endothelin-1 and Bronchiolitis-Obliterans

Topics: Bronchiolitis Obliterans; Bronchoalveolar Lavage; Cell Differentiation; Cell Movement; Cell Proliferation; Endothelin-1; Humans; Lung Transplantation; Mesenchymal Stem Cells; Yin-Yang

Bronchiolitis obliterans syndrome (BOS) has an incidence of 57% at 5 years after lung transplantation, accounts for 30% of all deaths 3 years posttransplant and because treatment options are extremely limited, it constitutes a significant health care problem. Adult mesenchymal stem cells (MSCs) play a role in lung turnover; however, their role in BOS remains unknown.. MSCs were isolated from bronchoalveolar lavage (BAL) in 101 lung allograft recipients. BAL was screened by protein array and MSCs were analyzed by real-time polymerase chain reaction, proliferation, migration, and enzyme linked immunosorbent assays.. Multipotent MSCs were isolated from BAL of lung recipients independent of BOS presence. However, MSCs from BOS patients proliferated at higher rates (P<0.001) and were associated with higher α-smooth muscle actin (P = 0.03) but lower surfactant protein B (P = 0.02) compared with those from no-BOS patients. Histological analysis revealed that MSCs are abundant in lung tissue of BOS patients. MSCs from BOS patients produced higher endothelin-1 (ET-1) amounts (P<0.001) compared with those from no-BOS; and ET-1 stimulated whereas ET-1 blockade suppressed MSC proliferation, migration, and differentiation.. These results indicate that MSCs are associated with BOS and are governed by ET-1. Targeting MSCs by ET-1 blockade might be useful in BOS treatment.

Topics: Actins; Adult; Aged; Bronchiolitis Obliterans; Bronchoalveolar Lavage; Cell Differentiation; Cell Movement; Cell Proliferation; Cohort Studies; Endothelin-1; Female; Humans; Lung; Lung Transplantation; Male; Mesenchymal Stem Cells; Middle Aged; Protein Precursors; Proteolipids

Bronchiolitis obliterans (BO) is a severe complication limiting long-term survival after lung transplantation. To date, no cure exists for BO, and the mechanisms leading to BO are not well understood. Endothelin-1 (ET-1) is a potent mitogenic and profibrotic peptide produced by pulmonary vascular endothelial cells that play a role in the pathophysiology of lung allograft dysfunction. Whether ET-1 could predict BO syndrome (BOS) development is unknown.. Transbronchial biopsy specimens and serum and bronchoalveolar lavage were obtained from 30 lung transplantation patients with and 30 without BOS at 3 points. The serum and bronchoalveolar lavage ET-1 concentrations were measured by enzyme-linked immunosorbent assay, and the ET-1 mRNA expression in the transbronchial biopsy specimens was examined using real-time polymerase chain reaction.. The pretransplant ET-1 serum concentrations were greater in the patients with BOS (P = .02); and ET-1 mRNA was significantly upregulated in the lung grafts of those with versus those without BOS at 3 and 12 months after transplant (P = .01). At 3 and 12 months after transplantation, the ET-1 concentrations were significantly elevated in the serum (P < .01 and P < .0001, respectively) and bronchoalveolar lavage (P < .01 and P = .02, respectively) of patients with compared with those without BOS. On logistic regression analysis, the pretransplant and 3-month post-transplant serum ET-1 level predicted for BOS (odds ratio, 1.01; 95% confidence interval, 1.004-1.025; P < .007; odds ratio, 2.9; 95% confidence interval, 1.01-8.52; P < .001). The serum ET-1 level at 12 months was diagnostic for BOS (odds ratio, 3.9; 95% confidence interval, 1.42-10.80; P = .008).. Elevated serum ET-1 concentrations were predictive of BOS, and the assessment of circulating ET-1 might be beneficial in diagnosing and monitoring BO.

