IL-10-592 polymorphism is associated with IL-10 expression and severity of enterovirus 71 infection in chinese children

BackgroundEnterovirus 71 (EV71) infection results in some severe complications with high mortality and disability in Hand, Foot and Mouth Disease (HFMD) in children. Recent studies have shown that cytokine genetic predispositions have associations with both the development of EV71 infection and severity of HFMD.ObjectiveThis study was designed to investigate whether the IL-10–592 polymorphism is associated with IL-10 levels and disease severity in Chinese children with EV71 infection.Study designIn patients selected, there were 378 cases with EV71 infection (including 291 mild cases, 70 severe cases and 17 critical cases), as well as 406 health controls. EV71 in serum was tested by RT-PCR, and IL-10-592 genotype was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis techniques.ResultThe IL-10-592C allele was observed with higher frequency in patients with critical EV71 infection (70.59%) compared with severe EV71 infection (41.43%, P < 0.01), mild EV71 infection (43.81%, P < 0.01) and healthy children (44.46%, P < 0.01). The blood IL-10 levels of critical cases were significantly higher than severe cases, mild cases, and healthy children. Among all of the four groups, IL-10 levels in patients with genotype AA were significantly lower than those with genotypes AC + CC (t = 4.86, P < 0.05; t = 2.30, P < 0.05; t = 3.44, P < 0.05; t = 5.58, P < 0.05).ConclusionIL-10-592C allele is associated with IL-10 expressions and the severity of EV71 infection in Chinese patients.     

细胞因子IL6、IL8及IL-10与变应性鼻炎的关系研究

目的探讨白细胞介素-6、8、10与变应性鼻炎（allergicrhinitis,AR）发病的相关性,为指导临床治疗提供实验依据。方法应用酶联免疫吸附测定（enzymelinkedimmu—nosorbentassay,ELISA）方法检测35例正常健康对照组以及60例变应性鼻炎患者脱敏治疗前后血清标本中IL-6、IL-8、IL-10的表达水平,并应用SPSSl3．0软件分析治疗前后各指标的表达差异,以及三者之间的相关性。结果IL-6、IL-8在AR患者血清中高表达,与健康对照组相比差异具有显著性（t=15．213,P〈0．01;t=12．231,P〈0．01）,脱敏治疗后表达均下降,治疗前后差异具有统计学意义（t=21．995,P〈0．01;t=19．766,P〈0．01）;IL-10在患者血清中表达低于对照组（t=7．446,P〈0．01）,脱敏治疗后其表达上升,与治疗前相比差异显著（t=10．228,P〈0．01）;IL-6、IL-8在AR患者中的表达呈正相关（r=0．523。P〈0．01）,而二者与IL-10的表达呈负相关（r=-0．482,P〈0．01）。结论IL-6、IL-8及IL-10与变应性鼻炎发病过程密切相关,相应的细胞因子或拮抗剂将可能成为候选的治疗药物。     

细胞因子IL6、IL8及IL-10与变应性鼻炎的关系研究

目的 探讨白细胞介素-6、8、10与变应性鼻炎( allergic rhinitis,AR)发病的相关性,为指导临床治疗提供实验依据.方法 应用酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)方法检测35例正常健康对照组以及60例变应性鼻炎患者脱敏治疗前后血清标本中IL-6、IL-8、IL-10的表达水平,并应用SPSS 13.0软件分析治疗前后各指标的表达差异,以及三者之间的相关性.结果 IL-6、IL-8在AR患者血清中高表达,与健康对照组相比差异具有显著性(t=15.213,P＜0.01;t=12.231,P＜0.01),脱敏治疗后表达均下降,治疗前后差异具有统计学意义(t=21.995,P＜0.01;t =19.766,P＜0.01);IL-10在患者血清中表达低于对照组(t=7.446,P＜0.01),脱敏治疗后其表达上升,与治疗前相比差异显著(t=10.228,P＜0.01);IL-6、IL-8在AR患者中的表达呈正相关(r=0.523,P＜0.01),而二者与IL-10的表达呈负相关(r=-0.482,P＜0.01).结论 IL-6、IL-8及IL-10与变应性鼻炎发病过程密切相关,相应的细胞因子或拮抗剂将可能成为候选的治疗药物.     

