Attenuated production of intracellular IL-10 and IL-12 in monocytes from patients with severe asthma

Interleukin (IL)-10 and IL-12 production is decreased in peripheral blood mononuclear cells of patients with mild asthma. Using whole blood culture and flow cytometry we examined whether monocyte heterogeneity influenced IL-10 and IL-12 production in subjects with severe asthma. We demonstrated that IL-10 release in LPS-stimulated whole blood culture was decreased in patients with severe persistent asthma compared to those with mild asthma and controls (P = 0.04 and P < 0.001, respectively). In asthmatic patients, the percentage of CD14(+)CD16(+) cells was higher than that from normal subjects (P = 0.04). Severe asthmatics showed significantly less positive staining for IL-10 and IL-12 (P < 0.001 and P = 0.02, respectively) after stimulation in monocytes, compared to mild asthmatics and controls in both CD14(+)CD16(+) and CD14(+)CD16(minus sign) cells. These results suggest that IL-10 synthesis is attenuated in severe persistent asthma compared to mild asthma and that this cannot be explained by the increase in the CD14(+)CD16(+) monocytes in asthma.     

Increase in the production of interleukin-6, interleukin-10, and interleukin-12 by lipopolysaccharide-stimulated peritoneal macrophages from women with endometriosis

PROBLEM: To verify whether the peritoneal macrophage (PM) is activated in endometriosis. METHOD OF STUDY: We examined the synthesis of nitric oxide (NO), total antioxidant, interleukin (IL)-6, IL-10, and IL-12 by cultured PMs, which were either unstimulated or stimulated with lipopolysaccharide (LPS), from women with endometriosis (early, n = 12; advanced, n = 11) or without endometriosis (n = 13). RESULTS: After stimulation with 2 ng/mL LPS for 24 hr, PMs from women with early-stage endometriosis secreted more NO, IL-6, and IL-10 than the controls. Higher IL-12 levels were noted in women with advanced endometriosis when compared with the controls. After 2 ng/mL-LPS stimulation for 24 hr, we also detected higher total antioxidant levels in the advanced-endometriosis group than those in the early-endometriosis group. CONCLUSION: The increased production of IL-6, IL-10, and IL-12 by stimulated PMs confirmed previous observations that the PM is the principle source of these cytokines in peritoneal fluid.     

类风湿性关节炎患者滑膜液中IL-10、IL-12和sFasL含量的检测

为了研究细胞因子和凋亡分子在类风湿性关节炎(RA)发病中的作用,用ELISA法分析了26例RA患者滑膜液和血清中IL-12、IL-10和可溶性FasL(sFasL)的含量。结果表明RA患者滑膜液中IL-12、IL-10含量分别为(419.9±89.2)pg/ml和(187.7±34.5)pg/ml,外周血中这两种细胞因子的含量均较低,分别为(65.3±34.2)pg/ml(IL-12)和(85.0±12.7)pg/ml(IL-10)。滑膜液中sFasL的含量为266pg/ml,明显高于血清含量(36pg/ml)。这一结果提示,RA患者滑膜液中IL-12含量和sFasL增高,这些炎性细胞因子增高可能参与了关节滑膜中的自身反应性T细胞的活化,继而造成免疫损伤。     

鼠感染Babesia microti和 Babesia rodhaini后早期 IL-12、IL-10诱生的研究

目的 研究Babesia microti 和Babesia rodhaini感染后早期IL-12和IL-10诱生情况,以探讨B. microti和 B. rodhaini不同种属感染的发病机制与免疫应答效应.方法 使用ELISA方法分别检测Babesia microti 和 Babesia rodhaini 感染鼠后的0、3、6、9、12、18、24、36、48、72、96 h血清中IL-12及IL-10的浓度.结果 B. microti感染后3 h,小鼠血液中IL-12产生达一个高峰,至感染后24 h达最高峰值,与对照组比较差异显著,同组鼠在感染的早期(96 h前)血液中IL-10水平无明显变化.而B. rodhaini感染鼠,在感染后3～72 h,血清中IL-10和IL-12水平与对照组比较均无明显差异.在感染后96 h,血清中IL-10和IL-12水平开始下降,与对照组比较差异显著.结论 B. microti 感染鼠早期诱生的细胞因子主要为IL-12,而B. rodhaini感染后早期IL-12和IL-10水平无明显变化.     

