corticosteroid‐resistant airway inflammation in chronic obstructive pulmonary disease (COPD) and leads to exacerbations. Anti‐inflammatory therapies other than corticosteroids are required and resveratrol is currently under discussion. Resveratrol is an activator of sirtuins, which are class III histone deacetylases (HDACs). We suggested that human airway smooth muscle cells (HASMCs) release COPD‐associated cytokines/chemokines in response to lipoteichoic acid (LTA), a major PAMP of gram‐positive bacteria and that resveratrol is superior to the corticosteroid dexamethasone in suppressing these cytokines/chemokines. Cultivated HASMCs of patients with COPD were pre‐incubated with resveratrol or dexamethasone before stimulation with LTA. CCL2, GM‐CSF, IL‐6 and IL‐8 were analysed in culture supernatants by enzyme‐linked immunosorbent assay. Drug effects were investigated in the absence and presence of trichostatin A (TSA), an inhibitor of class I/II HDACs, and EX527, an inhibitor of the sirtuin SIRT1. LTA induced robust cytokine/chemokine release. Resveratrol was superior to dexamethasone in reducing CCL‐2, IL‐6 and IL‐8 in LTA‐exposed HASMCs of patients with COPD. Both drugs were equally effective in reducing GM‐CSF. Resveratrol effects were partially reversed by EX527 but not by TSA. Dexamethasone effects were partially reversed by TSA but not by EX527. We conclude that HASMCs contribute to the increase in airway inflammation in COPD exacerbations caused by gram‐positive bacterial infections. Our data suggest resveratrol as an alternative anti‐inflammatory therapy in infection‐induced COPD exacerbations. Resveratrol and corticosteroids suppress cytokine/chemokine expression through activation of SIRT1 or interaction with class I/II HDACs, respectively, in HASMCs.     

Effects of IL-4 and IL-13 on adenosine receptor expression and responsiveness of the human mast cell line 1

BACKGROUND: Inhalation of adenosine-5'-monophosphate (AMP) causes bronchoconstriction in asthma but not in healthy subjects. Bronchoconstriction upon AMP inhalation is thought to occur by histamine release and subsequent binding to receptors on airway smooth muscle cells. METHODS: To explain enhanced sensitivity to AMP in asthma, mast cell expression of the adenosine A2A and A2B receptors and histamine release were measured after incubation of human mast cell line 1 (HMC-1) cells with AMP and the non-specific adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) for 1.5 and 6 h. To establish a Thelper-2 environment resembling the asthma phenotype, HMC-1 cells were additionally cultured with IL-4 and IL-13 alone or stimulated with the combination of both cytokines and AMP and NECA. To study effects of prolonged presence of the inflammatory environment, the cells were pre-incubated overnight (18 h) with IL-4 and IL-13 and additionally stimulated with AMP and NECA for 1.5 or 6 h. RESULTS: AMP and NECA hardly affected adenosine receptor expression but increased IL-8 secretion. Incubation with IL-4 and IL-13 for 6 h increased adenosine A2A receptor expression and histamine secretion, but decreased IL-8 secretion. The combination of IL-4, IL-13, and AMP/NECA for 6 h increased A2B receptor expression and IL-8 secretion. Overnight stimulation with IL-4, IL-13 and subsequent stimulation with AMP/NECA for 1.5 h decreased A2AR expression which was accompanied by increased histamine secretion. CONCLUSION: These results suggest a role for decreased A(2A)R expression in enhanced adenosine responsiveness as observed in asthma.     