IL-35 promoted STAT3 phosphorylation and IL-10 production in B cells,but its production was reduced in patients with coronary artery diseases

Interleukin (IL)-35 is a heterodimeric cytokine composed of the IL-12A subunit and the Epstein-Barr virus induced gene 3 (EBI3) subunit. Binding of IL-35 with IL-12 receptor subunit beta 2 (IL-12RB2) and IL-6 signal transducer (IL-6ST) occupies the binding sites of IL-6, IL-12, and IL-27 and prevents their signal transduction. IL-35 is also shown to promote the development of regulatory T cells (Tregs) and regulatory B cells (Bregs). In this study, we investigated B cell-mediated IL-35 production in patients with coronary artery disease (CAD). The expression levels of IL-35 subunits and IL-10 were significantly lower in B cells from CAD patients than in B cells from healthy control individuals. Exogenous IL-35 could effectively increase the IL-10 production by B cells in a concentration-dependent manner. IL-35 promoted the phosphorylation of STAT1 and STAT3 in B cells, and the inhibition of STAT3 phosphorylation suppressed IL-10 production. Raising the IL-35 concentration in cell culture eliminated the difference in IL-10 expression between CAD B cells and healthy B cells. We also demonstrated that B cells from CAD patients presented lower capacity to suppress interferon gamma (IFNG) and tumor necrosis factor (TNF) expression by T cells than B cells from healthy controls. Exogenous IL-35 could significantly improve the suppressive capacity of B cells in both healthy controls and CAD patients. Together, these results demonstrated that a reduction in IL-35 production was associated with Breg defects in CAD patients. IL-35 and IL-35 targets may serve as therapeutic candidates in the treatment of CAD and related diseases.     

慢性丙肝患者血清IL-6、IL-10、IL-32水平与HCV-RNA载量关系研究

目的探究慢性丙肝患者血清白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-32(IL-32)水平与丙型肝炎病毒RNA(HCV-RNA)载量的关系。方法选择我院于2015年3月至2018年3月收治的慢性丙型肝炎(CHC)患者90例为CHC组,同期以90例来我院进行健康体检的健康者为健康组,采用ELISA法检测所有受试者的血清IL-6、IL-10、IL-32水平,采用PE5700荧光PCR仪和实时荧光定量聚合酶链反应法检测CHC组患者的HCV-RNA载量,对比CHC组患者和健康组受试者的血清IL-6、IL-10、IL-32水平;对不同病毒载量的CHC患者血清IL-6、IL-10、IL-32水平进行对比,并分析病毒载量与检测指标之间的相关性。结果经对比,CHC组患者的血清IL-6、IL-10、IL-32水平均显著高于健康组受试者(P<0.05);随着病毒载量的不断增大,CHC患者的IL-6、IL-10、IL-32水平均显著升高(P<0.05);通过Pearson相关性分析结果显示,血清IL-6、IL-10、IL-32水平均与HCV-RNA载量显著相关,且为正相关。结论慢性丙肝患者血清IL-6、IL-10、IL-32水平与HCV-RNA载量呈正相关,可通过检测机体的血清IL-6、IL-10、IL-32水平了解丙型肝炎的发病机制和慢性化过程,对临床诊断慢性肝炎、了解疾病进展、评价治疗效果等意义重大。     

Chronic Colitis in IL-10-/- Mice: Insufficient Counter Regulation of a Th1 Response

IL-10-deficient (IL-10^-/-) mice, generated by a gene-targeted mutation, develop abnormal immune responses as a result of uncontrolled interactions between antigen presenting cells and lymphocytes. The studies reviewed herein have focused on the enterocolitis that spontaneously develops in IL-10^-/- mice. Not unexpectedly, heightened production of proinflammatory mediators accompanied pathologic changes in the gastrointestinal tract of young mutants. In a series of studies, the proinflammatory mediators responsible for initiating the pathogenic response were distinguished from those that were elicited as a consequence of persistent inflammation. We have also investigated the possibility that different mediators are involved in the inductive versus the maintenance phase of disease. The findings of these mechanistic studies as they relate to our understanding of progressive inflammatory disease and the role of IL-10 in controlling the acute and chronic stages are discussed.     

