Abstract PR012: Adoptive transfer of T cells surface-tethered with IL-12 activates a natural killer/dendritic cell axis to promote antigen spreading for enhanced anti-tumor efficacy
Authors: Stokes Kate L. , Jaehger Ditte E. , Pratama Alvin , Halldorsdottir Holmfridur R. , Ahmad Gulzar , Nardozzi Jon D. , Sackton Katharine L. , Andresen Thomas L. , Jones Douglas S. 
Published: February 2021

Abstract
Despite the promise of adoptive cell therapy (ACT) in the treatment of some cancer types, success against solid tumors remains elusive due to multiple immune escape pathways. T cell therapies that affect tumor microenvironment or provide adjuvant activity in addition to tumor cell lysis hold strong potential for overcoming immune escape in solid tumors. Interleukin-12 (IL-12) is a potent stimulator of innate and adaptive immune cells that holds strong potential for cancer immunotherapy, but its clinical utility has been limited by high systemic toxicities. We have previously shown that tethering an IL-12 immunocytokine to the surface of T cells prior to adoptive cell transfer (ACT) safely improves anti-tumor efficacy by promoting T cell function specifically in the tumor. Here, we demonstrate that tethered IL-12 further delivers adjuvant activity by initiating an immune cascade involving natural killer cell-mediated activation of cross-priming dendritic cells to resulting in broader anti-tumor T cell responses. In a preclinical cell therapy model tumor-specific T cells tethered with IL-12, but not the T cells alone, triggered proliferation of tumor-specific endogenous tumor infiltrating lymphocytes (TIL). This was accompanied by increased activation and infiltration of cross-presenting dendritic cells (cDC1) in tumor-draining lymph nodes (tdLN). Analyses of cDC1 subsets revealed upregulation of CD86 expression on both migratory and resident cDC1s within the tdLN, two cell types that are critical for both tumor antigen transport to tdLN and activation of naïve CD8 T cells, respectively. This activity was specific to cDC1s, as increased activation or accumulation of type 2 dendritic cells (cDC2) was not observed. Mechanistically, the IL-12 tethered T cells upregulated expression of FLT3 ligand on NK cells, a key cytokine for cDC1 recruitment and expansion. NK cell depletion ablated cDC1 tdLN infiltration and endogenous tumor infiltrating T cell expansion, indicating that recruitment and activation of endogenous tumor-specific T cells is at least partially driven by tethered IL-12-mediated activation of NK cells to recruit and expand the cross-priming dendritic cells. Together, this demonstrates that our surface tethered cytokine technology provides powerful adjuvant activity to adoptively transferred T cells and promotes activation of endogenous anti-tumor adaptive immune response, thus promoting a broad antigenic repertoire. This approach thus holds potential to help overcome challenges of antigen escape and tumor heterogeneity that limit efficacy of T cell therapies against solid tumors.
This abstract is also being presented as PO078.
Citation Format: Kate L. Stokes, Ditte E. Jaehger, Alvin Pratama, Holmfridur R. Halldorsdottir, Gulzar Ahmad, Jon D. Nardozzi, Katharine L. Sackton, Thomas L. Andresen, Douglas S. Jones. Adoptive transfer of T cells surface-tethered with IL-12 activates a natural killer/dendritic cell axis to promote antigen spreading for enhanced anti-tumor efficacy [abstract]. In: Abstracts: AACR Virtual Special Conference: Tumor Immunology and Immunotherapy; 2020 Oct 19-20. Philadelphia (PA): AACR; Cancer Immunol Res 2021;9(2 Suppl):Abstract nr PR012.