哮喘、慢性阻塞性肺病患者血清IL-18和总IgE水平的相关性

目的 探讨IL-18、总IgE、Th1和Th2类细胞因子在哮喘、慢性阻塞性肺病(COPD)发病时血清水平的变化及其相关性.方法 随机收集哮喘急性发作期患者50例(哮喘组),老年COPD急性加重期患者80例(COPD组),健康者50例(对照组)血清,采用双抗体夹心酶联免疫吸附试验检测血清IL-18、总IgE、IL-8、IL-4、IFN-γ水平.结果 CDPD组急性期IL-18、IL-8、IL-4、IFN-γ水平分别高于对照组1.8、25.2、116、150.8倍.哮喘组IL-18、IL-8、IL-4分别高于对照组29.9、3.4、5.2倍.哮喘组血清总IgE水平高于对照组223.2倍,而COPD组血清总IgE水平与对照组相比无显著性差异.COPD血清中IL-18与IL-8、IFN-γ,IL-8与IL-4呈正相关;哮喘组IL-18与IL-8、IL-4、总IgE,IL-4与总IgE呈正相关.结论 高血清IL-18提示细胞闪子参与哮喘及COPD的发病过程.哮喘发作时IL-18与总IgE正相关,而在COPD急性加重期无相关性,表明COPD和哮喘是两类有不同发生机制的气道炎症.     

Signal transduction of IL-13 and its role in the pathogenesis of bronchial asthma

Interleukin-13 (IL-13) is a Th2-type cytokine, secreted from CD4(+) T cells, mast cells, basophils and eosinophils. The human IL-13 gene locates at 5q31, generating a cluster with other Th2-type cytokines such as IL-4 and IL-5. Although the homology between IL-13 and IL-4 at the amino acid level is only about 25%, the IL-13 structure determined by NMR is very similar to that of IL-4. Both cytokines share their receptors and signal pathways, giving these two cytokines similar biological properties. However, the important role of IL-13 in the pathogenesis of bronchial asthma has recently been recognized, based mainly on analyses of mouse models. IL-13 and its signal pathway are thought to be promising targets to develop a therapeutic reagent for bronchial asthma. In this article, we summarize the signal transduction pathway of IL-13, the pathological roles of IL-13 in bronchial asthma and the reagents to inhibit IL-13 signals that are now under development.     

The expression of Foxp3 and ROR gamma t in lung tissues from normal smokers and chronic obstructive pulmonary disease patients

Foxp3- and ROR gamma t-expressing cells are involved in acquired immune responses. The change in Foxp3 and ROR gamma t expression in lung tissue and their role in emphysema has not been studied for COPD patients and normal smokers. In the present study, Foxp3 and ROR gamma t were assessed using real-time quantitative polymerase chain reaction and western blotting, and the expression and distribution of Foxp3, IL-17, IL-23R and CCR6 were measured by immunohistochemistry in peripheral lung tissue (10 smokers with COPD, 10 smokers and 10 nonsmokers with normal lung function). Foxp3 expression was lower and ROR gamma t expression was higher in COPD patients when compared with smokers and nonsmokers (all P values were less than 0.001). The ratios of Foxp3/ROR gamma t mRNA and protein were positively correlated to FEV1%pred and negatively correlated to the mean alveoli area. Foxp3⁺ cell numbers were decreased, while the number of IL-17⁺ cells, IL-23R⁺ cells and CCR6⁺ cells were increased in the lung alveolar walls of COPD patients compared with normal smokers and nonsmokers (all P values were less than 0.001). The IL-17⁺ cell numbers were positively correlated to both CCR6⁺ and IL-23R⁺ cells. Our data show a decreased Foxp3 expression and an increased ROR gamma t expression in COPD patients and normal smokers that parallels the aggravation of the disease. The IL-17⁺-cell-related cytokines receptors CCR6 and IL-23R had an association with the mechanism of IL-17⁺ cell number increasing, which will provide a new immuno-therapeutic target for COPD.     

