transforming-growth-factor-beta and Lung-Diseases--Interstitial

Idiopathic pulmonary fibrosis (IPF) and hypersensitivity pneumonitis (HP) belong to heterogenic group of interstitial lung diseases (ILD). For the reason that this group of diseases present with complex clinical non-specific features, they represent a diagnostic and therapeutic challenge. In this review we focus on several crucial signaling pathways participating in inflammation, fibrosis and EMT processes, so important in the course of ILD: TNF-α/NFκβ, TGF-β/SMAD, Wnt-β-catenin and PI3K-Akt signaling. Moreover, this review summarizes the role of selected signaling pathways and some miRNAs which are their regulators during development and progression of IPF and HP. Recent advances indicate the potential role of miRNAs as a molecular markers differentiating clinical course of ILD.

Topics: Biomarkers; Humans; Idiopathic Pulmonary Fibrosis; Lung Diseases, Interstitial; MicroRNAs; Phosphatidylinositol 3-Kinases; Signal Transduction; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Systemic sclerosis (scleroderma, SSc) is a heterogeneous autoimmune connective tissue disease of unknown etiology. Interstitial lung disease (ILD) is a frequent complication, and a significant contributor to morbidity and mortality among SSc patients. SSc-ILD most commonly occurs within 10 years of diagnosis, and may be seen in patients with either the limited or diffuse cutaneous subset of SSc. SSc-ILD is a multifaceted disease process in which different factors and pathways are involved. Aberrant function of a variety of lung cells, cytokines, growth factors, peptides, and bioactive proteins, in combination with genetic and epigenetic regulators, have crucial functions in the pathogenesis of this disease. Here we present our view on recent advances regarding the pathogenesis of SSc-ILD.

Topics: beta Catenin; Epigenomics; Genetic Predisposition to Disease; HLA-D Antigens; Humans; Lung Diseases, Interstitial; Scleroderma, Systemic; Thrombin; Transforming Growth Factor beta

Fibrosis is a state, in which excess amounts of extracellular matrix are deposited in the tissue. Fibrosis can occur in various organs, including the liver, lung, kidney and heart. The progression of fibrosis involves interstitial hypercellularity, accumulation of extracellular matrix, and atrophy of epithelial structures, resulting in a loss of normal function. Myofibroblasts play a crucial role in the development and progress of fibrosis. When stimulated, myofibroblasts actively synthesize connective tissue components and cause organ fibrosis. As a result, the process and the mechanism of myofibroblast activation represent a target for antifibrotic treatment. As yet, however, an effective treatment has not been developed, and new treatment modalities are expected. Because activation of myofibroblasts is a key event during fibrosis development, there is great interest in identifying and characterizing proteins whose expression is changed after this activation. In this review, fibrosis is outlined and the role of myofibroblasts in this disorder is described. Furthermore, the search for candidate proteins to target for treatment and the prospects of antifibrotic therapy are discussed.

Topics: Animals; Extracellular Matrix; Fibrosis; Humans; Kidney Diseases; Lung Diseases, Interstitial; Myofibroblasts; Proteome; Proteomics; Sclerosis; Signal Transduction; Transforming Growth Factor beta

Interstitial lung diseases (ILDs) in children represent a heterogeneous group of respiratory disorders characterized by derangements of the alveolar walls. The key pathologic feature of ILDs is the altered repair of the alveolar surface after injury with a marked disruption in the integrity of the epithelium and, consequently, a dysregulated communication between epithelial and mesenchymal pulmonary components. Concomitant to the loss of cell-cell contact, epithelial cells undergo a process called epithelial to mesenchymal transition and acquire a mesenchymal identity. Among the factors involved in disease progression, transforming growth factor-β has been identified as a master switch in the induction of fibrosis. This article reviews recent advances in the understanding of the mechanisms involved in the pathogenesis of ILDs, and provides information on their adaptation in the context of lung growth.

Topics: Airway Remodeling; Autophagy; Child; Disease Progression; Endoplasmic Reticulum; Humans; Lung; Lung Diseases, Interstitial; Pulmonary Alveoli; Respiratory Mucosa; Stress, Physiological; Transforming Growth Factor beta

PDGF functions as a primary mitogen and chemoattractant for cells of mesenchymal origin. Members of the PDGF family play an important role during embryonic development and contribute to the maintenance of connective tissue in adults. Deregulation of PDGF signalling has been linked to atherosclerosis, pulmonary hypertension and organ fibrosis. Elevated expression of PDGF and its receptors has been found in scleroderma skin and lung tissues. There is evidence for a TGF-beta and IL-1alpha-dependent autocrine PDGF-A/PDGFRalpha signalling loop in scleroderma skin and lung fibroblasts, suggesting that a cross-talk between TGF-beta and PDGF pathways may regulate chronic fibrosis in scleroderma.

Topics: Animals; Female; Fibroblasts; Humans; Lung; Lung Diseases, Interstitial; Platelet-Derived Growth Factor; Pregnancy; Receptors, Platelet-Derived Growth Factor; Receptors, Transforming Growth Factor beta; Scleroderma, Systemic; Signal Transduction; Skin; Transforming Growth Factor beta

Scleroderma-associated interstitial lung disease (SSc-ILD) occurs frequently and for many patients SSc-ILD is a significant complication of their disease. SSc-ILD is now one of the leading causes of death among patients with SSc. SSc-ILD, classified most often as a non-specific interstitial pneumonia, may culminate in interstitial pulmonary fibrosis and end-stage lung disease. Fibrosis in the lung is the net result of fibroblast proliferation and deposition of excessive amounts of extracellular matrix proteins. Among the many cytokines and growth factors involved in the pathogenesis of SSc-ILD, ET-1 may be a central mediator. In vitro and in vivo studies of animals and SSc patients support the notion that ET-1 can enhance the proliferation of pulmonary mesenchymal cells and may also enhance the fibrogenic effects of TGF-beta. Two well-designed randomized controlled trials of the dual ET receptor blocker bosentan were negative in their primary (and for SSc also secondary) endpoints, although there might be explanations for this apparent lack of efficacy.

Topics: Animals; Antihypertensive Agents; Bosentan; Endothelin Receptor Antagonists; Endothelin-1; Humans; Lung Diseases, Interstitial; Randomized Controlled Trials as Topic; Scleroderma, Systemic; Sulfonamides; Transforming Growth Factor beta; Treatment Failure

Systemic sclerosis (SSc) is a complex and heterogeneous chronic illness characterized by substantial patient to patient variability in clinical manifestations, internal organ involvement, and outcome. Genetic factors contribute to disease susceptibility, but environmental influences also play a significant role. The pathogenesis of SSc encompasses vascular, immunological, and fibrotic processes, which contribute to clinical manifestations and morbidity and must be addressed in the treatment plan. Although vascular interventions appear to reduce the frequency and severity of complications, such as scleroderma renal crisis and pulmonary hypertension, current therapies generally target the immune component of SSc in a non-selective fashion and have largely failed as diseases-modifying interventions. Newer insights into the mechanisms underlying autoimmunity, vascular injury and destruction, and particularly tissue fibrosis provide novel potential targets for therapy. Transforming growth factor-ss is a ubiquitous cytokine that appears to contribute to fibroblast activation, collagen overproduction, and pathological tissue fibrosis. Neutralizing antibodies and small molecules that block TGF-beta activation or function are effective in shutting down TGF-beta signaling and selectively inhibit the progression of fibrosis and may be entering clinical trials for the treatment of SSc.

Topics: Blood Vessels; Cardiovascular Agents; Fibrosis; Genetic Predisposition to Disease; Humans; Hypertension, Pulmonary; Immunosuppressive Agents; Inflammation; Kidney Diseases; Lung Diseases, Interstitial; Raynaud Disease; Scleroderma, Systemic; Skin; Transforming Growth Factor beta; Treatment Outcome

The requirement for precise geometric organization of endothelial cells and epithelial cells makes the gas-exchange region of the lung especially vulnerable to the adverse consequences of toxic products released from inflammatory cells. However, as a filter for large volumes of atmospheric gas, the lung is continually exposed to microorganisms and other toxic insults that require robust inflammatory defense. Enhanced production of extracellular matrix proteins is one important mechanism for restricting tissue damage, but excessive matrix production also has serious adverse effects on gas exchange. The amazing ability of the lung to recover from a barrage of environmental insults depends on precisely regulating both inflammation and extracellular matrix production in space and time. Below I review some of the evidence implicating members of the transforming growth factor beta family as critical mediators of this delicate dance and describe examples of how disruption of this balance by alterations in the magnitude of spatially restricted transforming growth factor beta activation can contribute to pathologic consequences of alveolar and airway injury and inflammation.

Topics: Animals; Bronchitis; Humans; Lung Diseases, Interstitial; Pulmonary Fibrosis; Transforming Growth Factor beta

Topics: Animals; Apoptosis; Humans; Inflammation; Lung Diseases, Interstitial; Models, Biological; Phagocytosis; Pulmonary Fibrosis; Transforming Growth Factor beta

Molecular investigation into the physiopathology of interstitial lung diseases has gained special interest through the trials carried out in the last decade. These trials seem to point at the role played by certain molecules, such as cytokines (transforming growth factor, platelet derived growth factor) and integrins, in the processes that lead to pulmonary fibrosis during the course of interstitial lung disease. They also demonstrate the important role that angiotensin II plays in increasing the secretion of transforming growth factor by several cells. The above-mentioned studies allow new therapeutic approaches to be considered which will possibly improve the serious prognosis of such diseases once they have reached the last stage of their course: pulmonary fibrosis.