Molecular basis of the synergistic production of IL-1 receptor antagonist by human neutrophils stimulated with IL-4 and IL-10

In this study, we report that the release of IL-1 receptor antagonist (IL-1ra) from IL-4-stimulated neutrophils is markedly enhanced in the presence of IL-10. We also show that up-regulation of IL-1ra release by IL-10 in IL-4-stimulated neutrophils takes place through IL-1ra mRNA stabilization and enhancement of IL-1ra de novo synthesis. Furthermore, we report that the ability of IL-10 to up-regulate IL-1ra mRNA expression in IL-4-treated neutrophils requires 5-6 h and it is preceded by the acquisition of the capacity to activate Stat3 tyrosine phosphorylation. This latter response to IL-10 was strictly dependent on the levels of expression of IL-10R1, which were in fact significantly increased by IL-4 in cultured neutrophils via a signaling pathway sensitive to the serine/threonine kinase inhibitor H-7. Collectively, our data emphasize the central role of IL-10R1 expression in regulating cell responsiveness to IL-10. In addition, the fact that IL-10 strongly up-regulates IL-1ra production in IL-4-activated neutrophils uncovers a novel mechanism whereby IL-10 and IL-4 cooperate to negatively modulate the inflammatory responses.     

IL-4, IL-10 and IL-13 down-regulate monocyte-chemoattracting protein-1 (MCP-1) production in activated intestinal epithelial cells

Several studies have demonstrated that intestinal epithelial cells play a major role in the initiation and perpetuation of intestinal inflammation by secreting proinflammatory cytokines and chemokines. MCP-1 is suggested to be a chemokine that plays a major part during intestinal inflammation in inflammatory bowel disease (IBD). Immunoregulatory cytokines such as IL-4, IL-10 and IL-13 have been described to exert anti-inflammatory properties on various cell types. The aim of our study was to determine the effect of Th2 cytokines on the production of MCP-1 by activated intestinal epithelial cells. We examined Caco-2 cells as well as intestinal epithelial cells which were isolated from surgical specimens. Production of the chemokine MCP-1 was determined under stimulated and non-stimulated conditions. IL-4, IL-10 and IL-13 were added to stimulated epithelial cells under various culture conditions. Supernatants were analysed for cytokine concentrations using ELISAs. Under stimulation with physiological agents like IL-1β or tumour necrosis factor-alpha (TNF-α), we observed markedly increased concentrations of MCP-1 in supernatants of Caco-2 cells and intestinal epithelial cells. IL-4, IL-10 and IL-13 all had the capacity to down-regulate the production of MCP-1 in Caco-2 cells as well as in freshly isolated epithelial cells. Caco-2 cells which were primed with Th2 cytokines 24 h before stimulation were subsequently decreased in their ability to be stimulated by IL-1β or TNF-α for MCP-1 production. As MCP-1 has been shown to play a major role during intestinal inflammation, the in vitro suppression of MCP-1 in enterocytes suggests the in vivo use of regulatory cytokines in patients with active IBD.     

