Generation of Autologous CMV and EBV-Specific T-Cell Lines from Seronegative Patients for Cell Therapy of Opportunistic Infections Following Kidney Transplantation.
C. Lamarche,1 T. Pincez,1 J. Orio,1 C. Carli,1 A. Humar,2 J.-S. Delisle.1
1Centre de Recherche de l'Hôpital Maisonneuve-Rosemont, Montreal, QC, Canada
2Toronto General Research Institute, Toronto, ON, Canada
Meeting: 2017 American Transplant Congress
Abstract number: B20
Keywords: Epstein-Barr virus (EBV), Kidney transplantation, T cells, Viral therapy
Session Information
Session Name: Poster Session B: Acute and Chronic Rejection
Session Type: Poster Session
Date: Sunday, April 30, 2017
Session Time: 6:00pm-7:00pm
Presentation Time: 6:00pm-7:00pm
Location: Hall D1
Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) reactivations after kidney transplant are associated with substantial morbidity, especially in seronegative patients. Adoptive immunotherapy using virus-specific T cells lines generated ex vivo could circumvent these limitations. In addition, autologous T-cell lines would probably be safer and durable relative to third party in the context of solid organ transplant. However, the optimal timing to manufacture these T-cell lines, before or after transplant, has never been studied. Our objective is to define the optimal timing to generate autologous CMV or EBV-specific T-cell lines from seronegative patients.
We adapted a protocol to obtain CMV or EBV-specific T-cell lines from seronegative CKD patients listed for transplant, KTR and healthy controls (HC). We used 15×106peripheral blood mononuclear cells (PBMC) stimulated twice with monocytes-derived dendritic cells (DC) pulsed with EBV or CMV-specific peptide libraries. At the end of the 14-days culture, we performed cell count, flow cytometry analysis and enzyme-linked immunospots (ELISpot) assay.
We found that CKD patients and KTR have less lymphocytes compared to HC. Also, there is more evidence of exhaustion in CD4+ and CD8+ T cells from CKD patients and their PBMC display lower survival leading to decreased monocyte-derived DC production. However, EBV and CMV-specific T-cell lines from seronegative CKD patients and KTR are comparable after a 14-days culture in term of expansion, antigen reactivity and exhaustion marker expression.
T cells from CKD patients seemed more exhausted and had a worse recovery post thawing that might be explained by a greater susceptibility of exhausted T cells to cell death. In conclusion, although T lymphocytes from both seronegative CKD and KTR are adequate to generate autologous T-cell lines for anti-viral adoptive immunotherapy, those from CKD patients were less resistant to freezing and thawing, thereby indicating that special care must be taken for the handling of these cells.
CITATION INFORMATION: Lamarche C, Pincez T, Orio J, Carli C, Humar A, Delisle J.-S. Generation of Autologous CMV and EBV-Specific T-Cell Lines from Seronegative Patients for Cell Therapy of Opportunistic Infections Following Kidney Transplantation. Am J Transplant. 2017;17 (suppl 3).
To cite this abstract in AMA style:
Lamarche C, Pincez T, Orio J, Carli C, Humar A, Delisle J-S. Generation of Autologous CMV and EBV-Specific T-Cell Lines from Seronegative Patients for Cell Therapy of Opportunistic Infections Following Kidney Transplantation. [abstract]. Am J Transplant. 2017; 17 (suppl 3). https://atcmeetingabstracts.com/abstract/generation-of-autologous-cmv-and-ebv-specific-t-cell-lines-from-seronegative-patients-for-cell-therapy-of-opportunistic-infections-following-kidney-transplantation/. Accessed December 4, 2024.« Back to 2017 American Transplant Congress