IL-18 enhances IFN-gamma-induced production of CXCL9, CXCL10, and CXCL11 in human keratinocytes

IL-18 is involved in the pathogenesis of atopic dermatitis, psoriasis, and allergic contact dermatitis. CXCL9, CXCL10, and CXCL11 recruit type 1 T cells, and the production of these chemokines by keratinocytes is enhanced in these dermatoses. We examined the in vitro effects of IL-18 on IFN-gamma-induced CXCL9, CXCL10, and CXCL11 production in human keratinocytes. IL-18 enhanced the IFN-gamma-induced secretion and mRNA expression of CXCL9, CXCL10, and CXCL11 in parallel to the activation of NF-kappaB, STAT1, and IFN-regulatory factor (IRF)-1. Antisense oligonucleotides against NF-kappaB p50, p65, or STAT1 suppressed CXCL9, CXCL10, and CXCL11 production, and antisense IRF-1 suppressed CXCL11 production. Inhibitors of PI3 K, p38 MAPK, and MEK suppressed IL-18 plus IFN-gamma-induced CXCL9, CXCL10, and CXCL11 production and NF-kappaB, STAT1, and IRF-1 activities. IL-18 induced phosphorylation of ERK and Akt, while IFN-gamma induced phosphorylation of p38 MAPK. These results suggest that IL-18 may potentiate IFN-gamma-induced CXCL9, CXCL10, and CXCL11 production in keratinocytes by activating NF-kappaB, STAT1, or IRF-1 through PI3 K/Akt and MEK/ERK pathways. These effects of IL-18 may promote the infiltration of type 1 T cells into lesions with inflammatory dermatoses and amplify the skin inflammation. IL-18 may act as a pro-inflammatory cytokine in these dermatoses and thus is a candidate therapeutic target.     

Alveolar macrophages from patients with bronchogenic carcinoma and sarcoidosis similarly express monocyte antigens.

It has been shown that bronchoalveolar lavages (BALs) from patients with sarcoidosis and other interstitial lung diseases contain abnormally increased numbers of alveolar macrophages (AM) expressing antigens found on monocytes. The aim of this study was to compare the phenotype of AM from patients with sarcoidosis with those from patients with non-interstitial lung disease, namely carcinoma. Using a panel of monoclonal antibodies against cells of the mononuclear phagocytic series and immunohistochemical staining, we have compared the expression of antigens on AM recovered at BAL and peripheral blood monocytes from patients with sarcoidosis, with similar cell preparations from bronchogenic carcinoma patients and normal volunteers. We have shown that CD14, CR1 (CD35) and CR3 (CD11b, CD18) are expressed on the majority of monocytes from all subjects, but on only a minority of normal AM. In both patients with sarcoidosis and patients with bronchogenic carcinoma increased proportions of AM expressed these monocyte-associated antigens. Since BALs from the carcinoma patients were derived from lung lobes which were radiologically free of tumour, the accumulation of AM expressing monocytic antigens is not a local response to the tumour. We conclude that infiltration of the lung with monocytes is a more general response to lung disease than has hitherto been reported.     

Evaluation of CXCL10, CXCL11, CXCL12 and CXCL13 chemokines in serum and cerebrospinal fluid in patients with tick borne encephalitis (TBE)

PurposeThe aim of the study was to assess the concentration of chemokines: CXCL10, XCL11, CXCL12, CXCL13 in serum and cerebrospinal fluid (CSF) in patients with tick-borne encephalitis (TBE) before and after treatment. We evaluated also the usefulness of these molecules in diagnosis and monitoring of inflammation in TBE.MethodsTwenty three patients hospitalized in The Department of Infectious Diseases and Neuroinfections of Medical University in Bia?ystok, Poland were included in the study. Patients were divided into 2 groups: TBE group-patients with confirmed TBE and control group (CG): patients with excluded TBE and other inflammatory diseases of CNS. Concentration of CXCL10/IP-10, CXCL11/I-TAC, CXCL12/SDF-1α, CXCL13/BLC/BCA-1 in serum and CSF were measured with ELISA kits (R&;D Systems, USA) according to the protocols.ResultsThe analysis of chemokines concentration in TBE patients before treatment and control group using ROC showed that serum CXCL10 and CXCL13 and CSF CXCL10, CXCL11, CXCL12 and CXCL13 differentiate both groups (p<0.05). The analysis of CXCL10, CXCL11, CXCL12 and CXCL13 before and after treatment showed that CXCL10 and CXCL11 in CSF and CXCL13 in serum differentiates both groups with p<0.05.ConclusionsConcentration of CSF CXCL10, CXCL11, CXCL12, CXCL13 and serum CXCL10, CXCL13 may be good biomarkers of CNS inflammation caused by TBEV. Moreover concentration of CXCL10 in CSF and CXCL13 in serum may be used as indicators of patients recovery.     

