Chronic rhinosinusitis with and without nasal polyps is associated with decreased expression of glucocorticoid-induced leucine zipper

Background Chronic rhinosinusitis without nasal polyps (CRSsNP) and with nasal polyps (CRSwNP) is characterized by persistent inflammation of sinonasal mucosa. Glucocorticoid-induced leucine zipper (GILZ) is a recently described anti-inflammatory mediator.
Objective Here we analysed the expression of GILZ in CRSsNP and CRSwNP, its association with response to surgery, and its cytokine-driven expression regulation in the upper airways.
Methods The messenger RNA (mRNA) and protein expression of GILZ in 33 CRSsNP, 32 CRSwNP, and 11 control samples was assessed by means of a quantitative RT-PCR and immunohistochemistry, respectively. Nasal explant culture was used to investigate the effect of IFN-γ, IL-4, IL-13, IL-1β, and TNF-α on GILZ mRNA expression in normal sinonasal mucosa.
Results The GILZ mRNA and protein expression was significantly suppressed in both CRSsNP and CRSwNP patients compared with controls. No significant difference in GILZ expression was found between CRSsNP and CRSwNP patients. Comparing patients responsive and patients recalcitrant to surgery, a significant further decrease of GILZ expression was found in recalcitrant patients both in the CRSsNP and in the CRSwNP group. IL-1β, TNF-α, IL-4, and IL-13 reduced, whereas IFN-γ enhanced GILZ mRNA levels in the sinonasal mucosa.
Conclusion Down-regulated expression of GILZ may contribute to the pathogenesis of CRSsNP and CRSwNP and associate with response to surgery. GILZ expression in the upper airways can be regulated differentially by different cytokines.     

Differentiation of chronic sinus diseases by measurement of inflammatory mediators

Background:  Chronic rhinosinusitis (CRS) clinically is a heterogeneous group of sinus diseases, which may cover different disease entities, or may represent a disease continuum. Studying inflammatory cells and mediators in clearly defined disease subgroups may lead to a better differentiation of chronic sinus diseases.
Methods:  Sinonasal mucosal tissue from 10 nasal polyp (NP) patients, 13 cystic fibrosis patients (CF-NP), eight CRS subjects without polyps, and nine control patients were stained for CD3, CD25, CD68, CD20, myeloperoxidase (MPO), CD138 and tissue homogenates were assayed for eotaxin, interleukin (IL)-1 β , IL-2sR α , IL-5, interferon (IFN)- γ , IL-8, transforming growth factor (TGF)- β 1, tumor necrosis factor- α , and MPO by enzyme-linked immunosorbent assay or UNICAP system.
Results:  Nasal polyp and CF-NP showed increased numbers and activation of T cells, while only NP displayed an increase in plasma cells. Nasal polyp had significantly higher levels of eosinophilic markers [eosinophils, eotaxin, and eosinophil cationic protein (ECP)] compared with CRS, controls and CF-NP. Chronic rhinosinusitis was characterized by a Th1 polarization with high levels of IFN- γ and TGF- β , while NP showed a Th2 polarization with high IL-5 and immunoglobulin (Ig) E concentrations. Nasal polyp and CF-NP were discriminated by edema from CRS and controls, with CF-NP displaying a very prominent neutrophilic inflammation.
Conclusion:  Based on cellular and mediator profiles, we suggest that CRS, NP, and CF-NP are distinct disease entities within the group of chronic sinus diseases.     