Association of Interleukin-10 promoter polymorphisms with neurosyphilis

Interleukin 10 (IL-10) is a potent anti-inflammatory cytokine. Increased production of IL-10 has been found in late syphilis, presumably creating favorable conditions for bacteria persistence. Single-nucleotide polymorphisms (SNPs) within the promoter of IL-10 gene have been found to influence IL-10 production. We investigated whether SNPs in the IL-10 gene promoter are associated with cerebrospinal fluid (CSF) levels of IL-10 and neurosyphilis.Polymorphisms in the gene for IL-10 (G → A mutation at the position −1084 and C → A mutation at the position −592) were sought in 35 patients with syphilis and 24 healthy volunteers. CSF examination (i.e. routine laboratory tests and IL-10 levels) was performed in all syphilis patients. Neurosyphilis was defined as reactive CSF VDRL test or CSF white blood cells ⩾ 5/μL and CSF protein concentration ⩾ 45 mg/dL.Overall, 31% of patients with syphilis had neurosyphilis. CSF IL-10 levels were significantly higher in patients with neurosyphilis when compared to those with syphilis but not neurosyphilis. −1082 GG and −592 CC genotypes were significantly associated with higher CSF IL-10 levels. Moreover, these genotypes were found to be more frequent in individuals with neurosyphilis in comparison to those without neurosyphilis.Anti-inflammatory immune response seems to be important in pathogenesis of neurosyphilis. Our data suggest that host-related factors, such as SNPs of immune regulatory genes may influence the susceptibility to neurosyphilis.     

IL-10与疾病关系的研究进展

IL-10是一种重要的抗炎细胞因子,对多种免疫细胞功能均有抑制作用.这种细胞因子的特殊生理意义在于防止和抑制强大的特异和非特异性免疫反应和由此导致的组织损伤,同时IL-10可增强“清道夫”功能,并有助于诱导免疫耐受.IL-10的这些作用与疾病的发生有密切的联系,因此研究其分子机制和生物学特征对IL-10相关性疾病的免疫治疗有重要意义.     

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。     

Interleukin-12p40 mediates transient protection against Mycobacterium avium infection in the absence of interleukin-12

Produced by macrophages and dendritic cells, interleukin (IL)-12 is composed of a p35 and a p40 subunit and promotes protection against intracellular pathogens through the development of interferon-gamma (IFNgamma) -producing T cells. The p40 subunit is also shared by the dimeric cytokines IL-12p40 homodimer and IL-23. In man, genetic defects in IL-12p40-mediated mechanisms are responsible for the familial occurrence of nontuberculous mycobacterial infections, the most common of which is infection with Mycobacterium avium. To experimentally differentiate the contribution of IL-12p40-containing cytokines in the outcome of M. avium infection, we studied wild-type, p35- and p35/p40 doubly deficient mice in an intravenous infection model which reflects many parameters of the disseminated infection in humans. Our study shows that in contrast to p35/p40 doubly deficient mice, p35-deficient mice mount a transient antibacterially protective response against M. avium although such animals were unable to produce detectable levels of IFNgamma or generate efficient granulomas. In conclusion, our results identify an antibacterial effector mechanism preserved in p35-deficient mice that is absent in mice devoid of p35 and p40. This phenotype probably reflects an IL-12p40-dependent effect on macrophage activation at the level of innate immunity.     