Immunophenotyping of macrophages in human pulmonary tuberculosis and sarcoidosis

Classic studies of tuberculosis (TB) revealed morphologic evidence of considerable heterogeneity of macrophages (MØs), but the functional significance of this heterogeneity remains unknown. We have used newly available specific antibodies for selected membrane and secretory molecules to examine the phenotype of MØs in situ in a range of South African patients with TB, compared with sarcoidosis. Patients were human immunodeficiency virus-negative adults and children, and the examined biopsy specimens included lung and lymph nodes. Mature pulmonary MØs (alveolar, interstitial, epithelioid and multinucleated giant cells) selectively expressed scavenger receptor type A and a novel carboxypeptidase-like antigen called carboxypeptidase-related vitellogenin-like MØ molecule (CPVL). CPVL did not display enhanced expression in sarcoidosis, vs. TB patients, as observed with angiotensin-converting enzyme (ACE), a related molecule. Immunocytochemical studies with surfactant proteins (SP)-A and -D showed that type II alveolar cells expressed these collectins, as did MØs, possibly after binding of secreted proteins. Studies with an antibody specific for the C-terminus of fractalkine, a tethered CX3C chemokine, confirmed synthesis of this molecule by bronchiolar epithelial cells and occasional endothelial cells. These studies provide new marker antigens and extend previous studies on MØ differentiation, activation and local interactions in chronic human granulomatous inflammation in the lung.     

Possible role of L-selectin in T lymphocyte alveolitis in patients with active pulmonary sarcoidosis

A number of adhesion molecules participate in the recruitment of inflammatory cells to the site of inflammation, and selectins together with their ligands are important in the early transient adhesion phase. In this study, we evaluated the role of L-selectin in T lymphocyte alveolitis in patients with active pulmonary sarcoidosis. We measured serum and bronchoalveolar lavage fluid (BALF) concentrations of soluble (s)L-selectin using an ELISA. Serum and BALF concentrations of sL-selectin were significantly elevated in patients with sarcoidosis compared with control healthy subjects and idiopathic pulmonary fibrosis (IPF) patients (P < 0.05 and P < 0. 01, respectively). The lymphocyte surface marker was also examined in peripheral blood and BALF by flow cytometric analysis. The percentage of CD3+CD62L+ cells (L-selectin-bearing T lymphocytes) was significantly lower in peripheral blood of sarcoidosis than in that of healthy subjects (P < 0.01). In contrast, the percentage of CD3+CD62L- cells (L-selectin-negative T lymphocytes) in BALF of patients with sarcoidosis was significantly higher than in healthy subjects (P < 0.05) and IPF patients (P < 0.01). Furthermore, there was a significant correlation between serum concentrations of sL-selectin and the number of L-selectin-negative T lymphocytes in BALF (r = 0.535, P < 0.01). Our results suggest that L-selectin may be involved in T lymphocyte alveolitis in patients with active pulmonary sarcoidosis.     