Intercellular adhesion molecule-1 on cultured human epithelial cell lines: influence of proinflammatory cytokines

Paolieri F, Battifora M, Riccio AM, Pesce G, Canonica GW, Bagnasco M. Intercellular adhesion molecule-1 on cultured human epithelial cell lines: influence of proinflammatory cytokines.
The expression of intercellular adhesion molecule-1 'CD54 or ICAM-1' on epithelial cells during acute or chronic inflammation may favor the interaction between epithelial cells and leukocytes expressing the natural ligands of ICAM-1, LFA-1 'CD11a/CD18', and Mac-1 'CD11b/CD18'. We have evaluated in vitro the expression of ICAM-1 by a conjunctival 'WK' and an intestinal '1407' human continuous epithelial cell line. Cells were cultured for 24 h in the presence or absence of IFN-γ, TNF-α, IL-1β, IL-4, IL-6, IL-8, IL-10, and TGF-Jβ1. Both epithelial cell lines showed a constitutive expression of ICAM-1. IFN-γ at 500 U/ml and TNF-α at 200 ng/ml upregulated ICAM-1 expression; IL-1β at 100 pg/ml upregulated ICAM-1 on WK cells only. Cells cultured in the presence of both IFN-γ and TNF-α exhibited a mean fluorescence intensity far greater than those cultured with IFN-γ or TNF-α alone. 1407 and WK cells were able to release soluble ICAM-1. IFN-γ and TNF-α enhanced the release of sICAM-1. IL-4, IL-6, IL-8, IL-10, and TGF-β1 did not affect either ICAM-1 expression or sICAM-1 release. In conclusion, continuously cultured human epithelial cells may express ICAM-1 on their surface and release it in culture medium. These phenomena are upregulated by proinflammatory cytokines.     

Counter regulation of the high affinity IgE receptor, FcεRI, on human airway dendritic cells by IL-4 and IL-10

Background:  Immunoglobulin E is a signalling molecule within the environment of the respiratory tract, the high affinity receptor for which, FcεRI, is expressed by dendritic cells (DC). Little is known, however, of the expression and function of FcεRI on DC in the human respiratory tract.
Methods:  CD1c⁺ DC were purified from surgically resected nasal turbinates of 11 atopic and 12 nonatopic patients with chronic rhinosinusitis. Expression of FcεRI was determined by flow cytometry. Cytokine production by DC was determined by cytometric bead array.
Results:  Expression of FcεRI was significantly elevated on respiratory tract dendritic cells (RTDC) from atopic as compared to nonatopic patients. Activation of RTDC through FcεRI induced production of the pro-inflammatory cytokines IL-6 and TNF-α, and the anti-inflammatory cytokine IL-10. The production of IL-6 and TNF-α was elevated in atopic compared to nonatopic patients studied. Conversely IL-10 production was elevated in nonatopic patients. Concomitant activation of FcεRI and stimulation of RTDC with IL-4 inhibited production of IL-10 by RTDC. Neutralization experiments with anti-IL-10 Ab enhanced whereas addition of exogenous IL-10 to RTDC inhibited FcεRI-mediated inflammatory cytokine production.
Conclusion:  The function of FcεRI on RTDC from patients with rhinosinusitis is susceptible to counter regulation by IL-4 and IL-10.     

Maternal Circulating TNF-α Levels are Highly Correlated with IL-10 Levels, but not IL-6 and IL-8 Levels, in Women with Pre-Eclampsia

Problem  Several lines of evidence have shown that maternal cytokine levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-10 were altered in women with pre-eclampsia (PE) compared to those from normal pregnancies. In this study, we determined whether these cytokine levels are correlated before and after delivery in patients with PE.
Method of study  Venous blood was obtained from 50 women diagnosed with severe PE at the time of admission and 24 hr after delivery. Plasma concentrations for TNF-α, IL-6, IL-8, and IL-10 were measured by ELISA.
Results  There were no statistical differences for maternal levels of TNF-α, IL-6, IL-8, and IL-10 before and 24 hr postpartum. TNF-α and IL-10, but not IL-6 and IL-8, levels were significantly correlated before and 24 hr after delivery: TNF-α: y  = 19.963 + 0.953* x ; r ² = 0.924; IL-10: y  = 10.521 + 1.113* x ; r ² = 0.984, P  < 0.001, respectively. Furthermore, TNF-α levels were correlated with IL-10 levels, but not with IL-6 and IL-8 levels.
Conclusion  The correlation patterns of TNF-α with IL-10 and TNF-α with IL-6 and IL-8 suggest disparity in functional regulations between these cytokines in maternal circulation in PE.     