Topics: Angiotensin II; Humans; Integrins; Lung Diseases, Interstitial; Pulmonary Fibrosis; Transforming Growth Factor beta

Topics: Animals; Disease Models, Animal; Environmental Exposure; Gene Expression Regulation; Humans; Lung Diseases, Interstitial; Mice; Mice, Knockout; Mice, Transgenic; Models, Biological; Platelet-Derived Growth Factor; Proteins; Transforming Growth Factor alpha; Transforming Growth Factor beta; Uteroglobin

Transforming growth factor-beta (TGF-beta), interleukin-8 (IL-8), and leukotrienes are potent neutrophil chemoattractants that are released in several lung diseases. There is limited information about the release of TGF-beta in bronchoalveolar lavage fluid (BALF) of patients with pneumonia. Furthermore, it is not clear if TGF-beta is differentially expressed in different lung diseases. The aim of our study was to compare the concentrations of TGF-beta1 and TGF-beta2 in the BALF of patients with pneumonia and other lung diseases. Furthermore, correlation of the TGF-beta levels with the concentration of chemoattractant mediators as well as with indicators of macrophage and granulocyte activation should be investigated. Patients with pneumonia, interstitial lung disease (ILD), or chronic obstructive pulmonary diseases (COPD) were included. Patients with ischemic heart disease without pulmonary involvement served as controls. The concentrations of TGF-beta1 and TGF-beta2, of the chemoattractant cytokine IL-8, of leukotriene B4, and of the leukotrienes C4, D4, and E4 were measured. Neutrophil elastase and granulocyte content (PMN) were used as markers for granulocyte activation, and neopterin was used as a marker for the activation of macrophages. Significantly elevated levels of TGF-beta1 (mean = 0.216 ng/ml, p < 0.01) were found in patients with microbiologically positive pneumonia but not in patients with ILD or COPD. A significant (p < 0.001) correlation was found between the TGF-beta1 concentrations and the IL-8 levels and the percentage of granulocytes (r = 0.76, and r = 0.44, respectively). Elevated TGF-beta2 concentrations were measured in the BALF of patients with pneumonia (mean = 1.4 ng/ml, p < 0.01) and with ILD. Pneumonia was also associated with increased concentrations of leukotrienes C4, D4, and E4 (mean = 91.61 pg/ml, p < 0.05) and leukotriene B4 (mean = 203.9 pg/ml, p < 0.01), significantly elevated levels of PMN elastase (mean = 2958.26 ng/ml, p < 0.01), and neopterin (mean = 0.42 nmol/L). Our results strongly suggest that different lung diseases do differ with regard to the released cytokines. TGF-beta1 probably plays a key role in regulation of pulmonary inflammation, particularly in pneumonia.

Topics: Bronchoalveolar Lavage Fluid; Granulocytes; Humans; Interleukin-8; Leukocyte Elastase; Leukotrienes; Lung Diseases, Interstitial; Lung Diseases, Obstructive; Macrophages; Monocytes; Neopterin; Pneumonia; Transforming Growth Factor beta

Pulmonary fibrosis involves various types of immune cells and soluble mediators, including TGF-β and IL-35, a recently identified heterodimeric cytokine that belongs to the IL-12 cytokine family. However, the effect of regulatory IL-35 may play an important role in fibrotic diseases. The aim of this paper is to explore the immunoregulatory role of IL-35 in the development of fibrosis in interstitial lung disease (ILD). To gain a better understanding of this issue, the concentrations of IL-35 and different profibrotic cytokines in fibrotic (F-ILD) and non-fibrotic (NF-ILD) patients by ELISA were compared to that of intracellular IL-35 and IL-17 on CD4+ T cells stimulated in the presence of BAL or with different ratios of recombinant IL-35 (rIL-35) and TGF-β (rTGF-β), which were evaluated by flow cytometry. We observed that BAL concentration of IL-35 was lower in F patients (p < 0.001) and was negatively correlated with concentrations of TGF-β (p < 0.001) and IL-17 (p < 0.001). In supplemented cell cultures, BAL from NF but not F patients enhanced the percentage of IL-35 + CD4+ T (p < 0.001) cells and decreased the percentage of IL-17 + CD4+ T cells (p < 0.001). The percentage of IL-35 + CD4+ T cells correlated positively with BAL concentration of IL-35 (p = 0.02), but correlated negatively with BAL concentrations of IL-17 (p = 0.007) and TGF-β (p = 0.01). After adjusting the concentrations of recombinant cytokines to establish a TGF-β: IL-35 ratio of 1:4, an enhanced percentage of IL-35 + CD4+ T cells (p < 0.001) but a decreased percentage of IL-17 + CD4+ T cells (p < 0.001) was observed. After adding recombinant IL-35 to the BAL from F patients until a 1:4 ratio of TGF-β: IL-35 was reached, a significantly increased percentage of IL-35 + CD4+ T cells (p < 0.001) and a decreased percentage of IL-17 + CD4+ T cells (p = 0.003) was found. These results suggest that IL-35 may induce an anti-fibrotic response, regulating the effect of TGF-β and the inflammatory response on CD4+ T cells. In addition, the TGF-β: IL-35 ratio in BAL has been shown to be a potential biomarker to predict the outcome of F patients with ILD.

Topics: Bronchoalveolar Lavage Fluid; Cytokines; Humans; Interleukin-17; Lung Diseases, Interstitial; Pulmonary Fibrosis; Transforming Growth Factor beta

A subset of severe COVID19 patients develop pulmonary fibrosis, but the pathophysiology of this complication is still unclear. We previously described the possibility to isolate lung mesenchymal cells (LMC) by culturing broncho-alveolar lavage (BAL) cells from patients with pulmonary fibrosis or chronic lung allograft dysfunction. Aim of this study was to investigate the possibility to isolate and characterize LMC from BAL of patients that, two months after discharge for severe COVID19, show CT signs of post-COVID19 fibrosis (Post-COVID) and in some cases has been considered transplant indication. Results were compared with those from BAL of patients with collagen tissue disease-associated interstitial fibrosis (CTD-ILD). BAL fluid levels of TGFβ, VEGF, TIMP2, RANTES, IL6, IL8, and PAI1 were assessed. LMC were cultured and expanded, phenotyped by flow cytometry, and tested for osteogenic and adipogenic differentiation. Finally, we tested immunomodulatory and proliferative capabilities, collagen I production + /- TGF-beta stimulation. BAL cytokine and growth factor levels were comparable in the two groups. Efficiency of isolation from BAL was 100% in post-COVID compared to 63% in CTD-ILD. LMC from post-COVID were positive for CD105, CD73, CD90, and negative for CD45, CD34, CD19 and HLA-DR as in CTD-ILD samples. Post-COVID LMC displayed higher collagen production with respect to CTD-ILD LMC. Immunomodulatory capacity towards lymphocytes was very low, while Post-COVID LMC significantly upregulated pro-inflammatory cytokine production by healthy PBMCs. Our preliminary data suggest that LMC from post-COVID19 fibrosis patients share several features with CTD-ILD ones but might have a higher response to fibrogenic signals and pro-inflammatory profile.

Topics: COVID-19; Cytokines; Fibrosis; Humans; Lung; Lung Diseases, Interstitial; Pulmonary Fibrosis; Transforming Growth Factor beta

Previous studies have suggested a correlation between uric acid (UA) and lung lesion in some diseases. However, it remains unknown whether UA contributes to the lung injury in rheumatoid arthritis (RA). Our study aimed to investigate the clinical value of the UA level in the severity of rheumatoid arthritis-associated interstitial lung disease (RA-ILD). We measured UA in serum and bronchoalveolar lavage fluid (BALF), and UA levels of subjects were compared. As for the role of UA on ILD, we incubated A549 cells with UA and the expression of EMT markers was measured by immunofluorescence staining. The concentrations and messenger RNA expression of IL-1, IL-6, and transforming growth factor-β (TGF-β) were measured by ELISA and RT-PCR, respectively. We observed that serum UA levels in RA were significantly higher than those in controls. And, higher UA was measured in both serum and BALF of patients with RA-ILD, particularly those with interstitial pneumonia (UIP) pattern. Additionally, the correlation of the serum and BALF UA levels with serum KL-6, a biomarker of ILDs, in RA was significant (r = 0.44, p < 0.01; r = 0.43, p < 0.01). And, the negative correlations of UA, in both serum and BALF, with forced vital capacity (r =  -0.61, p < 0.01; r =  -0.34, p < 0.01) and diffusing capacity for carbon monoxide (r =  -0.43, p < 0.01; r =  -0.30, p < 0.01) were measured in patients. In the ROC curve analysis, the AUC value of UA for RA-ILD was 0.744 (95% CI: 0.69-0.80; p < 0.01), and the AUC of serum UA for predicting UIP pattern of patients with RA-ILD was 0.845 (95% CI: 0.78-0.91; p < 0.01), which showed the significance of the UA in clinical settings. Also, the in vitro experiment showed that UA induced epithelial-to-mesenchymal transition (EMT) and production of IL-1, IL-6, and TGF-β in A549 cells. Therefore, the elevated UA levels may be a diagnostic marker in RA-ILD, particularly RA-UIP.

