Immunohistological detection of the ? subunit of prolyl 4-hydroxylase in rat and mini pig lungs with radiation-induced pulmonary fibrosis

Kasper, Michael; Sd, Fuller; Schuh, Dieter; Müller, Martin

doi:10.1007/bf00197555

Cited by 14 publications

(10 citation statements)

References 25 publications

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“…The immunohistochemical procedures have been described previously [16,18]. In brief, all sections (no enzymic or other pretreatment, consecutive serial sections) were treated with 0.3% hydrogen peroxide in methanol for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Loss of immunoreactivity for RTI40, a type I cell-specific protein in the alveolar epithelium of rat lungs with bleomycin-induced fibrosis

Koslowski

Dobbs²,

Wenzel³

et al. 1998

Eur Respir J

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The rat alveolar epithelium comprises three different epithelial cell types: the large squamous type I cells, the cuboidal type II pneumocytes and the bottle-shaped alveolar brush (type III) cells. Various biochemical markers have been used to distinguish type I from type II cells. Most of these markers are not unique for either cell type, but are helpful in defining the different phenotypes of type I and type II cells. Type II pneumocytes may be distinguished from type I cells by their cytokeratin pattern, by the presence of certain enzymes (e.g. alkaline phosphatase, carbonic anhydrase II, carbonyl reductase, protein disulphide isomerase and catalase), by the presence of surfactant apoproteins (SP-A, SP-B, SP-C and SP-D) and by different lectin binding (Maclura pomifera agglutinin (MPA)) (reviewed in [1,2]). Alveolar brush (type III) cells lack surfactant proteins and are selectively decorated with cytokeratin 18-and villin-specific antibodies [3,4]. Type I pneumocytes, in contrast to type II pneumocytes, bind various lectins (Lycopersicon esculentum agglutinin (LEA), Erythrina cristagalli agglutinin (ECA) or Bauhinia purpurea agglutinin (BPA)) [5][6][7], express the leukocytic intercellular adhesion molecule-1 (ICAM-1) [8,9] as well as caveolin, a structural protein of caveolae, and antigens recognized by other type I cell-specific monoclonal antibodies [10][11][12]. One of the type I cell-specific monoclonal antibodies recognizes a 40-42 kDa apical plasma membrane integral protein (RTI40) that was detected in the alveolar fluid of injured lungs [13]. Another type I cellspecific monoclonal antibody is .In the present study, the distribution of RTI40, MEP-1 antigen and the BPA lectin-binding glycoprotein was investigated in bleomycin-injured rat lungs and compared with that in normal adult and foetal lung tissue. The binding to the alveolar surface has been shown to be stable even under conditions of lung injury, compared with the immunoreactions of antibodies against type I cell-specific antigens [7,12]. Materials and methodsMale Wistar rats weighing 350-400 g were given a single dose of 7 units of bleomycin sulphate in phosphatebuffered saline (PBS)·kg body weight -1 by intratracheal instillation. Controls received PBS alone. Different animals were killed at 7, 14, 21, 28, 35 and 42 days after bleomycin application and the lungs were prepared and lavaged via the trachea with PBS [17].Sections of formalin-fixed, paraffin-embedded lung tissue (normal, n=5; injured tissue, 4 weeks after bleomycin administration, n=12) were dewaxed. In addition, sections These results suggest RTI40 as a tool for the evaluation of alveolar epithelial type I cell behaviour during re-epithelialization processes.

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Section: Methodsmentioning

confidence: 99%

Loss of immunoreactivity for RTI40, a type I cell-specific protein in the alveolar epithelium of rat lungs with bleomycin-induced fibrosis

Koslowski

Dobbs²,

Wenzel³

et al. 1998

Eur Respir J

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“…Five micrometer paraYn sections cut from formalinWxed material were deparaVinized and pretreated with microwave radiation as previously described (Kasper et al 1994b), washed in phosphate-buVered saline (PBS), and then incubated with normal horse serum followed by incubation with the primary antibody. All steps were performed at room temperature (RT).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Dual-immunohistochemistry provides little evidence for epithelial–mesenchymal transition in pulmonary fibrosis