Regulation of eosinophil migration and Th2 cell function by IL-5 and eotaxin

Asthma is characterized by elevated production of IgE, Th2 cytokines, chemokines, mucus hypersecretion, globlet cell metaplasia/hyperplasia, airway obstruction, eosinophilia and enhanced bronchial hyperresponsiveness. These hallmark features of asthma have all been linked to the effector functions of Th2 cytokines (e.g., interleukin-(IL)-4,5,9,10, and 13) in clinical and experimental investigations. This article will detail some of the pathogenic effects regulated by IL-13, IL-5 and the eotaxin subfamily of chemokines to regulate certain aspects of allergic disease. In particular, the potency of IL-13 in inducing enhanced bronchial responsiveness to spasmogenic stimuli and mucus hypersecretion suggests a key role of this molecule in the induction of airways obstruction. Recent studies also indicate that IL-5 and eotaxin, through eosinophils, may regulate Th2 cell function and IL-13 production from this lymphocyte. Therefore, IL-5 and IL-13 signaling systems are not necessarily mutually exclusive effector mechanisms, but may also be integrated through eosinophils to regulate certain aspects of allergic diseases. Blocking IL-13, or pathways that may promote IL-13-associated allergic lung responses (IL-5 and eotaxin) could provide an important strategy to improve the specificity of asthma therapy.     

Increased expression of CD4+IL-17+ cells in the lung tissue of patients with stable chronic obstructive pulmonary disease (COPD) and smokers

CD4⁺IL-17⁺ cells have an important role in controlling immune and inflammatory reactions. The authors of the present study hypothesize that these cells may be involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). To characterize the frequency of CD4⁺IL-17⁺ cells in the lung alveolar walls, small airways and muscular pulmonary arteries of nonsmokers, smokers with normal lung function and COPD patients, CD4⁺IL-17⁺ cell number was assessed using double immunofluorescence staining, and IL-17 and IL-21 expression were measured using real-time quantitative PCR in the peripheral lung tissues of 10 nonsmokers, 10 smokers with normal lung function and 10 smokers with stable COPD. In the lung alveolar walls, the number of CD4⁺IL-17⁺ cells was increased in COPD patients compared with nonsmokers and in normal smokers compared with nonsmokers. In the small airways, the CD4⁺IL-17⁺ cell numbers were higher in COPD patients than in normal smokers and nonsmokers. A positive correlation was observed between CD4⁺IL-17⁺ cell expression and pathological changes in the lung tissue. In the small airways, the number of CD4⁺IL-17⁺ cells was positively correlated with airflow limitations. The IL-17 mRNA levels in lung tissues were increased in COPD patients and normal smokers compared with nonsmokers. Increased CD4⁺IL-17⁺ cell number in lung tissue is involved in chronic inflammation of the lungs and parallels lung injury aggravation in COPD patients and in smokers without airway limitations. These findings contribute to a better understanding of CD4⁺ cell-related pathogenesis in COPD.     

Agents against cytokine synthesis or receptors

Various cytokines play a critical role in pathophysiology of chronic inflammatory lung diseases including asthma and chronic obstructive pulmonary disease (COPD). The increasing evidence of the involvement of these cytokines in the development of airway inflammation raises the possibility that these cytokines may become the novel promising therapeutic targets. Studies concerning the inhibition of interleukin (IL)-4 have been discontinued despite promising early results in asthma. Although blocking antibody against IL-5 markedly reduces the infiltration of eosinophils in peripheral blood and airway, it does not seem to be effective in symptomatic asthma, while blocking IL-13 might be more effective. On the contrary, anti-inflammatory cytokines themselves such as IL-10, IL-12, IL-18, IL-23 and interferon-gamma may have a therapeutic potential. Inhibition of TNF-alpha may also be useful in severe asthma or COPD. Many chemokines are also involved in the inflammatory response of asthma and COPD through the recruitment of inflammatory cells. Several small molecule inhibitors of chemokine receptors are now in development for the treatment of asthma and COPD. Antibodies that block IL-8 reduce neutrophilic inflammation. Chemokine CC3 receptor antagonists, which block eosinophil chemotaxis, are now in clinical development for asthma therapy. As many cytokines are involved in the pathophysiology of inflammatory lung diseases, inhibitory agents of the synthesis of multiple cytokines may be more useful tools. Several such agents are now in clinical development.     