Common variants of chemokine receptor gene CXCR3 and its ligands CXCL10 and CXCL11 associated with vascular permeability of dengue infection in peninsular Malaysia

Dengue causes significantly more human disease than any other arboviruses. It causes a spectrum of illness, ranging from mild self-limited fever, to severe and fatal dengue hemorrhagic fever, as evidenced by vascular leakage and multifactorial hemostatic abnormalities. There is no specific treatment available till date. Evidence shows that chemokines CXCL10, CXCL11 and their receptor CXCR3 are involved in severity of dengue, but their genetic association with the susceptibility of vascular leakage during dengue infection has not been reported. We genotyped 14 common variants of these candidate genes in 176 patients infected with dengue. rs4859584 and rs8878 (CXCL10) were significantly associated with vascular permeability of dengue infection (P < 0.05); while variants of CXCL11 showed moderate significance of association (P = 0.0527). Haplotype blocks were constructed for genes CXCL10 and CXCL11 (5 and 7 common variants respectively). Haplotype association tests performed revealed that, “CCCCA” of gene CXCL10 and “AGTTTAC” of CXCL11 were found to be significantly associated with vascular leakage (P = 0.0154 and 0.0366 respectively). In summary, our association study further strengthens the evidence of the involvement of CXCL10 and CXCL11 in the pathogenesis of dengue infection.     

Silica-induced pulmonary fibrosis involves the reaction of particles with interstitial rather than alveolar macrophages

Macrophage-derived products have been implicated in fibroblast stimulation following particle deposition in the lung. To assess the role of macrophages in the alveolus versus those in the interstitium in the induction of pulmonary fibrosis, we compared the pulmonary response to silica when phagocytosis occurred predominantly in each of these compartments. One group of mice received intratracheal silica which was phagocytosed largely by alveolar macrophages (AM). A second group was exposed to whole body irradiation prior to receiving the same dose of silica. This prevented the usual efflux of PMN and monocytes into the air sacs, allowing passage of silica particles across the alveolar epithelium to reach the interstitial macrophages (IM). In the irradiation plus silica group, many large interstitial granulomas were formed at 2-4 weeks, and collagen levels were significantly greater than in all other groups at 16 weeks. More silica was found in a lung tissue residue and in lymph nodes of these animals. Pulmonary fibrosis was limited to interstitial areas where there was a high level of retained silica, whereas peripheral regions of the lung, where free AM containing silica were found, did not show fibrosis of the alveolar walls. The results suggest that factors secreted by IM in response to silica are more effective in stimulating fibrogenesis than secretions made by the AM into the alveolar space.     

CXCL10 induces the recruitment of monocyte-derived macrophages into kidney,which aggravate puromycin aminonucleoside nephrosis

The mechanism responsible for trafficking of monocyte-derived macrophages into kidney in the puromycin aminonucleoside model of nephrotic syndrome in rats (PAN-NS), and the significance of this infiltration, remain largely unknown. CXCL10, a chemokine secreted in many T helper type 1 (Th1) inflammatory diseases, exhibits important roles in trafficking of monocytes and activated T cells. We hypothesized that induction of circulating interferon (IFN)-γ and glomerular tumour necrosis factor (TNF)-α during PAN-NS would stimulate the release of CXCL10 by podocytes, leading to infiltration of activated immune cells and greater glomerular injury. We found that serum IFN-γ, glomerular Cxcl10 mRNA and intra- and peri-glomerular macrophage infiltration were induced strongly during the late acute phase of PAN-NS in Wistar rats, but not in nude (Foxn1^rnu/rnu) rats lacking functional effector T lymphocytes. Wistar rats also developed significantly greater proteinuria than nude rats, which could be abolished by macrophage depletion. Stimulation of cultured podocytes with both IFN-γ and TNF-α markedly induced the expression of Cxcl10 mRNA and CXCL10 secretion. Together, these data support our hypothesis that increased circulating IFN-γ and glomerular TNF-α induce synergistically the production and secretion of CXCL10 by podocytes, attracting activated macrophages into kidney tissue. The study also suggests that IFN-γ, secreted from Th1 lymphocytes, may prime proinflammatory macrophages that consequently aggravate renal injury.     

