Development of next generation car's targeting the lewis y antigen for the treatment of cancer

ConclusionWe successfully generated a stably expressing packaging cell line for this anti-LeY-CAR-CD34 containing retroviral vector. Retroviral supernatant derived from these packaging cells was then used to successfully transduce primary human T cells with the anti-LeY CAR and positive cells were sorted by flow cytometry using the CD34 marker. After sorting by flow cytometry, the majority of the T cells (>90%) were double positive for the anti-LeY CAR and CD34 marker. Antigen-specific function of the gene-modified T cells was then examined in cytokine and cytotoxicity assays to examine whether the presence of the CD34 marker may have compromised CAR function. The anti-LeY-CD34 CAR-T cells were found to secrete high levels of interferon γ, TNFα and IL-2 following direct stimulation through the CAR using immobilized plate bound anti-idiotype antibody or LeY+ tumor targets. The CAR T cells also specifically killed LeY+ tumor cells but not LeY− targets cells indicating that functional responses were not affected by the presence of the CD34 marker. Our results suggest that these new hu-CD34 CARs against LeY tumor antigen could offer new-targeted therapy with acceptable toxicity for patients with solid cancers given the low expression of this antigen on normal tissues. Furthermore this innovative tracking system with the CD34 marker will allow better characterisation and monitoring of the infused CAR-T cells in vivo in preclinical mouse models and in patients in future trials.
Source: Cytotherapy - Category: Cytology Source Type: research