Identification of a meningococcal adhesin that inhibits epithelial cell migration

10.26686/wgtn.17144027.v1 ◽

2021 ◽

Author(s):

◽

Gabrielle Greig

Keyword(s):

Cell Migration ◽

Epithelial Cell ◽

Epithelial Cells ◽

Host Cell ◽

Wound Repair ◽

Invasive Disease ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Household Contacts ◽

Cell Association

<p>Neisseria meningitidis virulence is polygenic, therefore comparing many genomes may not yield strictly disease-associated virulence factors. An alternative approach is comparing closely related isolates, such as those from household contacts. Disease isolates have been shown to inhibit epithelial cell wound repair, while many carriage isolates do not. In this study, bacteria collected from disease patients and healthy household contacts were compared to identify the meningococcal factor responsible for wound repair inhibition and investigate how it contributes to invasive disease. Host cell wound repair inhibition was compared between disease-associated meningococcal isolate, NZ97/052, and isolates NZCM111 and NZCM112, from asymptomatic household contacts. Migrating bronchial airway cells were infected with meningococcal isolates and wound closure was evaluated by microscopy. NZ97/052 and NZCM111 both inhibited wound repair, whereas NZCM112 did not. To investigate if this was due to bacterial consumption of an important nutrient, infected cells were supplemented with nutrients known to be important for meningococcal growth and cell migration. Iron supplementation resulted in carriage associated isolates gaining the ability to inhibit wound repair. Genome and transcriptome comparisons were completed between NZ97/052 and NZCM112, which differ in wound repair inhibition. This analysis identified a frameshift mutation in NZCM112 in the haptoglobin-haemoglobin utilization gene, hpuA, which caused complete loss of expression. The hpuA gene was deleted by allelic replacement from NZ97/052, nullifying its ability to inhibit host cell migration. Furthermore, bacterial association and fluorescence microscopy assays suggested that HpuA contributes to meningococcal attachment to bronchial epithelial cells, as the hpuA mutant had significantly lower cell association. Heterologous expression of HpuA in E. coli resulted in higher levels of cell association, indicating that HpuA is sufficient to mediate bacterial adhesion to human bronchial epithelial cells. This work reveals novel roles for HpuA as a meningococcal adhesin and a bacterial factor that inhibits host cell migration.</p>

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cAMP-dependent protein kinase activation decreases cytokine release in bronchial epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00373.2013 ◽

2014 ◽

Vol 307 (8) ◽

pp. L643-L651 ◽

Cited By ~ 15

Author(s):

Todd A. Wyatt ◽

Jill A. Poole ◽

Tara M. Nordgren ◽

Jane M. DeVasure ◽

Art J. Heires ◽

...

Keyword(s):