Inverse modulation of IL-10 and IL-12 in the blood of women with preneoplastic lesions of the uterine cervix

It has been postulated that an impaired immune response may contribute to the progression of human papillomavirus (HPV)-associated preneoplastic lesions. Based on this hypothesis, we evaluated the cytokine production in the blood of patients with squamous intraepithelial lesions (SIL) of the uterine cervix. The levels of type-1 (interferon-gamma (IFN-γ) and IL-12) and type-2 (IL-4 and IL-10) cytokines were measured in whole blood culture supernatants of patients with low- and high-grade SIL and control women. There was no difference in IL-4 and IFN-γ levels between patients with SIL and the control group. In contrast, the ratio of IL-12/IL-10 levels was significantly lower in patients with SIL compared with the control group. A lower IL-12/IL-10 ratio in women with SIL was also observed when peripheral blood mononuclear cell (PBMC) culture supernatants and plasma samples were analysed. In patients, neither the lower expression of the CD3ε chain nor the higher frequency of HLA-DRB1*1501 expression could be correlated with abnormal cytokine production. These results suggest that a part of the cytokine network, namely IL-10 and IL-12, is perturbed in patients with SIL. A better knowledge of the role of these cytokines in regulating the growth of HPV-associated SIL might have practical implications for the development of vaccines or immunomodulatory strategies in the treatment of cervical cancers.     

Interleukin-12: Clinical usage and molecular markers of cancer susceptibility

The last decade has seen the emergence of immunomodulators as therapeutic agents in cancer treatment. Interleukins (ILs) are a category of small cell-signaling molecules that organize communication and interaction between immune cells and therefore they could be used as perfect immunomodulators. IL-12 is a promising candidate for cancer immunotherapy since it plays a major role in development of antitumor immune response. Numerous studies report that IL-12 promotes an effective destruction of cancer cells both in vivo and in vitro. In addition, IL-12 has anti-angiogenic activity and it is able to dramatically decrease tumor-supportive activities of tumor-associated macrophages. The first part of the review is devoted to immunobiology of IL-12. Signaling pathways of IL-12 as well as clinical trials of this cytokine are discussed. The second part of the review is concerned on the inherited variations in IL-12A and IL-12B genes that could modulate cancer susceptibility, and as a consequence, possess predictive, therapeutic, or prognostic significance. It is known that functional single nucleotide polymorphisms (SNPs) in IL-12A and IL-12B genes may dramatically affect on protein expression level, or alter its functions, which may lead to immune disorders, autoimmune diseases, and eventually contribute to cancer occurrence. The list of genetic polymorphisms for further investigations might include the following: IL-12B_+1188A/C (rs3212227), IL-12A_+277G/A (rs568408), IL-12A_-798T/A (rs582054), IL-12A_-504T/G (rs190533), IL-12A_-1148T/C (rs2243123), and IL-12B_+16974 A/C. Perhaps, some of these SNPs may become an attractive target for oncogenomics and possibly could be used in programs of early cancer diagnosis as well as cancer prevention in the nearest future.     

The Peritoneal Fluid Levels of Interleukin-12 in Women with Endometriosis

PROBLEM: Interleukin-12 (IL-12) is produced mainly by monocytes/macrophages, and it induces proliferation and cytotoxicity of T-cells and natural killer cells. In women with endometriosis, natural killer cell activity in the peritoneal fluid is significantly decreased. We aimed to measure the peritoneal fluid level of IL-12 in endometriosis. METHOD OF STUDY: We measured IL-12 levels in peritoneal fluid samples from women with or without endometriosis and in supernatants from endometrial stromal, ovarian stromal, and mesothelial cell cultures, using a high-sensitivity enzyme-linked immunosorbent assay. RESULTS: The median concentration of IL-12 in the peritoneal fluid of women with endometriosis was 1.1 pg/ml (range, 0.2–5.5) and was 1.6 pg/ml (range, 0.4-2.8) in women without endometriosis, not a statistically significant difference. IL-12 was not detected in the supernatants of endometrial stromal, ovarian stromal, and mesothelial cell cultures. CONCLUSION: Concentrations of IL-12 in the peritoneal fluid of women with or without endometriosis are low, but they are detectable and are not affected significantly by the presence of endometriosis.     

Interleukin-4 and Interleukin-10 Gene Polymorphisms in Patients with Inflammatory Bowel Disease

Background: Changes in cytokine expression have been frequently found in patients with inflammatory bowel disease (IBD). Cytokine values outside the normal range may be somewhat related to common polymorphisms within cytokine genes.

