bromochloroacetic-acid and Pulmonary-Fibrosis

The usefulness of selected serum biomarkers (KL-6, SP-A, SP-D, IL-8, MCP-1, CYFRA-21) in diagnosis, monitoring and prognosis prediction of idiopathic pulmonary fibrosis is discussed in this paper.

Topics: Antigens, Neoplasm; Biomarkers; Carrier Proteins; Chemokine CCL2; Humans; Interleukin-8; Keratin-19; Keratins; Mucin-1; Pulmonary Fibrosis; Pulmonary Surfactant-Associated Protein D

This review discusses current knowledge of the involvement of the alveolar epithelium in tissue remodelling during fibrogenesis. The purpose of the present paper is to give an overview, including the authors' own results, of knowledge of ultrastructural alterations, proliferation kinetics and phenotypic changes of pneumocytes in experimental and clinical pathology of pulmonary fibrosis. After lung injury, the alveolar epithelial cells show ultrastructural alterations, hypertrophy and hyperplasia, and a modulation of a series of structural and membrane proteins such as cytoskeletal changes, loss or de novo expression of epithelial adhesion molecules, and altered lectin binding. Furthermore, enhanced secretion of proteases, of cytokines and other soluble factors can be observed in the alveolar epithelium. These findings suggest the contribution of the epithelium in the remodelling process to be greater than expected. Estimations of the cell kinetics show that type II pneumocytes have the proliferative capacity to restore high proportions of damaged type I cells within few hours. In fibrosis this capacity also seems to be affected seriously, resulting in transitional phenotypes between type II and type I cells. Additionally, in the light of the detection of CD44 type of adhesion molecules at the foot processes of type II pneumocytes, some aspects of epithelial-fibroblast interaction are described.

Topics: Animals; Cadherins; Carcinoembryonic Antigen; Cell Cycle; Cytokines; Epithelium; Humans; Hyaluronan Receptors; Immunohistochemistry; Integrins; Intercellular Adhesion Molecule-1; Keratins; Models, Biological; Pulmonary Alveoli; Pulmonary Fibrosis

Fibrosing alveolitis is a usually chronic pulmonary disease affecting middle-aged men and women and causing progressive dyspnea. This review discusses the nosologic, etiologic, immunopathogenic, histologic, immunohistochemical and ultrastructural features of this condition. A hypothesis is presented suggesting microvascular damage as the primary pathologic change in cases associated with collagen vascular diseases and viral pneumonia.

Topics: Acquired Immunodeficiency Syndrome; Biopsy; Female; Fibronectins; Growth Substances; Histocytochemistry; Humans; Interferon Type I; Keratins; Lung; Male; Microcirculation; Microscopy, Electron; Molecular Weight; Pleura; Pulmonary Fibrosis; T-Lymphocytes

The serum concentration of several markers in patients with collagen disease was studied to evaluate the useful indices for the diagnosis of interstitial pneumonia/pulmonary fibroses (pneumonitis). Procollagen N-terminal type III peptides, type IV collagen and monoamine oxidase were measured as the fibrosing markers. Squamous cell related antigen (SCC) and soluble cytokeratin 19 fragments (CYFRA21-1) were measured as the tumor markers. Hyaluronic acid, ESR and CRP were measured as the inflammation markers. The 119 patients with collagen disease (71 patients with RA, 16 with SSc, 9 with SLE, 8 with PM/DM, 6 with MCTI) and 9 with other collagen diseases) who have been followed in our hospital were studied. Of 119 patients, 23 patients (RA14, SSc7, PM/DM2) were complicated with pneumonitis. The results were as follows. 1. All the serum markers except CYFRA21-1 had no significantly difference between with and without pneumonitis. The serum CYFRA21-1 level in the pneumonitis group was higher than that of the non pneumonitis group (1.38 vs 0.66 ng/ml, P < 0.001). 2. The cut-off value was set at 2.0 ng/ml, corresponding to 26.1% sensitivity and 97.9% specificity for pneumonitis complicated with collagen disease. 3. The CYFRA21-1 level in early stage of pneumonitis group (from onset to measurement < 1 year) was significantly higher than that of late stage group (from onset to measurement > 1 year). And there are 75.0% sensitivity in early stage of pneumonitis group. 4. Case study was suggested that CYFRA21-1 had a potential as a diagnostic and monitoring marker for pneumonitis.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Collagen Diseases; Female; Humans; Keratin-19; Keratins; Lung Diseases, Interstitial; Male; Middle Aged; Pulmonary Fibrosis; Sensitivity and Specificity

Idiopathic pulmonary fibrosis (IPF) is the most common of interstitial lung diseases and is characterised by a significant mortality rate. That is way both clinicists and patients are interested to identify factors that may influence outcome of disease. This factors are named biological markers or biomarkers. Their usefulness in diagnostic, monitoring and prognosis of interstitial pneumonia, and idiopathic pulmonary fibrosis was estimated in many researches. The most of them was concerned to serum biomarkers, such as surfactant proteins, mucin-connected proteins, Clara cells proteins, cytoceratines and cytokines.