Increased Expression of Glutathione by Estradiol, Tumor Necrosis Factor-Alpha, and Interleukin 1-Beta in Endometrial Stromal Cells

Problem  The intracellular antioxidant system, based on glutathione (GSH), plays a key role in endometrial detoxification reactions and has been proposed to be involved in the pathogenesis endometriosis. This study was designed to evaluate whether estradiol (E₂) and proinflammatory cytokines have any effects on expression of glutathione in endometrial stromal cells (ESCs).
Method of study  Glutathione levels were measured utilizing high-performance liquid chromatography following in vitro culture and treatment of ESCs with estradiol, tumor necrosis factor-alpha (TNF-α) and interleukin 1-beta (IL-1β).
Results  The GSH level in E₂ (10⁻⁸  m ) treatment group was significantly higher than in the control group at 48 h ( P  < 0.05). In vitro treatment of ESCs with TNF-α 10 ng/mL as well as E₂ (10⁻⁸  m ) plus TNF-α 10 ng/mL for 48 hr also led to a significant increase in GSH level ( P  < 0.05; P  < 0.05, respectively). Both IL-1β 10 ng/mL and E₂ (10⁻⁸  m ) plus IL-1β 10 ng/mL for 48 hr increased GSH level significantly ( P  < 0.05; P  < 0.05, respectively) as well.
Conclusions  These findings might suggest that increased production of estradiol and proinflammatory cytokines in the peritoneal cavity possibly leads to the establishment of endometriosis through increased level of GSH.     

Contribution of Intestinal Epithelial Cells to Innate Immunity of the Human Gut – Studies on Polarized Monolayers of Colon Carcinoma Cells

The aim was to establish an in vitro model for studies of innate defence mechanisms of human intestinal epithelium. Ultrastructural characterization and determination of mRNA expression levels for apical glycocalyx and mucous components showed that polarized, tight monolayers of the colon carcinoma cell lines T84 and Caco2 acquire the features of mature- and immature columnar epithelial cells, respectively. Polarized monolayers were challenged with non-pathogenic Gram+ and Gram− bacteria from the apical side and the proinflammatory cytokines interferon-γ (IFN-γ), tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) from the basolateral side. Immune responses were estimated as changes in mRNA expression levels for the mucous component mucin-2 (MUC2), the glycocalyx components carcinoembryonic antigen (CEA), CEA-related cell adhesion molecule-1 (CEACAM1), CEACAM6, CEACAM7 and MUC3, the antimicrobial factors human β-defensin-1 (hBD1), hBD2, hBD3 and lysozyme, the chemokine IL-8 and the cytokines IL-6 and TNF-α. Tight monolayer cells were generally unresponsive to bacterial challenge, but increased their hBD2 levels when challenged with Bacillus megaterium. T84 cells also increased their TNF-α levels upon bacterial challenge. Tight monolayer cells responded to cytokine challenge suggesting awareness of basolateral attack. TNF-α induced significantly increased levels of IL-8 and TNF-α itself in both cell lines suggesting recruitment and activation of immune cells in the underlying mucosa in vivo . Cytokine challenge also increased levels of CEACAM1, which includes two functionally different forms, CEACAM1-L and CEACAM1-S. In T84 cells, IFN-γ was selective for CEACAM1-L while TNF-α upregulated both forms. Increased CEACAM1 expression may influence epithelial function and communication between epithelial cells and intraepithelial lymphocytes.     

