B195
GMP production of therapeutic T-cells for a Cancer Research UK phase I clinical trial of autologous tumour antigen-specific T-cells for patients with advanced CEA positive tumours
Ryan Guest1, Natalia Kirillova1, Joely Irlam-Jones1, Lidan Christie1, Eric Austin1, Keith Smith3, Robert Hawkins2
1NHS Blood and Transplant, Manchester, UK, 2University of Manchester, Manchester, UK, 3NHS Blood and Transplant, Cambridge, UK
The paucity of an autologous T-cell response to tumour associated antigens can be due to multiple factors such as: abnormal HLA expression; depletion of self re-acting T-cells during thymic selection; and expresion of immune inhibitory factors such as TGF-beta and IL-10. In contrast some tumour associated antigens have been shown to be good targets for antibody recognition. MFEz consists of the CEA recognition domains of an antibody (scFv) fused directly to the T-cell receptor signalling domain (CD3z). T-cells engineered to express MFEz have shown anti-tumour responses in model systems. Here we describe the production of the MFEz T-cells to GMP for use in a clinical Phase I trial.
Systems were developed and validated to activate, transfect, expand, and finally wash and concentrate the cells prior to infusion. This required the use of: GMP sourced reagents; culture and transfection bags; sterile connecting devices; and a Cytomate for the final product processing.
Using these systems we were able to obtain a GMP product: that had expanded 100-300 fold over 14 days; had an average transfection frequency of 28±10%; with a final T-cell viability of 85%±5; and was negative by gram stain, mycoplasma and BacT/ALERT testing. The Cytomate process allowed the volume to be reduced from upto 20L to 250ml with no obvious loss of cells or viability.
This fully validated GMP system has therefore generated a gene modified T-cell product expressing MFEz which meets both the functional and regulatory requirements for infusion.