Topics: Arthritis, Rheumatoid; Biomarkers; Humans; Interleukin-1; Interleukin-6; Lung Diseases, Interstitial; Transforming Growth Factor beta; Uric Acid

The interstitial lung diseases ILDs are a heterogeneous group of diseases that often lead to progressive fibrosis of the lungs with corresponding functional impairment. With nintedanib, a tyrosinkinase inhibitor and angiokinase inhibitor, as well as pirfenidone, which unfolds its effect among other things by inhibiting the transforming growth factor β, there are currently 2 approved antifibrotic drugs. In the rapidly progressing idiopathic pulmonary fibrosis IPF, the antifibrotic drugs nintedanib and pirfenidone have been established and approved in therapy for several years. The initiation of antifibrotic therapy should be carried out early after diagnosis by multidisciplinary discussion (MDD). In systemic scleroderma with lung involvement nintedanib should be used in the case of relevant fibrosis in addition to immunosuppressive therapy. Recently, nintedanib has also become a new option for the treatment of progressive fibrosing ILDs (PF-ILDs). This describes the course of various disease entities such as connective tissue disease associated ILDs (CTD-ILDs), fibrosing hypersensitivity pneumonitis or fibrosing courses of non-IPF idiopathic interstitial pneumonitis (non-IPF IIPs) that have a corresponding fibrose-related worsening of respiratory symptoms, a deterioration of lung-functioning parameters or a disease progression in CT. Although pirfenidone also shows positive signals for this group of patients in some selected studies, its use in PF-ILD is not yet recommended. In particular, gastrointestinal side effects can occur under therapy with antifibrotic drugs and require a long-term close interdisciplinary connection of patients.

Topics: Disease Progression; Fibrosis; Humans; Idiopathic Pulmonary Fibrosis; Lung; Lung Diseases, Interstitial; Transforming Growth Factor beta

Dysregulation of the prostaglandin E2 (PGE2) signaling pathway has been implicated in interstitial pneumonia (IP) pathogenesis. Due to the unstable nature of PGE2, available detection methods may not precisely reflect PGE2 levels. We explored the clinical usefulness of measuring stable prostaglandin E-major urinary metabolite (PGE-MUM) with respect to pathogenesis and extent of chronic fibrosing IP (CFIP), including idiopathic pulmonary fibrosis (IPF), as PGE-MUM is reflective of systemic PGE2 production.. PGE-MUM was measured by radioimmunoassay in controls (n = 124) and patients with lung diseases (bronchial asthma (BA): n = 78, chronic obstructive pulmonary disease (COPD): n = 33, CFIP: n = 44). Extent of lung fibrosis was assessed by fibrosing score (FS) of computed tomography (CT) (FS1-4). Immunohistochemical evaluation of COX-2 was performed to find PGE2 producing cells in IPF. Human bronchial epithelial cells (HBEC) and lung fibroblasts (LFB) were used in in vitro experiments.. Compared to control, PGE-MUM levels were significantly elevated in CFIP. PGE-MUM levels were positively correlated with FS, and inversely correlated with %DLCO in IP (FS 1-3). COX-2 was highly expressed in metaplastic epithelial cells in IPF, but lower expression of EP2 receptor was demonstrated in LFB derived from IPF. TGF-β induced COX-2 expression in HBEC.. PGE-MUM, elevated in CFIP, is a promising biomarker reflecting disease activity. Metaplastic epithelial cells can be a source of elevated PGE-MUM in IPF.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Cyclooxygenase 2; Epithelial Cells; Female; Fibroblasts; Humans; Idiopathic Pulmonary Fibrosis; Japan; Lung; Lung Diseases, Interstitial; Male; Middle Aged; Prostaglandins; Prostanoic Acids; Transforming Growth Factor beta; Urine

 Previous studies have suggested that hepatocyte growth factor (HGF) inhibits lung fibrosis as an antagonist of transforming growth factor β (TGF‑β)..  We assessed HGF expression levels in the lower airways of patients with selected interstitial lung diseases..  HGF levels were examined by an enzyme‑linked immunosorbent assay in bronchoalveolar lavage (BAL) fluid supernatants from patients with pulmonary sarcoidosis (PS, n = 52), idiopathic pulmonary fibrosis (IPF, n = 23), nonspecific interstitial pneumonia (NSIP, n = 14), extrinsic allergic alveolitis (EAA, n = 6), bronchiolitis obliterans organizing pneumonia (BOOP, n = 8), chronic eosinophilic pneumonia (EP, n = 6), and in control subjects (n = 13). Intracellular HGF expression in BAL cells was evaluated by flow cytometry..  HGF concentrations were elevated in BAL fluid from nonsmokers with IPF (261 ±204 pg/ml, P <0.02), smokers with IPF (220 ±13 pg/ml, P <0.001), and smokers with PS (172 ±33 pg/ml, P <0.02), as compared with controls (148 ±17 pg/ml for nonsmokers; 137 ±9 pg/ml for smokers). HGF levels were positively correlated with TGF‑β concentrations in BAL fluid (r = 0.3; P = 0.02) and negatively-with vital capacity (r = -0.2; P = 0.02). BAL neutrophils, and, for the first time, BAL lymphocytes, were identified as intracellular HGF‑positive cells..  Our results do not support evidence for strong antifibrotic HGF activity. The highest HGF concentrations were observed in BAL fluid from patients with IPF, and they were also positively correlated with TGF‑β levels. Thus, although the local protective mechanisms such as the HGF expression are upregulated in chronic interstitial lung diseases, they are not enough to prevent lung fibrosis.

Topics: Adult; Aged; Bronchoalveolar Lavage Fluid; Female; Hepatocyte Growth Factor; Humans; Lung Diseases, Interstitial; Male; Middle Aged; Transforming Growth Factor beta; Young Adult

Interstitial lung disease (ILD) is a serious side-effect of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI) treatment. Therefore, it is necessary to study underlying mechanisms for the development of pulmonary fibrosis induced by EGFR-TKI and potential approaches to attenuate it. Metformin is a well-established and widely prescribed oral hypoglycemic drug, and has gained attention for its potential anticancer effects. Recent reports have also demonstrated its role in inhibiting epithelial-mesenchymal transition and fibrosis. However, it is unknown whether metformin attenuates EGFR-TKI-induced pulmonary fibrosis. The effect of metformin on EGFR-TKI-induced exacerbation of pulmonary fibrosis was examined in vitro and in vivo using MTT, Ki67 incorporation assay, flow cytometry, immunostaining, Western blot analysis, and a bleomycin-induced pulmonary fibrosis rat model. We found that in lung HFL-1 fibroblast cells, TGF-β or conditioned medium from TKI-treated lung cancer PC-9 cells or conditioned medium from TKI-resistant PC-9GR cells, induced significant fibrosis, as shown by increased expression of Collegen1a1 and α-actin, while metformin inhibited expression of fibrosis markers. Moreover, metformin decreased activation of TGF-β signaling as shown by decreased expression of pSMAD2 and pSMAD3. In vivo, oral administration of gefitinib exacerbated bleomycin-induced pulmonary fibrosis in rats, as demonstrated by HE staining and Masson staining. Significantly, oral co-administration of metformin suppressed exacerbation of bleomycin-induced pulmonary fibrosis by gefitinib. We have shown that metformin attenuates gefitinib-induced exacerbation of TGF-β or bleomycin-induced pulmonary fibrosis. These observations indicate metformin may be combined with EGFR-TKI to treat NSCLC patients.

Topics: Animals; Antineoplastic Agents; Bleomycin; Blotting, Western; Epithelial-Mesenchymal Transition; Flow Cytometry; Gefitinib; Humans; Immunohistochemistry; Lung Diseases, Interstitial; Male; Metformin; Pulmonary Fibrosis; Quinazolines; Rats; Rats, Sprague-Dawley; Signal Transduction; Transforming Growth Factor beta

Interstitial lung disease (ILD) is the leading cause of death in patients with systemic sclerosis (SSc; scleroderma). Although SSc-related ILD is more common and severe in African Americans than in Caucasians, little is known about factors underlying this significant health disparity. The aim of this study was to examine the role that low expression of caveolin-1 might play in susceptibility to ILD among African Americans.. Assays of monocyte migration toward stromal cell-derived factor 1 (SDF-1) were performed using monocytes from Caucasian and African American healthy donors and patients with SSc. For fibrocyte differentiation studies, total peripheral blood mononuclear cells were incubated on fibronectin-coated plates. Protein expression was evaluated by immunohistochemistry and Western blotting.. Monocytes from healthy African American donors and those from patients with SSc had low caveolin-1 levels, enhanced migration toward the CXCR4 ligand SDF-1, and enhanced differentiation to fibrocytes. Enhanced migration and differentiation of monocytes from African Americans and patients with SSc appeared to be attributable to the lack of caveolin-1, because restoring caveolin-1 function using a caveolin-1 scaffolding domain peptide inhibited these processes. Although they differed from monocytes from Caucasians, monocytes from both African Americans and patients with SSc were not identical, because SSc monocytes showed major increases from baseline in ERK, JNK, p38, and Smad2/3 activation, while monocytes from African Americans showed only limited ERK activation and no activation of JNK, p38, or Smad2/3. In contrast, SDF-1 exposure caused no additional ERK activation in SSc monocytes but did cause significant additional activation in monocytes from African Americans.. African Americans may be predisposed to SSc-related ILD due to low baseline caveolin-1 levels in their monocytes, potentially affecting signaling, migration, and fibrocyte differentiation. The monocytes of African Americans may lack caveolin-1 due to high levels of transforming growth factor β in their blood.