Preferential production of IgG1, IL-4 and IL-10 in MuSK-immunized mice

Myasthenia gravis (MG) is an autoimmune disease characterized by muscle weakness associated with acetylcholine receptor (AChR), muscle-specific receptor kinase (MuSK) or low-density lipoprotein receptor-related protein 4 (LRP4)-antibodies. MuSK-antibodies are predominantly of the non-complement fixing IgG4 isotype. The MuSK associated experimental autoimmune myasthenia gravis (EAMG) model was established in mice to investigate immunoglobulin (Ig) and cytokine responses related with MuSK immunity. C57BL/6 (B6) mice immunized with 30 μg of recombinant human MuSK in incomplete or complete Freund's adjuvant (CFA) showed significant EAMG susceptibility (> 80% incidence). Although mice immunized with 10 μg of MuSK had lower EAMG incidence (14.3%), serum MuSK-antibody levels were comparable to mice immunized with 30 μg MuSK. While MuSK immunization stimulated production of all antibody isotypes, non-complement fixing IgG1 was the dominant anti-MuSK Ig isotype in both sera and neuromuscular junctions. Moreover, MuSK immunized IgG1 knockout mice showed very low serum MuSK-antibody levels. Sera and MuSK-stimulated lymph node cell supernatants of MuSK immunized mice showed significantly higher levels of IL-4 and IL-10 (but not IFN-γ and IL-12), than those of CFA immunized mice. Our results suggest that through activation of Th2-type cells, anti-MuSK immunity promotes production of IL-4, which in turn activates anti-MuSK IgG1, the mouse analog of human IgG4. These findings might provide clues for the pathogenesis of other IgG4-related diseases as well as development of disease specific treatment methods (e.g. specific IgG4 inhibitors) for MuSK-related MG.     

The IL1A genotype is associated with nasal polyposis in asthmatic adults

BACKGROUND: Nasal polyposis (NP) is a chronic inflammatory disease often found coexisting with asthma. As this disorder tends to cluster in families, a genetic predisposition has been suggested. Interleukin-1 (IL-1) has been proposed to play a role in the pathogenesis of NP. METHODS: We analysed the single G-to-T base exchange polymorphism in exon 5 at +4845 of the gene encoding IL-1alpha (IL1A) and the C-to-T base exchange polymorphism at -511 of the gene encoding IL-1beta (IL1B) in a population-based sample of adult asthma patients (n = 245). The data were assessed for correlation with data on history of NP and other phenotype-related characteristics. RESULTS: The prevalence of NP in our study group was 14.3%. The distribution of the IL1A genotype differed significantly between asthmatics with and without NP (P = 0.005). The risk of NP was markedly increased in allele G homozygous subjects (OR = 2.73; 95%CI = 1.40-5.32). In the case of IL1B we found no significant associations. Asthmatics with NP had more symptoms than others, but lung function and blood eosinophil counts were similar. CONCLUSIONS: Our study demonstrates an association of IL1A with NP inasthmatic patients and addresses the role of IL-1alpha as an inflammatory modulator in the pathogenesis of this disease.     

Chinese herbal medicinal ingredients affect secretion of NO,IL-10, ICAM-1 and IL-2 by endothelial cells

Abstract

The aim of this study was to investigate the anti-endotoxin effects of sinomenine, fangchinoline, stachydrine, chuanxionggzine, oxymartrine and evodiamine alkaloids commonly found in Chinese herbal medicines. Porcine endothelial cells were challenged with 1?μg LPS/ml for 3?h and then treated with one of the six alkaloids at three concentrations (1, 5 or 10?μg/ml) for a further 21?h. The supernatants of the cultures were then collected and analyzed for levels of nitric oxide (NO), interleukin (IL)-10, intercellular cell adhesion molecule-1 (ICAM-1) and IL-2 using ELISA kits. The results revealed that sinomenine, stachydrine and chuanxionggzine inhibited production of NO; stachydrine and evodiamine inhibited secretion of IL-10; sinomenine and chuanxionggzine down-regulated ICAM-1 expression; oxymartrine and evodiamine decreased production of IL-2 by the LPS-stimulated endothelial cells. Overall, the data from these studies suggested to us that these six alkaloids might effectively reduce inflammatory responses in situ via changes in the formation of these key regulatory molecules/proteins.     

