scholarly journals Bone Marrow Cells Expressing Clara Cell Secretory Protein Increase Epithelial Repair After Ablation of Pulmonary Clara Cells

2013 ◽  
Vol 21 (6) ◽  
pp. 1251-1258 ◽  
Author(s):  
Martha L Bustos ◽  
Marco Mura ◽  
Paula Marcus ◽  
David Hwang ◽  
Olga Ludkovski ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Secretory Protein ◽  
Clara Cell ◽  
Clara Cells ◽  
Epithelial Repair ◽  
Clara Cell Secretory Protein ◽  
Marrow Cells

Clara cell secretory protein deficiency increases oxidant stress response in conducting airways

1998 ◽  
Vol 275 (2) ◽  
pp. L348-L356 ◽  
Author(s):  
Gregory W. Mango ◽  
Carl J. Johnston ◽  
Susan D. Reynolds ◽  
Jacob N. Finkelstein ◽  
Charles G. Plopper ◽  
...  
Keyword(s):  
Stress Response ◽  
Mrna Expression ◽  
Molecular Basis ◽  
Oxidant Stress ◽  
Secretory Protein ◽  
Protective Role ◽  
Clara Cell ◽  
Clara Cells ◽  
Clara Cell Secretory Protein ◽  
Oxidant Sensitivity

Little is known about the molecular basis for differential pulmonary oxidant sensitivity observed between genetically disparate members of the same species. We have generated mice that are deficient in Clara cell secretory protein (CCSP −/−) and that exhibit an oxidant-sensitive phenotype. We characterized the kinetics and distribution of altered stress-response [interleukin-6 (IL-6) and metallothionein (MT)] and epithelial cell-specific [cytochrome P-450 2F2 (CYP2F2)] gene expression to further understand the cellular and molecular basis for altered oxidant sensitivity in 129 strain CCSP −/− mice. Increases in IL-6 and MT mRNA abundance were detected by 2 h of exposure to 1 part/million ozone and preceded reductions in Clara cell CYP2F2 mRNA expression. Despite being qualitatively similar, increases in IL-6 and MT mRNA expression were enhanced in CCSP −/− mice with respect to coexposed 129 strain wild-type mice. Increased MT mRNA expression, indicative of the stress response, localized to the airway epithelium, surrounding mesenchyme, and endothelium of blood vessels. These results demonstrate a protective role for Clara cells and their secretions and indicate potential genetic mechanisms that may influence susceptibility to oxidant stress.

Homing and Engraftment of Bone Marrow Derived Cells in Irradiated Mouse Lungs.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4600-4600
Author(s):  
Umar Salimi ◽  
Malolan Rajagopalan ◽  
Jean-Claude Rwigema ◽  
Tracy Dixon ◽  
Hongmei Shen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Bone Marrow Cells ◽  
Conflicts Of Interest ◽  
Subcutaneous Administration ◽  
Clara Cell ◽  
Cell Potential ◽  
Hematopoietic Stem ◽  
Osmotic Pumps ◽  
Marrow Cells

Abstract Abstract 4600 To determine the degree to which murine bone marrow migrates to and can repopulate damaged lung, we studied lung stem cell expression of the Clara Cell Specific Protein (CCSP). Several different models have been used to deplete the lung stem cells. We used CCtk mice which expresses the HSV-tk under control of the CCSP promoter. Subcutaneous administration of gancicylovir (50 mg/ml) over a 7 day period using Alzet osmotic pumps results in the reduction of CCSP RNA and CCSP positive cells. To determine if a reduced dose of gancicylovir reduced CCSP cells to a nonfatal degree and opened a stem cell niche for homing of hematopoietic cells, CCtk mice had osmotic pumps placed subcutaneously containing a lower dose of 25 mg/ml of gancicylovir. The lungs were explanted at serial times up to 7 days after pump placement. RNA was extracted and real time polymerase chain reaction (RT-PCR) using primers specific for CCSP was used to quantitate CCSP expression. CCtk mice treated with gancicylovir showed a 70% reduction of lung CCSP expression. CCtk mice implanted with osmotic pumps containing 25 mg/ml gangcicylovir and injected intravenously with bone marrow from sex-mismatched GFP+ transgenic FVB male mouse 2 days later were followed for thirty days. The mice were sacrificed, lungs removed, single cell suspensions prepared, stained with a PE labeled anti-CD45 monoclonal antibody and analyzed by flow cytometry for GFP positive, CD45 negative cells. No GFP+, CCSP+ cells cells were detected. Injection of FVB mice with naphthalene (which kills differentiated but not stem CCSP+ cells) at 200 mg/kg intraperitoneally resulted in an 87% decrease in CCSP expression. When injected with bone marrow cells from GFP+ transgenic mice at serial times to 7 days and sacrificed at day 30, again no GFP+ CCSP+ cells were identified. In a third attempt to remove additional CCSP cells and prepare sites for homing of marrow cells, FVB mice were treated with naphthalene (200 mg/kg) followed by 20 Gy irradiation to the lungs 48 hr later, then injected with GFP+ bone marrow, 24 hr later. The mice were sacrificed 30 days later, and examined for GFP+, CD45- cells. Low numbers of GFP+ CCSP+ cells were detected. The data indicates a low baseline level of hematopoietic stem cell potential to home and differentiate to CCSP positive lung cells. Supported by NIH-2R01CA119927-08A1. Disclosures: No relevant conflicts of interest to declare.