Topics: Black or African American; Caveolin 1; Cell Differentiation; Cell Movement; Cytoskeleton; Fibroblasts; Humans; In Vitro Techniques; Lung Diseases, Interstitial; MAP Kinase Signaling System; Monocytes; Receptors, CXCR4; Risk Factors; Scleroderma, Systemic; Transforming Growth Factor beta; White People

To determine serum and sputum Caveolin-1 (Cav-1) levels and their associations with transforming growth factor- ß (TGF-ß) and interstitial lung disease (ILD) in systemic sclerosis (SSc).. Serum and induced sputum samples from 55 patients with SSc, 25 asthma patients and 16 healthy volunteers (HC) were tested for Cav-1 and TGF-ß by the ELISA technique. As a possible downstream signaling regulator of TGF-ß, Endothelin-1 (ET-1), a potent profibrotic protein, was also measured in all serum and sputum samples and relations with Cav-1 and TGF-ß were sought. All scleroderma patients were evaluated for their clinical and laboratory parameters. Pulmonary function tests (PFT) and high resolution computerized tomography (HRCT) were performed for the diagnosis of ILD. The alveolitis-fibrosis index and the SSc disease severity scores were noted for each patient.. Serum Cav-1 levels were lower in SSc compared to HC (p<0.01). Cav-1 levels were significantly lower in the sputum of SSc patients compared to both control groups (p<0.001). It was also found significantly lower in SSc-ILD compared to those without ILD (0.19±0.04 vs 0.25±0.07, respectively, p<0.01). Although no difference was found in the serum TGF-ß levels among the groups, sputum TGF-ß levels correlated positively with the alveolitis index (r=0.34) and correlated inversely with FVC measurements (r=-0.44, p<0.05) among SSc patients. Serum ET-1 was significantly higher in SSc patients (p<0.01) but no association was found between ET-1 and Cav-1 or TGF-ß.. These results suggest that decreased sputum Cav-1 levels is associated with SSc related-ILD and may be used as a marker for the detection of SSc-ILD.

Topics: Adult; Aged; Case-Control Studies; Caveolin 1; Endothelin-1; Female; Humans; Lung Diseases, Interstitial; Male; Middle Aged; Prospective Studies; Scleroderma, Systemic; Sputum; Transforming Growth Factor beta

Idiopathic interstitial lung diseases (iILDs) are characterized by inflammation, hyperplasia of Type-II alveolar epithelial cells (AECs) and lung remodelling often with progressive fibrosis. It remains unclear which signals initiate iILD and/or maintain the disease processes. Using real-time RT-PCR and immunohistochemistry on archival biopsies of three patterns of iILD (usual interstitial pneumonitis/UIP, non-specific interstitial pneumonitis/NSIP and cryptogenic organizing pneumonia/COP) we investigated whether hedgehog signalling (previously associated with lung damage and repair) was functional and whether the damage associated extracellular matrix protein tenascin-C was present in activated Type-II AECs in all three iILDs. Using tissue culture, protein and mRNA detection we also determined how two signals (oxidative damage and TGF-β) associated with iILD pathogenesis affected Sonic hedgehog (SHH) and tenascin-C production by a Type-II AEC cell line. We report that SHH pathway and tenascin-C mRNA and proteins were found in UIP, NSIP and COP. SHH signalling was most active at sites of immature organizing fibrous tissue (fibroblastic foci) in UIP. In vitro Type-II AECs constitutively secrete SHH but not tenascin-C. Oxidative injury stimulated SHH release whereas TGF-β inhibited it. TGF-β and oxidative damage both upregulated tenascin-C mRNA but only TGF-β induced synthesis and release of a distinct protein isoform. SHH signalling is active in Type-II AECs from three types of ILD and all three express tenascin-C.

Topics: Adult; Aged; Blotting, Western; Epithelial Cells; Female; Hedgehog Proteins; Humans; Immunohistochemistry; Lung Diseases, Interstitial; Male; Middle Aged; Oxidative Stress; Respiratory Mucosa; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Tenascin; Transforming Growth Factor beta

Interstitial lung disease (ILD) is an intractable disease induced by various factors in humans. However, there is no universally effective treatment for ILD. In this study, we investigated the role of transforming growth factor (TGF)-beta signalling in the pathogenesis of ILD by using model mice. Injection of interleukin (IL)-18 plus IL-2 in C57BL6 (B6) mice resulted in acute ILD by infiltration of natural killer (NK) cells and a significant increase of TGF-beta mRNA in the lung. To examine the pathogenetic role of TGF-beta in ILD mice, we used SB-431542 (4-[4-(1,3-benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]-benzamide), which is a potent and selective inhibitor of TGF-beta receptor I (TbetaRI), also known as activin receptor-like kinase 5 (ALK5). Treatment of B6-ILD mice with SB-431542 resulted in improvement of ILD, delay in mortality, reduction of the expression of interferon (IFN)-gamma and IL-6 in the lungs. The same treatment also decreased significantly the percentage of natural killer (NK) cells in the lungs (P < 0.05) and mRNA expression levels of certain chemokines such as CCL2, CCL3, CCL4, CCL5 and CXCL10 in B6-ILD. These findings were confirmed by IL-18 plus IL-2 treatment of Smad3-deficient (Smad3(-/-)) mice (P < 0.05). Our results showed that inhibition of TGF-beta signalling reduced the percentage of NK cells and the expression of certain chemokines in the lungs, resulting in improvement of ILD. The findings suggest that TGF-beta signalling may play an important role in the pathogenesis of IL-18 plus IL-2-induced ILD in mice.

Topics: Acute Disease; Animals; Antineoplastic Agents; Benzamides; Chemokines; Dioxoles; Humans; Interferon-gamma; Interleukin-18; Interleukin-2; Interleukin-6; Killer Cells, Natural; Lung Diseases, Interstitial; Mice; Mice, Knockout; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; RNA, Messenger; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta

Angiotensin-(1-7) [Ang-(1-7)] is an endogenous 7-amino acid peptide hormone of the renin-angiotensin system that has antiproliferative properties. In this study, Ang-(1-7) inhibited the growth of cancer-associated fibroblasts (CAF) and reduced fibrosis in the tumor microenvironment. A marked decrease in tumor volume and weight was observed in orthotopic human breast tumors positive for the estrogen receptor (BT-474 or ZR-75-1) and HER2 (BT-474) following Ang-(1-7) administration to athymic mice. Ang-(1-7) concomitantly reduced interstitial fibrosis in association with a significant decrease in collagen I deposition, along with a similar reduction in perivascular fibrosis. In CAFs isolated from orthotopic breast tumors, the heptapeptide markedly attenuated in vitro growth as well as reduced fibronectin, transforming growth factor-β (TGF-β), and extracellular signal-regulated kinase 1/2 kinase activity. An associated increase in the mitogen-activated protein kinase (MAPK) phosphatase DUSP1 following treatment with Ang-(1-7) suggested a potential mechanism by which the heptapeptide reduced MAPK signaling. Consistent with these in vitro observations, immunohistochemical analysis of Ang-(1-7)-treated orthotopic breast tumors revealed reduced TGF-β and increased DUSP1. Together, our findings indicate that Ang-(1-7) targets the tumor microenvironment to inhibit CAF growth and tumor fibrosis.

Topics: Angiotensin I; Animals; Antihypertensive Agents; Blotting, Western; Breast Neoplasms; Carcinoma, Ductal, Breast; Dual Specificity Phosphatase 1; Female; Fibronectins; Fibrosis; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Lung Diseases, Interstitial; Mice; Mice, Nude; Mitogen-Activated Protein Kinase 3; Peptide Fragments; Phosphorylation; Transforming Growth Factor beta; Tumor Cells, Cultured

To observe the clinical feature of rheumatoid arthritis associated interstitial lung disease (RA-ILD) patients and changes of serum cytokines tumor growth factor (TGF)-beta 1, tumor necrosis factor (TNF)-alpha, insulin-like growth factor (IGF)-1, and platelet derived growth factor (PDGF)-AB.. The clinical manifestations, lung high resolution CT (HRCT), lung functions, blood gas and other relative laboratory findings of 30 RA-ILD patients and 35 RA patients were observed. ELISA was used to detect the levels of TGF-beta 1, TNF-alpha, IGF-1, and PDGF-AB. Thirty healthy volunteers were observed too as controls.. The clinical manifestations of RA-ILD patients were more serious than those of the RA patients. The ESR was faster, the serum C-reactive protein, rheumatoid factor (RF), and globulin levels higher, and pulmonary arterial pressure higher too in the RA-ILD patients than in the RA patients (all P<0.01). The main respiratory manifestations of the RA-ILD patients were cough, expectoration, chest distress, short breath, chest pain, change of breath sounds, Velcro râles, and dyspnea. The main lung HRCT findings included thickening of interlobular septum and bronchial wall, pachynsis pleurae, mosaic sign, bronchiectasis, emphysema, patching shadow, honeycombing, fibrous scar, etc. Pulmonary function test showed that the levels of vital capacity, forced vital capacity, maximum midexpiratory flow, and diffusing capacity of the lung for carbon monoxide of the RA-ILD patients were all significantly lower than those of the RA patients (all P<0.01). Arterial gas test showed that the PO2 of the RA-ILD patients was significantly lower than that of the RA patients (P<0.01). The TGF-beta 1; TNF-alpha, IGF-1, and PDGF-AB of both the RA-ILD and RA patients were all significantly higher than those of the healthy volunteers (all P<0.01), and the levels of these cytokines of the RA-ILD patients were all higher than those of the RA patients (all P<0.01).. The symptoms and signs of the RA-ILD patients are more serious, the lung HRCT changes more obvious, lung function decreases, and the levels of TGF-beta 1, TNF-alpha, IGF-1, and PDGF-AB increase.