HIV-induced IL-6/IL-10 dysregulation of CD4 cells is associated with defective B cell help and autoantibody formation against CD4 cells

To analyse CD4 cell cytokine secretion and helper/suppressor function at a clonal level we established 446 CD4⁺ T cell clones (TCC) in four healthy controls, three HIV^? haemophilia patients, four CDC II,III and four CDC IV patients. Spontaneous TCC secretion of Th1 cytokines (IL-2, interferon-gamma (IFN-γ)) and Th2 cytokines (IL-4, IL-6, IL-10) was determined by ELISA. TCC helper and suppressor functions were tested in a pokeweed mitogen (PWM)-stimulated allogeneic co-culture system using a reverse haemolytic plaque assay for assessment of B cell responses. There was a significant association of TCC surface marker expression (Leu-8, CD45RA) with TCC IL-6 secretion in healthy controls (P < 0.01), HIV^? patients (P 0.001) and CDC II,III patients (P 0.01) but not in CDC IV patients. Likewise, TCC expression of Leu-8 and CD45RA was significantly associated with TCC suppressor function in healthy controls (P 0.0005) but not in HIV-infected patients. A reduced TCC helper frequency (10% of TCC) and an enhanced TCC suppressor frequency (> 80% of TCC) were detected only in those HIV-infected patients who showed an excessively increased TCC IL-6 secretion (> 70% of TCC) together with a significantly diminished TCC IL-10 secretion (10% of TCC). CD4 cell autoantibodies also were found only in patients with this type of cytokine dysregulation. These data indicate that CD4 cell surface markers lose their functional relevance in HIV-infected patients. HIV-induced IL-6/IL-10 dysregulation of CD4⁺ T cells, i.e. the up-regulation of spontaneous IL-6 and down-regulation of spontaneous IL-10 secretion, appears to be involved in inducing CD4 helper defects and may promote autoantibody formation against CD4 cells.     

慢性肝炎患者血清中降钙素原、C-反应蛋白和IL-10的相关性研究

目的 探讨降钙素原(procalcitonin, PCT)、C-反应蛋白(C-reactive protein,CRP)和白细胞介素-10(interleukin-10,IL-10)在慢性肝炎患者血清中的表达及意义.方法 收集慢性肝炎患者血清并检测血清中降钙素原、C-反应蛋白和IL-10的含量.结果 与对照组相比,慢性活动性肝炎(chronic active hepatitis,CAH) 组和慢性迁移性肝炎(chronic persistent hepatitis,CPH) 组患者血清内降钙素原、C-反应蛋白和IL-10水平显著升高,且CAH组显著高于CPH组,差异具有统计学意义(P<0.05).患者血清降钙素原、C-反应蛋白和IL-10联合检出率显著高于各指标单独检出率.结论 降钙素原、C-反应蛋白和IL-10在慢性肝炎患者的血清中呈过表达状态;且降钙素原、C-反应蛋白和IL-10联合检测对于了解慢性肝炎患者的疾病状态具有重要的临床意义.     

Topical glucocorticoids application induced an augmentation in the expression of IL-1α while inhibiting the expression of IL-10 in the epidermis in murine contact hypersensitivity

