Rapid generation of genetically engineered T cells for the treatment of virus‐related cancers

Adoptive transfer of T cell receptor (TCR)‐engineered T cells targeting viral epitopes represents a promising approach for treating virus‐related cancers. However, the efficient identification of epitopes for T cells and the corresponding TCR remains challenging. Here, we report a workflow permitting the rapid generation of human papillomavirus (HPV)‐specific TCR‐T cells. Six epitopes of viral proteins belonged to HPV16 or HPV18 were predicted to have high affinity to A11:01 according to bioinformatic analysis. Subsequently, CTL induction were performed with these six antigen peptides separately, and antigen‐specific T cells were sorted by FACS. TCR clonotypes of these virus‐specific T cells were determined using next‐generation sequencing. To improve the efficiency of TCRαβ pair validation, a lentiviral vector library containing 116 TCR constructs was generated that consisted of predominant TCRs according to TCR repertoire analysis. Later, TCR library transduced T cells were simulated with peptide pool‐pulsed antigen‐presenting cells, then CD137‐positive cells were sorted and subjected to TCR repertoire analysis. The top‐hit TCRs and corresponding antigen peptides were deduced and validated. Through this workflow, a TCR targeting the E6(92–101) of HPV16 was identified. These HPV16‐specific TCR‐T cells showed high activity towards HPV16‐positive human cervical cancer cells in vitro and efficiently repressed tumor growth in a murine model. This study provides a HPV16‐specific TCR fitted to the HLA‐A11:01 population, and exemplifies an efficient approach that can be applied in large‐scale screening of virus‐specific TCRs, further encouraging researchers to exploit the therapeutic potential of the TCR‐T cell technique in treating virus‐related cancers.