Humoral Reactivity of Renal Transplant-Waitlisted Patients to Cells From GGTA1/CMAH/B4GalNT2, and SLA Class I Knockout Pigs
Top Cited Papers
- 1 April 2017
- journal article
- research article
- Published by Ovid Technologies (Wolters Kluwer Health) in Transplantation
- Vol. 101 (4), e86-e92
- https://doi.org/10.1097/tp.0000000000001646
Abstract
Antipig antibodies are a barrier to clinical xenotransplantation. We evaluated antibody binding of waitlisted renal transplant patients to 3 glycan knockout (KO) pig cells and class I swine leukocyte antigens (SLA). Peripheral blood mononuclear cells from SLA identical wild type (WT), α1, 3-galactosyltransferase (GGTA1) KO, GGTA1/ cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) KO, and GGTA1/ CMAH /b1,4 N-acetylgalactosaminyl transferase (B4GalNT2) KO pigs were screened for human antibody binding using flow cytometric crossmatch (FCXM). Sera from 820 patients were screened on GGTA1/CMAH/B4GalNT2 KO cells and a subset with elevated binding was evaluated further. FCXM was performed on SLA intact cells and GGTA1/SLA class I KO cells after depletion with WT pig RBCs to remove cell surface reactive antibodies, but leave SLA antibodies. Lastly, human and pig reactive antibodies were eluted and tested for cross-species binding and reactivity to single-antigen HLA beads. Sequential glycan KO modifications significantly reduce antibody binding of waitlisted patients. Sera exhibiting elevated binding without reduction after depletion with WT RBCs demonstrate reduced binding to SLA class I KO cells. Human IgG, eluted from human and pig peripheral blood mononuclear cells, interacted across species and bound single-antigen HLA beads in common epitope-restricted patterns. Many waitlisted patients have minimal xenoreactive antibody binding to the triple KO pig, but some HLA antibodies in sensitized patients cross-react with class I SLA. SLA class I is a target for genome editing in xenotransplantation.Keywords
This publication has 26 references indexed in Scilit:
- Immunogenicity of Renal Microvascular Endothelial Cells From Genetically Modified PigsTransplantation, 2016
- Recent investigations into pig antigen and anti-pig antibody expressionInternational Journal of Surgery, 2015
- Pre‐transplant antibody screening and anti‐CD154 costimulation blockade promote long‐term xenograft survival in a pig‐to‐primate kidney transplant modelXenotransplantation, 2015
- Evaluation of human and non‐human primate antibody binding to pig cells lacking GGTA1/CMAH/β4GalNT2 genesXenotransplantation, 2015
- Creating Class I MHC–Null Pigs Using Guide RNA and the Cas9 EndonucleaseThe Journal of Immunology, 2014
- Potential deleterious role of anti‐Neu5Gc antibodies in xenotransplantationXenotransplantation, 2014
- Identification of New Carbohydrate and Membrane Protein Antigens in Cardiac XenotransplantationTransplantation, 2011
- Xenoantigens and xenoantibodiesXenotransplantation, 1998
- CARBOHYDRATE ANTIGENS OF PIG TISSUES REACTING WITH HUMAN NATURAL ANTIBODIES AS POTENTIAL TARGETS FOR HYPERACUTE VASCULAR REJECTION IN PIG-TO-MAN ORGAN XENOTRANSPLANTATION1Transplantation, 1993
- Identification of T-Cell EpitopesJournal of Immunotherapy, 1993