Lung fibroblasts produce chemotactic factors for bronchial epithelial cells

1989 ◽  
Vol 257 (2) ◽  
pp. L71-L79 ◽  
Author(s):  
S. Shoji ◽  
K. A. Rickard ◽  
R. F. Ertl ◽  
J. Linder ◽  
S. I. Rennard
Keyword(s):  
Epithelial Cells ◽  
Tissue Repair ◽  
Bronchial Epithelial Cell ◽  
Bronchial Epithelial Cells ◽  
Chemotactic Activity ◽  
Lung Fibroblasts ◽  
Molecular Weight Range ◽  
Bronchial Epithelial ◽  
Chemotactic Factors ◽  
Two Peaks

The interaction between the epithelial cells and the subjacent mesenchymal cells in the airway is thought to play a major role during tissue repair and morphogenesis. To evaluate this interaction, we cultured human lung fibroblasts and bovine bronchial epithelial cells and determined that fibroblast-conditioned medium has chemotactic activity for bronchial epithelial cells. This activity was nondialyzable, heat labile, pepsin labile, acid stable, lipid inextractable, and eluted from Sephadex G-150 column chromatography in the high-molecular-weight range. DEAE-Sephacyl ion exchange and gelatin-Sepharose affinity chromatography revealed two peaks containing chemotactic activity, one of which may be fibronectin, since it binds to gelatin, reacts in a specific immunoassay, and is inhibited of chemotactic activity by anti-fibronectin antiserum, and another of which does not appear to be fibronectin, since it does not bind to gelatin nor react in the immunoassay. Thus lung fibroblasts can produce at least two chemotactic factors for bronchial epithelial cells that may play a role during lung tissue repair and morphogenesis by modulating bronchial epithelial cell migration.

Bronchial epithelial cells release chemoattractant activity for monocytes

1989 ◽  
Vol 257 (2) ◽  
pp. L130-L136 ◽  
Author(s):  
S. Koyama ◽  
S. I. Rennard ◽  
S. Shoji ◽  
D. Romberger ◽  
J. Linder ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Epithelial Cells ◽  
Bronchial Epithelial Cells ◽  
Chemotactic Activity ◽  
High Power Field ◽  
Local Immunity ◽  
Bronchial Epithelial ◽  
Cellular Environment ◽  
Weight Protein ◽  
Chemotactic Factors

Lung macrophages are monocyte-derived cells that play a central and essential part in local immunity. Because the bronchial epithelial cells that line the airway can modulate their local cellular environment by releasing chemotactic factors for neutrophils, lymphocytes, and fibroblasts, we postulated that the bronchial epithelial cells might release chemotactic activity for monocytes. To test this hypothesis, bovine bronchial epithelial cells were isolated and cultured. The supernatant fluids were collected at 12, 24, 36, 48, 72, 96, and 120 h and evaluated for monocyte chemotactic activity, using a blind-well chamber technique. The supernatant fluids possessed significantly greater chemotactic activity than medium alone, with optimal migration contained in supernatant fluids harvested at 72 h (5.8 +/- 2.3 vs. 39.8 +/- 2.8 cells/high-power field, P less than 0.001). Partial characterization of the released monocyte chemotactic activity revealed that the cells released a low-molecular-weight lipid-soluble chemotactic factor after 24 h in culture, but in contrast, a high-molecular-weight protein chemokinetic factor was released after 72 h in culture. These findings suggest that bronchial epithelial cells may release chemotactic activity for, and thus may modulate the recruitment of, monocytes into bronchial passages.