The Role of Cytokine Expression in Different Subgroups of Subfertile Men

Problem  The aim of this study was to evaluate the levels of seminal plasma cytokines interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10), interleukin 11 (IL-11), interleukin 12 (IL-12), tumour necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) in male subfertility.
Method of study  A total of 73 male partners of an infertile couple attending a regional andrology unit were recruited into this prospective study and subdivided into the various groups based on semen analysis. Concentrations of cytokines such as IL-6, IL-8, IL-10, IL-11, IL-12, TNF-α and IFN-γ in the seminal plasma were determined using enzyme linked immunosorbent assay (ELISA).
Results  Significant higher concentrations ( P  < 0.05) of IL-6 in the mild and severe oligospermic group, IL-8 and IL-10 in the asthenospermic group and IL-6, IL-10, TNF-α and IFN-γ in the obstructed azoospermic group were determined. IL-10 concentrations correlated significantly with other cytokines in the obstructed azoospermic group and the asthenospermic group.
Conclusion  Our study confirms that cytokines rarely act in isolation, but rather in a network of other cytokines and may affect sperm function directly or indirectly. The presence of increased levels of cytokines in the obstructed azoospermic group suggests that the cytokines may not originate from the testis.     

Effect of an Extract Based on the Medicinal Mushroom Agaricus blazei Murill on Release of Cytokines, Chemokines and Leukocyte Growth Factors in Human Blood Ex Vivo and In Vivo

An immunostimulatory extract based on the medicinal mushroom Agaricus blazei Murill (AbM) has been shown to stimulate mononuclear phagocytes in vitro to produce pro-inflammatory cytokines, and to protect against lethal peritonitis in mice. The present aim was to study the effect of AbM on release of several cytokines in human whole blood both after stimulation ex vivo and in vivo after oral intake over several days in healthy volunteers. The 17 signal substances examined were; T helper 1 (Th1) cytokines [interleukin (IL)-2, interferon (IFN)-γ and IL-12], T helper 2 cytokines (IL-4, IL-5 and IL-13), pleiotropic (IL-7, IL-17), pro-inflammatory [IL-1β, IL-6, tumour necrosis factor (TNF)-α (mainly produced by Th1 cells)] – and anti-inflammatory (IL-10) cytokines, chemokines [IL-8, macrophage inhibitory protein (MIP)-1β and monocyte chemoattractant protein (MCP)-1] and leukocyte growth factors [granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony stimulating factor]. After stimulation of whole blood ex vivo with 0.5–5.0% of a mushroom extract, AndoSan™ mainly containing AbM , there was a dose-dependent increase in all the cytokines studied, ranging from two to 399-fold (TNF-α). However, in vivo in the eight volunteers who completed the daily intake (60 ml) of this AbM extract for 12 days, a significant reduction was observed in levels of IL-1β (97%), TNF-α (84%), IL-17 (50%) and IL-2 (46%). Although not significant, there was a trend towards reduced levels for IL-8, IFN-γ and G-CSF, whilst those of the remaining nine cytokines tested, were unaltered. The discrepant results on cytokine release ex vivo and in vivo may partly be explained by the antioxidant activity of AbM in vivo and limited absorption of its large, complex and bioactive β-glucans across the intestinal mucosa to the reticuloendothelial system and blood.     

Human Brain Endothelial Cells and Astrocytes Produce IL-1β but not IL-10

The ability of human brain endothelial cells to produce mRNA for interleukin-10, and release IL-10 in culture supernatants after in vitro stimulation with LPS, TNF-α and γ-IFN was assessed and compared to that of astrocytes, peripheral blood mononuclear cells and human umbilical vein endothelial cells. IL-1β and β₂-microglobulin release were also analysed. IL-10 and TNF-α mRNA presence was investigated in normal brain as well as in three plaques from two multiple sclerosis patients. While increased IL-1β and β₂-microglobulin release in the supernatants of stimulated cells could be detected in all the studied cell lineages, IL-10 mRNA and protein release was only seen in LPS-stimulated PBMNCs. Similarly, mRNA for IL-10 was not detected in CNS tissues, while TNF-α was present in all plaques. The lack of production of significant amounts of IL-10 by astrocytes and human brain endothelial cells suggests that these cells may not be the primary source of in vivo IL-10-mediated down-regulation of immune reactions within the central nervous system.     

