Étude du potentiel inflammatoire et immunitaire des vésicules extracellulaires dérivées des plaquettes
|Abstract:||Platelets are very abundant in the blood where they play a role in hemostasis, inflammation and immunity. Activated, platelets will undergo a change of conformation which allows the release of numerous effector molecules as well as the production of extracellular vesicles (EVs). EVs are formed by the budding of the platelet and bring some of its contents, including nucleic acids, surface proteins and organelles. Heterogeneous, the diverse content of platelet EVs suggests that they can perform many functions. As part of this thesis, we first looked at the use of the SPADE algorithm coupled with flow cytometry to improve the detection of EVs and allow a better appreciation of their heterogeneity in platelet concentrates (PC) used for transfusion. We also evaluated the use of this approach for the development of biomarkers in the analysis of EVs in the synovial fluid of arthritis patients. This study revealed that the use of algorithms such as SPADE coupled with flow cytometry could facilitate the understanding of the functions of EVs and the development of their studies as biomarkers. We then looked at a first subcategory of platelet EV, those that contain mitochondria (mito⁺EVs). We hypothesized that these mito⁺EVs represent a reservoir of mitochondrial DNA, a damage-associated molecular pattern (DAMP) recognized and present in many inflammatory conditions, and that they could be involved in transfusion reactions. We observed that mito⁺EVs were more abundant in PCs involved in transfusion reactions and that they correlate significantly with mitochondrial DNA. As the majority of mitochondrial DNA is encapsulated in EVs, this study suggests that EVs may be a useful biomarker for predicting the potential risk of transfusion reactions, although further investigation is needed to determine if there is a pathogenic role of mitochondrial DAMP encapsulated in EVs as opposed to mitochondrial DNA in solution. We finally focused on a second subcategory of platelet EVs, those that contain proteasome. Platelets contain an active proteasome and present antigens through the major histocompatibility complex I (MHC I), which gives them an important function in adaptive immunity. Whether the proteasome is also transferred into the v platelet EVs has never been examined. We hypothesized that a functioning machinery for the processing and presentation of the antigen is transferred to the platelet EVs. Using a combination of molecular and functional assays, we have demonstrated the presence of an active 20 S proteasome under various conditions where platelets are activated, as well as MHC I and lymphocyte coactivation molecules. Demonstrated by activation and proliferation of ovalbumin (OVA) antigen specific CD8⁺ T lymphocytes, EVs from platelets incubated with OVA can efficiently prepare and present antigen. These results suggest that platelet EVs contribute to adaptive immunity. Overall, we have shown that platelet EVs have a role in inflammation and immunity with their mitochondria and proteasome content. They are heterogeneous and can be used as biomarkers in different contexts.|
|Document Type:||Thèse de doctorat|
|Open Access Date:||7 June 2021|
|Collection:||Thèses et mémoires|
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