An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry.
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 t...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4619545?pdf=render |
| _version_ | 1828744282591199232 |
|---|---|
| author | Maximilian Schmid Bianca Dufner Julius Dürk Konstanze Bedal Kristina Stricker Lukas Ali Prokoph Christoph Koch Anja K Wege Henner Zirpel Ger van Zandbergen Rupert Ecker Bogdan Boghiu Uwe Ritter |
| author_facet | Maximilian Schmid Bianca Dufner Julius Dürk Konstanze Bedal Kristina Stricker Lukas Ali Prokoph Christoph Koch Anja K Wege Henner Zirpel Ger van Zandbergen Rupert Ecker Bogdan Boghiu Uwe Ritter |
| author_sort | Maximilian Schmid |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T01:54:20Z |
| format | Article |
| id | doaj.art-b91e8f9a406c4f4ca4716de6abac82c6 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T01:54:20Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-b91e8f9a406c4f4ca4716de6abac82c62022-12-22T03:07:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011010e013986610.1371/journal.pone.0139866An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry.Maximilian SchmidBianca DufnerJulius DürkKonstanze BedalKristina StrickerLukas Ali ProkophChristoph KochAnja K WegeHenner ZirpelGer van ZandbergenRupert EckerBogdan BoghiuUwe RitterCharacterization 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.http://europepmc.org/articles/PMC4619545?pdf=render |
| spellingShingle | Maximilian Schmid Bianca Dufner Julius Dürk Konstanze Bedal Kristina Stricker Lukas Ali Prokoph Christoph Koch Anja K Wege Henner Zirpel Ger van Zandbergen Rupert Ecker Bogdan Boghiu Uwe Ritter An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry. PLoS ONE |
| title | An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry. |
| title_full | An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry. |
| title_fullStr | An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry. |
| title_full_unstemmed | An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry. |
| title_short | An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry. |
| title_sort | emerging approach for parallel quantification of intracellular protozoan parasites and host cell characterization using tissuefaxs cytometry |
| url | http://europepmc.org/articles/PMC4619545?pdf=render |
| work_keys_str_mv | AT maximilianschmid anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT biancadufner anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT juliusdurk anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT konstanzebedal anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT kristinastricker anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT lukasaliprokoph anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT christophkoch anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT anjakwege anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT hennerzirpel anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT gervanzandbergen anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT rupertecker anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT bogdanboghiu anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT uweritter anemergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT maximilianschmid emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT biancadufner emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT juliusdurk emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT konstanzebedal emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT kristinastricker emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT lukasaliprokoph emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT christophkoch emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT anjakwege emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT hennerzirpel emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT gervanzandbergen emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT rupertecker emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT bogdanboghiu emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry AT uweritter emergingapproachforparallelquantificationofintracellularprotozoanparasitesandhostcellcharacterizationusingtissuefaxscytometry |