Topics: Biomarkers; Cytokines; Humans; Keratins; Mucins; Pulmonary Fibrosis; Pulmonary Surfactants; Uteroglobin

To analyze the clinical characteristics of interstitial pulmonary fibrosis (IPF) in patients with Rheumatoid arthritis (RA).. We retrospectively analyzed 198 RA patients with or without IPF. Characteristics of RA-IPF in clinical and lab data were analyzed. Age, duration of disease, clinical and laboratory parameters, history of smoking and medicine were compared between the patients with and without IPF.. (1) Among the 198 RA patients, 15.2% (30/198) were found with IPF. 100% RA-IPF patients had HRCT findings. However, 63.3% (19/30) had positive findings in chest X-ray, and only 46.7% (14/30) had the complaints of cough and short breath. Velcro rales were found in 50.0% (15/30) patients with IPF and no acropachy occurred. Only one patient suffered from hypoxemia. IPF presented after the joint symptoms in most patients. (2) RA-IPF patients were older than those without IPF [(65.50 +/- 9.71) vs (55.22 +/- 12.98) years, P<0.01]; Higher positivity of anti-keratin antibodies (AKA) were found in RA-IPF compared to patients without IPF (61.5% vs 35.9%, P=0.014). Furthermore, the levels of anti-cyclic citrullinated peptide (CCP) antibody were significantly higher in RA-IPF [(4.38 +/- 2.08) vs (3.20 +/- 2.12), P=0.01]. No differentiation of duration of disease, history of smoking and medicine, IgM rheumatoid factor, IgG rheumatoid factor, anti-nuclear antibody, anti-SSA antibody and levels of immunoglobins and complements were found between the two groups of RA patients with and without IPF.. The clinical symptoms of IPF in RA patients are mild and more common in older patients. AKA and anti-CCP antibody might be important antibodies associated with RA-IPF.

Topics: Aged; Antibodies; Arthritis, Rheumatoid; Female; Humans; Keratins; Male; Middle Aged; Peptides, Cyclic; Pulmonary Fibrosis; Retrospective Studies

Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first manifested in humans in November 2002. The SARS-associated coronavirus (SARS-CoV) has been identified as the causal agent, but the pathology and pathogenesis are still not quite clear.. Post-mortem lung samples from six patients who died from SARS from April to July 2003 were studied by light and electron microscopy, Masson trichromal staining and immunohistochemistry. Evidence of infection with the SARS-CoV was determined by reverse-transcription PCR (RT-PCR) , serological examination and electron microscopy.. Four of six patients had serological and RT-PCR evidence of recent infection of SARS-CoV. Morphologic changes are summarized as follows: (1) Diffuse and bilateral lung consolidation was seen in all patients (6/6) with increasing lung weight. (2) Diffuse alveolar damage was universal (6/6) with hyaline membrane formation (6/6), intra-alveolar edema/hemorrhage (6/6), fibrin deposition (6/6), pneumocyte desquamation (6/6). A marked disruption in the integrity of the alveolar epithelium was confirmed by immunostaining for the epithelial marker AE1/AE3 (6/6). (3) Type II pneumocytes, with mild hyperplasia, atypia, cytomegaly with granular amphophilic cytoplasm and intracytoplasmic lipid accumulation (5/6). (4) Giant cells in the alveoli were seen in five of 6 patients (5/6) , most of which were positive for the epithelial marker AE1/AE3 (5/6), but some cells were positive for the macrophage marker CD68(2/6). (5) A pronounced increase of macrophages were seen in the alveoli and the interstitium of the lung (6/6), which was confirmed by histological study and immunohistochemistry. (6) Haemophagocytosis was present in five of the 6 patients(5/6). (7) Lung fibrosis was seen in five patients(5/6), with alveolar septa and interstitium thickening(5/6), intraalveolar organizing exudates (6/6) and pleura thickening (4/6). Proliferation of collagen was confirmed by Masson trichromal staining, most of which was type III collagen by immunostaining. The formation of distinctive fibroblast/myofibroblast foci was seen in five patients (5/6) by light microscopy and immunochemistry. (8) Squamous metaplasia of bronchial mucosa was seen in five patients(5/6). (9) Thrombi was seen in all patients(6/6). (10) Accompanying infection was present in two patients, one was bacteria, the other was fungus. In addition, electron microscopy revealed viral particles in the cytoplasm of alveolar epithelial cells and endothelial cells corresponding to coronavirus.. Direct injury of SARS-CoV on alveolar epithelium, prominent macrophage infiltration and distinctive fibroblast/myofibroblast proliferation may play major roles in the pathogenesis of SARS.

