Bead-based flow-cytometry for semi-quantitative analysis of complex membrane vesicle populations released by bacteria and host cells
| Autor: | Birke J. Benedikter, Gert Grauls, Charlotte Volgers, Paul H. M. Savelkoul, Frank R. M. Stassen |
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| Přispěvatelé: | Med Microbiol, Infect Dis & Infect Prev, RS: NUTRIM - R2 - Liver and digestive health, Promovendi NTM, RS: NUTRIM - R3 - Chronic inflammatory disease and wasting, MUMC+: DA Medische Microbiologie en Infectieziekten (5), RS: CAPHRI - R4 - Health Inequities and Societal Participation |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Vesicle fusion 030106 microbiology BIOGENESIS Colony Count Microbial Outer membrane vesicles Microbiology Exocytosis Antibodies Flow cytometry Cell Line 03 medical and health sciences Epitopes Bead-based flow-cytometry medicine Humans Transport Vesicles Screening procedures Secretory pathway EXOSOMES medicine.diagnostic_test biology Bacteria Vesicle Macrophages Cell Membrane Extracellular vesicles biology.organism_classification Flow Cytometry Microvesicles Cell biology Membrane vesicles 030104 developmental biology Membrane vesicle quantification Pseudomonas aeruginosa Moraxella catarrhalis |
| Zdroj: | Microbiological Research, 200, 25-32. Elsevier GMBH, Urban & Fischer Verlag |
| ISSN: | 0944-5013 |
| DOI: | 10.1016/j.micres.2017.04.003 |
| Popis: | During infection, the release of nano-sized membrane vesicle is a process which is common both for bacteria and host cells. Host cell-derived membrane vesicles can be involved in innate and adaptive immunity whereas bacterial membrane vesicles can contribute to bacterial pathogenicity. To study the contribution of both membrane vesicle populations during infection is highly complicated as most vesicles fall within a similar size range of 30-300 nm. Specialized techniques for purification are required and often no single technique complies on its own. Moreover, techniques for vesicle quantification are either complicated to use or do not distinguish between host cell-derived and bacterial membrane vesicle subpopulations. Here we demonstrate a bead-based platform that allows a semi-quantitatively analysis by flow-cytometry of bacterial and host-cell derived membrane vesicles. We show this method can be used to study heterogeneous and complex vesicle populations composed of bacterial and host-cell membrane vesicles. The easy accessible design of the protocol makes it also highly suitable for screening procedures to assess how intrinsic and environmental factors affect vesicle release. |
| Databáze: | OpenAIRE |
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