Abstract 1385: The influence of MHC class I and II on T cell responses in mouse melanoma
Authors: Erbe Amy K. , Feils Arika , Heck Mackenzie , VandenHeuvel Sabrina , Castillo Julianna , Hampton Alina , Frankel Lizzie , Hoefges Anna , Carlson Peter , Pieper Alex A. , Aiken Taylor , Zebertavage Lauren , Komjathy David , Spiegelman Dan , Tsarovsky Noah , Morris Zachary S. , Patel Ravi , Rakhmilevich Alexander , Sondel Paul M. 
Published: June 2022

Introduction: Using an in situ vaccine (ISV) regimen consisting of radiation combined with immunocytokine (tumor-targeting mAb linked to IL2), we can cure mice of well established B78 melanoma tumors (B78s). Mice cured of their B78s with ISV demonstrate long-term immune memory, evidenced by rejection of engraftment of a tumor rechallenge &gt;180 days post initial cure of tumor. Traditionally, immaune memory is thought to be mediated via CD8+ T cells, which require antigen presentation via MHC Class I (MHCI). However, B78s express little to no MHCI but do express MHCII when stimulated with IFNγ. While not commonly expressed on solid tumors, MHCII is expressed on 50-60% of melanomas in humans. Here we explored implications of MHCI and MHCII expression in both the primary and long-term memory anti-tumor responses generated with ISV.

Methods: CD4+ or CD8+ T cells were depleted in mice during the primary anti-tumor response (i.e. B78 tumor bearing mice receiving the ISV regimen) or during tumor rechallenge experiments (i.e. B78-cured mice rechallenged with B78s). Tumor growth was monitored. In separate studies, tumors and tumor draining lymphnodes (TDLNs) were harvested during the primary antitumor response and analyzed via flow cytometry to assess T cell activation and immune infiltrate. Finally, TDLNs were harvested from B78-cured mice, 7 days after tumor rechallenge, to define memory T cell subsets.

Results: Depletion studies revealed CD4+ T cells are required for both the antitumor response to ISV and the long-term memory response in B78-cured mice, but CD8+ T cells are not required for either of these responses. Increased CD8+ and CD4+ T cell infiltrates are observed in the tumor microenvironment during the primary anti-tumor response. In B78-cured mice, though not required for memory responses, CD8+ central memory T cells are significantly increased in the TDLNs compared to naïve or primary tumor bearing mice. The amount of CD4+ effector memory T cells are significantly increased in the TDLN of B78-cured mice compared to CD8+ effector memory T cells.

Conclusion: Often not expressed on solid tumors, MHCII is expressed on some melanoma tumors, and its expression has been correlated with a positive response to immunotherapies. CD4+ cytotoxic T cells can directly engage MHCII on tumors, suggesting this interaction has an important role in the response to immunotherapy for MHCII expressing tumors. Our data suggest that CD4+ T cells drive both the primary anti-tumor and long-term immune memory responses in the B78 model when treating with this effective ISV. We are continuing our efforts to understand the characteristics of the B78 cell line that may be relevant to its response to immunotherapy and its resistance to single agent checkpoint blockade. Understanding the cellular and molecular mechanisms involved in ISV-induced immune recognition and destruction of B78s may guide future improvements of this clinically-relevant immunotherapy regimen.

Citation Format: Amy K. Erbe, Arika Feils, Mackenzie Heck, Sabrina VandenHeuvel, Julianna Castillo, Alina Hampton, Lizzie Frankel, Anna Hoefges, Peter Carlson, Alex A. Pieper, Taylor Aiken, Lauren Zebertavage, David Komjathy, Dan Spiegelman, Noah Tsarovsky, Zachary S. Morris, Ravi Patel, Alexander Rakhmilevich, Paul M. Sondel. The influence of MHC class I and II on T cell responses in mouse melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1385.