Topics: Adult; Antibodies, Monoclonal; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Epithelium; Female; Humans; Keratins; Lung; Male; Middle Aged; Pulmonary Alveoli; Pulmonary Fibrosis; Severe Acute Respiratory Syndrome; Severe acute respiratory syndrome-related coronavirus

Cytokeratin 19 fragment (CYFRA) is a specific tumor marker in patients with lung cancer; however, it has been reported that serum CYFRA levels are elevated in some patients with nonmalignant respiratory diseases such as interstitial pulmonary fibrosis (IPF) and collagen disease-associated pulmonary fibrosis (CDPF). To investigate the serum CYFRA levels in nonmalignant respiratory diseases in detail, we studied 413 patients with respiratory diseases.. Retrospective study.. University hospital.. Four hundred thirteen patients with nonmalignant respiratory diseases and lung cancer.. Serum CYFRA levels were measured with a commercially available enzyme immunoassay kit. Immunohistochemical study was performed using monoclonal antibody Ks 19.1 (Rosch Diagnosica; Bern, Switzerland) on surgically resected or autopsied lung specimens. Gel electrophoresis and immunoblotting was performed with serum samples. In 149 patients with nonmalignant diseases except IPF and CDPF, the ratio of patients with > 3.5 ng/mL of serum CYFRA was 13.4%. In 13 of 30 patients (43.3%) with IPF and CDPF, the serum CYFRA levels were abnormally elevated. The 95th percentile serum CYFRA level of the patients with nonmalignant respiratory diseases was 6.2 ng/mL, and none of them had CYFRA levels > 20.3 ng/mL. Survival in patients with IPF and CDPF with elevated serum CYFRA levels were significantly lower than in those with normal range (p = 0.0335). Western blotting using serum from nonmalignant lung diseases and patients with lung cancer showed no apparent difference between them. An immunohistochemical study indicated CYFRA was selectively expressed in the pulmonary epithelial cells that covered the remodeling alveolar septi in nonmalignant respiratory disease.. Serum CYFRA was elevated in some nonmalignant respiratory diseases, especially in IPF and CDPF. The value of serum CYFRA would reflect the severity of lung injury in nonmalignant respiratory diseases and might be related to the prognosis in patients with IPF and CDPF.

Topics: Adult; Biomarkers, Tumor; Collagen Diseases; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Female; Humans; Immunoblotting; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Male; Pulmonary Fibrosis; Respiratory Tract Diseases; Retrospective Studies

Cytokeratin 19 (CK19) is a specific cytoskeletal structure for simple epithelia, including bronchial and alveolar epithelial cells (BAEC). Since CK19 is abundant in alveolar epithelial cells, and could be released from injured alveolar epithelium in idiopathic pulmonary fibrosis (IPF), we investigated the levels of CK19 fragments in the bronchoalveolar lavage fluids (BALF) of 16 patients with idiopathic pulmonary fibrosis (IPF) and 12 patients with sarcoidosis using enzyme-linked immunoassay. There were also 19 control subjects (10 asymptomatic smokers and nine non-smokers). BALF from the non-smokers as well as the asymptomatic smokers contained few CK19 fragments (0.2+/-0.2, 1.3+/-0.5 pg ml(-1) respectively). There were significantly high levels of CK19 in the BALF from patients with IPF (7.3+/-1.4 pg/ml; P<0.01 vs. control non-smoker). Even if the levels of CK19 were expressed as relative to the albumin concentration, significantly increased levels of CK19 fragments were noted in BALF from patients with IPF. However, these levels were not found in BALF from patients with sarcoidosis. Importantly, levels of CK19 fragment in BALF were significantly correlated to the number of neutrophils (r = 0.791, P<0.001) and eosinophils (r = 0.771, P<0.001) but not to that of macrophages or lymphocytes in BALF from IPF patients. Our results suggest the usefulness of CK19 measurement in BALF for assessing the presence of bronchiolo-alveolar epithelial injuries in idiopathic pulmonary fibrosis.

Topics: Aged; Bronchoalveolar Lavage Fluid; Eosinophils; Female; Forced Expiratory Volume; Humans; Keratins; Male; Middle Aged; Neutrophils; Pulmonary Fibrosis; Vital Capacity

There are several unsolved clinical findings in patients with idiopathic pulmonary fibrosis (IPF); (i) predominance of fibrosis in the lower lung fields, (ii) digital clubbing, and (iii) patchy distribution of pulmonary fibrosis. To explain these unsolved problems, we hypothesized that regenerated or premature bronchoepithelial cells may circulate in the blood in patients with IPF. To prove this, we performed the reverse transcriptase-polymerase chain reaction (RT-PCR) for cytokeratin 19 (CK19) and pulmonary surfactant protein A (SPA) in peripheral blood in patients with IPF and pulmonary fibrosis associated with collagen vascular disorders. In addition, 20 patients with chronic pulmonary emphysema as a disease control and 19 normal volunteers were also evaluated for the existence of circulating bronchoepithelial cells. RT-PCR analysis showed that CK19 was expressed in 12 of 38 blood samples (31.6%) of IPF and pulmonary fibrosis associated with collagen vascular disorders, seven of 20 (35.0%) blood samples of chronic pulmonary emphysema, and four of 19 (21.1%) blood samples of normal volunteers. mRNA for SPA was positive in eight of 38 (21.1%) blood samples of IPF. In contrast, SPA expressing cells were not detected in any blood samples obtained from patients with chronic pulmonary emphysema or normal volunteers. This evidence suggests that there were some circulating bronchoepithelial cells expressing mRNA for SPA in peripheral blood of patients with IPF and pulmonary fibrosis associated with collagen vascular disorders.

