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Titre : | Validation de tests de suppression pour étudier lactivité immunosuppressive des MDSCs humainesStress oxydatif et inflammatoire induits par le H2O2 et le LPS: les effets dune cyclodextrine (HPbCD) et dun anti-inflammatoire glucocorticoïde (Budésonid) |
Auteurs : | Hortense KEMPEN, Auteur ; Pierre Van der 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 aim of the project carried out at the Institut De Duve in the laboratory of Prof. Van der Bruggen is to validate a micro-proliferation test using human MDSCs on T lymphocytes. MDSCs are part of a population of heterogeneous cells with high immunosuppressive capacities, present in the blood and on the tumours of cancer patients.
This assay will establish the immunosuppressive profile of MDSCs and ultimately lead to the establishment of an immunotherapy treatment targeting this cell population in cancer patients. In the micro-proliferation test, T cells isolated from the blood of a patient with hematochromatosis are cultured for four days with MDSCs isolated from the blood or tumor of a cancer patient. Mature dendritic cells autologous to T cells are added to the culture to enhance their proliferation because T cells do not tolerate freezing well. The proliferative profile of the T cells is established using CMFDA, a fluorescent tracking dye whose intensity decreases with each cycle of cell division. The results are analyzed at FACS and processed on the "FlowJo" program. Before performing the micro-proliferation test with MDSCs, cells are generated and must be validated. We have generated T cells from blood bags from donors with hematochromatosis. Their ability to proliferate is studied in the presence of proliferative and suppressive control cells. We have therefore differentiated blood monocytes into mature dendritic cells with poly-IC and TNF-, which serve as proliferation controls. Blood monocytes were also differentiated into DC-10 with IL-10 and DC PGE2 with PGE2, which serve as a suppressor control. The phenotype of the generated cells was analyzed at FACS. We showed that the cells were pure and can be used for the micro-proliferation control test. For the test, T cells are cultured in the presence of control cells for four days. The suppressive or proliferative character of the cells must be validated as well as the ability of the T cells to proliferate correctly. In this project, we also wanted to compare the behaviour of DC-10 and DC-PGE2 after freezing. DC-10 cells are less and less efficient after freezing in Pr Van der Bruggen's laboratory, so we generated DC-PGE2 cells that have the same suppressive character as DC-10 cells. Due to lack of time, we were unable to set up this experiment. |
Accès : | Identifiez-vous avant d'accéder au document électronique |
Disponible en ligne : | Oui |
Lieu du stage : | UCL-Institut De Duve |
Département : | Biologie médicale |
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TFE biologie médicale Adobe Acrobat PDF |