Building and Optimizing a Virus-specific T Cell Receptor Library for Targeted Immunotherapy in Viral Infections

Banu, Nasirah; Chia, Adeline; Ho, Zi Zong; Garcia, Alfonso Tan; Paravasivam, Komathi; Grotenbreg, Gijsbert M.; Bertoletti, Antonio; Gehring, Adam J.

As a final characterization of their function, and to determine whether culturing in different conditions altered transduced T cell cytotoxic function, we compared the ability of T cells transduced with the HBV core antigen-specific TCR to lyse hepatocyte-like cell lines (HepG2) expressing HBV antigens. We found that T cells cultured under any of the conditions exhibited efficient and similar lysis of targets expressing the HBV core antigen (HG2cAg). We observed 95% specific lysis at a 155 E5T ratio and 80% specific lysis at a 1510 E5T ratio (Fig. 6G) . Co-culture with HepG2 cells expressing the irrelevant HBV surface antigen (HG2sAg) did not result in any significant lysis. We were unable to titrate the cytotoxicity experiment for sensitivity as the antigen in HepG2 cells is produced from a CMV promoter following transduction with a lentiviral vector. However, this data demonstrated that transduced T cells cultured under any condition tested are specific and highly cytotoxic.