Topics: Adult; Aged; Aged, 80 and over; Female; Humans; Keratins; Male; Middle Aged; Pulmonary Emphysema; Pulmonary Fibrosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Staphylococcal Protein A

In this study, we hypothesize that anti-cytokeratin 18 (CK18) antibody and CK18:anti-CK18 immune complex increase in sera in patients with idiopathic pulmonary fibrosis (IPF). To prove the existence of anti-CK18 antibodies in patients' sera, bovine CK18 was stained with patients' sera using a Western blotting. In patients with IPF, anti-CK18 antibodies were clearly demonstrated in sera by Western blotting. Then, we tried to establish an enzyme-linked immunosorbent assay (ELISA) to quantify anti-CK18 antibodies and CK18:anti-CK18 immune complexes in sera of patients with IPF. Levels of anti-human CK18 antibodies in sera of patients with IPF (0.81 +/- 0.31, mean +/- SD) measured by ELISA were significantly high compared with that of normal volunteers (0.45 +/- 0.06, p < 0.01). In addition, levels of CK18:anti-CK18 antibody complexes in patients' sera (0.64 +/- 0.35, man +/- SD) significantly increased compared with those of normal subjects (0.40 +/- 0.10, p < 0.01). These results suggest that anti-CK18 antibody and its immune complex may have played a role in the process of lung injury in IPF.

Topics: Adult; Aged; Antigen-Antibody Complex; Autoantibodies; Biomarkers; Blotting, Western; Cell Division; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Middle Aged; Prognosis; Pulmonary Alveoli; Pulmonary Fibrosis; Retrospective Studies; Severity of Illness Index

It has been suggested that the humoral immune system plays a role in the pathogenesis of idiopathic pulmonary fibrosis (IPF) and pulmonary fibrosis associated with collagen vascular disorders (PF-CVD). Although circulating immune complexes in patients' sera have been suggested, none of the antigens have been characterized.. The purpose of this study is to characterize the antigen of the immune complexes in patients' sera of pulmonary fibrosis.. As we previously established that one of the antibodies against A549 cells (lung alveolar type II cells) was anti-cytokeratin 8 (CK8), we confirmed the existence of anti-CK8 antibody in patients' sera by Western immunoblot. In addition, we tried to demonstrate circulating CK8:anti-CK8 immune complexes in patients' sera by Western immunoblot. Furthermore, we established an enzyme-linked immunosorbent assay to quantitate CK8:anti-CK8 immune complexes.. In patients with pulmonary fibrosis, anti-CK8 antibodies were clearly demonstrated in sera by Western immunoblot. In addition, circulating CK8:anti-CK8 immune complexes were also clearly demonstrated by Western immunoblot. It was possible to establish ELISA to quantitate CK8:anti-CK8 immune complexes. If the cutoff value, which was determined based on the highest value of normal volunteers, was introduced, high CK8:anti-CK8 antibody complexes were demonstrated in 9 of 31 patients (29.0%) with IPF and PF-CVD.. This is the first study to clarify the antigen of the circulating immune complex in sera of patients with IPF. These results suggest that circulating CK8:anti-CK8 immune complexes may have played a role in the process of lung injury in pulmonary fibrosis.

Topics: Aged; Aged, 80 and over; Antibodies; Antigen-Antibody Complex; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Humans; Keratins; Middle Aged; Pulmonary Fibrosis

Topics: Animals; Epithelial Cells; Humans; Immunohistochemistry; Keratins; Pulmonary Alveoli; Pulmonary Fibrosis; Swine; Swine, Miniature

It has been suggested that cytokeratin 19 is expressed in regenerated bronchoepithelial cells in patients with pulmonary fibrosis, and serum cytokeratin 19 fragment is elevated in patients with pulmonary fibrosis. We hypothesized that serum antibodies to cytokeratin 19 may be formed in patients with pulmonary fibrosis. To prove the existence of anti-cytokeratin 19 antibodies in patients' sera, human recombinant cytokeratin 19 was stained with patients' sera by a Western immunoblot. Then, we tried to establish an enzyme-linked immunosorbent assay to quantitate anti-cytokeratin 19 antibody in the sera of patients with idiopathic pulmonary fibrosis (IPF) and pulmonary fibrosis associated with collagen vascular disorders (PF-CVD). We demonstrated the anti-cytokeratin 19 antibody in patient' sera by a Western immunoblot. In patients with IPF and PF-CVD, significantly high anti-cytokeratin 19 antibody was demonstrated compared with normal volunteers, patients with chronic bronchitis, and patients with pneumonia. These results suggest that anti-cytokeratin 19 antibody may have played a role in the process of lung injury in pulmonary fibrosis.