Cell Migration ◽

Protein Kinase ◽

Epithelial Cells ◽

Wound Repair ◽

Bronchial Epithelial Cells ◽

Dependent Protein Kinase ◽

Bronchial Epithelial ◽

Cyclic Monophosphate ◽

Tnf Α ◽

Dependent Protein

Lung injury caused by inhalation of dust from swine-concentrated animal-feeding operations (CAFO) involves the release of inflammatory cytokine interleukin 8 (IL-8), which is mediated by protein kinase C-ε (PKC-ε) in airway epithelial cells. Once activated by CAFO dust, PKC-ε is responsible for slowing cilia beating and reducing cell migration for wound repair. Conversely, the cAMP-dependent protein kinase (PKA) stimulates contrasting effects, such as increased cilia beating and an acceleration of cell migration for wound repair. We hypothesized that a bidirectional mechanism involving PKA and PKC regulates epithelial airway inflammatory responses. To test this hypothesis, primary human bronchial epithelial cells and BEAS-2B cells were treated with hog dust extract (HDE) in the presence or absence of cAMP. PKC-ε activity was significantly reduced in cells that were pretreated for 1 h with 8-bromoadenosine 3′,5′-cyclic monophosphate (8-Br-cAMP) before exposure to HDE ( P < 0.05). HDE-induced IL-6, and IL-8 release was significantly lower in cells that were pretreated with 8-Br-cAMP ( P < 0.05). To exclude exchange protein activated by cAMP (EPAC) involvement, cells were pretreated with either 8-Br-cAMP or 8-(4-chlorophenylthio)-2'- O-methyladenosine-3',5'-cyclic monophosphate (8-CPT-2Me-cAMP) (EPAC agonist). 8-CPT-2Me-cAMP did not activate PKA and did not reduce HDE-stimulated IL-6 release. In contrast, 8-Br-cAMP decreased HDE-stimulated tumor necrosis factor (TNF)-α-converting enzyme (TACE; ADAM-17) activity and subsequent TNF-α release ( P < 0.001). 8-Br-cAMP also blocked HDE-stimulated IL-6 and keratinocyte-derived chemokine release in precision-cut mouse lung slices ( P < 0.05). These data show bidirectional regulation of PKC-ε via a PKA-mediated inhibition of TACE activity resulting in reduced PKC-ε-mediated release of IL-6 and IL-8.

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Activation of protein kinase A accelerates bovine bronchial epithelial cell migration

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00148.2001 ◽

2002 ◽

Vol 282 (5) ◽

pp. L1108-L1116 ◽

Cited By ~ 32

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.

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Cloning of a Novel Protein Interacting with BRS-3 and Its Effects in Wound Repair of Bronchial Epithelial Cells

PLoS ONE ◽

10.1371/journal.pone.0023072 ◽

2011 ◽

Vol 6 (8) ◽

pp. e23072 ◽

Cited By ~ 9

Author(s):

Hui Jun Liu ◽

Yu Rong Tan ◽

Meng Lan Li ◽

Chi Liu ◽

Yang Xiang ◽

...

Keyword(s):

Epithelial Cells ◽

Wound Repair ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Novel Protein

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IL-4 induces ICAM-1 expression in human bronchial epithelial cells and potentiates TNF-α

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1999.277.1.l58 ◽

1999 ◽

Vol 277 (1) ◽

pp. L58-L64 ◽

Cited By ~ 9

Author(s):

Ilja Striz ◽

Tadashi Mio ◽

Yuichi Adachi ◽

Peggy Heires ◽

Richard A. Robbins ◽

...

Keyword(s):

Epithelial Cell ◽

Epithelial Cells ◽

Adhesion Molecule ◽

Airway Epithelial Cells ◽

Bronchial Epithelial Cells ◽

Intercellular Adhesion Molecule ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Macrophage Cells ◽

Tnf Α

Interleukin (IL)-4 is thought to contribute to the Th2 type of immune response and hence the development of allergic reactions such as asthma. In asthmatic patients, the airway epithelium expresses increased amounts of the cell surface adhesion molecule intercellular adhesion molecule (ICAM)-1 (CD54). One cytokine capable of inducing ICAM-1 in airway epithelial cells, tumor necrosis factor-α (TNF-α), is present in asthma. This study evaluated if IL-4 either alone or together with TNF-α costimulation might modulate CD54 expression by human bronchial epithelial cells (HBECs). CD54 positivity increased in response to IL-4 (16 ± 2% positive vs. 3 ± 1%, P < 0.01); greater induction of CD54 resulted from TNF-α (45 ± 2%, P < 0.001). Costimulation with TNF-α plus IL-4 further augmented expression (56 ± 1%, P < 0.05). Immunoperoxidase results were confirmed by flow cytometry. RT-PCR revealed no increase in ICAM-1 mRNA expression under control conditions or after stimulation with IL-4 alone. TNF-α increased IL-4 mRNA, and IL-4 potentiated this. Functionally, IL-4 augmented the adhesion of THP-1 monocyte/macrophage cells to monolayers of HBECs both alone and in the presence of TNF-α. We conclude that 1) IL-4 augments epithelial cell ICAM-1 expression, 2) IL-4 potentiates the adhesion of THP-1 monocyte/macrophage cells to epithelial cells, and 3) modulation of epithelial cell ICAM-1 expression by IL-4 may play a role in the immunopathology of bronchial asthma.