IL-10对小鼠肺泡巨噬细胞清道夫受体及CD14表达的影响

本研究旨在观察IL-10对肺泡巨噬细胞（AM），清道夫受体（SR），CD14表达的影响。探讨IL-10在内毒素血症时防止AM由免疫防御向炎症效应转变中的作用。分离培养小鼠AM，以不同剂量（0,0.01,0.1,1,10,100μg/L）IL-10刺激细胞16h或以100μg/LIL-10刺激细胞不同时间（0,2,4,6,8,12,16h)，采用免疫细胞化学及RT-PCR方法观察SR，CD14表达变化。结果显示低至10μg/LIL-10刺激16h或100μg/LIL-10刺激12-24h能显著增强SRmRNA并抑制CD14mRNA表达，SR蛋白表达也显著增强，但CD14蛋白表达无明显变化，IL-10刺激下对SR蛋白表达的增强能降低AM对LPS的反应性，在内毒素血症时防止AM由免疫防御型向炎症效应型转变中发挥重要作用。     

巨噬细胞在人类肺组织纤维化中的作用

目的：探讨肺泡巨噬细胞在纤维性间质性肺病胶原纤维异常沉积中的作用。方法：应用ＨＥ 染色、Ｍａｓｓｏｎ 三色染色和免疫组织化学染色。结果：在纤维性间质性肺病组和疾病对照组之间,肺泡巨噬细胞ＴＧＦβ１ 和ｂＦＧＦ阳性表达率差异均有显著性（ Ｐ＜ ０－０５） ;肺泡巨噬细胞ＰＤＧＦα受体阳性率差异亦有显著性（Ｐ＜０－０５）。在增生性肺泡Ⅱ型上皮细胞,ＰＤＧＦα受体阳性表达率,无胶原沉积组高于胶原沉积组（ Ｐ＜ ０－０５） ;而ＴＧＦβ１ 阳性表达率胶原沉积组高于无胶原沉积组（ Ｐ＝ ０－０１）。结论：肺泡巨噬细胞参与了纤维性间质性肺病及其胶原纤维异常沉积的发生发展过程。     

Increased expression of adhesion molecules (ICAM-1 and LFA-1) on alveolar macrophages from asthmatic patients

In the airways inflammation observed in asthma, activated macrophages are present in increased numbers. Adhesion molecules are required for the cell: cell contacts between leukocytes and endothelial cells or other leukocytes, and they are induced by inflammatory stimuli. We studied the expression of two adhesion molecules (ICAM-1 and LFA-1) on alveolar macrophages recovered by bronchoalveolar lavage from 11 normal subjects and 13 asthmatic patients by using immunocytochemistry. Two specific monoclonal antibodies were used, and the reaction was revealed by the alkaline phosphatase-antialkaline phosphatase (APAAP) method. The percentage of cells expressing ICAM-1 or LFA-1 was significantly increased in asthmatic patients, as compared with normal subjects ( P < 0.001 and P < 0.002, respectively; Mann-Whitney U test), and there was a significant correlation with the percentage of cells expressing both markers in asthma ( P < 0.03, Spearman rank test). This study highlights the importance of macrophages in the inflammation of asthma and suggests that macrophage interactions with other cells play a role in this inflammation.     

Dose- and time-dependent activation of rat alveolar macrophages by glucocorticoids

Effects of glucocorticoids on immune functions are generally thought to be suppressive and anti-inflammatory. However, most reports dealing with this issue describe effects of long-term treatment with high doses of glucocorticoids on immune functions. In the present study we have investigated both dose and timing effects of exposure of alveolar macrophages with dexamethasone on lipopolysaccharide (LPS)-induced IL-1β and nitric oxide secretion. For this purpose, alveolar macrophages were preincubated with various doses of dexamethasone during varying intervals, followed by stimulation of the cells with endotoxin, either in the absence or presence of dexamethasone. Subsequently, the effects of this treatment on IL-1β and nitric oxide secretion were measured. It was shown that both short-term incubation of alveolar macrophages with high doses of dexamethasone and long-term incubation with low doses of dexamethasone lead to enhanced nitric oxide and enhanced IL-1β secretion upon subsequent stimulation of the cells with LPS. In contrast, long-term incubation of alveolar macrophages with high-dose dexamethasone leads to decreased IL-1β and nitric oxide secretion upon subsequent stimulation. Thus, it is concluded that the effects of dexamethasone on rat alveolar macrophages are both time- and dose-dependent. It is therefore argued that effects of glucocorticoids on immune functions are not a priori suppressive, but that both dose and timing effects should be taken into account.     