Topics: Adult; Arthritis, Rheumatoid; Female; Humans; Insulin-Like Growth Factor I; Lung Diseases, Interstitial; Male; Middle Aged; Platelet-Derived Growth Factor; Rheumatoid Factor; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

There has been ongoing controversy related to what differentiates normal lung repair and fibrosis. For example, the current prevailing concept has been that idiopathic forms of pulmonary fibrosis are due only to epithelial injury in response to some unknown cause that results in persistent evolving fibrosis without preceding inflammation. This concept would suggest that the lung responds to injury in a different manner than other organs, such as the liver, kidney, and heart. However, that would seem to contradict known established pathological concepts. To address this controversy, concepts were presented as follows: (1) loss of basement membrane integrity is critical in determining the "point of no return," and contributes to the inability to reestablish normal lung architecture with promotion of fibrosis; (2) loss of epithelial cells, endothelial cells, and basement membrane integrity in usual interstitial pneumonia associated with idiopathic pulmonary fibrosis leads to destroyed lung architecture and perpetual fibrosis; (3) transforming growth factor-beta is necessary, but not entirely sufficient, to promote permanent fibrosis; (4) persistent injury/antigen/irritant is critical for the propagation of fibrosis; (5) idiopathic pulmonary fibrosis is an example of a process related to the persistence of an "antigen(s)," chronic inflammation, and fibrosis; and (6) unique cells are critical cellular players in the regulation of fibrosis. In keeping with the theme of the Aspen Lung Conference, it is hoped that more questions are raised than answered in this presentation, in support of the continued need for research in this area to address these important concepts.

Topics: Basement Membrane; Endothelial Cells; Humans; Inflammation; Lung; Lung Diseases, Interstitial; Macrophages, Alveolar; Pulmonary Fibrosis; Respiratory Distress Syndrome; Respiratory Mucosa; Transforming Growth Factor beta; Wound Healing

Myofibroblasts are primary effector cells in idiopathic pulmonary fibrosis (IPF). Defining mechanisms of myofibroblast differentiation may be critical to the development of novel therapeutic agents.. To show that myofibroblast differentiation is regulated by phosphatase and tensin homolog deleted on chromosome 10 (PTEN) activity in vivo, and to identify a potential mechanism by which this occurs.. We used tissue sections of surgical lung biopsies from patients with IPF to localize expression of PTEN and alpha-smooth muscle actin (alpha-SMA). We used cell culture of pten(-/-) and wild-type fibroblasts, as well as adenoviral strategies and pharmacologic inhibitors, to determine the mechanism by which PTEN inhibits alpha-SMA, fibroblast proliferation, and collagen production.. In human lung specimens of IPF, myofibroblasts within fibroblastic foci demonstrated diminished PTEN expression. Furthermore, inhibition of PTEN in mice worsened bleomycin-induced fibrosis. In pten(-/-) fibroblasts, and in normal fibroblasts in which PTEN was inhibited, alpha-SMA, proliferation, and collagen production was upregulated. Addition of transforming growth factor-beta to wild-type cells, but not pten(-/-) cells, resulted in increased alpha-SMA expression in a time-dependent fashion. In pten(-/-) cells, reconstitution of PTEN decreased alpha-SMA expression, proliferation, and collagen production, whereas overexpression of PTEN in wild-type cells inhibited transforming growth factor-beta-induced myofibroblast differentiation. It was observed that both the protein and lipid phosphatase actions of PTEN were capable of modulating the myofibroblast phenotype.. The results indicate that in IPF, myofibroblasts have diminished PTEN expression. Inhibition of PTEN in vivo promotes fibrosis, and PTEN inhibits myofibroblast differentiation in vitro.

Topics: Actins; Animals; Antibiotics, Antineoplastic; Bleomycin; Cell Culture Techniques; Cell Differentiation; Fibroblasts; Fibrosis; Humans; Lung Diseases, Interstitial; Mice; Mice, Inbred C57BL; PTEN Phosphohydrolase; Pulmonary Fibrosis; Transforming Growth Factor beta

Polymyositis and dermatomyositis (PM/DM) are often complicated by interstitial pneumonitis (IP), which is an important cause of death. It has been reported that blood concentration of transforming growth factor-beta (TGF-beta), which is produced by a wide range of cells including endothelial cells and enhances the fibrotic changes in various tissues, is increased in PM/DM with IP. Endothelial damage is likely to exist in PM/DM. We studied the relationship between endothelial damage and IP in PM/DM.. Blood levels of sialylated carbohydrate antigen KL-6, TGF-beta, endothelin-1 (ET-1), thrombomodulin (TM), and plasminogen activator inhibitor-1 (PAI-1) were determined in 43 patients with PM or DM with or without IP, and the relationship between these measures was analyzed.. Blood levels of KL-6 and TGF-beta were higher in the patients with IP than those without, and these measures were well correlated with each other. Levels of ET-1, TM, and PAI-1, all known to reflect the extent of endothelial damage, were also increased in patients with IP, and these measures correlated well with TGF-beta.. Our data suggest that endothelial damage might play an important role through the production of fibrosis-enhancing factors such as TGF-beta or ET-1 in PM/DM.

Topics: Adult; Aged; Antigens, Neoplasm; Blood Cell Count; Dermatomyositis; Endothelin-1; Endothelium, Vascular; Humans; Lung Diseases, Interstitial; Middle Aged; Mucin-1; Mucins; Plasminogen Activator Inhibitor 1; Polymyositis; Thrombomodulin; Transforming Growth Factor beta

Pigment epithelium-derived factor (PEDF) is a 50-kD protein with angiostatic and neurotrophic activities that regulates vascular development within the eye. PEDF expression was increased in the lungs of patients with idiopathic pulmonary fibrosis (IPF) based on microarray analyses. Angiogenesis has been implicated in the pathogenesis of fibrotic lung diseases, we therefore hypothesized that regional abnormalities in vascularization occur in IPF as a result of an imbalance between PEDF and vascular endothelial growth factor. We demonstrated that vascular density is regionally decreased in IPF within the fibroblastic foci, and that within these areas PEDF was increased, whereas vascular endothelial growth factor was decreased. PEDF colocalized with the fibrogenic cytokine, transforming growth factor (TGF)-beta 1, particularly within the fibrotic interstitium and the fibroblastic focus, and prominently within the epithelium directly overlying the fibroblastic focus. This suggested that TGF-beta 1 might regulate PEDF expression. Using 3T3-L1 fibroblasts and human lung fibroblasts, we showed that PEDF was indeed a TGF-beta 1 target gene. Collectively, our findings implicate PEDF as a regulator of pulmonary angiogenesis and an important mediator in IPF.

Topics: Biopsy; Bronchoalveolar Lavage Fluid; Eye Proteins; Fibroblasts; Humans; Lung; Lung Diseases, Interstitial; Neovascularization, Pathologic; Nerve Growth Factors; Proteins; Pulmonary Fibrosis; Serpins; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

Montelukast, a potent cysteinyl receptor antagonist, may be an antifibrotic therapeutic agent for lung fibrosis. Seven sarcoidosis patients and 10 with unusual interstitial pneumonia underwent conventional bronchoalveolar lavage, from which myofibroblasts were recovered. Myofibroblast proliferation was assayed, alpha smooth muscle actin levels were measured, TGFbeta mRNA RT-PCR transcripts were semiquantitated, and secretion was evaluated in myofibroblast supernatants. Montelukast at 10(-8) M concentration had a suppressive effect on cell proliferation (31 +/- 18%), which was significantly enhanced by LTD4 10(-8) M. No differences were found between sarcoidosis (31.28 +/- 15.9%) and unusual interstitial pneumonia (30.56 +/- 24.3%) lines. Fetal calf serum (20%) produced an enhancing effect (29.8 +/- 21.6%) in all lines. Myofibroblasts recovered from sarcoidosis patients showed lower alpha-smooth muscle actin contents than unusual interstitial pneumonia lines (0.09 +/- 0.02 vs. 0.34 +/- 0.16, p =0.039, respectively). Montelukast suppressed alpha-actin in short-term cultures in sarcoidosis myofibroblasts and in long-term unusual interstitial pneumonia myofibroblasts. Montelukast at 10(-6) M concentratin decreased the TGFbeta-induced alpha-actin expression in all lines tested. Montelukast decreased mRNA expression of TGFbeta. Montelukast may be a therapeutic agent in pathological conditions involving fibrotic and remodeling processes.