Bronchial epithelial cells release chemotactic activity for lymphocytes

1989 ◽  
Vol 257 (2) ◽  
pp. L109-L115
Author(s):  
R. A. Robbins ◽  
S. Shoji ◽  
J. Linder ◽  
G. L. Gossman ◽  
L. A. Allington ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
B Lymphocytes ◽  
High Power ◽  
Bronchial Epithelial Cells ◽  
Chemotactic Activity ◽  
T Helper ◽  
High Power Field ◽  
Bronchial Epithelial ◽  
T Helper Lymphocytes ◽  
Chemotactic Factors

Lymphocytes can frequently be observed in association with bronchial tissues. One mechanism that might account for this association is that bronchial epithelial cells might release chemotactic factors for lymphocytes. To test this hypothesis, bovine bronchial epithelial cells were cultured in serum-free media, and the supernatant fluids were harvested and evaluated for lymphocyte chemotactic activity using a blind-well chamber technique. Media alone attracted few lymphocytes (12 +/- 2 cells/high power field), but in contrast, there was a significant increase in the number of cells attracted by supernatant fluids obtained from bronchial epithelial cell cultures (40 +/- 6 cells/high power field, P = 0.002). The activity was dose dependent and was demonstrated to be chemotactic activity by checkerboard analysis. Partial characterization of the activity revealed it was not extractable into ethyl acetate but was partially inactivated by trypsin and heat (100 degrees C, 15 min). The responding cells were predominantly T-helper lymphocytes as shown by monoclonal antibody staining, with a smaller proportion being B-lymphocytes. Molecular sieve column chromatography revealed multiple peaks of lymphocyte chemotactic activity, with three of the peaks preferentially attracting T-helper lymphocytes and one of the peaks preferentially attracting B-lymphocytes. These data demonstrate that bronchial epithelial cells can release chemotactic factors for lymphocytes and suggest that bronchial epithelial cells may modulate their local population of immune effector cells.

scholarly journals A Tissue-Engineered Tracheobronchial In Vitro Co-Culture Model for Determining Epithelial Toxicological and Inflammatory Responses

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 631
Author(s):  
Luis Soriano ◽  
Tehreem Khalid ◽  
Fergal J. O'Brien ◽  
Cian O'Leary ◽  
Sally-Ann Cryan
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Responses ◽  
Bronchial Epithelial Cells ◽  
Lung Fibroblasts ◽  
Drug Induced ◽  
Bronchial Epithelial ◽  
Culture Model ◽  
Epithelial Cultures

Translation of novel inhalable therapies for respiratory diseases is hampered due to the lack of in vitro cell models that reflect the complexity of native tissue, resulting in many novel drugs and formulations failing to progress beyond preclinical assessments. The development of physiologically-representative tracheobronchial tissue analogues has the potential to improve the translation of new treatments by more accurately reflecting in vivo respiratory pharmacological and toxicological responses. Herein, advanced tissue-engineered collagen hyaluronic acid bilayered scaffolds (CHyA-B) previously developed within our group were used to evaluate bacterial and drug-induced toxicity and inflammation for the first time. Calu-3 bronchial epithelial cells and Wi38 lung fibroblasts were grown on either CHyA-B scaffolds (3D) or Transwell® inserts (2D) under air liquid interface (ALI) conditions. Toxicological and inflammatory responses from epithelial monocultures and co-cultures grown in 2D or 3D were compared, using lipopolysaccharide (LPS) and bleomycin challenges to induce bacterial and drug responses in vitro. The 3D in vitro model exhibited significant epithelial barrier formation that was maintained upon introduction of co-culture conditions. Barrier integrity showed differential recovery in CHyA-B and Transwell® epithelial cultures. Basolateral secretion of pro-inflammatory cytokines to bacterial challenge was found to be higher from cells grown in 3D compared to 2D. In addition, higher cytotoxicity and increased basolateral levels of cytokines were detected when epithelial cultures grown in 3D were challenged with bleomycin. CHyA-B scaffolds support the growth and differentiation of bronchial epithelial cells in a 3D co-culture model with different transepithelial resistance in comparison to the same co-cultures grown on Transwell® inserts. Epithelial cultures in an extracellular matrix like environment show distinct responses in cytokine release and metabolic activity compared to 2D polarised models, which better mimic in vivo response to toxic and inflammatory stimuli offering an innovative in vitro platform for respiratory drug development.