Probiotic Bacteria Induce Regulatory Cytokine Production via Dendritic Cells

Probiotic bacteria, e.g. Lactobacillus spp., may improve diseases such as chronic inflammatory bowel disease. We examined cytokine production and phenotypic change after in vitro stimulation of T cells from healthy volunteers using different probiotic strains.
Methods:  T cells were cultured from colonic biopsies from eight healthy volunteers (Agnholt and Kaltoft, Exp Clin Immunogenet 2001;18:213–25), and dendritic cells were matured from their peripheral blood mononuclear cells. T-cell cultures were stimulated with autologous bacterial sonicate or strains of Lactobacillus spp., with and without the addition of dendritic cells. Cytokine levels (TNF-α, IFN-γ, IL-10 and GM-CSF) and phenotype (CD3, CD4, CD25 and CD69) were measured on day 4.
Results:  Lactobacillus spp. induced higher productions of TNF-α and IL-10 than did autologous bacteria. In presence of dendritic cells, the production of all cytokines increased. However, the increases of IFN-γ and TNF-α were more pronounced in wells with autologous bacteria than in wells with Lactobacillus spp. The addition of dendritic cells upregulated CD25 expression without simultaneous upregulation of CD69. The upregulation was pronounced after stimulation with Lactobacillus rhamnosus GG compared with autologous bacteria and other lactobacilli.
Discussion:  In presence of dendritic cells, autologous bacteria induced inflammatory cytokines, while probiotics mainly induced regulatory cytokines. Lactobacillus rhamnosus GG induced a regulatory phenotype (cd25⁺), in part mediated by dendritic cells. Future studies will address whether this shift to a CD25⁺ phenotype represents a differentiation into competent regulatory T cells. In a clinical context, such cells might be used for treatment of inflammatory diseases.     

Constitutive and Inducible Cytokine mRNA Expression in the Human Mast Cell Line HMC-1

The discovery that mast cells are a potential source of cytokines has suggested new ways in which mast cells can act in immunological and inflammatory responses. In this study we have used the HMC-1 cell line as a model for human mast cells to study the constitutive and inducible mRNA expression of interleukins, colony-stimulating factors, interferons, tumour necrosis factors α and β, tumour growth factor β and platelet-derived growth factor A and B. We found that HMC-1 cells constitutively expressed mRNA for TNF-α and TGF-β, and a low level of M-CSF. After treatment with the phorbol ester TPA or the calcium ionophore ionomycin expression of several cytokines, i. e. IL-1β, IL-3, IL-6, GM-CSF, TNF-β and PDGF-A, could be detected. Both TPA and ionomycin induced the same set of cytokines, but the effect of TPA was more prominent. The relative induction was calculated to be 70X for IL-1β and IL-3, 30X for GM-CSF and PDGF-A and 3 - 10X for IL-6, M-CSF and TNF-β. This study shows that human mast cells have the capacity to express not only cytokines mediating an immune response but also cytokines affecting other cell types, e. g. fibroblasts and endothelial cells, involved in later steps of the inflammatory response.     

