건강인 말초혈액으로부터 항원 특이적 T 세포 분석 및 개발

Abstract

Adoptive cell therapy using antigen-specific T cells is a promising treatment mo- dality for cancer patients. Therefore, novel methods for isolating antigen-specific T cells and identifying corresponding T cell receptors are needed to find effective targets for adoptive cell therapy. We evaluated the proliferation of antigen-specific T cells in PBMC by staining them with CFSE dye and stimulating them with antigen peptides to isolate antigen-specific T cells. In addition, we analyzed CMV and EBV-specific TCR sequences and transduction of specific TCRs to T cells and validation of TCR-T. Isolated CD8+ cells from healthy donor PBMC exhibited the most proliferation in response to CMV and EBV at day 7, under G2 conditions using minimal IL-2. TCR sequence analysis also demon- strated responsiveness to CMV and EBV peptides in vitro. Reactivity was confirmed in all 10 types of CMV-specific TCR candidates and in EBV TCR 3. In conclusion, this methodology can be a cost-effective and time-efficient way to identify neoantigen-spe- cific T cells and TCRs, which may serve as novel potential targets for immunotherapy.