1. CRISPR/Cas9-Engineered HLA-Deleted Glomerular Endothelial Cells as a Tool to Predict Pathogenic Non-HLA Antibodies in Kidney Transplant Recipients
- Author
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Marion Rabant, Jean Luc Taupin, Aniela Zablocki, Tifanie Blein, Soëli Charbonnier, Dany Anglicheau, Charlotte Leclaire, Baptiste Lamarthée, Carole Burger, Julien Zuber, Fabiola Terzi, Mélanie Hardy, Lise Morin, Xavier Lebreton, Béatrice Charreau, Claire Tinel, Christophe Legendre, Renaud Snanoudj, Emilie Lebraud, Morgan Gallazzini, Simon C. Satchell, Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre de Recherche en Transplantation et Immunologie - Center for Research in Transplantation and Translational Immunology (U1064 Inserm - CR2TI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Nantes Université - UFR de Médecine et des Techniques Médicales (Nantes Univ - UFR MEDECINE), Nantes Université - pôle Santé, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Nantes Université - pôle Santé, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ), Centre européen des sciences de la transplantation et de l'immunologie [CHU Nantes] (IHU CESTI), Centre hospitalier universitaire de Nantes (CHU Nantes)-Institut de transplantation urologie-néphrologie (ITUN), LabEX IGO Immunothérapie Grand Ouest, Nantes Université (Nantes Univ), Ecotaxie, microenvironnement et développement lymphocytaire (EMily (UMR_S_1160 / U1160)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Hopital Saint-Louis [AP-HP] (AP-HP), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Immunologie humaine, physiopathologie & immunothérapie (HIPI (UMR_S_976 / U976)), University of Bristol [Bristol], and Le Bihan, Sylvie
- Subjects
Adult ,Graft Rejection ,Male ,Reoperation ,CRISPR-Cas systems ,Non hla antibodies ,[SDV]Life Sciences [q-bio] ,Kidney Glomerulus ,Bristol Heart Institute ,kidney transplantation ,Human leukocyte antigen ,Kidney transplant ,kidney rejection ,Clinical Research ,HLA Antigens ,Isoantibodies ,endothelial ,Medicine ,CRISPR ,Humans ,antibodies ,Cells, Cultured ,Aged ,Retrospective Studies ,business.industry ,Nuclear Proteins ,General Medicine ,Middle Aged ,Tissue Donors ,endothelial cells ,non-HLA antibodies ,[SDV] Life Sciences [q-bio] ,Nephrology ,inflammation ,Immunology ,Trans-Activators ,antibody-mediated rejection ,Female ,endothelial inflammation ,business ,beta 2-Microglobulin ,Gene Deletion - Abstract
International audience; Background After kidney transplantation, donor-specific antibodies against human leukocyte antigen donor-specific antibodies (HLA-DSAs) drive antibody-mediated rejection (ABMR) and are associated with poor transplant outcomes. However, ABMR histology (ABMRh) is increasingly reported in kidney transplant recipients (KTRs) without HLA-DSAs, highlighting the emerging role of non-HLA antibodies (Abs). Methods W e designed a non-HLA Ab detection immunoassay (NHADIA) using HLA class I and II–deficient glomerular endothelial cells (CiGEnC Δ HLA) that had been previously generated through CRISPR/Cas9-induced B2M and CIITA gene disruption. Flow cytometry assessed the reactivity to non-HLA antigens of pretransplantation serum samples from 389 consecutive KTRs. The intensity of the signal observed with the NHADIA was associated with post-transplant graft histology assessed in 951 adequate biopsy specimens. Results W e sequentially applied CRISPR/Cas9 to delete the B2M and CIITA genes to obtain a CiGEnC Δ HLA clone. CiGEnC Δ HLA cells remained indistinguishable from the parental cell line, CiGEnC, in terms of morphology and phenotype. Previous transplantation was the main determinant of the pretransplantation NHADIA result ( P
- Published
- 2021
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