Clinical evaluation of proinflammatory cytokine inhibitors (sTNFR I, sTNFR II, IL-1 ra), anti-inflammatory cytokines (IL-10, IL-13) and activation of neutrophils after burn-induced inflammation

The study was aimed at evaluating the involvement of sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 and reactive oxygen species (ROS) in systemic inflammatory response syndrome (SIRS) development in severely burned children and at assessing the prognostic value of the immunological markers studied. The study comprised 37 patients (17 burned children and 20 controls). Serum levels of the markers determined by means of ELISA and respiratory burst of neutrophils as well as p55 and p75 tumour necrosis factor-α (TNF-α) receptor expression using flow cytometry were evaluated twice. The burned children presented significantly higher levels of IL-10 and cytokine inhibitors within the first 6–24 h after injury compared with controls ( P  < 0.05). The decreased oxygen metabolism of neutrophils and increased TNF-α receptor expression were found on admission. Moreover, a significant decrease in initially high sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 concentrations ( P  < 0.05) and reduced expression of TNF-α receptors ( P  < 0.05) were observed after burn therapy, whereas ROS generation evidently augmented ( P  < 0.05). Four of our children who developed hypovolaemic shock revealed a significantly lower ROS generation and higher concentrations of soluble TNF-α receptors and IL-1 ra together with IL-10, IL-13 compared with children with good outcome ( P  < 0.05). Our results revealed the involvement of both ROS, soluble TNF-α receptors and IL-1 ra in the development of SIRS in burned children; their monitoring allows for an assessment of the systemic inflammatory reaction activity. The neutrophil BURSTTEST and IL-1 ra might have been clinically helpful markers of SIRS prognosis.     

Protection Against Concanavalin A-Induced Murine Liver Injury by the Organic Germanium Compound, Propagermanium

Propagermanium (3-oxygermylpropionic acid polymer) is an organic germanium compound that activates the immune system. In this study, we investigated the action of propagermanium on T-cell-mediated murine hepatic injury induced by concanavalin A (Con A). Oral administration of propagermanium inhibited the development of liver injury about 10 h after Con A injection. Histological analysis demonstrated that propagermanium attenuated the extent of liver damage compared with controls, reducing infiltration by leucocytes, especially CD11b-positive cells. Infiltration by CD4-positive cells was not affected. Tumour necrosis factor (TNF)-α and interferon (IFN)-γ are crucial for the development of hepatitis in this model. Propagermanium treatment induced significant inhibition of subsequent TNF-α production about 10 h after Con A injection, without affecting IFN-γ, interleukin (IL)-10, IL-4 and IL-12 production. This effect on TNF-α production coincided with the inhibition of aminotransferase activity late in the progression of Con A-induced liver injury. These facts suggest that this compound affects the macrophages (Mφ) function in the liver sinusoid. Therefore, Mφ were cultured with liver sinusoidal endothelial cells (SEC) and the effect of propagermanium on TNF-α production in the presence of IFN-γ was determined. TNF-α production was reduced significantly in the coculture of Mφ and SEC when Mφ was treated with propagermanium. These results might explain the mechanisms by which propagermanium inhibits Con-A-induced liver injury. That is, propagermanium improves hepatitis through mechanisms including the reduced production of TNF-α, without modification of Th1- and Th2-cell function.     

Role of Tyrosine Kinase Enzymes in TNF-α and IL-1 Induced Expression of ICAM-1 and VCAM-1 on Human Umbilical Vein Endothelial Cells

The aim of this study was to analyse the potential roles of protein kinase enzymes in tumour necrosis factor-α (TNF-α) and interleukin-1 (IL-1) induced expression of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on human umbilical vein endothelial cells (HUVEC). The authors observed a marked increase in ICAM-1 and VCAM-1 expression on HUVEC stimulated for 24 h by TNF-α (10 ng/ml) or IL-1 (20 ng/ml). Pre-treatment of HUVEC for 30 min with protein tyrosine kinase (PTK) inhibitors genistein and herbimycin A (10 μg/ml and 0.5 μg/ml, respectively) before stimulation with IL-1 did not affect the expression of these molecules. Similar results were observed with respect to VCAM-1 expression on HUVEC stimulated by TNF-α. In contrast, pre-incubation of HUVEC with PTK inhibitors prior to the addition of TNF-α significantly enhanced subsequent expression of ICAM-1, although spontaneous expression of ICAM-1 on unstimulated HUVEC was unaffected. Western blot analysis demonstrated a significant increase in phosphorylated tyrosine protein levels in HUVEC stimulated by TNF-α , and significantly lower levels of these proteins in TNF-α stimulated HUVEC pre-treated with PTK inhibitors. These results demonstrate that IL-1 induced ICAM-1 and VCAM-1 expression does not result from activation of PTK-dependent pathways. In the case of TNF-α induced responses, the selective co-stimulatory effect of this cytokine in combination with PTK inhibitors on ICAM-1 expression suggests a complicated intracellular pathway of TNF-α induced ICAM-1 expression, possibly involving down-modulation of increases in ICAM-1 by PTK enzymes.     

