Login
Communauté Vinci
Extérieur
Si votre nom d'utilisateur ne se termine pas par @vinci.be ou @student.vinci.be, utilisez le formulaire ci-dessous pour accéder à votre compte de lecteur.
Titre : | La réponse immunitaire cellulaire et le cancer |
Auteurs : | Deborah MATUMIKINA VUMUNA, Auteur ; Pierre Vander Bruggen, Promoteur |
Type de document : | Travail de fin d'études |
Editeur : | Bruxelles : Institut Paul Lambin, 2020 |
Langues: | Français |
Index. décimale : | TFE - Biologie médicale |
Résumé : |
The laboratory wants to understand and identify the mechanisms of inhibition of T cell function by MDSCs.
During my first project, we wish to study the immunosuppressive potential of MDSCs on T lymphocytes (T-cell). To achieve this, T-cells are activated and are cultured in the absence or presence of MDSCs and their capacity to proliferate is analysed. T cells are labelled with CMFDA, an intracellular tracer dye whose fluorescence intensity decreases with each division. This makes it possible to follow by flow cytometry the proliferation of T lymphocytes during the test. For suppression assay, T cells are co-incubated with MDSC. As T cells alone show a low proliferation capacity after thawing autologous dendritic cells (DCs) are added in the test to improve proliferation of T-cell. The cells used for the proliferation test must be validated in blank proliferation test before the real suppression assay with MDSC. We isolated T-cells and differentiated DCs from blood bags of 3 donors. We also tried to generate proliferative and suppressive controls by differentiating mature DC and DC-10 from monocytes respectively. We showed that the cells we isolated from proliferation assays can be used in a real suppressive test with MDSCs. We successfully used two different approaches to generated controls mature DC for proliferation. However, we were unable to generate immunosuppressive DC-10 as controls of suppression. For my second project, we wanted to study the recognition of tumour antigens by T-cells, in the context of the generation of a therapeutic vaccine against tumour with microsatellite instabilities. To fulfil this task, we had a colorectal cancer cell line (HCT-116) which naturally expresses mutated peptides that are encoded by the vaccine and CD8 T-cell clones recognizing these antigens. More specifically, we wanted to know if the expression level of the one mutated peptide in the HCT-116 cell line was high enough to be recognized by two clones specific for this peptide. We used IFNγ secretion assay to monitor T-cell activation after co-culture with cells presenting the peptide of interest. We showed that the endogenous processing of this peptide is not dependant on the proteasome complex expressed as expression of both standard and immuno-proteasome allowed peptide presentation to CD8 T-cells. However, we saw no recognition of the peptide when T-cells were co-cultured with HCT-116, even after increasing T-cell/HCT-116 ratios. These results suggest that the level of expression of the mutated peptide by HCT-116 is not high enough to allow a correct presentation to effector T-cell. |
Accès : | Identifiez-vous avant d'accéder au document électronique |
Disponible en ligne : | Oui |
Lieu du stage : | UCL-Institut de Duve-Immunologie des tumeurs, TILS |
Département : | Biologie médicale |
Documents numériques (1)
Ce document n'est visible qu'après identification
TFE biologie médicale Adobe Acrobat PDF |