Quantification of bacterial invasion into adherent cells by flow cytometry
| Autor: | Christof R. Hauck, Stefan Pils, Tim Schmitter, Florian Neske |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Microbiology (medical)
Integrins Staphylococcus aureus media_common.quotation_subject Cellular microbiology Biology medicine.disease_cause Kidney Intracellular bacteria Microbiology Flow cytometry Cell Line Mice Antigens CD ddc:570 medicine Extracellular Animals Humans Internalization Molecular Biology media_common medicine.diagnostic_test Intracellular parasite Epithelial Cells Fibroblasts Flow Cytometry Neisseria gonorrhoeae Carcinoembryonic Antigen Bacterial invasion Fluorescence labelling Cell Adhesion Molecules Intracellular Bacterial Outer Membrane Proteins |
| Zdroj: | Journal of microbiological methods. 65(2) |
| ISSN: | 0167-7012 |
| Popis: | Quantification of invasive, intracellular bacteria is critical in many areas of cellular microbiology and immunology. We describe a novel and fast approach to determine invasion of bacterial pathogens in adherent cell types such as epithelial cells or fibroblasts based on flow cytometry. Using the CEACAM-mediated uptake of Opa-expressing Neisseria gonorrhoeae as a well-characterized model of bacterial invasion, we demonstrate that the flow cytometry-based method yields results comparable to a standard antibiotic protection assay. Furthermore, the quantification of intracellular bacteria by the novel approach is not biased by intracellular killing of the microbes and correctly discriminates between cell-associated extracellular and bona fide intracellular bacteria. As flow cytometry-based quantification is also applicable to other pathogen-host interactions such as the integrin-mediated internalization of Staphylococcus aureus, this approach provides a fast and convenient alternative for the quantification of bacterial uptake and should be particularly useful in elucidating the molecular mechanisms of pathogen-triggered host cell invasion. |
| Databáze: | OpenAIRE |
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