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).

« Back to 2017 American Transplant Congress