Chapter title |
Measurement of Salmonella enterica Internalization and Vacuole Lysis in Epithelial Cells.
|
---|---|
Chapter number | 19 |
Book title |
Phagocytosis and Phagosomes
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6581-6_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6579-3, 978-1-4939-6581-6
|
Authors |
Jessica A. Klein, TuShun R. Powers, Leigh A. Knodler |
Editors |
Roberto Botelho |
Abstract |
Establishment of an intracellular niche within mammalian cells is key to the pathogenesis of the gastrointestinal bacterium, Salmonella enterica serovar Typhimurium (S. Typhimurium). Here we will describe how to study the internalization of S. Typhimurium into human epithelial cells using the gentamicin protection assay. The assay takes advantage of the relatively poor penetration of gentamicin into mammalian cells; internalized bacteria are effectively protected from its antibacterial actions. A second assay, the chloroquine (CHQ) resistance assay, can be used to determine the proportion of internalized bacteria that have lysed or damaged their Salmonella-containing vacuole and are therefore residing within the cytosol. Its application to the quantification of cytosolic S. Typhimurium in epithelial cells will also be presented. Together, these protocols provide an inexpensive, rapid and sensitive quantitative measure of bacterial internalization and vacuole lysis by S. Typhimurium. |
X Demographics
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United States | 1 | 50% |
United Kingdom | 1 | 50% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 27 | 100% |
Demographic breakdown
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Student > Ph. D. Student | 7 | 26% |
Student > Master | 4 | 15% |
Student > Bachelor | 3 | 11% |
Professor | 1 | 4% |
Student > Doctoral Student | 1 | 4% |
Other | 2 | 7% |
Unknown | 9 | 33% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 6 | 22% |
Agricultural and Biological Sciences | 4 | 15% |
Immunology and Microbiology | 4 | 15% |
Nursing and Health Professions | 1 | 4% |
Environmental Science | 1 | 4% |
Other | 2 | 7% |
Unknown | 9 | 33% |