Yamada

Kuwano

Maeyama

et al. 2008

Histochem Cell Biol

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Epithelial-mesenchymal transition (EMT) has been considered to be involved in organ fibrogenesis. However, there is few direct evidence of this process in the pathophysiology of pulmonary fibrosis in vivo. Therefore, we tried to verify the involvement of this process in the development of pulmonary fibrosis. Since the co-expressions of epithelial and mesenchymal markers are thought to be a marker of EMT, we performed dual-immuunohistochemistry to assess the co-expressions of these proteins in lung tissues from bleomycin-induced pulmonary fibrosis in mice, and from patients with idiopathic pulmonary fibrosis, and nonspecific interstitial pneumonia. Double positive cells for epithelial markers including E-cadherin, T1alpha, or aquaporin 5, and a mesenchymal markers alpha-smooth muscle actin or vimentin were not found in bleomycin-induced pulmonary fibrosis in mice. Double positive cells for E-cadherin, ICAM-1, LEA, CD44v9, or SP-A and alpha-smooth muscle actin or vimentin were not found in lung tissues from normal lung parenchyma, idiopathic pulmonary fibrosis and nonspecific interstitial pneumonia. These results offer at least two possibilities. One is that EMT does not occur in IPF or bleomycin-induced pulmonary fibrosis in mice. Another is that EMT may occur in pulmonary fibrosis but the time during this transition in which cells express detectable levels of epithelial and mesenchymal markers is too small to be detected by double immunohistochemistry.

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“…For the indirect immunoperoxidase technique, dewaxed paraffin sections were microwaved as described previously [22] and incubated with primary polyclonal rabbit antisera for 1 h at 37uC. The following antisera were used: rabbit antihuman cathepsin D (Dianova, Hamburg, Germany; 1:20 dilution); rabbit antirat cathepsin B (provided by H. Kirschke, University of Turku, Finland; 1:800 dilution); and rabbit antirat cathepsin H (H. Kirschke; 1:1,000 dilution).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Cathepsins in bleomycin-induced lung injury in rat

Koslowski¹,

Knoch²,

Kuhlisch³

et al. 2003

Eur Respir J

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Cathepsins in bleomycin-induced lung injury in rat. R. Koslowski, K. Knoch, E. Kuhlisch, D. Seidel, M. Kasper. #ERS Journals Ltd 2003. ABSTRACT: Endogenous inhibitors tightly control the activity of proteinases in the extracellular space. Proteinase/antiproteinase imbalance may be caused by predominance of proteinases, resulting in severe tissue damage or abundance of proteinase inhibitors, leading to a shift in the balance of synthesis and degradation of extracellular matrix proteins and accumulation of these matrix components. Lung fibrosis is characterised by accumulation of fibrous matrix proteins in the alveolar interstitium.The activity of cathepsin D and amounts of cathepsins D and B in bleomycin-injured rat lung tissue and alveolar macrophages were examined. In addition, the activities of cathepsins and cysteine proteinase inhibitors (CPIs) in bronchoalveolar lavage fluid (BALF) were determined.No cathepsin but high CPI activity and large amounts of procathepsin B were detected in the BALF. In the alveolar lumen, the disturbed proteinase/antiproteinase balance for cysteine proteinases was clearly dominated by CPIs. In alveolar macrophages, the main source of increased cathepsin levels, large changes in cathepsin B and D content were observed during the inflammatory phase, corresponding to the occurrence of procathepsin B in BALF. With the end of the phase of tissue remodelling, imbalances in cathepsin and CPI activities were largely eliminated. Immunoblot data, revealing an increase in cathepsin D levels in myofibroblast-like cells compared to fibroblasts and in resting fibroblasts compared to proliferating cells, implicate this proteinase in the differentiation and conversion processes occurring at the beginning of the fibrotic phase of lung injury.The results show that cathepsin amounts and activities are increased transiently in lung tissue during regeneration processes in bleomycin-induced lung injury. Imbalances of cathepsin and cysteine proteinase inhibitors activities are also a phenomenon of the phase of tissue remodelling initiated by lung injury.

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Immunohistological detection of the ? subunit of prolyl 4-hydroxylase in rat and mini pig lungs with radiation-induced pulmonary fibrosis

Cited by 14 publications

References 25 publications

Loss of immunoreactivity for RTI40, a type I cell-specific protein in the alveolar epithelium of rat lungs with bleomycin-induced fibrosis

Loss of immunoreactivity for RTI40, a type I cell-specific protein in the alveolar epithelium of rat lungs with bleomycin-induced fibrosis

Dual-immunohistochemistry provides little evidence for epithelial–mesenchymal transition in pulmonary fibrosis

Cathepsins in bleomycin-induced lung injury in rat

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