Topics: Acetates; Actins; Adult; Cell Proliferation; Cells, Cultured; Cyclopropanes; Female; Fibroblasts; Fibrosis; Humans; Lung Diseases, Interstitial; Male; Middle Aged; Quinolines; RNA, Messenger; Sarcoidosis; Sulfides; Transforming Growth Factor beta

The pathogenesis of interstitial lung diseases (ILDs) is known to be associated with reactive oxygen and nitrogen metabolites and increased oxidant stress. One of the major antioxidants in human lung is glutathione (GSH) and enzymes linked to its synthesis. The rate-limiting enzyme of GSH synthesis is gamma-glutamylcysteine synthetase (gamma-GCS) containing catalytically active heavy (gamma-GCSh) and regulatory light (gamma-GCSl) subunits. It can be hypothesized that gamma-GCS is the major determinant in explaining reduced GSH levels in fibrotic lung disorders. We investigated the regulation of gamma-GCS by transforming growth factor beta(1) (TGF-beta(1)) and tumor necrosis factor alpha (TNF-alpha) in human lung cells and its expression and distribution in fibrotic (biopsy-proven idiopathic pulmonary fibrosis, for instance, usual interstitial pneumonia, UIP, n = 15), inflammatory, and granulomatous diseases of human lung parenchyma (desquamative interstitial pneumonia, n = 10; ILD associated with collagen diseases, n = 10; sarcoidosis, n = 19 and allergic alveolitis, n = 8). In human lung alveolar epithelial cells, gamma-GCSh was decreased by TGF-beta(1), whereas TNF-alpha caused a transient enzyme induction. In normal lung, gamma-GCS was mainly localized to the bronchiolar epithelium. In UIP, the highest immunoreactivities were observed in the airway epithelium and metaplastic alveolar epithelium, but fibroblastic foci were negative. In sarcoidosis, the highest reactivities were detected in the epithelium, alveolar macrophages and pulmonary granulomas. gamma-GCS was ultrastructurally localized to the cytoplasm of regenerating type II pneumocytes and macrophages. In conclusion, gamma-GCS is widely expressed in sarcoidosis and regenerating epithelium but is low in the fibrotic areas of usual interstitial pneumonia, probably because of enzyme down-regulation.

Topics: Adult; Aged; Blotting, Western; Cell Line; Female; Glutamate-Cysteine Ligase; Humans; Immunohistochemistry; Lung Diseases, Interstitial; Male; Microscopy, Immunoelectron; Middle Aged; Pulmonary Alveoli; Respiratory Function Tests; Respiratory Mucosa; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Glutaredoxins (Grx) are thiol-disulfide oxidoreductases with antioxidant capacity and catalytic functions closely associated with glutathione, an antioxidant abundantly present in human lung. The present study investigated the expression of both human glutaredoxins in cultured human lung cells and lung homogenates by reverse-transcription polymerase chain reaction and Western blotting. Immunohistochemical studies were conducted with 38 human lung specimens, including healthy lung, parenchymal sarcoidosis, extrinsic allergic alveolitis, and usual interstitial pneumonia (UIP). The ultrastructural localization of Grx1 was assessed by immunoelectron microscopy. In addition, cultured airway epithelial cells were exposed to tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta. Both Grx1 and Grx2 could be detected at the mRNA and protein level in cultured human lung cells, but only Grx1 was prominently expressed in lung homogenates and alveolar macrophages. Immunohistochemically, Grx1 was highly concentrated to alveolar macrophages and weakly positive in the bronchial epithelium. Grx1 was ultrastructurally localized to the plasma membrane, cytoplasmic vacuoles, and nucleus. The expression of Grx1 decreased in alveolar macrophages of sarcoidosis and allergic alveolitis compared with the case for controls (P < 0.001), and bronchial epithelium of these diseases revealed no Grx1 immunoreactivity. Fibroblast foci and other fibrotic areas in UIP were mainly negative. In A549 cells, Grx1 was down-regulated by TGF-beta, whereas TNF-alpha caused no clear effect. Overall, high expression of Grx1 in alveolar macrophages suggests its importance in the primary defense of human lung. Decreased expression of Grx1 further suggests the impairment of this system both in inflammatory and fibrotic lung diseases, consistent with the down-regulation of Grx1 by TGF-beta in vitro.

Topics: Blotting, Western; Cell Line, Transformed; Epithelial Cells; Female; Fluorescent Antibody Technique, Indirect; Glutaredoxins; Humans; Lung; Lung Diseases, Interstitial; Macrophages, Alveolar; Male; Microscopy, Immunoelectron; Middle Aged; Oxidoreductases; Respiratory Mucosa; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Decorin, a small leucin-rich proteoglycan, is a negative regulator of transforming growth factor-beta, but the antifibrotic effect of decorin gene transfer has not been examined in a mouse model of usual interstitial pneumonia (UIP). We constructed a replication-defective recombinant adenovirus harboring human decorin gene (AdCMV.DC) and administered 1 x l0(9) plaque-forming units of AdCMV.DC intratracheally or intravenously to C57BL/6 mice with intraperitoneal injection of bleomycin, which induces a subpleural fibroproliferation, mimicking UIP, by day 28. Only intratracheal administration of AdCMV.DC increased decorin mRNA expression in the lung and decreased the hydroxyproline content augmented in bleomycin-induced pulmonary fibrosis (1.13 +/- 0.02 to 0.96 +/- 0.02, P = 0.006). In contrast, intravenous administration of AdCMV.DC increased the decorin expression only in the liver, but not in the lung, and without reducing lung fibrosis. These results indicate that adenoviral decorin gene transfer is effective only by direct administration to fibrosing lungs.

Topics: Adenoviridae; Animals; Antimetabolites, Antineoplastic; Bleomycin; Cell Line; Decorin; Disease Models, Animal; Extracellular Matrix Proteins; Female; Gene Transfer Techniques; Genes, Reporter; Genetic Therapy; Genetic Vectors; Humans; Injections, Intravenous; Instillation, Drug; Liver; Lung; Lung Diseases, Interstitial; Mice; Mink; Proteoglycans; Pulmonary Fibrosis; RNA, Messenger; Trachea; Transfection; Transforming Growth Factor beta

Cytokine levels in bronchoalveolar lavage fluid from patients with eosinophilic pneumonia (n = 7), allergic alveolitis (n = 11), (cryptogenic) fibrosing alveolitis (n = 8), sarcoidosis (n = 10) were determined, as well as levels in control samples from healthy non-smoking volunteers (n = 11). Fibronectin levels were increased in all the patient categories, the highest absolute levels of fibronectin (100-fold increase) being found in eosinophilic pneumonia and allergic alveolitis. TGF-beta (transforming growth factor-beta) was significantly elevated in allergic alveolitis only. There was a significant difference between allergic alveolitis on the one hand and both sarcoidosis and fibrosing alveolitis on the other. Tumour necrosis factor-alpha (TNF-alpha) was significantly increased in eosinophilic pneumonia and allergic alveolitis; allergic alveolitis and fibrosing alveolitis differed significantly in this respect. Platelet-derived growth factor-BB (PDGF-BB) levels were significantly elevated in allergic alveolitis and fibrosing alveolitis. It was found that the level of PDGF-BB was significantly decreased in the case of sarcoidosis, with no overlapping with allergic alveolitis or fibrosing alveolitis. Interferon-gamma (IFN-gamma) was decreased in all patient categories. A significant difference in extent of the decrease was found between allergic alveolitis and sarcoidosis. The interstitial lung diseases thus differed in the pattern of cytokines expressed, indicating that these cytokines could well be a part of the pathogenic process, and also that the measurement of cytokine levels could be diagnostically useful.

Topics: Adult; Becaplermin; Bronchoalveolar Lavage Fluid; Cytokines; Female; Fibronectins; Humans; Interferon-gamma; Lung Diseases, Interstitial; Male; Middle Aged; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-sis; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To study the distribution, the expression and the significance of TGF-beta(1), b-FGF, IL-8, IL-13 and IFN-gamma in different lung tissue compartments in usual interstitial pneumonia/idiopathic pulmonary fibrosis (UIP/IPF) and nonspecific interstitial pneumonia (NSIP).. Specimens were obtained by open or video-assisted thoracoscopic lung biopsy from patients with UIP (n = 5) and NSIP (n = 8). Control specimens were obtained by surgical lobectomy from patients with primary lung cancer (n = 5). The distribution of these cytokines in lung tissues was observed by semi-quantitative method using immunohistochemical staining.. TGF-beta(1), IL-8 and b-FGF were localized in alveolar epithelial cells, alveolar macrophages, and the bronchial epithelium. Overall intensity of TGF-beta(1), IL-8 and b-FGF expression in UIP was stronger in comparison with NSIP. IL-13 was distributed in alveolar epithelial cells, alveolar macrophages and interstitial mononuclear cells. Its expression in UIP was similar to that in NSIP. IFN-gamma was expressed mainly in interstitial mononuclear cells. Its expression in NSIP was stronger than that in UIP. The ratio of IL-13 to IFN-gamma in UIP (2.18 +/- 0.76) was significantly higher than that in NSIP (0.95 +/- 0.28) or that in the control (0.91 +/- 0.16) (P < 0.05, UIP versus NSIP or control), whereas the ratio of IL-13 to IFN-gamma in NSIP was similar to that in the control. In normal lungs, only alveolar macrophages expressed these cytokines.. The different expression of TGF-beta(1), IL-8 and b-FGF in UIP and NSIP and the balance of IL-13/IFN-gamma may be involved in the different pathogenesis in these two diseases.

