Inhibiting Protein Kinase D Promotes Airway Epithelial Barrier Integrity in Mouse Models of Influenza A Virus Infection
| Autor: | Janelle M. Veazey, Sophia I Eliseeva, Sara E. Hillman, Kristie Stiles, Timothy R. Smyth, Charlotte E. Morrissey, Erika J. Tillotson, Dave J. Topham, Timothy J. Chapman, Steve N. Georas |
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| Rok vydání: | 2020 |
| Předmět: |
lcsh:Immunologic diseases. Allergy
0301 basic medicine Sp1 Transcription Factor medicine.medical_treatment Immunology respiratory tract infections Respiratory Mucosa Biology medicine.disease_cause Virus Mice 03 medical and health sciences 0302 clinical medicine Immune system Orthomyxoviridae Infections Influenza Human medicine Influenza A virus Animals Humans Immunology and Allergy RNA Small Interfering Protein Kinase Inhibitors Cells Cultured Protein Kinase C Original Research Protein Kinase D Mice Knockout Antiviral immune response Sp1 transcription factor medicine.diagnostic_test Dextrans Airway epithelial barrier musculoskeletal system Mice Inbred C57BL innate immuity Disease Models Animal 030104 developmental biology Bronchoalveolar lavage Cytokine cardiovascular system Cancer research Signal transduction lcsh:RC581-607 Viral load Signal Transduction 030215 immunology |
| Zdroj: | Frontiers in Immunology Frontiers in Immunology, Vol 11 (2020) |
| ISSN: | 1664-3224 |
| DOI: | 10.3389/fimmu.2020.580401 |
| Popis: | RationaleProtein kinase D (PKD) is a serine/threonine kinase family that is involved in a wide array of signaling pathways. Although PKD has been implicated in immune responses, relatively little is known about the function of PKD in the lung or during viral infections.ObjectivesWe investigated the hypothesis that PKD is involved in multiple aspects of host response to viral infection.MethodsThe selective PKD inhibitor CRT0010166 was administered to C57BL/6 mice prior to and during challenge with either inhaled double-stranded RNA or Influenza A Virus. PKD signaling pathways were investigated in human bronchial epithelial cells treated with CRT0010166, double-stranded RNA, and/or infected with Influenza A Virus.MeasurementsTotal protein and albumin accumulation in the bronchoalveolar fluid was used to asses inside/out leak. Clearance of inhaled FITC-dextran out of the airspace was used to assess outside/in leak. Cytokines and neutrophils in bronchoalveolar lavage were assayed with ELISAs and cytospins respectively. Viral RNA level was assessed with RT-PCR and protein level assessed by ELISA.Main ResultsPKD inhibition prevented airway barrier dysfunction and pro-inflammatory cytokine release. Epithelial cells express PKD3, and PKD3 siRNA knock-down inhibited polyI:C induced cytokine production. Lung epithelial-specific deletion of PKD3 (CC10-Cre x PKD3-floxed mice) partially attenuated polyI:C-induced barrier disruption in vivo. Mechanistically, we found that PKD promoted cytokine mRNA transcription, not secretion, likely through activating the transcription factor Sp1. Finally, prophylactic CRT treatment of mice promoted barrier integrity during influenza virus infection and reduced viral burden.ConclusionsInhibiting PKD promotes barrier integrity, limit pathogenic cytokine levels, and restrict Influenza A Virus infection. Therefore, PKD is an attractive target for novel antiviral therapeutics. |
| Databáze: | OpenAIRE |
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