scholarly journals Activation of protein kinase A accelerates bovine bronchial epithelial cell migration

2002 ◽  
Vol 282 (5) ◽  
pp. L1108-L1116 ◽  
Author(s):  
John R. Spurzem ◽  
Jitendrakumar Gupta ◽  
Thomas Veys ◽  
Kristen R. Kneifl ◽  
Stephen I. Rennard ◽  
...  
Keyword(s):  
Cell Migration ◽  
Protein Kinase ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Protein Kinase A ◽  
Bronchial Epithelial Cell ◽  
Bronchial Epithelial Cells ◽  
Bronchial Epithelial ◽  
Epithelial Cell Migration ◽  
Kinase A

Bronchial epithelial cell migration is required for the repair of damaged airway epithelium. We hypothesized that bronchial epithelial cell migration during wound repair is influenced by cAMP and the activity of its cyclic nucleotide-dependent protein kinase, protein kinase A (PKA). We found that, when confluent monolayers of bronchial epithelial cells are wounded, an increase in PKA activity occurs. Augmentation of PKA activity with a cell-permeable analog of cAMP, dibutyryl adenosine 3′,5′-cyclic monophosphate, isoproterenol, or a phosphodiesterase inhibitor accelerated migration of normal bronchial epithelial cells in in vitro wound closure assays and Boyden chamber migration assays. A role for PKA activity was also confirmed with a PKA inhibitor, KT-5720, which reduced stimulated migration. Augmentation of PKA activity reduced the levels of active Rho and the formation of focal adhesions. These studies suggest that PKA activation modulates Rho activity, migration mechanisms, and thus bronchial epithelial repair mechanisms.

Fibronectin supports bronchial epithelial cell adhesion and survival in the absence of growth factors

1997 ◽  
Vol 273 (3) ◽  
pp. L684-L693 ◽  
Author(s):  
K. Aoshiba ◽  
S. I. Rennard ◽  
J. R. Spurzem
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Focal Adhesion ◽  
Bronchial Epithelial Cell ◽  
Bronchial Epithelial Cells ◽  
Peptide Sequence ◽  
Human Bronchial Epithelial Cells ◽  
Bronchial Epithelial

Cell-extracellular matrix interactions support the ability of cells to migrate into areas of inflammation and injury. The present study evaluated the ability of different matrix proteins to support bronchial epithelial cell attachment and survival. Collagens were able to support attachment and survival of normal cultured human bronchial epithelial cells but only in the presence of added soluble growth factors such as insulin, epidermal growth factor, platelet-derived growth factor, and bovine pituitary extract. In contrast, fibronectin was able to support attachment and survival of normal human bronchial epithelial cells in growth factor-deficient medium. In addition, fibronectin, in the absence of added growth factors, was able to induce integrin clustering, focal adhesion formation, and phosphorylation of focal adhesion kinase. A 120-kDa chymotryptic fragment of fibronectin containing the Arg-Gly-Asp peptide sequence was able to reproduce the effects of the whole fibronectin molecule. This study supports the concept that fibronectin has specialized roles in injury and repair.

Bronchial epithelial cells release neutrophil chemotactic activity in response to tachykinins

1992 ◽  
Vol 263 (2) ◽  
pp. L226-L231 ◽  
Author(s):  
S. G. Von Essen ◽  
S. I. Rennard ◽  
D. O'Neill ◽  
R. F. Ertl ◽  
R. A. Robbins ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Proteinase Inhibitors ◽  
Bronchial Epithelial Cells ◽  
Chemotactic Activity ◽  
Induced Response ◽  
Neurokinin A ◽  
Maximal Effect ◽  
Neutrophil Chemotaxis ◽  
Bronchial Epithelial ◽  
Neutrophil Chemotactic Activity