Topics: Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Autoantibodies; Blotting, Western; Collagen Diseases; Dermatomyositis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Pulmonary Fibrosis; Sjogren's Syndrome

Cytokeratin 19 fragment (CK19) levels in serum have already been documented as a useful tumour marker for lung cancer. In the present study, it was hypothesized that CK19 may be increased in the serum and epithelial lining fluid of the respiratory tract from patients with pulmonary fibrosis. CK19 was measured in the serum and bronchoalveolar lavage fluid (BALF) of patients with pulmonary fibrosis and the correlation between CK19 levels and clinical parameters evaluated. Nineteen patients diagnosed with idiopathic pulmonary fibrosis (IPF), eight with pulmonary fibrosis associated with a collagen vascular disorder (PF-CVD), seven patients with acute interstitial pneumonia (AIP), and 10 normal smokers as a control group were studied. CK19 levels in sera of patients with IPF and patients with PF-CVD were significantly increased compared to those of normal smokers. CK19 levels in sera of patients with AIP were significantly increased compared to those of other groups. CK19 values in the BALF of patients with pulmonary fibrosis were significantly elevated compared to those of normal smokers. CK19 values in sera charged according to the progression or improvement of the acute lung injury. Immunohistochemical study using pulmonary tissues obtained from patients with AIP demonstrated that the hyaline membrane and proliferating type II pneumocytes were stained by anti-human cytokeratin 19 antibody. These data demonstrated that the measurement of cytokeratin 19 fragment is a useful parameter to evaluate the activity of lung epithelial cell damage and repair.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Biomarkers; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Collagen Diseases; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Diseases, Interstitial; Male; Middle Aged; Pulmonary Fibrosis; Vascular Diseases

1. It has been suggested that CYFRA21-1, a cytokeratin subunit 19 fragment, is potentially useful for diagnosis and monitoring of lung carcinoma. However, serum levels of CYFRA21-1 are also increased in a high proportion of patients with interstitial lung disease. In this study we measured CYFRA21-1 levels in bronchoalveolar lavage fluid from 10 normal subjects, 18 patients with idiopathic pulmonary fibrosis and 14 patients with sarcoidosis, and determined whether any relationship exists between CYFRA21-1 levels in bronchoalveolar lavage fluid and clinical parameters. 2. CYFRA21-1 levels in bronchoalveolar lavage fluid were significantly higher in patients with sarcoidosis (mean value 8.3 ng/ml, P < 0.01) and idiopathic pulmonary fibrosis (42.5 ng/ml, P < 0.005) than in normal controls (1.0 ng/ml). Moreover, higher CYFRA21-1 levels in bronchoalveolar lavage fluid were found in sarcoidosis patients in radiological stage 2 or 3 than in those in stage 1. In patients with idiopathic pulmonary fibrosis, there was a significant correlation between CYFRA21-1 levels, and percentage of inflammatory cells in bronchoalveolar lavage fluid (r = 0.56, P < 0.05) and the magnitude of the alveolar--arterial oxygen pressure difference [P(A-a)O2] gradient (r = 0.66, P < 0.01). 3. Serial bronchoalveolar lavage samples were obtained from six patients with clinically active pneumonitis after they had undergone systemic corticosteroid therapy. CYFRA21-1 levels were significantly lower after these patients exhibited clinical improvement (P < 0.05). 4. These findings suggest that the level of CYFRA21-1 in bronchoalveolar lavage fluid is a useful marker for the clinical diagnosis of pneumonitis, and is also adequate for the evaluation of disease activity, especially over the course of treatment.

Topics: Adrenal Cortex Hormones; Antigens, Neoplasm; Biomarkers; Bronchoalveolar Lavage Fluid; Female; Humans; Keratin-19; Keratins; Lung Diseases, Interstitial; Male; Middle Aged; Prospective Studies; Pulmonary Fibrosis; Sarcoidosis; Statistics, Nonparametric

It has been suggested that the humoral immune system plays a role in the pathogenesis of cryptogenic fibrosing alveolitis (CFA). Although circulating autoantibodies to lung protein(s) have been suggested, none of the lung proteins have been characterised. The purpose of this study was to determine the antigen to which the serum from patients with pulmonary fibrosis reacted.. The anti-A549 cell antibody was characterised in a patient with CFA using Western immunoblotting and immunohistochemical staining of A549 cells. As we identified that one of the antibodies against A549 cells was anti-cytokeratin 8, the expression of mRNA of cytokeratin 8 in A549 cells was evaluated. In addition, we attempted to establish an enzyme linked immunosorbent assay to measure the levels of anti-cytokeratin 8 antibody in the serum of patients with CFA and pulmonary fibrosis associated with collagen vascular disorders (PF-CVD).. Initially two anti-A549 cell antibodies were detected in the serum of patients with pulmonary fibrosis, one of which was characterised as anticytokeratin 8 antibody by Western immunoblotting. We were able to establish an ELISA to measure anti-cytokeratin 8 antibody and found significantly higher levels in patients with CFA and PF-CVD than in normal volunteers, patients with sarcoidosis, pneumonia, and pulmonary emphysema.. One of the anti-A549 cell antibodies in the serum of patients with CFA was against cytokeratin 8. The serum levels of anti-cytokeratin 8 antibody were increased in patients with CFA and PF-CVD. These results suggest that anticytokeratin 8 antibody may be involved in the process of lung injury in pulmonary fibrosis.

