Small-molecules as regulatory T cell modulators: A pilot screen
SIRKIÄ, MARKO (2008)
SIRKIÄ, MARKO
2008
Biokemia - Biochemistry
Lääketieteellinen tiedekunta - Faculty of Medicine
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Hyväksymispäivämäärä
2008-04-25
Julkaisun pysyvä osoite on
https://urn.fi/urn:nbn:fi:uta-1-18166
https://urn.fi/urn:nbn:fi:uta-1-18166
Tiivistelmä
Backround and aims: Regulatory T cells are widely recognized as key players of immune control and have been shown to function in many immune related diseases. This has lead to intensive research efforts and high hopes for novel therapeutic and diagnostic measures. We aimed to develop methods suitable for high-throughput expression analysis from low cell counts and to discover small-molecules and key signalling pathways critical to regulatory T cell function.
Methods: Solid-phase mRNA extraction, quantitative real-time PCR and flow cytometric methods were developed, optimized and applied using automated workstations to a screening assay for small-molecules able to modulate ex vivo human T cells.
Results: A functional protocol for high-throughput capable gene expression analysis in low cell counts was produced. We were able to show that the selected methods were applicable and well suited for the planned screening purpose, producing reliable and reproducible data from a complex model. As a result we identified several small-molecule compounds of interest able to modify regulatory T cell phenotype.
Conclusions: Regulatory T cell modifiers can be identified using the methods described in this work, and the results presented here indicate that large scale screening of such compounds is feasible. Suitability of the discovered compounds for potential diagnostic or therapeutic purposes need to be assessed in further studies.
Keywords: Regulatory T cell, FOXP3, high-throughput screening, chemical genomics
Methods: Solid-phase mRNA extraction, quantitative real-time PCR and flow cytometric methods were developed, optimized and applied using automated workstations to a screening assay for small-molecules able to modulate ex vivo human T cells.
Results: A functional protocol for high-throughput capable gene expression analysis in low cell counts was produced. We were able to show that the selected methods were applicable and well suited for the planned screening purpose, producing reliable and reproducible data from a complex model. As a result we identified several small-molecule compounds of interest able to modify regulatory T cell phenotype.
Conclusions: Regulatory T cell modifiers can be identified using the methods described in this work, and the results presented here indicate that large scale screening of such compounds is feasible. Suitability of the discovered compounds for potential diagnostic or therapeutic purposes need to be assessed in further studies.
Keywords: Regulatory T cell, FOXP3, high-throughput screening, chemical genomics