慢性阻塞性肺疾病急性期血清白细胞介素-6、白细胞介素-13和白细胞介素-17A水平与GOLD分级的关系

目的 通过检测慢性阻塞性肺疾病(COPD)急性加重期和稳定期前后血清白细胞介素6(IL-6)、白细胞介素13(IL-13)和白细胞介素17A(IL-17A)的水平变化,探讨IL-6、IL-13和IL-17A在COPD急性加重期中的作用.方法 选取2017年1月至2018年6月上海市静安区市北医院呼吸内科收治的COPD急...     

Allergic inflammation: role of cytokines with special emphasis on IL-4

This review examines recent articles on the relationship of cytokines to allergy and inflammation with particular emphasis on interleukin (IL)-4. The objective of this article is therefore to review published studies to identify cytokines consistently involved in allergic inflammation. Proinflammatory cytokines, including IL-4, IL-5, IL-13 and GM-CSF along with TNF-alpha play a role in allergen-induced airway leukocyte recruitment and these cytokines can be generated by T mast cells and other cells. In addition, IL-9, IL-25, IL-33, IL-17, IL-27 and IFN-gamma are deeply involved in the regulation of asthma. Blocking the effect of these proinflammatory cytokines might provide new therapeutic approaches for the control of allergy and inflammation.     

Reassessing the Th2 cytokine basis of asthma

T helper (Th) 2 cytokines, particularly interleukin 4 (IL-4), IL-5 and IL-13, might be important in the development of allergic asthma. Humanized monoclonal antibodies (hMAbs) against IL-5, and a recombinant soluble human IL-4 receptor have been developed as possible treatments for this disorder. However, these approaches have not yet proven to be successful in the treatment of persistent asthma, suggesting that neither IL-4 nor IL-5 is important in asthma pathogenesis. Indeed, there is insufficient information about the efficacy of soluble IL-4 receptor and the anti-IL-5 hMAbs in the treatment of asthma to draw firm conclusions about the importance of these Th2 cytokines. Nevertheless, because IL-4 is required for IgE production and IL-5 is required for eosinophilopoesis, these Th2 cytokines must remain important candidates for a role in the pathogenesis of allergic asthma.     

丙泊酚全麻对胆道手术患者血清IL-2和IL-6的影响

目的 探讨七氟烷吸入全麻或丙泊酚全凭静脉全麻对胆道手术患者围术期血清细胞因子IL-2和IL-6的影响。方法 30例择期行开腹胆道手术患者,随机分入七氟烷吸入全麻组(S组)或丙泊酚全凭静脉全麻组(P组),每组各15例。分别于麻醉诱导前(T₀)、术毕(T₁)、术后24h(T₂)、术后72h(T₃)采集静脉血3ml,酶标法测定血清IL-2、IL-6水平。结果 与T₀时段相比,两组T₁时段血清IL-2显著下降(P＜0.05),而两组T₁、T₂时段血清IL-6显著升高(P＜0.05)。T₁时段S组与P组比较,血清IL-6的显著升高(P＜0.05)。结论 七氟烷和丙泊酚对血清IL-2水平变化影响无显著差别。丙泊酚全凭静脉全麻显著抑制血清IL-6水平升高,抑制手术麻醉应激引起的免疫反应。     

Intranasal IL-12 produces discreet pulmonary and systemic effects on allergic inflammation and airway reactivity