Topics: Adult; Aged; Aged, 80 and over; Autoantibodies; Blotting, Western; Collagen Diseases; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Middle Aged; Pulmonary Emphysema; Pulmonary Fibrosis; RNA, Messenger; Sarcoidosis, Pulmonary

It is generally recognized that epithelial cytokeratins (CKs) are expressed in tissue-specific patterns and reflect differentiation, functional specialization, and pathological alterations of the cells. Differential epithelial cell types can thus be distinguished from each other by their selective expression of particular sets of CKs. To determine the characteristics of metaplastic and hyperplastic changes of alveolar-lining epithelial cells in the lungs of idiopathic pulmonary fibrosis (IPF), the expression of individual CKs was studied immunohistochemically using monospecific anti-CK monoclonal antibodies (anti-CKs 7, 8, 10, 13, 14, 16, 17, 18, 19). Biopsy specimens from 17 patients with IPF and normal lung tissues (NL) from seven patients with lung cancer were studied. In the IPF specimens, several kinds of altered epithelial cells were observed, which showed characteristic changes in CK expression compared with NL, especially CKs 8, 14, and 17. Hyperplastic type II cells expressed increased CKs 7, 8, and 19, but not CK 17; flattened or stratified squamous metaplastic cells expressed increased CKs 17 and 14, co-expressed with CKs 7, 8, and 19; bronchiolar metaplastic cells expressed increased CKs 7, 8, and 19, co-expressed with CKs 14 and 17; cuboidal metaplastic cells expressed increased CKs 7, 8, 17, and 19. The quantification of individual CKs in the tissues by enzyme-linked immunosorbent assay revealed increased expression of CKs 8, 14, and 17 in IPF lung tissues compared with NL. These results were consistent with the immunohistochemical observations. The hyperplastic and bronchiolar metaplastic phenotypes were characterized by their increased expression of simple CKs without CK alteration. The squamous metaplastic phenotype showed CK alterations, with the appearance of CKs 17 and 14. Epithelial cells are thus altered not only in shape, but possibly also in differentiation and function, with potential implications for the pathogenesis of IPF.

Topics: Aged; Enzyme-Linked Immunosorbent Assay; Epithelium; Female; Humans; Hyperplasia; Immunohistochemistry; Keratins; Male; Metaplasia; Middle Aged; Pulmonary Alveoli; Pulmonary Fibrosis

Cytokeratins (CK) are one of the main families of intermediate filaments which make up the cytoskeleton. CK19 is strongly expressed by normal simple bronchial and respiratory epithelium as well as by their malignant counterpart. Although CK19 is a part of the cytoskeleton, a soluble fragment of this polypeptide can be released and assayed in the blood as CYFRA 21-1, new sensitive and valuable marker of non small cell lung cancer. In some cases, however, discrepancies between the serum level of CYFRA 21-1 and presence of tumor and its histological type have been observed. We studied immunohistochemically tissue localization of CK19 in tumors and non invaded lung parenchyma in a series of 34 patients surgically treated due to lung cancer. CK 19 was detected in cancer cells as well as in non neoplastic epithelium covering bronchial tree and alveolar surfaces. We found a different expression of CK19 in different histological type of tumors. The most intensive expression revealed squamous cell carcinomas and adenocarcinomas. Small cell cancer revealed poor expression of CK19. In non invaded parts of the resected lungs we found the strong expression of CK19 in the cytoplasm of regenerative II type pneumocytes occurring in large quantity in the cases of interstitial lung fibrosis concomitant with some tumors. We suggest it may be a cause of unexpectedly elevated serum levels of CYFRA 19-21 in some not oncological patients or patients with small cell lung cancer.

Topics: Adenocarcinoma; Adult; Aged; Antibodies, Monoclonal; Biomarkers, Tumor; Bronchi; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cytoplasm; Epithelium; Female; Humans; Infant; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Pulmonary Alveoli; Pulmonary Fibrosis

Despite extensive research, the role of the commonly employed tumour markers in the diagnosis of lung carcinoma is yet to be clarified. The utility of a new marker, CYFRA 21-1, in the preoperative evaluation of patients with bronchogenic carcinoma was investigated. CYFRA 21-1 was determined with a radiometric assay in serum of 280 patients with lung cancer and 208 patients with various nonmalignant lung diseases. The levels of the marker were significantly higher in lung cancer patients. Among benign lung diseases, elevated CYFRA 21-1 levels were found in pulmonary fibrosis. Using a cut-off of 3.2 ng.ml-1 (95th percentile of levels obtained in benign lung disease), the total sensitivity of the marker was 48%. The best sensitivity was obtained in squamous cell lung cancer (60%). The highest values of CYFRA 21-1 were found in metastatic lung cancer, and the marker sensitivity was more elevated in stage IIIb and IV. On the other hand, 40% of patients with surgically resectable lung cancer had CYFRA 21-1 levels above the cut-off. We conclude that CYFRA 21-1 may be satisfactorily employed in the differential diagnosis between malignant and benign lung diseases in association with other clinical and radiological data.

