Integrating High-Content Imaging and Chemical Genetics to Probe Host Cellular Pathways Critical for Yersinia Pestis Infection
| Autor: | Brian D. Peyser, J. Jaissle, Douglas Lane, Rekha G. Panchal, Brett P. Eaton, Camenzind G. Robinson, Melanie P. Ulrich, Ricky L. Ulrich, Krishna P. Kota, Sina Bavari, Gianluca Pegoraro |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Bacterial Diseases
Applied Microbiology Drug Evaluation Preclinical lcsh:Medicine IκB kinase Wortmannin chemistry.chemical_compound Mice Phosphatidylinositol 3-Kinases Drug Discovery Gram Negative lcsh:Science Immune Response Protein Kinase C 0303 health sciences Multidisciplinary Kinase NF-kappa B 3. Good health Cell biology Bacterial Pathogens Molecular Imaging Host-Pathogen Interaction Chemistry Protein Transport Infectious Diseases Medicine Signal transduction Chemical genetics Research Article Biotechnology Signal Transduction Yersinia Pestis Immune Cells Immunology Biology Microbiology Cell Line Immunomodulation Small Molecule Libraries 03 medical and health sciences Phagocytosis Microbial Control Chemical Biology Animals Parthenolide Microbial Pathogens Protein kinase C 030304 developmental biology 030306 microbiology Macrophages lcsh:R biology.organism_classification chemistry Yersinia pestis Small Molecules Immune System lcsh:Q |
| Zdroj: | PLoS ONE PLoS ONE, Vol 8, Iss 1, p e55167 (2013) |
| ISSN: | 1932-6203 |
| Popis: | The molecular machinery that regulates the entry and survival of Yersinia pestis in host macrophages is poorly understood. Here, we report the development of automated high-content imaging assays to quantitate the internalization of virulent Y. pestis CO92 by macrophages and the subsequent activation of host NF-κB. Implementation of these assays in a focused chemical screen identified kinase inhibitors that inhibited both of these processes. Rac-2-ethoxy-3 octadecanamido-1-propylphosphocholine (a protein Kinase C inhibitor), wortmannin (a PI3K inhibitor), and parthenolide (an IκB kinase inhibitor), inhibited pathogen-induced NF-κB activation and reduced bacterial entry and survival within macrophages. Parthenolide inhibited NF-κB activation in response to stimulation with Pam3CSK4 (a TLR2 agonist), E. coli LPS (a TLR4 agonist) or Y. pestis infection, while the PI3K and PKC inhibitors were selective only for Y. pestis infection. Together, our results suggest that phagocytosis is the major stimulus for NF-κB activation in response to Y. pestis infection, and that Y. pestis entry into macrophages may involve the participation of protein kinases such as PI3K and PKC. More importantly, the automated image-based screening platform described here can be applied to the study of other bacteria in general and, in combination with chemical genetic screening, can be used to identify host cell functions facilitating the identification of novel antibacterial therapeutics. |
| Databáze: | OpenAIRE |
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