Topics: Adult; Aged; Cytokines; Female; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-13; Interleukin-8; Lung; Lung Diseases, Interstitial; Male; Middle Aged; Transforming Growth Factor beta

The epithelial alteration in interstitial pneumonias is one of the repair processes at the sites of disease activity. Regenerative epithelial cells may participate in remodeling of the lung. To determine the phenotype of regenerative epithelial cells in usual interstitial pneumonia (UIP) and nonspecific interstitial pneumonia (NSIP), the expression of Clara cell 10KD protein (CC10), cytokeratin (CK) 14 and 17, surfactant apoprotein (SP)-A, KL-6/MUC1, transforming growth factor (TGF) beta2 were examined in 25 patients with UIP, 9 patients with NSIP and normal lung tissues from 10 patients with lung cancer. In honeycomb lesions of UIP, non-ciliated columnar cells mainly expressed CC10, cuboidal cells expressed CC10, CK17, CK14 and SP-A in descending order. Fibroblastic foci are covered by CK17, CK14, CC10, and a few SP-A positive flattened or cuboidal cells. Regenerative epithelium in NSIP mainly comprised cuboidal cells expressing SP-A, CC10 and CK17. KL-6 was more remarkably expressed in cuboidal and non-ciliated columnar cells both in UIP and NSIP. Expression of TGFbeta2 was observed in cuboidal and flattened epithelium. In severe fibrotic areas, CC10 expressing cells were more prominent, while SP-A positive cells were more prominent in less fibrotic areas. Regenerative epithelial cells in remodeling area in UIP may be derived from bronchiolar basal cells and Clara cells, while most of those in NSIP may be derived from type II pneumocytes. The different origin of regenerative epithelium may reflect the severity and extent of the injury and the degree of consequent fibrosis in UIP and NSIP.

Topics: Antigens; Antigens, Neoplasm; Apoproteins; Bronchi; Enzyme Inhibitors; Epithelial Cells; Female; Fibroblasts; Fibrosis; Glycoproteins; Humans; Keratins; Lung; Lung Diseases, Interstitial; Lung Neoplasms; Male; Middle Aged; Mucin-1; Mucins; Phenotype; Phospholipases; Proteins; Pulmonary Surfactant-Associated Proteins; Regeneration; Transforming Growth Factor beta; Transforming Growth Factor beta2; Uteroglobin

Successful repair after tissue injury and inflammation requires resolution of the inflammatory response and removal of extracellular matrix breakdown products. We have examined whether the cell-surface adhesion molecule and hyaluronan receptor CD44 plays a role in resolving lung inflammation. CD44-deficient mice succumb to unremitting inflammation following noninfectious lung injury, characterized by impaired clearance of apoptotic neutrophils, persistent accumulation of hyaluronan fragments at the site of tissue injury, and impaired activation of transforming growth factor-beta1. This phenotype was partially reversed by reconstitution with CD44+ cells, thus demonstrating a critical role for this receptor in resolving lung inflammation.

Topics: Animals; Apoptosis; Bleomycin; Bone Marrow Transplantation; Bronchoalveolar Lavage Fluid; Cell Count; Chemokines; Chimera; Humans; Hyaluronan Receptors; Hyaluronic Acid; Lung; Lung Diseases, Interstitial; Macrophages, Alveolar; Mice; Mice, Inbred C57BL; Neutrophil Infiltration; Neutrophils; Phagocytosis; Phenotype; Pulmonary Alveoli; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

An association between Epstein-Barr virus (EBV) infection and fibroblast proliferation in the interstitial spaces of the lung has been suggested in idiopathic interstitial pneumonia. In this study we show that EBV can infect human lung fibroblasts in vitro. A primary-cultured human lung fibroblast cell line, designated CCD-32Lu, expressed EBV nuclear antigen 1 after coculture with lethally irradiated EBV producing cells. The infection further induced CCD-32Lu cells to produce the fibrogenic cytokines basic fibroblast growth factor (bFGF) and interleukin-1beta. These findings indicate that lung fibroblasts may be a target for EBV infection and suggest that EBV may play a role in increased production of these cytokines and induce fibroblast proliferation in idiopathic interstitial pneumonia.

Topics: Antibodies, Blocking; Antibodies, Viral; Blotting, Western; Cell Division; Cell Line; Coculture Techniques; Enzyme-Linked Immunosorbent Assay; Epstein-Barr Virus Infections; Epstein-Barr Virus Nuclear Antigens; Fibroblast Growth Factor 2; Fibroblasts; Flow Cytometry; Fluorescent Antibody Technique; Herpesvirus 4, Human; Humans; Interleukin-1; Lung; Lung Diseases, Interstitial; Neutralization Tests; Transforming Growth Factor beta; Transforming Growth Factor beta1

The aim of this study was to compare the function of lung fibroblasts obtained from surgically biopsied specimens of patients with idiopathic pulmonary fibrosis/usual interstitial pneumonia (UIP; n = 5), nonspecific interstitial pneumonia (NSIP; n = 5), and normal parts of surgically resected lungs (control; n = 5). The results showed that (1) fibroblasts obtained from UIP showed increased contractility compared with those obtained from NSIP or controls (UIP, 72.7 +/- 6.21%; NSIP, 32.8 +/- 5.46; controls, 28.5 +/- 3.51, p < 0.01 in UIP versus NSIP or control); (2) this increase in contractility was consistent with enhanced F-actin content in fibroblasts; (3) conditioned media from UIP fibroblast cultures enhanced control fibroblast contractility, whereas those obtained from NSIP or controls did not; (4) the 180 and 25 kD products representing the contractility in conditioned media were identified as fibronectin (ED-A domain) and TGF-beta1 by immunoblots, respectively; (5) the UIP-conditioned media contained higher amounts of fibronectin or TGF-beta 1 (fibronectin: UIP 289 +/- 47.1 ng/ml, NSIP 121 +/- 23.0, control 118 +/- 16.0; TGF-beta1: UIP 798 +/- 119 pg/ml, NSIP 246 +/- 69.1, control 247 +/- 53.6, p < 0.01 in UIP versus NSIP or control); () the contractility positively correlated with the amount of either fibronectin (r = 0.867, p < 0.001, n = 15) or TGF-beta 1 (r = 0.939, p < 0.001, n = 15), respectively. Thus, UIP fibroblasts showed greater contractility than did NSIP fibroblasts and up-regulated control fibroblasts.

Topics: Actins; Adenocarcinoma; Analysis of Variance; Biopsy; Cells, Cultured; Culture Media, Serum-Free; Female; Fibroblasts; Gels; Hamartoma; Humans; Lung; Lung Diseases; Lung Diseases, Interstitial; Lung Neoplasms; Male; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1

The involvement of the insulin-like growth factor (IGF) system in lung growth and repair following injury is sustained by a number of studies. Based on this knowledge, the aim of the present work was to document the expression of the IGFs and their binding proteins by alveolar cells obtained by bronchoalveolar lavage (BAL). Two groups were investigated: a control group of five children and a group of 11 children referred to the department for exploration of interstitial lung disease (ILD). Components of the IGF system studied included IGF-I, IGF-II and IGF-binding proteins (IGFBP). Expression of these factors was analysed at the level of messenger ribonucleic acid (mRNA) (by semi-quantitative reverse transcription polymerase chain reaction techniques), and of protein for the IGFBPs. In addition, expression of two major cytokines associated with the inflammatory process, tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta), was also documented. In children without parenchymal disease, the growth factor expressed was IGF-I, in association with the presence of mRNA for IGFBP-2 in all cases. In children with ILD, expression of IGF-I was observed in nine patients and of IGF-II in three patients, and the presence of IGFBP-2 was found in all extracts analysed (mRNA and proteins). Evaluation of IGFBP-2 expression indicated an increase in the group of children with ILD. Interestingly, a significant association was observed between the increase in IGFBP-2 expression and TGF-beta expression. The present data emphasize the presence on insulin-like growth factor-binding protein-2 in the BAL of all patients, and suggest that this protein may be an important factor of the injury/repair processes during the progression of alveolar inflammation.

Topics: Adolescent; Bronchoalveolar Lavage Fluid; Cell Count; Child; Child, Preschool; Female; Humans; Insulin-Like Growth Factor Binding Protein 2; Insulin-Like Growth Factor Binding Proteins; Insulin-Like Growth Factor I; Lung Diseases, Interstitial; Male; Polymerase Chain Reaction; Pulmonary Alveoli; RNA, Messenger; Somatomedins; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The case of a 3-month-old boy with lymphoid interstitial pneumonia (LIP) is reported. He had cough and tachypnea, his weight gain was poor and a chest radiograph showed microgranular shadows in almost all lung areas. Histological investigations revealed severe cellular infiltration by a variety of lymphoid and plasma cells with lymphoid follicle formation in the alveolar walls and also around the bronchioles. Foamy macrophages, a few lymphocytes and exudate filled the alveolar spaces. Epithelial cells lining the air spaces expressed human leukocyte antigen (HLA)-DR. Lymphocytes and macrophages in the alveolar spaces expressed transforming growth factor (TGF)-beta strongly. Serum TGF-beta 1 concentrations were measured eight times during the course of his illness. They exceeded the upper end of the normal range in four samples and were within it in the others. These results suggested that dysfunction of the immune system, especially abnormal expression of HLA-DR in non-immune cells and exaggerated production of TGF-beta played important roles in the pathogenesis of LIP in this patient.

