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.

Addition of TLR agonists promotes Th1 polarization. To investigate the discrepancy between HLA-pentamer positive cells and IFN-c positive cells in cultures with polyI:C or ssRNA40 (Fig. 4B -D) we analyzed the supernatant taken from cultures after the 48 h incubation with anti-CD3, IL-2 and TLR ligands. We tested for the production of 42 different cytokines in cultures derived from 3 separate healthy donors and measured significant production of 27 different cytokines. We found that addition of TLR ligands generally reduced the production of immunoregulatory/Th2 cytokines compared to IL-2 alone (Fig. 5A) . This was especially true for ssRNA40 in all cases except the increased production in IL-10; observed with both polyI:C and ssRNA40. In coordination with the reduced production of regulatory cytokines we observed increased production of inflammatory cytokines. In particular, the Th1 polarizing cytokine IL-12p70 was significantly increased with ssRNA40 and mildly increased with polyI:C (Fig. 5B) .