The purpose of this study was to determine whether substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) induce the release of neutrophil chemotactic activity (NCA) from bovine bronchial epithelial cells (BBEC) and whether neutral endopeptidase (NEP), a membrane-bound metalloenzyme that hydrolyzes tachykinins, modulates these effects. BBEC monolayers were exposed to SP, NKA, and NKB in the absence or presence of phosphoramidon (10(-6) M), a selective NEP inhibitor, for 72 h. Using a modified blind-well in vitro neutrophil chemotaxis assay, we found that tachykinin-exposed BBEC culture supernatant fluids induced significant neutrophil chemotaxis compared with supernatants obtained from unstimulated BBEC. Maximal effect was observed after 48 h of incubation and at SP concentration of 10(-13) M [92 +/- 3 (SP) vs. 64 +/- 2 (media) cells/high-power field (HPF), mean +/- SE, n = 7, P less than 0.05]. Release of NCA was mediated by the COOH-terminal of the SP molecule. The rank order of potency of tachykinins in inducing release of NCA was SP greater than NKA = NKB. SP-induced response was significantly potentiated by phosphoramidon (109 +/- 3 vs. 92 +/- 3 cells/HPF, n = 7, P less than 0.05), whereas other proteinase inhibitors had no effect. The released NCA was composed of protein and lipid-soluble components. These data indicate that mammalian tachykinins induce the release of NCA from BBEC and that NEP modulates these effects. We suggest that tachykinins regulate neutrophil recruitment into the lower respiratory tract, in part, by inducing the release of NCA from airway epithelial cells.

Bronchial epithelial cells exposed to isocyanates potentiate activation and proliferation of T-cells

1990 ◽  
Vol 259 (4) ◽  
pp. L320-L327 ◽  
Author(s):  
S. Mattoli ◽  
S. Miante ◽  
F. Calabro ◽  
M. Mezzetti ◽  
A. Fasoli ◽  
...  
Keyword(s):  
T Cell ◽  
Epithelial Cells ◽  
Inflammatory Responses ◽  
Cell Damage ◽  
Bronchial Epithelial Cells ◽  
Bronchial Epithelial ◽  
Hla Dr ◽  
Immunological Mechanisms ◽  
Chemotactic Factors

Bronchial epithelial cells release chemotactic factors for lymphocytes and express HLA-DR antigens. Thus they may contribute to the T-cell-mediated inflammatory responses involved in a number of pulmonary diseases such as asthma. In this study, the in vitro exposure of human bronchial epithelial cells to toluene 2,4-diisocyanate (TDI), an inflammatory and asthmogenic stimulus presumed to act at least in part through immunological mechanisms, provoked cell damage followed by proliferation of the cells that survived the injury. At the time of the proliferative response, epithelial cells released factors that upregulated the activation and proliferation of T lymphocytes presensitized by antigen receptor triggering. The T-cell activating factors were interleukin (IL) 1- and 6-like substances, as demonstrated by the ability of specific antisera to inhibit most of the biological effect, and by the ability of recombinant IL-1 and IL-6 to reproduce it. Appreciable amounts of immunoreactive IL-1 and IL-6 were indeed recovered in the supernatants of TDI-exposed epithelial cells. The release of these cytokines may represent an important mechanism by which epithelial cells respond to some environmental stimuli and contribute to the persistence of inflammatory responses in the airways.

Cigarette Smoke Stimulates Release of Neutrophil Chemotactic Activity from Cultured Bovine Bronchial Epithelial Cells

1995 ◽  
Vol 88 (3) ◽  
pp. 337-344 ◽  
Author(s):  
Shunsuke Shoji ◽  
Ronald F. Ertl ◽  
Sekiya Koyama ◽  
Richard Robbins ◽  
George Leikauf ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cyclic Amp ◽  
Cigarette Smoke ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Bronchial Epithelial Cells ◽  
Chemotactic Activity ◽  
Bronchial Epithelial ◽  
Lipoxygenase Inhibitors ◽  
Neutrophil Chemotactic Activity