Topics: Biomarkers; Biopsy; Bronchiolitis Obliterans; Bronchoalveolar Lavage; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunosuppressive Agents; Logistic Models; Lung Transplantation; Male; Middle Aged; Predictive Value of Tests; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Endothelin-1 (ET-1) expression is increased after lung transplantation in association with ischemia reperfusion injury and acute rejection. However, little is known of the role of ET-1 during the development of obliterative bronchiolitis. In this study, we investigated the biological significance of ET-1 in obliterative airway disease development using a rat tracheal allograft model. Immunoreactivity of ET-1 and its receptors ET-RA and ET-RB was increased four-fold in allografts compared with syngrafts and localized to mononuclear cells and smooth muscle cells of the myofibroproliferative lesion and airway wall, indicating that ET-1 may mediate its effects in both a paracrine and autocrine manner in smooth muscle cells. Inhibition of ET-1 action by a nonselective ET-1 receptor antagonist, bosentan, significantly decreased tracheal occlusion, which was linked to delayed epithelial necrosis, suppressed smooth muscle cell proliferation, and a marked reduction in the number of interleukin-1beta and interleukin-2 immunoreactive cells. Our findings show that endogenous ET-1 activation is associated with obliteration of the airway wall, and blocking signaling downstream of ET-1 receptors leads to attenuation of obliterative airway disease. The results suggest that ET-1 has a proproliferative and proinflammatory role in the development of obliterative bronchiolitis.

Topics: Airway Obstruction; Animals; Antihypertensive Agents; Bosentan; Bronchiolitis Obliterans; Endothelin-1; Lung Transplantation; Male; Rats; Receptors, Endothelin; Sulfonamides; Trachea

Endothelin-1 (ET-1) has fibrogenic and inflammatory properties. Its pathogenic role in pulmonary fibrosis and certain inflammatory airway diseases is now well known. Its production is, in part, triggered by infectious processes. Episodes of infection are suspected to be involved in the development of bronchiolitis obliterans syndrome (BOS), which is the main feature of chronic lung rejection and the major factor limiting the long-term survival of transplanted patients. We postulated that ET-1 is upregulated during infectious complications arising from the graft and that this could partly explain the remodeling of airway structures observed in BOS. We, therefore, set up this study to assess ET-1 expression in relation to complications of the graft in human lung transplant recipients.. ET-1 mRNA was quantified by reverse transcription-competitive polymerase chain reaction in cells from 119 samples of bronchoalveolar lavage (BAL) fluid from 17 lung transplant recipients. ET-1 and big ET-1 proteins were assessed in BAL cell culture supernatants by enzyme immunoassay. Transbronchial biopsies (n=21) were stained immunohistochemically for ET-1 receptors.. Episodes of bacterial infection strongly correlated with increased ET-1 mRNA and protein expression. ET-1 receptors were also upregulated during these episodes, especially on endothelial and smooth muscle cells. Five of the seven patients with the highest ET-1 levels subsequently developed BOS.. These results raise the possibility that ET-1, part of whose production is triggered by infectious postgraft complications, might play a role in the development of BOS through its potential effects on airway remodeling.

Topics: Adult; Bacterial Infections; Bronchi; Bronchiolitis Obliterans; Bronchoalveolar Lavage Fluid; Endothelin-1; Female; Humans; Lung Diseases; Lung Transplantation; Male; Receptor, Endothelin A; Receptors, Endothelin; RNA, Messenger; Tissue Distribution; Up-Regulation

Overproduction and overexpression of endothelin-1 (ET-1) have been reported to contribute to the pathophysiology of pulmonary diseases, including pulmonary fibrosis, obliterative bronchiolitis, and primary pulmonary hypertension. To determine whether ET-1 contributes to the pathogenesis of pulmonary disease, we locally overexpressed ET-1 using an in vivo UV-inactivated hemagglutinating virus of Japan (HVJ) liposome-mediated gene transfer system. Plasmid DNA of ET-1 (pME18fc preproET-1) and high mobility group 1 (HMG1) protein were co-encapsulated in liposomes. Then the plasmid DNA and liposome complexes were introduced into the lung via the trachea in Wistar rats, using HVJ-mediated membrane fusion. Control animals received instillation of HVJ liposome with an empty cassette. Two weeks after in vivo transfection of the preproET-1 gene, hyperplastic connective tissue plaques were seen in the alveolar duct and small conducting airways, indicating histologically distinctive obliterative bronchiolitis. No histopathologic changes were seen in the control animals. These results suggested that local overexpression of ET-1 may play an important role in the pathogenesis of obliterative bronchiolitis.