The repeated application of glucocorticoids (GC) on the skin augmented the inflammatory response of both allergic and irritant contact dermatitis in our studies. In order to further clarify the mechanism of such an augmentation of contact hypersensitivity (CHS), we investigated the modulatory effects of cytokines in the epidermis after the administration of GC at challenged sites in CHS. Diflucortolone valerate was applied to BALB/c mice on alternate days for a total of nine times. On day 12, they were contact sensitized with dinitrofluorobenzene (DNFB). Next, on day 17, one day after the last application of GC, they were challenged with DNFB on the ear. The whole challenged ear lobes were removed after a hapten challenge and then were analysed by the RT-PCR method or underwent an immunohistochemical analysis. To clarify the modulatory effects of cytokines in vivo, DNFB sensitized mice pre-treated with GC were injected with rIL-10, IL-1 receptor antagonist (ra) and anti-IL-1alpha monoclonal antibody (mAb) and thereafter were challenged with DNFB. A RT-PCR analysis has demonstrated IL-10 mRNA to be detected in the challenged skin of non-GC-pretreated mice but not in that of GC-pre-treated mice after challenge. On the other hand, the expression of IL-1alpha mRNA in the challenged skin of mice pretreated with GC was more strongly detected that that in mice without GC-pretreatment. Furthermore, an immuno-histochemical analysis in the challenge showed the expression of IL-10 in the skin showed the expression of IL-10 in the challenged epidermis of the non-GC-pretreated mice but not in the GC-pretreated mice and IL-1alpha was also strongly expressed in the epidermis of the GC-pretreated mice. A subcutaneous injection of anti-IL-1alpha mAb or IL-1 ra inhibited the augmented CHS reaction in the GC-pretreated mice. A subcutaneous injection of rIL-10 also inhibited the augmentation of the CHS reaction in the GC-pretreated mice; however, no such inhibition was observed in the non-GC-pretreated mice. These results indicated that both an up-regulation of IL-1alpha production and the inhibition of the IL-10 production in the epidermis at the challenged skin sites in the GC-pretreated mice appear to play a critical role in the GC-induced augmentation of murine CHS.     

Differential production of IL-10 by T cells and monocytes of HIV-infected individuals: association of IL-10 production with CD28-mediated immune responsiveness

Immune unresponsiveness in HIV-1 infection can result from impaired signals delivered by the costimulatory CD28-B7 pathway and the altered production of immunoregulatory cytokines, in particular IL-10, whose production is altered in HIV-1 infection. In this study we investigate IL-10 regulation in T cells and monocytes from HIV⁺ individuals, and its association with CD28-mediated T cell proliferation. IL-10 production as analysed in T cell- and monocyte-depleted peripheral blood mononuclear cells (PBMC), and by intracellular staining at the single-cell level, reveals a defect in IL-10 production by CD4⁺ and CD8⁺ T cells, whereas monocytes constitute the major IL-10-producing cell type. To investigate the impact of IL-10 on immune responsiveness, CD28-mediated proliferative responses in HIV⁺ individuals were correlated with PHA-induced IL-10 production. CD4⁺ T cells expressed CD28, yet exhibited markedly reduced CD28-mediated cell proliferation. This CD28-mediated CD4⁺ T cell proliferation was found to be inversely associated with the levels of PHA-induced IL-10 production and could be restored, at least in part, by anti-IL-10 antibodies. These results suggest that IL-10 production is differentially regulated in T cells and monocytes of HIV⁺ individuals, and that IL-10 may have a role in inducing immune unresponsiveness by modulating the CD28-B7 pathway.     

丙型肝炎患者血清细胞因子水平与肝功能损伤的相关性探讨

目的：检测血清可溶性L-selectin（L-选择素）、sICAM-1（细胞粘附因子-1）、IL-6（白细胞介素-6）、IL-10（白细胞介素-10）、IL-18（白细胞介素-18）在丙型肝炎病毒感染造成肝功能损害过程中的作用。方法：应用酶联免疫吸附法（ELISA）对62例丙型肝炎患者进行了血清中L-selectia、sICAM-1、IL-6、IL-10、IL-18水平检测及相关的肝功能、生化指标进行相关分析,并与36例正常健康人作比较。结果：丙型肝炎患者血清L-selectin、sICAM-1、IL-6、IL-10、IL-18水平均非常显著地高于正常人组（P〈0．01）,且与ALT（谷丙转氨酶）、TBIL（总胆红素）水平呈明显的正相关。结论：检测丙型肝炎患者血清中L-selectin、sI—CAM-1、IL-6、IL-10、1L-18水平在一定程度上反映了机体的免疫功能状态和肝细胞损伤的程度,具有临床应用价值。