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PLoS One. 2015 Oct 21;10(10):e0139866. doi: 10.1371/journal.pone.0139866. eCollection 2015.

An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry.

Author information

1
Institute of Immunology, University of Regensburg, Regensburg, Germany.
2
Department of Gynecology and Obstetrics, University of Regensburg, Regensburg, Germany.
3
Division of Immunology, Paul-Ehrlich-Institute, Langen, Germany.
4
Department of Gynecology and Obstetrics, University of Regensburg, Regensburg, Germany; Institute of Immunology, University Medical Center of the Johannes Gutenberg University of Mainz, Mainz, Germany.
5
TissueGnostics GmbH, Vienna, Austria.

Abstract

Characterization of host-pathogen interactions is a fundamental approach in microbiological and immunological oriented disciplines. It is commonly accepted that host cells start to change their phenotype after engulfing pathogens. Techniques such as real time PCR or ELISA were used to characterize the genes encoding proteins that are associated either with pathogen elimination or immune escape mechanisms. Most of such studies were performed in vitro using primary host cells or cell lines. Consequently, the data generated with such approaches reflect the global RNA expression or protein amount recovered from all cells in culture. This is justified when all host cells harbor an equal amount of pathogens under experimental conditions. However, the uptake of pathogens by phagocytic cells is not synchronized. Consequently, there are host cells incorporating different amounts of pathogens that might result in distinct pathogen-induced protein biosynthesis. Therefore, we established a technique able to detect and quantify the number of pathogens in the corresponding host cells using immunofluorescence-based high throughput analysis. Paired with multicolor staining of molecules of interest it is now possible to analyze the infection profile of host cell populations and the corresponding phenotype of the host cells as a result of parasite load.

PMID:
26488169
PMCID:
PMC4619545
DOI:
10.1371/journal.pone.0139866
[Indexed for MEDLINE]
Free PMC Article

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