Lease of EVs per cell, higher purity EVs.OF11.Prolongation of allograft survival via donor MHC chimerism

Lease of EVs per cell, higher purity EVs.OF11.Prolongation of allograft survival via donor MHC chimerism induced by extracellular vesicles Bruno Adonai Gonzalez Nolascoa, Mengchuan Wanga, William Orenta, Aurore Prunevieillea, Jane Oa, Kaitlan Ahrensa, Joren C Madsenb and Gilles BenichouaISEV2019 ABSTRACT BOOKa Division of Gastrin Proteins supplier Surgery, Center for Transplantation Sciences, Massachusetts General Hospital and Harvard Healthcare School, Boston, USA; bDepartment of Surgery, Center for Transplantation Sciences and Division of Cardiac Surgery, Massachusetts Basic Hospital and Harvard Medical College, Boston, USAOF11.Proteomic and transcriptomic characterization of exosomes-mimetic nanovesicles reveals their relevance as a therapeutic delivery system Amirmohammad Nasiri Kenaria, Kenneth Kastaniegaardb, Mitch C. Shambrooka, David Greeninga, Allan Stensballeb, Lesley Chenga and Andrew HillcaIntroduction: Achieving robust and durable host immune tolerance of allogeneic transplants is the ultimate purpose in clinical transplantation. Mixed chimerism induced by means of donor bone marrow transplantation and host non-myeloablative conditioning has reliably accomplished tolerance of allogeneic organ transplants in mice and humans. Tolerance within this model is believed to rely primarily on the presentation of donor MHC molecules within the host’s thymus. Within this study, we investigated whether or not donor MHC chimerism could possibly be achieved by means of donor extracellular vesicles (EVs) injections and subsequent cross-dressing of recipient cells inside the host’s thymus. Solutions: Conditioned SJL (CD45.1+, H2-Ks+) recipient mice received a single IV dose of purified bone marrow derived exosome-enriched EVs (BM-EVs) isolated from C57BL/6 (CD45.2+, H2-Kb+) donors by means of sequential centrifugation or utilizing a commercially offered exosome isolation kit. Nanoparticle tracking showed vesicles of roughly 100nm in size in the BM-EVs preparation and Western Blot showed the presence of MHCI. Image flow cytometry was applied to detect the presence of cross-dressed cells from day ten through one hundred immediately after exosome injection. For NHP studies, MHC class I H38+ BM-EVs have been injected into a H38- conditioned cynomolgus macaque before a combined heart and kidney transplant. PBMCs, thymus, spleen and mesenteric lymph nodes have been collected for image flow cytometry. Outcomes: Intravenous injection of BM-EVs into conditioned mice resulted within the presentation of donor MHC and CD45.1 molecules by host’s thymic and splenic cells. Similarly, H38+cross-dressed cells have been detected at D33 after exosome injection in all of the NHP recipient tissues collected. In mice, donor but not syngeneic or third-party BM-EVs drastically prolonged skin allograft survival (median survival = 17 VS 11 days, p 0.001). Summary/Conclusion: These outcomes show that delivery of donor-derived extracellular vesicles can induce donor MHC chimerism by means of cross-dressing of recipient APCs with allogeneic MHC molecules in the host’s thymus. This suggests that donor EVs may be utilized in spot of bone marrow cells to induce chimerism and allograft survival with minimal conditioning and no threat of graft versus host CD160 Proteins Molecular Weight illness (GVHD). Funding: NIH R01DK115618.bDepartment of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia; Department of Overall health Science and Technology, Faculty of Medicine, Aalborg University, Denmark, Aalborg, Denmark; cThe Division of Biochemistry and Genetics, La Trobe Institute for Molec.