Secretory product expression during Clara cell differentiation in the rabbit and rat

1993 ◽  
Vol 264 (6) ◽  
pp. L543-L552 ◽  
Author(s):  
W. V. Cardoso ◽  
L. G. Stewart ◽  
K. E. Pinkerton ◽  
C. Ji ◽  
G. E. Hook ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Secretory Granules ◽  
Close Association ◽  
Secretory Protein ◽  
Clara Cell ◽  
Secretory Proteins ◽  
Secretory Product ◽  
Secretory Function ◽  
Clara Cells ◽  
Clara Cell Secretory Protein

One function of the nonciliated (Clara) cells of bronchiolar epithelium is to synthesize, store, and release small-molecular-mass (6–12 kDa) secretory proteins or Clara cell secretory protein (CCSP). This study compares the emergence of this secretory function during Clara cell differentiation in rabbits and rats. Lungs of fetal and postnatal animals were evaluated by ultrastructural morphometry and immunohistochemistry. Secretory granules were rarely seen in perinatal animals and increased to adult levels of abundance earlier in rats (1 wk postnatal) than in rabbits (3–4 wk). In contrast, rough endoplasmic reticulum was abundant in perinatal animals and decreased with age. Antibodies raised against CCSP revealed little CCSP in fetal animals; however, after birth CCSP increased to adult levels earlier in rats (1 wk postnatal) than in rabbits (3 wk). We conclude that the maturation of Clara cell secretory function 1) occurs postnatally, 2) involves a decrease in biosynthetic organelles, 3) shows close association between CCSP expression and secretory granule abundance, and 4) varies by species in timing and cellular abundance of biosynthetic machinery.

Targeted cell replacement with bone marrow cells for airway epithelial regeneration

2007 ◽  
Vol 293 (3) ◽  
pp. L740-L752 ◽  
Author(s):  
Amy P. Wong ◽  
Andre E. Dutly ◽  
Adrian Sacher ◽  
Haeyul Lee ◽  
David M. Hwang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Targeted Delivery ◽  
Fluorescent Protein ◽  
Bone Marrow Cells ◽  
Conditioning Regimen ◽  
Clara Cell ◽  
Epithelial Regeneration ◽  
Airway Injury ◽  
Over Time ◽  
Marrow Cells

It has been suggested that some adult bone marrow cells (BMC) can localize to the lung and develop tissue-specific characteristics including those of pulmonary epithelial cells. Here, we show that the combination of mild airway injury (naphthalene-induced) as a conditioning regimen to direct the site of BMC localization and transtracheal delivery of short-term cultured BMC enhances airway localization and adoption of an epithelial-like phenotype. Confocal analysis of airway and alveolar-localized BMC (fluorescently labeled) with epithelial markers shows expression of the pulmonary epithelial proteins, Clara cell secretory protein, and surfactant protein C. To confirm epithelial gene expression by BMC, we generated transgenic mice expressing green fluorescent protein (GFP) driven by the epithelial-specific cytokeratin-18 promoter and injected BMC from these mice transtracheally into wild-type recipients after naphthalene-induced airway injury. BMC retention in the lung was observed for at least 120 days following cell delivery with increasing GFP transgene expression over time. Some BMC cultured in vitro over time also expressed GFP transgene, suggesting epithelial transdifferentiation of the BMC. The results indicate that targeted delivery of BMC can promote airway regeneration.

Clara cell secretory protein gene expression in bronchiolar epithelium

1992 ◽  
Vol 262 (4) ◽  
pp. L399-L404 ◽  
Author(s):  
B. P. Hackett ◽  
N. Shimizu ◽  
J. D. Gitlin
Keyword(s):  
Gene Expression ◽  
Lung Tissue ◽  
Prolonged Exposure ◽  
Secretory Protein ◽  
Clara Cell ◽  
Fetal Life ◽  
Clara Cells ◽  
Adult Rats ◽  
Clara Cell Secretory Protein ◽  
Bronchiolar Epithelium

To determine the mechanisms of Clara cell secretory protein (CCSP) gene expression, a cDNA clone was isolated and used in RNA blot analysis. A single 600 bp CCSP specific transcript was detected in the developing rat lung on fetal day 18. This transcript increased in abundance during late fetal life such that adult levels were attained within 2 wk postpartum. CCSP gene expression was tissue specific, being confined to lung and trachea at all developmental stages. The abundance of CCSP mRNA in lung tissue was unchanged after the induction of lung injury in adult rats either with lipopolysaccharide or prolonged exposure to hyperoxia. In situ hybridization of lung tissue revealed that CCSP gene expression is localized to the nonciliated epithelial (Clara) cells of the bronchiolar epithelium throughout fetal and postnatal development. Taken together the results indicate that the gene for CCSP is abundantly expressed in a cell-specific fashion in the lung and suggest that analysis of such expression will be useful in elucidating the role of Clara cells in the growth and development of the bronchiolar epithelium.

