TREG INVIVO_IMAGING · Cellular interactions in peripheral immune regulation
6РП — Действия „Мария Кюри“
- Период
- 2004-01-05 → 2006-01-04
- Финансиране от ЕС
- 139 948 €
- Участници
- 1
- Схема
- EIF
Линиите свързват координатора с партньорите.
Накратко на български
Взаимодействията между регулаторните Т-клетки и други имунни клетки се наблюдават в реално време в тъкани като кожата и лимфните възли. Това помага за разбирането на процесите по регулиране на имунния отговор и възпаленията в организма.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - TREG INVIVO_IMAGING (Cellular interactions in peripheral immune regulation)
Live imaging of lymphocyte motility by intravital microscopy This aim of this research project is to characterise CD4+Foxp3+ Regulatory T cell (Treg) -related cellular interactions occurring during immune regulatory events. Conventional and multi-photon confocal microscopy will be applied to visualise interactions of APCs with Treg and naïve (effector) T cells both in vitro (suppression assay system) and in vivo by analysing intact organs of different animal models of immune regulation. Here, we present our intravital multi-photon microscopy system that was set-up to visualise concomitantly 3 different cellular subsets during (I) in vitro cultures and (II) real-time in vivo images. We successful managed not only to obtain kinetics and aggregation patterns that are being used to test the prediction of the models in vitro but also we managed develop the conditions to test the models in vivo. The system allows us to perform kinetics of cellular movements and interactions, in various tissues for each single mouse, namely in inguinal, popliteal and cervical LN's and in the skin. In addition, the analysis can be performed at various time points during the course of an immune response/inflammation. We have also developed a semi-automated program to analyse and illustrate cell-velocities, cell-trajectories and cellular interactions occurring between 3 different cell-types within live tissues and during in vitro cultures. Generation of Treg and development of Animal models A limiting step in the study of Treg mediated immune regulation in vivo is the low number of Treg available. To perform in vivo real-time imaging of Treg and their target cell population/s, several millions of monoclonal, antigen-specific Treg are required for each mouse. We developed several experimental strategies to promote the generation of Treg in mice that are Transgenic for a T-cell Receptor (TCR) specific for a non-self antigen. These TCR Tg mice are normally devoid of CD4+Foxp3+ Treg. We demonstrate that by altering the cellular composition of the thymus (cells expressing or not expressing the nominal antigen), and thus altering the conditions for negative and positive selection, we induce Treg in high number. We also demonstrate that a single immunisation of the specific peptide emulsified in Complete Freund's Adjuvant (CFA) into the foot-pad of the corresponding TCR transgenic mouse generates high numbers of Treg and prevents spontaneous development of EAE in MBP TCR transgenic mic. These findings are original per se and two manuscripts are being prepared. In addition, these tools allow the purification of peptide-specific Treg and their real-time imaging during active immune and autoimmune reactions.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
The immune response is a potent defensive reaction within the organism that can lead to deleterious self-damaging effects in absence of control mechanisms. The modulation of the immune response is exerted at many different levels including T cell-mediated immuno-regulation. Regulatory T cells (TR cells) have recently been identified which have the capacity to control the activity of effectors T cells. Operationally, TR cells are defined by the ability to suppress the expansion of pathogenic naive T cells in vitro and to inhibit inflammatory reactions in mouse experimental systems. However, the mechanisms by which TR cells mediate immune-regulatory events are poorly described. In fact, it is not known whether this TR cell mediated suppression requires a direct cell-to-cell contact between TR cells, Antigen Presenting Cells (APCs) and target Tcells. Furthermore, the stochiometry of the cellular interactions, the dynamics of such interactions and the site(s) of in vivoimmune regulation are largely unknown. The aim of this project is to characterise the cellular interactions occurring during immune regulation events in vitro using the suppression assay system, and in vivo by analysing intact organs of different animal models of immune regulation. Conventional and multi-photon confocal microscopy will be applied to visualise invitro and in vivo interactions of APCs with CD4+CD25+ TR cells and naïve (effector) T cells previously sorted by flowcytometry, from Yellow Fluorescent Protein and Cyan Fluorescent Protein transgenic mice, respectively. These experiments will be designed to assess (1) whether CD4+CD25+ TR cells suppress naïve T cells by direct interaction or through an indirect effect via APCs, (2) whether the CD4+CD25+ TR cell-mediated immune regulation occurs at the site of tissue inflammation or in draining lymph nodes and (3) whether the kinetics of cell migration to the sites of immune regulation and tissue inflammation.
Оригинален текст от CORDIS (на английски).
Участници
- FUNDAÇÃO CALOUSTE GULBENKIAN · LISBOAКоординаторПортугалия
Връзки
Данни: CORDIS, © Европейски съюз