Production and titration of African swine fever virus in porcine alveolar macrophages

The broncho-alveolar lavage of a pig (20–40 kg) contains about 1.6 × 10⁹ alveolar cells, half of which were macrophages. The number of cells in the lavage of bacille Calmette Guerin (BCG)-treated pigs increased about 4-fold. Both African swine fever virus-infected porcine alveolar macrophages and blood monocytes produced about 1000 hemadsorption units/cell, a value 10-fold larger than that obtained in virus-infected Vero cells. Porcine alveolar cells could be stored frozen and, after thawing, they could be infected with African swine fever virus, producing the same amount of virus as the unfrozen cells. With the number of alveolar macrophages obtained from a single pig it is possible to titer about 3000 virus samples with the same stock of alveolar macrophages.     

Low-dose theophylline does not exert its anti-inflammatory effects in mild asthma through upregulation of interleukin-10 in alveolar macrophages

BACKGROUND: There is accumulating evidence that theophylline has anti-inflammatory or immunomodulatory effects. This may be, in part, mediated via an upregulation in the production of the anti-inflammatory cytokine interleukin (IL)-10. We determined whether low-dose theophylline (LDT) would increase the production of IL-10, and attenuate the production of proinflammatory cytokines by alveolar macrophages. METHODS: In a double-blind, placebo-controlled, crossover study involving 15 steroid-free patients with mild asthma, fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) were performed at the end of the treatment and placebo periods. Alveolar macrophages were cultured in vitro, and we measured their release of IL-10, GM-CSF, and TNF-alpha. We also measured IL-10 production in whole blood together with the number of monocytes and T cells expressing intracellular IL-10 by flow cytometry. RESULTS: LDT did not increase the production of IL-10, or attenuate the production of GM-CSF or TNF-alpha by alveolar macrophages. However, after theophylline treatment, there was a significant reduction in mean (SD) (95% CI) BAL eosinophil number from 3.4 (1.7)% (95% CI 2.4-4.4) to 1.7 (1.0)% (95% CI 1.1-2.3) compared with placebo (P<0.05). Similarly, there was no increase in whole-blood IL-10 release or in the number of monocytes and T cells expressing intracellular IL-10 after treatment. CONCLUSIONS: LDT has an anti-inflammatory effect in asthma; however, this effect is not mediated via the production of IL-10 or the attenuation of GM-CSF or TNF-alpha. The mechanisms of theophylline activity remain to be determined.     

Glucocorticoid sensitivity of lipopolysaccharide‐stimulated chronic obstructive pulmonary disease alveolar macrophages

It has been reported that alveolar macrophages from patients with chronic obstructive pulmonary disease (COPD) display glucocorticoid (Gc) resistance. The Gc sensitivity of inflammatory mediators released by COPD macrophages may vary. The objective of this study was to identify Gc‐insensitive inflammatory mediators produced by lipopolysaccharide (LPS)‐stimulated alveolar macrophages from COPD patients. LPS‐stimulated alveolar macrophages from 15 COPD patients, nine smokers (S) and nine healthy non‐smokers (HNS) were stimulated with LPS with or without dexamethasone (100 and 1000 nM). Luminex and enzyme‐linked immunosorbent assay were used to measure 23 inflammatory mediators. After LPS stimulation there were lower levels of inflammatory mediators in COPD patients and S compared to HNS. There was no difference between groups for the effects of dexamethasone at either concentration (P > 0·05 for all comparisons). Tumour necrosis factor (TNF)‐α, interleukin (IL)‐6 and growth‐related oncogene (GRO)‐α displayed the greatest sensitivity to dexamethasone in COPD patients, while IL‐8, granulocyte–macrophage colony‐stimulating factor (GM‐CSF) and granulocyte colony‐stimulating factor (G‐CSF) were the least sensitive. COPD macrophages have a reduced response to LPS. Gc sensitivity was similar in COPD macrophages compared to controls. We identify some Gc‐insensitive cytokines, including GM‐CSF, G‐CSF and IL‐8, that may be involved in the progression of airway inflammation in COPD patients.