Objective: The present study was designed to investigate the possible association between polymorphisms within Interleukin IL-4 and IL-10 genes and susceptibility to and clinical features of IBD.

Methods: The study population was composed of 140 healthy controls and 75 patients with IBD (40 patients with Crohn’s disease (CD) and 35 patients with ulcerative colitis (UC)). Genotyping was performed using polymerase chain reaction with sequence-specific primers.

Results: Higher frequencies for the C allele of IL-4–590 polymorphism (P < 0.0001; odds ratio [OR], 5.68; 95% confidence interval [95% CI], 3.28–9.83) and for the T allele of IL-4–1098 polymorphism (P = 0.016; OR, 1.83; 95% CI, 1.11–3.02) were observed in the whole group of IBD patients. The IL-4–590 C allele was also significantly overrepresented when IBD patients were subdivided into CD and UC (P < 0.0001; OR, 5.2–6.28). While the IL-4–1098 T allele was present at higher frequencies in patients with UC (P = 0.05; OR, 1.95), but not in CD (P = 0.09). Multiple pairwise comparisons indicated that genotypes of all polymorphisms investigated within IL-4 gene are correlated with IBD, CD, and UC. Haplotype analysis showed that the IL-4–1098/-590 TC haplotype might predispose individuals to IBD, CD, and UC whereas the IL-4–1098/-590 TT and GC haplotypes have a protective effect. On the contrary, neither allele nor genotype frequencies of IL-10 polymorphisms (IL-10–1082 A > G, IL-10–592 A > C, and IL-10–819 T > C) were associated with IBD, CD, or UC.

Conclusions: The present study suggests that IL-4 polymorphisms might play a role in susceptibility to IBD and its major subtypes in the Iranian population.     

Plasma Interleukin-12 Is Elevated in Patients with Preeclampsia

PROBLEM: Abnormal immune activation has been suggested as a contributor to the development of preeclampsia. We hypothesized that intact interleukin (IL)-12 directly, or through its main mediator, interferon (IFN)-γ, contributes to the altered immune response observed in preeclampsia. METHOD OF STUDY: Plasma samples were collected from 20 patients with preeclampsia and 20 normotensive patients with uncomplicated pregnancies who were matched with the preeclamptic patients by age, gestational age, and parity. Samples were collected before the onset of labor, induction, or medical intervention. The samples were assayed for IL-12 and IFN-γ by specific enzyme-linked immunoassays. RESULTS: IL-12 was detected in 35% of the preeclamptic patients and in 5% of the patients with normal pregnancies (P < 0.01). The detection rate and mean concentration of IFN-γ were comparable in both groups. CONCLUSION: Intact plasma IL-12 is detected more frequently in preeclamptic patients, suggesting the involvement of this cytokine in the enhanced immune response observed in preeclampsia.     

The combined effect of interleukin (IL)-10 and IL-12 polymorphisms on induced cytokine production

Interleukin (IL)-12 and IL-10 are immunoregulatory cytokines with an antagonistic effect of the T-helper (Th)1/Th2 cytokine balance and provide a functional link between innate and adaptive immune responses. The aim of the study was to investigate the combined effect of -1082A*G in IL10 and +16974A*C in IL12B single nucleotide polymorphisms (SNPs) on induced cytokine production by stimulated peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. The presence of the high-producer IL-12p40 genotype led to diminished production of IL-10 as determined by the -1082*G allele of SNP in IL10. Significantly decreased IL-10 production was detected in AA+AG/GG in comparison with the low-producer IL-12p40 (AC/CC+AG/GG) genotype combination after stimulation with C3bgp (2 +/- 4 vs. 29 +/- 14.2 pg/ml; p = 0.0003) and LPS (33.4 +/- 13.5 vs. 93.3 +/- 59.6 pg/ml; p = 0.019). IL-12p40 production was independent of IL10 genotype. The present results demonstrated that the production of IL-10 from PBMC depended on both -1082A*G in IL10 and +16974A*C in IL12B polymorphisms.     

