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Development of a 'Contact Allergen Activated T-Cell (CAATC)-Assay' using dendritic cells from skin: characterization of the sensitizing potency of chemicals via dendritic cell-induced expression of lineage specific T cell transcription factors

Project

Food and consumer protection

This project contributes to the research aim 'Food and consumer protection'. Which funding institutions are active for this aim? What are the sub-aims? Take a look:
Food and consumer protection


Project code: BfR-PRS-08-1334-202
Contract period: 01.06.2010 - 31.07.2012
Purpose of research: Applied research

We propose to develop an in vitro testing system capable of characterizing the sensitizing potency of contact allergens through measurement of the intensity of induced immune cell responses via certain biological endpoints, i.e. expression of lineage specific transcription factors in T cell subpopulations. This testing system would allow to detect cell proliferation and differentiation of the true effector cell populations responsible for hypersensitivity reactions, T-helper(TH) cells and cytotoxic T cells (CTLs). To simulate the chemically induced allergy type IV-reaction of human skin in vitro, we will apply dendritic cells as they occur in human epidermis, so called Langerhans cells (LCs), in a T cell activation assay (Contact Allergen Activated T Cell (CAATC)-Assay).In individual experiments, monocyte-derived Langerhans Cell-like Cells (MoLCs) will be stimulated with seven sensitizers (classified as strong and less strong) and five non-sensitizers. These cells can be considered as surrogates for LCs in vivo that migrate to the regional lymph nodes as activated cells. In cocultures of LCs together with CD4+ T cells, polarization of naive T cells toward specific effector T cells will be induced after subsequent interaction. The following biological endpoints will be explored in the CAATC-Assay: 1. Chemotactic migration of LCs through a 8 µm porus of a Transwell-system; 2. Flow cytometric characterization of cell surface markers like costimulatory and adhesion molecules on LCs; 3. Expression of transcription factors specific for effector T cell subpopulations by quantitative real-time-PCR.In our in vitro allergy testing system the transcription factors Runx3, T-bet, GATA-3, RORC2, and FOXP3 will be evaluated for the first time according to their eligibility to serve as functional markers for induction of CTLs, TH1, TH2, TH17, or regulatory T cells (TReg). qRT-PCR analyses of the expression of effector T cells are expected to point to specific surface markers

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Subjects

Framework programme

BMEL Frameworkprogramme 2008

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