Abstract

Background: Haemophilus influenzae infection of the nasal epithelium has long been associated with observations of decreased nasal ciliary beat frequency (CBF) and injury to the ciliated epithelium. Previously, we have reported that several agents that slow CBF also have the effect of activating protein kinase C epsilon (PKCε{lunate}) activity in bronchial epithelial cells. The subsequent auto-downregulation of PKCε{lunate} or the direct inhibition of PKCε{lunate} leads to the specific detachment of the ciliated cells. METHODS: Primary cultures of ciliated bovine bronchial epithelial cells were exposed to filtered conditioned media supernatants from non-typeable H. influenzae (NTHi) cultures. CBF and motile points were measured and PKCε{lunate} activity assayed.Results: NTHi supernatant exposure significantly and rapidly decreased CBF in a dose-dependent manner within 10 minutes of exposure. After 3 hours of exposure, the number of motile ciliated cells significantly decreased. Direct measurement of PKCε{lunate} activity revealed a dose-dependent activation of PKCε{lunate} in response to NTHi supernatant exposure. Both CBF and PKCε{lunate} activity changes were only observed in fresh NTHi culture supernatant and not observed in exposures to heat-inactivated or frozen supernatants.Conclusions: Our results suggest that CBF slowing observed in response to NTHi is consistent with the stimulated activation of PKCε{lunate}. Ciliated cell detachment is associated with PKCε{lunate} autodownregulation.

Original languageEnglish (US)
Article number49
JournalRespiratory Research
Volume13
DOIs
StatePublished - Jun 19 2012

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Protein Kinase C-epsilon
Cilia
Haemophilus influenzae
Epithelial Cells
Haemophilus Infections
Nasal Mucosa
Conditioned Culture Medium
Nose
Down-Regulation
Epithelium
Hot Temperature
Wounds and Injuries

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Non-typeable Haemophilus influenzae decreases cilia beating via protein kinase C epsilon. / Bailey, Kristina L; LeVan, Tricia D; Yanov, Daniel A.; Pavlik, Jaqueline A.; DeVasure, Jane M.; Sisson, Joseph Harold; Wyatt, Todd A.

In: Respiratory Research, Vol. 13, 49, 19.06.2012.

Research output: Contribution to journalArticle

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author = "Bailey, {Kristina L} and LeVan, {Tricia D} and Yanov, {Daniel A.} and Pavlik, {Jaqueline A.} and DeVasure, {Jane M.} and Sisson, {Joseph Harold} and Wyatt, {Todd A}",
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AU - Bailey, Kristina L

AU - LeVan, Tricia D

AU - Yanov, Daniel A.

AU - Pavlik, Jaqueline A.

AU - DeVasure, Jane M.

AU - Sisson, Joseph Harold

AU - Wyatt, Todd A

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N2 - Background: Haemophilus influenzae infection of the nasal epithelium has long been associated with observations of decreased nasal ciliary beat frequency (CBF) and injury to the ciliated epithelium. Previously, we have reported that several agents that slow CBF also have the effect of activating protein kinase C epsilon (PKCε{lunate}) activity in bronchial epithelial cells. The subsequent auto-downregulation of PKCε{lunate} or the direct inhibition of PKCε{lunate} leads to the specific detachment of the ciliated cells. METHODS: Primary cultures of ciliated bovine bronchial epithelial cells were exposed to filtered conditioned media supernatants from non-typeable H. influenzae (NTHi) cultures. CBF and motile points were measured and PKCε{lunate} activity assayed.Results: NTHi supernatant exposure significantly and rapidly decreased CBF in a dose-dependent manner within 10 minutes of exposure. After 3 hours of exposure, the number of motile ciliated cells significantly decreased. Direct measurement of PKCε{lunate} activity revealed a dose-dependent activation of PKCε{lunate} in response to NTHi supernatant exposure. Both CBF and PKCε{lunate} activity changes were only observed in fresh NTHi culture supernatant and not observed in exposures to heat-inactivated or frozen supernatants.Conclusions: Our results suggest that CBF slowing observed in response to NTHi is consistent with the stimulated activation of PKCε{lunate}. Ciliated cell detachment is associated with PKCε{lunate} autodownregulation.

AB - Background: Haemophilus influenzae infection of the nasal epithelium has long been associated with observations of decreased nasal ciliary beat frequency (CBF) and injury to the ciliated epithelium. Previously, we have reported that several agents that slow CBF also have the effect of activating protein kinase C epsilon (PKCε{lunate}) activity in bronchial epithelial cells. The subsequent auto-downregulation of PKCε{lunate} or the direct inhibition of PKCε{lunate} leads to the specific detachment of the ciliated cells. METHODS: Primary cultures of ciliated bovine bronchial epithelial cells were exposed to filtered conditioned media supernatants from non-typeable H. influenzae (NTHi) cultures. CBF and motile points were measured and PKCε{lunate} activity assayed.Results: NTHi supernatant exposure significantly and rapidly decreased CBF in a dose-dependent manner within 10 minutes of exposure. After 3 hours of exposure, the number of motile ciliated cells significantly decreased. Direct measurement of PKCε{lunate} activity revealed a dose-dependent activation of PKCε{lunate} in response to NTHi supernatant exposure. Both CBF and PKCε{lunate} activity changes were only observed in fresh NTHi culture supernatant and not observed in exposures to heat-inactivated or frozen supernatants.Conclusions: Our results suggest that CBF slowing observed in response to NTHi is consistent with the stimulated activation of PKCε{lunate}. Ciliated cell detachment is associated with PKCε{lunate} autodownregulation.

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