The Elevated Interferon Gamma Production is an Important Immunological Marker in HAM/TSP Pathogenesis

Human T-lymphotropic virus type 1 (HTLV-1) is the agent of the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), which may occur in >5% of patients during their lifetime. HTLV-1-infection causes disturbances in the immune system, and the viral load may also play an important role in the pathogenesis of HAM/TSP. Some cytokines are involved in the pathogenesis of this disorder. We have determined IL-2, IL-4, IL-10, IL-12 p70, IFN-γ and TNF-α production among HTLV-1-infected subjects from our HTLV-out Clinic in Institute of Infectious 'Emílio Ribas' in Sao Paulo city, Brazil. PBMC obtained from healthy controls ( n  = 32), asymptomatic HTLV-1 carriers ( n  = 68) and HAM/TSP patients ( n  = 44) were grown in the absence and in the presence of phytohaemagglutinin (PHA), and the supernatants' fluids were measured for cytokines production. IL-2 levels were increased in the asymptomatic HTLV-1 carriers, and IFN-γ was increased in both groups of patients (asymptomatic HTLV-1 carriers and more significantly among HAM/TSP patients). IL-4, IL-10, TNF-α and IL-12 p70 levels were not significantly increased on both groups of patients, as compared with controls. The major finding of this study is that IFN-γ was an important cytokine for the HAM/TSP pathogenesis. Therefore, immune modulation of IFN-γ may be critical to treat of HAM/TSP patients.     

Epithelium-derived chemokines induce airway smooth muscle cell migration

Background The remodelling of airway smooth muscle (ASM) associated with asthma severity may involve the migration of ASM cells towards the epithelium. However, little is known about the mechanisms of cell migration and the effect of epithelial-derived mediators on this process.
Objective The main objective of the current study is to assess the effects of epithelial-derived chemokines on ASM cell migration.
Methods Normal human ASM cells were incubated with supernatants from cells of the bronchial epithelial cell line BEAS-2B and normal human bronchial epithelial (NHBE) cells. To induce chemokine production, epithelial cells were treated with TNF-α. Chemokine expression by epithelial cells was evaluated by quantitative real-time PCR, ELISA and membrane antibody array. To identify the role of individual chemokines in ASM cell migration, we performed migration assays with a modified Boyden chamber using specific neutralizing antibodies to block chemokine effects.
Results Supernatants from BEAS-2B cells treated with TNF-α increased ASM cell migration; migration was increased 1.6 and 2.5-fold by supernatant from BEAS-2B cells treated with 10 and 100 ng/mL TNF-α, respectively. Protein levels in supernatants and mRNA expression by BEAS-2B cells of regulated on activation, normal T cell expressed and secreted (RANTES) and IL-8 were significantly increased by 100 ng/mL TNF-α treatment. The incubation of supernatant with antibodies to RANTES or IL-8 significantly reduced ASM cell migration, and the combined antibodies further inhibited the cell migration. The migratory effects of supernatants and inhibiting effects of RANTES and/or IL-8 were confirmed also using NHBE cells.
Conclusion The results show that chemokines from airway epithelial cells cause ASM cell migration and might potentially play a role in the process of airway remodelling in asthma.