scholarly journals Maternal exposure to environmental tobacco smoke alters Clara cell secretory protein expression in fetal rat lung

1998 ◽  
Vol 275 (5) ◽  
pp. L870-L876 ◽  
Author(s):  
Chun-Mei Ji ◽  
Fred H. Royce ◽  
Uyen Truong ◽  
Charles G. Plopper ◽  
Gurmukh Singh ◽  
...  
Keyword(s):  
Secretory Protein ◽  
Maternal Exposure ◽  
Clara Cell ◽  
Developmental Expression ◽  
Clara Cells ◽  
Rat Lung ◽  
Differentiation Markers ◽  
Clara Cell Secretory Protein ◽  
Fetal Rat ◽  
Fetal Rat Lung

We have previously demonstrated that aged and diluted sidestream cigarette smoke (ADSS) alters the development of bronchiolar epithelial cells in postnatal animals (C. M. Ji, C. G. Plopper, H. P. Witschi, and K. E. Pinkerton. Am. J. Respir. Cell Mol. Biol. 11: 312–320, 1994). This study was designed to examine the effects of maternal exposure to ADSS on the development of fetal Clara cells in rats with Clara cell 10-kDa protein (CC10; also designated Clara cell secretory protein) and CC10 mRNA as differentiation markers. Immunohistochemistry, Northern blots, and in situ hybridization were used to determine the abundance and distribution of CC10 at gestational days 14, 18, and 21. CC10 and CC10 mRNA were absent at gestational day 14 but were detectable at gestational day 18 and further increased by gestational day 21. Maternal exposure to ADSS was found to significantly increase fetal expression of CC10 and CC10 mRNA by gestational day 21 but not by gestational day 14 or 18. These findings demonstrate that in utero exposure to ADSS alters the normal developmental expression of CC10 in the fetal rat lung.

scholarly journals Purification, characterization and proteinase-inhibitory activity of a Clara-cell secretory protein from the pulmonary extracellular lining of rabbits

1987 ◽  
Vol 248 (2) ◽  
pp. 337-344 ◽  
Author(s):  
R P Gupta ◽  
S E Patton ◽  
A M Jetten ◽  
G E R Hook
Keyword(s):  
Inhibitory Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Secretory Protein ◽  
Clara Cell ◽  
Gel Chromatography ◽  
Secretory Product ◽  
Clara Cells ◽  
Rabbit Lung ◽  
Clara Cell Secretory Protein ◽  
Reducing Conditions

A low-Mr Clara-cell secretory protein, CCSP, previously shown to be a major secretory product of Clara cells, was isolated from rabbit lung lavage effluents. CCSP accounted for 4.4 +/- 0.5% of the protein in the soluble phase of cell- and surfactant-free pulmonary lavage effluents (LE). Purification of this protein from LE was achieved in two steps. First, the LE was acidified with HCIO4 and, secondly, CCSP was isolated by gel-exclusion chromatography on Sephadex G-50. Purified CCSP was homogeneous by SDS/polyacrylamide-gel electrophoresis (PAGE), consisting of a single major isoform with a pI of 6.0. The Mr of CCSP was 5800 according to SDS/PAGE under reducing conditions and 12,600 under non-reducing conditions. However, by gel chromatography the Mr of the protein under non-reducing conditions was 12,400 and under reducing conditions it increased to 15,000. The discrepancy obtained by using these two techniques was attributed to anomalous electrophoretic mobilities of the protein in its reduced state. The molecule contained three half-cystine residues, but no free thiol groups, and tryptophan was not detectable. The first seven N-terminal amino acid residues were Gly-Ile-Xaa-Pro-Arg-Phe-Ala-. The third residue was not identified. CCSP showed inhibitory activity against the thiol proteinase papain (50% inhibition at 4 microM-CCSP), but only weak activities against human polymorphonuclear-leucocyte elastase, and bovine trypsin. The molecule was not digested by, and did not complex with, trypsin. CCSP was immunochemically different from surfactant apoprotein B (Mr 10,000) present in rabbit lung surfactant. This study is the first partial characterization of the major secretory protein of rabbit lung Clara cells.

scholarly journals Exacerbation of Lung Radiation Injury by Viral Infection: The Role of Clara Cells and Clara Cell Secretory Protein

2013 ◽  
Vol 179 (6) ◽  
pp. 617-629 ◽  
Author(s):  
Casey M. Manning ◽  
Carl J. Johnston ◽  
Eric Hernady ◽  
Jen-nie H. Miller ◽  
Christina K. Reed ◽  
...  
Keyword(s):  
Viral Infection ◽  
Radiation Injury ◽  
Secretory Protein ◽  
Clara Cell ◽  
Clara Cells ◽  
Clara Cell Secretory Protein
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