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Human Bronchial Epithelial Cells Secrete Laminin 5, Express Hemidesmosomal Proteins, and Assemble Hemidesmosomes

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215540004800411 ◽

2000 ◽

Vol 48 (4) ◽

pp. 535-544 ◽

Cited By ~ 24

Author(s):

Peter H. Michelson ◽

Margaret Tigue ◽

Jonathan C.R. Jones

Keyword(s):

Epithelial Cell ◽

Epithelial Cells ◽

Basement Membrane ◽

Cell Monolayer ◽

Bronchial Epithelial Cells ◽

Basal Cells ◽

Bronchial Epithelial ◽

Laminin 5 ◽

Α6β4 Integrin

Epithelial cells attach to the basement membrane through adhesive contacts between the basal cells of the epithelium and the proteins of the extracellular matrix (ECM). The hemidesmosome (HD) is a specialized cell-ECM contact, that mediates the attachment of the epithelial cell basal surface to the ECM. In bronchial epithelial cells, the protein components that constitute the HD have not been demonstrated. Using immunohistochemical techniques, we determined that normal human bronchial epithelial (NHBE) cells express the HD cell surface integrin α6β4 and produce laminin 5, the ECM protein associated with HDs. Furthermore, expression of the HD-associated structural proteins, bullous pemphigoid antigens 1 (BPAG 1) and 2 (BPAG 2), was demonstrated in NHBE cells by immunofluorescence microscopy and immunoblot analyses. In addition, we confirmed the presence of laminin 5 in the basement membrane (BM) of bronchial epithelial biopsy specimens and of BP230, BP180, and the α6β4 integrin heterodimer at the site of bronchial epithelial cell-ECM interaction in vivo. Finally, using electron microscopy, we were able to demonstrate intact HDs in a glutaraldehyde-fixed NHBE cell monolayer. These findings suggest that bronchial epithelium forms HDs and that the laminin 5-α6β4 integrin interaction may be important in stabilizing epithelial cell adhesion to the BM in the lung.

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Airway Epithelial Repair by a Prebiotic Mannan Derived from Saccharomyces cerevisiae

Journal of Immunology Research ◽

10.1155/2017/8903982 ◽

2017 ◽

Vol 2017 ◽

pp. 1-7 ◽

Cited By ~ 2

Author(s):

Christie F. Michael ◽

Christopher M. Waters ◽

Kim S. LeMessurier ◽

Amali E. Samarasinghe ◽

Chi Y. Song ◽

...

Keyword(s):

Epithelial Cells ◽

Wound Repair ◽

Epithelial Mesenchymal Transition ◽

Bronchial Epithelial Cells ◽

Current Therapy ◽

Airway Epithelial ◽

Coated Pits ◽

Bronchial Epithelial ◽

Mesenchymal Transition ◽

Mannose Receptors

In asthmatic airways, repeated epithelial damage and repair occur. No current therapy directly targets this process. We aimed to determine the effects of mannan derived from S. cerevisiae (SC-MN) on airway epithelial wound repair, in vitro. The presence of functional mannose receptors in bronchial epithelial cells was shown by endocytosis of colloidal gold-Man BSA via clathrin-coated pits in 16HBE cells. In primary normal human bronchial epithelial cells (NHBEC), SC-MN significantly facilitated wound closure. Treatment with SC-MN stimulated cell spreading as indicated by a significant increase in the average lamellipodial width of wound edge 16HBE cells. In addition, NHBEC treated with SC-MN showed increased expression and activation of Krüppel-like factors (KLFs) 4 and 5, transcription factors important in epithelial cell survival and regulation of epithelial-mesenchymal transition. We conclude that SC-MN facilitates wound repair in human bronchial epithelium, involving mannose receptors.

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