Th141 - Single Cell Isolation of CD4+ T Cells Specific for NY-ESO-1 Presented on HLA-DRB3*02:02

Andrea Aran – FUNDACIÓ DE RECERCA CLÍNIC BARCELONA-INSTITUT D’INVESTIGACIONS BIOMÈDIQUES AUGUST PI I SUNYER.; Ariadna Bartoló – FUNDACIÓ DE RECERCA CLÍNIC BARCELONA-INSTITUT D’INVESTIGACIONS BIOMÈDIQUES AUGUST PI I SUNYER.; Mariona Pascal – Hospital Clínic de Barcelona (HCB); Europa Azucena González-Navarro – Hospital Clínic de Barcelona (HCB); Manel Juan – Hospital Clínic de Barcelona (HCB)

Abstract Text: Natural transgenic T-cell receptors (tTCR) represent an emergent type of cell therapy against cancer, recognizing peptides expressed both on the intracellular and extracellular level. Recognizing intracellular antigens constitutes a significant advantage in comparison to CAR therapy that can only recognize extracellular surface proteins, which are difficult to define as Tumor Associated Antigens (TAA), especially on solid tumors. However, it must be taken into consideration that TCR therapies against TAAs could potentially generate off-tumor on-target toxicity. In order to prevent this risk, this work is focused on NY-ESO-1, from the cancer/testis antigen family, expressed on several types of tumors but not relevantly on healthy tissues besides testicles. Our work is focused on class II-presented peptides, namely NY-ESO-1119-143 on HLA-DRB3*02:02 (DR52b), expressed approximately by half of the Caucasian population.
Specific CD4+ CD25- T cells against NY-ESO-1119-143 presented on HLA-DRB3*02:02 were obtained after 30 days of cell culture and two rounds of cell sorting at days 15 and 30. T cell purity and specificity were assessed using CD4 and CD154 markers, respectively. Purity was maintained at 97%, and specificity increased significantly in the second cell sorting in comparison with the first one. Single cell sequencing by Rhapsody® system was performed to obtain TCR sequences as well as RNA and protein expression profiling.