Topics: Biomarkers, Tumor; Carcinoma, Bronchogenic; Diagnosis, Differential; Female; Humans; Immunoradiometric Assay; Keratins; Lung Diseases; Lung Neoplasms; Male; Pulmonary Fibrosis; Sensitivity and Specificity

We compared lectin staining patterns in rat and mini pig tissues of normal and fibrotic (irradiation-induced) lungs. Two lectins were studied: Dolichos biflorus (DBA) and Soybean (SBA). Both lectins strongly stained a subpopulation of alveolar macrophages. In the rat, DBA positive macrophages were a subpopulation of the SBA binding cells. In mini pig lungs, a further specific binding of DBA and SBA was observed: DBA reacted with endothelia, and SBA stained the alveolar type I cells. Double immunofluorescence experiments using a type II cell-specific cytokeratin antibody confirmed the selective reactivity of SBA with type I cells, which was also present in fibrotic areas with epithelial cell proliferation.

Topics: Animals; Binding Sites; Epithelial Cells; Epithelium; Female; Fluorescent Antibody Technique; Glycine max; Keratins; Lectins; Lung; Macrophages, Alveolar; Male; Plant Lectins; Pulmonary Alveoli; Pulmonary Fibrosis; Rats; Rats, Inbred F344; Soybean Proteins; Swine; Swine, Miniature

The expression of pan-cadherin was investigated immunohistochemically in normal and irradiated mini pig lungs. Using a double labelling immunofluorescence technique, pan-cadherin expression was also compared with that of cytokeratin 18, which is selectively present in type II cells. In untreated animals pan-cadherin was detected in type I pneumocytes, in alveolar macrophages and in endothelial cells of larger blood vessels. Radiation caused increased pan-cadherin immunoreactivity of bronchial epithelial cells and interstitial cells. In addition, pan-cadherin expression was induced in type II pneumocytes. These changes in the distribution of epithelial cadherin molecules may implicate a role of cadherins for epithelial remodeling in response to radiation-induced lung injury.

Topics: Animals; Bronchi; Cadherins; Endothelium, Vascular; Epithelium; Fluorescent Antibody Technique; Keratins; Lung; Macrophages, Alveolar; Pulmonary Alveoli; Pulmonary Fibrosis; Swine; Swine, Miniature; Tissue Fixation

The majority of fibroblasts in alveolar septa are characterized by the presence of cytoplasmic bundles of microfilaments that contain cytoplasmic actin isoforms; these cells have been named contractile interstitial cells or V-type myofibroblasts. In the rat, they express desmin as intermediate filament protein. In this study, we explored the possibility that modulation and replication of such septal fibroblasts result in the appearance of alpha-smooth muscle (alpha-SM) actin-positive myofibroblasts, typical of lung fibrosis. Experimental pulmonary fibrosis was produced by a unique intratracheal instillation of bleomycin to 28 rats. Eight additional rats used as controls received the equivalent volume of saline. Paraffin and frozen sections of lungs were examined at days 1, 3, 5 and 7 after treatment. Microfilaments and intermediate filaments were stained using antibodies against total actin, alpha-SM actin, desmin, vimentin, keratin, and SM myosin. Electron microscopic labeling of desmin and alpha-SM actin using immunogold technique was done on Lowicryl K4M resin-embedded specimens. alpha-SM actin appeared in desmin-positive alveolar fibroblasts as early as 24 hours after intratracheal bleomycin instillation; the modulation of alpha-SM actin in these cells was preceded by a lymphomonocytic infiltration of alveolar septa. Twenty-four hours to 3 days after bleomycin administration, a proliferation of alveolar myofibroblasts occurred. Fibrosis with laying down of collagen fibers took place after the above mentioned cellular modifications. Our results support the view that septal fibroblastic cells can modulate into typical alpha-SM actin-containing myofibroblasts during experimental bleomycin-induced pulmonary fibrosis. In such a modulation a possible role of cytokines, particularly of transforming growth factor-beta, is considered.

Topics: Actin Cytoskeleton; Actins; Animals; Antibodies, Monoclonal; Bleomycin; Desmin; Female; Fibroblasts; Fluorescent Antibody Technique; Immunohistochemistry; Intermediate Filaments; Isomerism; Keratins; Microscopy, Electron; Myosins; Pulmonary Alveoli; Pulmonary Fibrosis; Rats; Rats, Wistar; Vimentin

The expression of cytokeratins, desmoplakin and vimentin has been studied immunohistochemically in the rat lung injured by x-irradiation using 14 well characterized monoclonal antibodies. A time-dependent relationship between the cytokeratin expression pattern and the morphological alterations observed was apparent. A cytokeratin 8 and 18 expression in normally cytokeratectable even at 3-6 h after irradiation. Between 14 days and 2 months, a remarkable heterogeneity in the epithelial cell cytokeratin pattern and an increasing immunoreaction for desmoplakin was found. In terminal bronchial epithelial cells, a heterogeneous CK8, 18 and 19 staining and a neoexpression of cytokeratins 4 and 7 was detected. Finally, peribronchiolar and vascular smooth muscle cells were cytokeratin-positive. At 6 months after irradiation, cytokeratin 13 and vimentin were focally present in bronchial epithelial cells and atypical type I and II pneumocytes as well as scattered epithelioid cell complexes were noted. During the course of injury, a loss of type III alveolar epithelial cells was found, which was characterized in the rat by a specific globular cytokeratin pattern and restricted immunoreactivity with cytokeratin-specific antibodies. These results show that the expression pattern of cytokeratins is a sensitive marker in monitoring epithelial alterations during lung injury.