Topics: Antigens, CD; Antigens, CD20; Biopsy; CD79 Antigens; HLA-DR Antigens; Humans; Immunohistochemistry; Infant; Lung; Lung Diseases, Interstitial; Lymphocytes; Macrophages; Male; Radiography; Receptors, Antigen, B-Cell; Transforming Growth Factor beta

Although it is recognized that three isoforms of transforming growth factor-beta (TGF-beta) exist in mammals, their expression, distribution, and function in injury and repair are not well characterized. Using immunohistochemistry and antibodies to synthetic peptides of TGF-beta 1, TGF-beta 2, and TGF-beta 3, we determined the distribution of TGF-beta isoforms in lung sections with acute and chronic lesions of idiopathic pulmonary fibrosis (IPF), chronic asbestosis and hypersensitivity pneumonitis, as well as non-specific pneumonitis. In lung sections with advanced pulmonary fibrosis and honeycombing, irrespective of the diagnosis, TGF-beta 1 was prominently expressed in epithelial cells and macrophages and was found to be associated with the extracellular matrix. In lungs with early lesions of IPF and only inflammatory changes, TGF-beta 1 was present in alveolar macrophages but TGF-beta 1 was not present in epithelial cells. Small amounts of matrix-associated TGF-beta 1 were present subepithelially in areas of lung sections from patients with IPF with minimal inflammation and no fibrosis. In normal lungs with no evidence of inflammation or fibrosis TGF-beta 1 was not seen in alveolar macrophages, epithelial cells, or extracellularly. TGF-beta 2 and TGF-beta 3 were expressed in alveolar macrophages, epithelial cells, and smooth muscle cells of vessels and bronchi of normal lungs and lungs with both inflammatory and fibrotic changes. Our findings suggest that while TGF-beta 2 and TGF-beta 3 are ubiquitously expressed in the lung, TGF-beta 1 is expressed in epithelial cells of fibrotic lungs where the presence of TGF-beta 1 is not disease-specific but an indication of the chronicity of the injury.

Topics: Adult; Aged; Antibody Specificity; Asbestosis; Biopsy; Cysts; Epithelium; Humans; Immunohistochemistry; Lung; Lung Diseases, Interstitial; Lung Neoplasms; Macrophages; Middle Aged; Muscle, Smooth, Vascular; Pulmonary Fibrosis; Transforming Growth Factor beta

Topics: Acquired Immunodeficiency Syndrome; Animals; Cytokines; Female; HIV-1; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-2; Interleukin-4; Lung Diseases, Interstitial; Lymphocyte Activation; Mice; Mice, Inbred C57BL; RNA, Messenger; T-Lymphocytes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Virus Replication

T lymphocytes expressing the alpha E beta 7 integrin are localized and selectively retained in mucosal tissues. To investigate a potential relationship between alpha E beta 7 expression and pulmonary inflammation, the distribution of alpha E beta 7-bearing CD4+ and CD8+ T cells in peripheral blood and bronchoalveolar lavage (BAL) fluids obtained from patients with allergic asthma, sarcoidosis, hypersensitivity pneumonitis, and idiopathic pulmonary fibrosis (IPF) was determined. In contrast to the distribution in peripheral blood, BAL fluid from these patients contained high number of cells expressing alpha E beta 7 with markedly different expression patterns on CD4 or CD8 cells as well as among the various diseases. Despite similar numbers of activated CD4 cells, alpha E beta 7+CD4+ T cells ranged from 15% in asthmatics to 70% in IPF. In contrast, even in normal individuals, 60% to 90% of BAL fluid CD8+ T cells express alpha E beta 7, suggesting differential induction mechanisms on CD4 and CD8 cells. In vitro experiments revealed that a substantial proportion of peripheral blood CD+ T cells express alpha E beta 7 after stimulation with anti-CD3 antibodies, and up to 80% positive cells were found after the addition of TGF-beta. In contrast, less than 10% of CD4 cells express this particular integrin after in vitro stimulation, and the presence of TGF-beta only increased the number to 30%. Supernatants from in vitro-activated BAL cells as well as concentrated BAL fluid from patients with high alpha E beta 7 expression had no further enhancing effect. However, crosslinking of alpha 4 beta 1-, but not beta 2-integrins, significantly increased the number of alpha E beta 7 expressing CD4+ and CD8+ T cells, even in the absence of TGF-beta. These data indicate that in addition to TGF-beta, the interaction of particular T-cell subsets with specific endothelial cell and extracellular matrix proteins may upregulate alpha E beta 7 integrin expression and thereby contribute to the selective accumulation of these cells in inflammatory lung diseases.

Topics: Adult; Alveolitis, Extrinsic Allergic; Antibodies; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cross-Linking Reagents; Eosinophils; Female; HLA-DR Antigens; Humans; Integrin alpha4beta1; Integrins; Leukocyte Common Antigens; Lung Diseases, Interstitial; Lymphocyte Activation; Lymphocyte Subsets; Macrophages, Alveolar; Male; Middle Aged; Neutrophils; Pulmonary Fibrosis; Receptors, Interleukin-2; Receptors, Lymphocyte Homing; Sarcoidosis; T-Lymphocytes; Transforming Growth Factor beta

Interstitial pneumonia is characterized by alveolitis with resulting fibrosis of the interstitium. To determine the relevance of humoral factors in the pathogenesis of interstitial pneumonia, we introduced expression vectors into Wistar rats via the trachea to locally overexpress humoral factors in the lungs. Human interleukin (IL) 6 and IL-6 receptor genes induced lymphocytic alveolitis without marked fibroblast proliferation. In contrast, overexpression of human transforming growth factor beta 1 or human platelet-derived growth factor B gene induced only mild or apparent cellular infiltration in the alveoli, respectively. However, both factors induced significant proliferation of fibroblasts and deposition of collagen fibrils. These histopathologic changes induced by the transforming growth factor beta 1 and platelet-derived growth factor B gene are partly akin to those changes seen in lung tissues from patients with pulmonary fibrosis and markedly contrast with the changes induced by overexpression of the IL-6 and IL-6 receptor genes that mimics lymphocytic interstitial pneumonia.

Topics: Adenoviridae; Animals; Antigens, CD; Becaplermin; Genetic Vectors; Humans; Interleukin-6; Lung; Lung Diseases, Interstitial; Male; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-sis; Pulmonary Fibrosis; Rats; Rats, Wistar; Receptors, Interleukin; Receptors, Interleukin-6; Recombinant Proteins; Transforming Growth Factor beta

To study the role of transforming growth factor-beta 1 (TGF-beta 1) in the pathogenesis of pulmonary fibrosis we have examined lung biopsies from nine patients with systemic sclerosis and interstitial lung disease, eight with 'lone' cryptogenic fibrosing alveolitis, two with cystic fibrosis, two with extrinsic allergic alveolitis, two with Langerhans' cell histiocytosis, one with lymphangioleiomyomatosis, one with giant cell interstitial pneumonia, and one adenocarcinoma of the lung. In cryptogenic fibrosing alveolitis, both 'lone' and associated with systemic sclerosis alveolar macrophages, bronchial epithelium and hyperplastic type II pneumonocytes expressed intracellular TGF-beta 1. Extracellular TGF-beta 1 was found in the fibrous tissue immediately beneath the bronchial and hyperplastic alveolar epithelium. In normal lung, however, the alveolar epithelium and alveolar interstitium were negative for both forms of TGF-beta 1. There was strong expression of TGF-beta 1 in hyperplastic mesothelium and its underlying connective tissue and in Langerhans' cells in the two cases of histiocytosis. In the organizing pneumonia in cystic fibrosis, the intraalveolar buds of granulation tissue reacted strongly for the extracellular form of TGF-beta 1 and the overlying hyperplastic epithelium expressed the intracellular form. In lymphangioleiomyomatosis, the aberrant smooth muscle cells strongly expressed intracellular TGF-beta 1 and the extracellular form was expressed in the adjacent connective tissue. In giant cell interstitial pneumonia, the numerous alveolar macrophage including the multinucleate forms, expressed intracellular TGF-beta 1, as did the hyperplastic alveolar epithelium.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Humans; Immunohistochemistry; Lung Diseases, Interstitial; Pulmonary Fibrosis; Scleroderma, Systemic; Transforming Growth Factor beta

We have studied the gene expression of ten growth factors in bronchoalveolar lavage (BAL) cells from patients with lung fibrosis by using a reverse-transcription-DNA polymerase chain reaction (RT-PCR) technique. IL-1 beta mRNA was detected in almost all of the samples, and TGF beta and IGF-I mRNA were detected in some. The BAL supernatant from the patients was found to have mitogenic activity for lung fibroblasts. Moreover, human recombinant IL-1 beta, TGF beta and IGF-I were found to promote the proliferation of lung fibroblasts. These data suggest that IL-1 beta, TGF beta and IGF-I gene expression in BAL cells might be involved in the development of lung fibrosis.

Topics: Adult; Base Sequence; Bronchoalveolar Lavage Fluid; Bronchoscopy; Gene Expression; Humans; Insulin-Like Growth Factor I; Interleukin-1; Lung Diseases, Interstitial; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; Transforming Growth Factor beta