充血性心力衰竭患者血清白介素-10水平变化及意义

目的通过检测充血性心力衰竭（CHF）患者血清白介素-10（IL-10）水平，探讨其与CHF的关系及临床意义。方法采用双抗体夹心法测定30例CHF息者治疗前后及25例健康人血清IL-10水平。结果CHF患者血清IL-10水平显著高于时照组（P〈0．01），随CHF加重，IL-10水平升高越明显（心功能Ⅱ级与对照组比较差异不显著P〉0．05，Ⅲ级及Ⅳ级与对照组比较P〈0.01）：心衰好转后明显下降（P〈0．01）。结论 IL-10可能参与了CHF的发生和发展，其在临床方面的意义有待于进一步探讨。     

Production of interleukins 10 and 12 by peripheral blood mononuclear cells (PBMC) in chronic hepatitis C virus (HCV) infection

We previously reported that interferon-gamma (IFN-γ) production by PBMC in response to HCV core protein was increased in patients with type C chronic liver disease. To understand better the pathophysiology of this disease, we evaluated production of IL-10 and IL-12 by PBMC from 41 patients with chronic HCV infection, including asymptomatic HCV carriers with persistently normal serum ALT values. IL-10 is known to inhibit many effector functions of the immune system, suppressing Th1-type cell development, while IL-12 stimulates differentiation of Th1-type cells, facilitating cell-mediated immunity. IL-10 production was determined by culturing lymphocytes with concanavalin A (Con A), while IL-12 was produced by monocytes in the presence of Staphylococcus aureusCowan 1 (SAC) with or without recombinant HCV core protein, respectively. The cytokine levels in culture supernatants were measured by ELISA. Spontaneous IL-10 production was greater in patients with chronic hepatitis (CH) (229±119 pg/ml, P<0.01) and liver cirrhosis (LC) (185±88 pg/ml, P<0.05) than in controls (119±27 pg/ml), while it was decreased during IFN treatment (70±25 pg/ml). Both HCV core protein and Con A enhanced IL-10 production by cells from HCV-infected patients. IL-12 was not detectable in medium alone cultures, and SAC-induced IL-12 production did not differ between various patient groups and controls. Simultaneous addition of HCV protein resulted in an increase of IL-12 production in chronic liver disease compared with SAC-alone cultures. Addition of IL-10 to the cultures equally suppressed IFN-γ production for both controls and patient groups, but the enhancing effect of IL-12 on IFN-γ production was significantly less in LC than in controls and other patient groups. The findings suggest that secretion of IL-10/IL-12 by cells from control individuals and various patient groups may be different, and that the cytokines might show different effects on IFN-γ production by some cells.     

Natural killer cells in cross-regulation of IL-12 by IL-10 in Leishmania antigen-stimulated blood donor cells.

We have previously shown that natural killer (NK) cells play a role in protection against leishmaniasis. Furthermore, we have shown that NK cells in mononuclear cells derived from unexposed donors are induced to proliferate in vitro in response to leishmanial antigens. Since interleukin (IL)-12, a strong inducer of NK cells, acts on the early events in NK cells and T-cells, and is considered as an adjuvant for use in a potential antileishmaniasis antigen, we wished to investigate how this cytokine influences the in vitro Leishmania induced proliferative and cytokine response in healthy donors. We demonstrate that in an innate response to Leishmania antigen involving NK cells, a critical level of IL-12 is required to induce interferon (IFN)-gamma secretion below which, IL-10 is released in amounts which apparently inhibit IFN-gamma secretion and cellular proliferation. However, at higher IL-12 levels, there is simultaneous secretion of IFN-gamma and IL-10 as well as proliferation of cells. In a similar vein, exogenous IL-10 in turn inhibited IFN-gamma secretion as well as proliferation when used at low/medium concentrations, but at high concentrations this effect was abolished and replaced by the simultaneous detection of IFN-gamma, IL-10 and proliferation. The contribution of NK cells in cross regulation of these two very important immuneregulatory cytokines and the effect of exogenous IL-12 in a Leishmania driven response are discussed.