IL-12 modulates T cell responses between helper T cells Th2 and Th1; however, the therapeutic potential of IL-12 for allergic diseases either directly or as an adjuvant in allergen therapy has been controversial. The role of intranasal IL-12 as an adjuvant in modulating the grass pollen allergen (GAL) therapy-induced systemic immune response and lung-specific inflammation and airway reactivity was examined in this study using a mouse model of established allergic asthma. The effects of intranasal or nebulized IL-12 with or without intranasal anti-IFN-gamma antibody were examined in groups of control and allergen-sensitized or -challenged mice. T cell cytokine patterns, antibody response profiles, pulmonary inflammation and airway reactivity were examined. Intranasal IL-12 was found to be more effective in the Th2-Th1 shifting of immune response and anti-inflammatory activity in the lung compared to nebulized IL-12 at the given doses. Intranasal IL-12 significantly decreased production of IFN-gamma, eotaxin and LTC4/D4/E4 in the lung and decreased eosinophil infiltration, resulting in attenuated airway hyper-responsiveness in GAL-sensitized (GS) mice. In contrast, intranasal IL-12 significantly increased IFN-gamma production in the thoracic lymph node cultures and decreased the IL-5/IFN-gamma ratio, suggesting a Th2-Th1 shift. Also, intranasal IL-12 increased GAL-specific IgG2a antibody response, while the IgE response remained unaffected. The systemic effects of IL-12 were IFN-gamma dependent. IL-12 induces differential expression of its own receptor beta1 and beta2 subunits in the lung tissues to augment IL-12 responsiveness. Together, these results demonstrate that intranasal IL-12 is effective in shifting the systemic immune response in the direction of Th1 in IFN-gamma-dependent manner, while decreasing pulmonary inflammation and airway reactivity independent of IFN-gamma. Thus, intranasal delivery of IL-12 may provide an approach for the treatment of asthma and may be useful as an adjuvant in local nasal immunotherapy (IT) and in asthma.     

Attenuation of the ovalbumin-induced allergic airway response by cannabinoid treatment in A/J mice

T cells are sensitive to modulation by cannabinoids as evidenced by their ability to inhibit expression of cytokines, including interleukin (IL)-2 and IL-4. Because T cells play a key role in the pathophysiology of allergic asthma by expressing T helper cell (Th)2 cytokines, the objective of the present studies was to examine the effect of cannabinoids on immunologic and pathologic features associated with the allergic airway response induced by ovalbumin (Ova). A/J mice were systemically sensitized with Ova and subsequently challenged with aerosolized Ova. The steady-state mRNA expression of IL-2 and Th2 cytokines (IL-4, IL-5, and IL-13) was markedly increased in the lungs of Ova-sensitized mice 24 h after a single Ova challenge. Concordantly, the level of total and Ova-specific serum immunoglobulin (Ig)E and intraepithelial mucosubstances in the axial intrapulmonary airway of Ova-sensitized mice was robustly elevated 96 h after the second Ova challenge. Cannabinol (CBN) or Delta(9)-tetrahydrocannabinol (Delta(9)-THC; 50 mg/kg, ip), administered daily for 3 consecutive days before sensitization and then before challenge, significantly attenuated the elevation of IL-2, IL-4, IL-5, and IL-13 steady-state mRNA expression elicited by Ova challenge in the lungs. In addition, the elevation of serum IgE and the mucus overproduction induced by Ova challenge was also markedly attenuated by CBN or Delta(9)-THC administration in Ova-sensitized mice. These results suggest that plant-derived immunomodulatory cannabinoids exhibit potential therapeutic utility in the treatment of allergic airway disease by inhibiting the expression of critical T cell cytokines and the associated inflammatory response.     

哮喘患儿sIL-2R与细胞免疫功能检测的临床意义

目的探讨儿童支气管哮喘（简称哮喘）发作期淋巴细胞免疫功能和白细胞介素-2（IL-2）、可溶性白介素2受体（sIL-2R）检测的临床意义。方法应用流式细胞术和酶联免疫吸附试验（ELISA）方法对25例支气管哮喘患儿和21例支气管哮喘缓解期患儿以及20例正常儿童外周血T细胞亚群及血清IL-2和sIL-2R水平进行测定。结果支气管哮喘患儿发作期血清sIL-2R水平明显高于缓解期和对照组，缓解期sIL-2R水平仍高于对照组。IL-2水平明显低于缓解期和对照组。外周血总CD3^＋、CD4^＋、CD4^＋／CD8^＋细胞比值均降低，CD8^＋细胞增高。结论动态监测IL-2和sIL-2R水平含量，观察T细胞亚群变化可以判断哮喘发作的程度、指导治疗和评估预后。