Topics: Animals; Bronchi; Cytoskeletal Proteins; Desmoplakins; Epithelium; Fluorescent Antibody Technique; Keratins; Lung; Muscle, Smooth, Vascular; Pulmonary Alveoli; Pulmonary Fibrosis; Radiation Injuries; Rats; Rats, Inbred F344; Vimentin

Interstitial pneumonia unrelated to Pneumocystis carinii or other infections was observed histopathologically in 5 of 25 rhesus monkeys infected with simian immunodeficiency virus (SIV). The predominant lesion was lymphocytic infiltration of interalveolar septa and hyperplasia of peribronchial and perivascular lymphoid tissue. Immunohistochemical staining using a panel of antibodies against human T and B lymphocytes, macrophages, and immunoglobulins showed that peribronchial aggregates and interstitial infiltrates were predominantly B cells, whereas perivascular masses consisted mainly of T cells. One animal with a primary B-cell lymphoma of the spinal cord had secondary plasmacytoid lymphomatous nodules throughout the lung which were accompanied locally by reactive B-cell lymphoid follicles. Another animal also had large areas of diffuse alveolar fibrosis and epithelial metaplasia to a bronchiolar type. In two monkeys, branches of the pulmonary arteries showed intimal proliferation and organizing occlusive thrombi, some of which were mineralized.

Topics: Animals; B-Lymphocytes; Immunoglobulin Heavy Chains; Immunoglobulin Light Chains; Immunohistochemistry; Keratins; Lung; Macaca mulatta; Macrophages; Pulmonary Fibrosis; Simian Acquired Immunodeficiency Syndrome; T-Lymphocytes

Sequential histologic, ultrastructural, immunohistochemical and morphometric studies were made of the evolutional changes of metaplastic and regenerating alveolar epithelial cells in monkeys from 3 days to 8 weeks after paraquat administration. In the early proliferative phase, many alveoli were lined by single-layered and stratified squamous epithelium and bronchiolized epithelium (i.e., presumably derived from bronchi and bronchioles). The regenerating epithelial cells had well developed bundles of actin-like filaments, which were arranged parallel to the basal surfaces of the cells and were associated with zonulae adherentes; these cells also had intermediate filaments and some desmosomes, but lacked basement membranes, hemidesmosomes and anchoring fibrils. They covered either denuded, wavy and disrupted original epithelial basement membranes or areas of developing intraalveolar fibrosis. In zones of squamous epithelial cell metaplasia associated with intraalveolar fibrosis, fibronexus-like structures appeared to be responsible for the initial adhesion of the cells to the underlying connective tissue. In later phases, single-layered and stratified squamous epithelial cells disappeared, and only bronchiolized epithelial cells, with hemidesmosomes and anchoring fibrils on their basal surfaces, were found in fibrotic alveoli. Although bronchiolized and squamous metaplastic epithelial cells are generally thought to be formed as late events in pulmonary damage, such cells play an important role in early, temporary repair of damaged alveoli.

Topics: Animals; Epithelium; Immunohistochemistry; Keratins; Macaca fascicularis; Metaplasia; Microscopy, Electron; Paraquat; Pulmonary Alveoli; Pulmonary Fibrosis; Time Factors

There is substantial tissue reorganization in the interstitium in pulmonary fibrosis. In order to determine the origin of cells in the interstitium of normal or fibrotic rat lung, we stained lung tissue immunohistochemically with antiserums directed against intermediate filament proteins specific for cells of mesenchymal or muscle origin. Vimentin, specific for mesenchymal cells, was isolated from cultured 3T3 fibroblasts, and desmin, specific for muscle cells, was isolated from chicken gizzard and antiserums prepared in guinea pigs. Paraffin-embedded sections of normal or fibrotic rat lung from rats 28 days after treatment with bleomycin sulfate were reacted with each antiserum and stained by the peroxidase-antiperoxidase technique. Antivimentin antiserum stained discrete interstitial cells of both control and fibrotic lung. Smooth muscle surrounding large airways and vascular smooth muscle was not significantly stained with this antiserum but was heavily stained with antidesmin. However, antidesmin did not stain cells in the interstitium of normal or fibrotic rat lung. These results suggest that most cells in the normal as well as in the fibrotic interstitium, including contractile interstitial cells, are probably of mesenchymal (fibroblastic) rather than of smooth muscle origin.

Topics: Actins; Animals; Bleomycin; Desmin; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Fluorescent Antibody Technique; Immune Sera; Immunoenzyme Techniques; Keratins; Lung; Male; Muscle, Smooth; Pulmonary Fibrosis; Rats; Rats, Inbred F344; Vimentin