1. Recruitment of neutrophils into the airway is a prominent feature of chronic bronchitis, a syndrome often associated with exposure to cigarette smoke. Since bronchial epithelial cells are the ‘first’ lung cells to come into contact with smoke, these cells may be responsible for neutrophil recruitment into the airway by release of neutrophil chemotactic activity in response to cigarette smoke. 2. To evaluate this hypothesis, we prepared bovine bronchial epithelial cells and measured their ability to release neutrophil chemotactic activity following exposure to cigarette smoke. Bronchial epithelial cells were prepared by overnight digestion with protease, filtered through 100-μm Nitex mesh and resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum and antibiotics and cultured at 2×106 cells in 2 ml of medium in 35-mm culture dishes. After 4 days, dishes were rinsed and refed with medium without fetal calf serum and incubated with and without 1:10 diluted smoke extract for 6 h at 37°C. The neutrophil chemotactic activity of the supernatant fluids was measured by the blindwell chamber technique. 3. Cigarette smoke itself added to medium did not stimulate chemotaxis. In contrast, cigarette smoke did stimulate the release of neutrophil chemotactic activity from bovine bronchial epithelial cells [15 ± 3 (control) versus 74 ± 5 (smoke), P < 0.01]. 4. This neutrophil chemotactic activity was dialysable, pepsin and acid stable, heat sensitive and lipid extractable. Sephadex G-75 column chromatography demonstrated two peaks of neutrophil chemotactic activity. 5. The lipoxygenase inhibitors diethylcarbamazine and nordihydroguaratic acid both diminished the release of chemotactic activity, suggesting that the activity may be a lipoxygenase product(s). 6. Dibutyryl cyclic AMP also blocked the smoke-stimulated release of neutrophil chemotactic activity, suggesting that its release may be regulated by intracellular cyclic AMP. 7. Thus, bovine bronchial epithelial cells release neutrophil chemotactic activity in response to cigarette smoke, suggesting that bronchial epithelial cells may be modulators of the airway inflammatory response caused by cigarette smoke.

Acetylcholine and substance P stimulate bronchial epithelial cells to release eosinophil chemotactic activity

1998 ◽  
Vol 84 (5) ◽  
pp. 1528-1534 ◽  
Author(s):  
Sekiya Koyama ◽  
Etsuro Sato ◽  
Hiroshi Nomura ◽  
Keishi Kubo ◽  
Sonoko Nagai ◽  
...  
Keyword(s):  
Substance P ◽  
Epithelial Cells ◽  
Receptor Antagonist ◽  
Platelet Activating Factor ◽  
Bronchial Epithelial Cells ◽  
Chemotactic Activity ◽  
Dependent Manner ◽  
Bronchial Epithelial ◽  
Lipoxygenase Inhibitors

We investigated a role of neuroregulation in the release of eosinophil chemotactic activity (ECA) from bovine bronchial epithelial cells (BBEC). BBEC were stimulated with acetylcholine (ACh) and substance P (SP), and the supernatant fluids were tested for ECA by a blind-well chemotactic chamber technique. BBEC released ECA in response to ACh and SP in a dose- and time-dependent manner. Checkerboard analysis showed that ECA in regard to ACh and SP was chemotactic rather than chemokinetic. Partial characterization revealed that ECA involved both lipids and peptides. The release of ECA in response to ACh and SP was inhibited by nonspecific and 5-specific lipoxygenase inhibitors and by cycloheximide ( P < 0.01). Molecular-sieve column chromatography revealed that these mediators induced three molecular mass peaks (near 25 kDa, 9 kDa, and 400 Da, respectively). The lowest peak, which represented the predominant activity, was blocked by leukotriene B4-receptor antagonist ( P < 0.01) but not by platelet-activating factor-receptor antagonist. The release of leukotriene B4 in the supernatant fluids was increased in response to ACh and SP stimulation ( P < 0.01). Platelet-activating factor was not detected. These results raise the possibility of a role of neuroregulation for the elaboration of ECA in the airway.

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