Topics: Animals; Bronchiolitis Obliterans; DNA, Viral; Endothelin-1; Gene Transfer Techniques; Liposomes; Lung; Lung Diseases; Male; Rats; Rats, Wistar; Respirovirus; Respirovirus Infections

This study was designed to assess which cells in transplanted lungs express endothelin-1 (ET-1) and if expression of the peptide can be used to discriminate between rejection and infection in transplanted lungs.. Transbronchial biopsies (n=104) from 29 human lung transplant recipients were stained immunohistochemically for ET-1. Cells expressing ET-1 (pneumocytes, endothelial cells, airway epithelial cells, lymphocytes, and macrophages) were quantified and correlated with clinical histopathology findings.. ET-1 was expressed in airway epithelial cells (93% of the biopsies), infiltrating macrophages (86%), and lymphocytes (19%) but not in endothelial cells or pneumocytes. ET-1 expression did not vary with rejection, obliterative bronchiolitis, or infection. ET-1 expression did not correlate with age, grade of rejection, pulmonary function, or time after transplantation.. In transplanted human lungs, ET-1 is expressed in airway epithelial cells and infiltrating macrophages, and expression does not vary with pathological processes. Therefore, immunostaining for ET-1 probably cannot be used to discriminate between rejection and infection in transplanted lungs.

Topics: Acute Disease; Adult; Aged; Bronchiolitis Obliterans; Diagnosis, Differential; Endothelin-1; Epithelial Cells; Female; Graft Rejection; Humans; Lung; Lung Transplantation; Male; Middle Aged; Tissue Distribution

Bronchiolitis obliterans (OB) is a lesion that results when injury to small conducting airways is repaired by a proliferation of fibrous granulation tissue. Bronchiolitis obliterans has emerged as a main cause of morbidity and mortality in the setting of lung and heart-lung transplantation. Endothelin-1 (ET-1), initially discovered as a vasoconstrictive peptide, has a mitogenic activity on vascular smooth cells and airway epithelial cells. Overproduction of endothelin has been reported in patients with OB or chronic rejection after lung transplantation. It is still undetermined whether locally overexpressed ET-1 has a potential impact in the pathogenesis of OB.. We locally overexpressed ET-1 using ultraviolet irradiation-inactivated hemagglutinating virus of Japan (HVJ)-liposome-mediated in vivo gene transfer. Plasmid DNA of prepro-ET-1 and high mobility group 1 protein were coencapsulated in liposomes, and were introduced into airway epithelial cells by HVJ-mediated membrane fusion. Control animals received instillation of HVJ-liposome with an empty expression cassette. To confirm the efficiency of transfection, HVJ liposome with beta-galactosidase gene was introduced. The expression of ET-1 and beta-galactosidase was assessed by immunohistochemistry.. Bronchial epithelium alveolar cells and alveolar macrophage were stained blue (X-Gal) 1 week after in vivo gene transfer of beta-galactosidase gene, indicating beta-gal activity. In animals 1 to 2 weeks after in vivo transfection of prepro-ET-1 gene, hyperplastic connective tissue plaque was seen in the alveolar duct and small conducting airway, indicating histologically distinctive bronchiolitis obliterans. Strong ET-1-like immunoactivities were seen in the airway epithelial, hyperplastic connective tissue, and alveolar cells. No histopathologic changes were seen in the control animals.. These results suggested that ET-1 may play an important role in the pathogenesis of OB. The effective pharmacologic antagonist or inhibitor may possibly control the progression of disease in patients of OB.

Topics: Animals; beta-Galactosidase; Bronchi; Bronchiolitis Obliterans; Disease Models, Animal; Endothelin-1; Endothelium, Vascular; Epithelium; Gene Transfer Techniques; Genetic Vectors; Liposomes; Macrophages, Alveolar; Male; Plasmids; Pulmonary Alveoli; Rats; Rats, Wistar; Respirovirus; Transfection