Ibrutinib, but not acalabrutinib, inhibits anti-lymphoma T cell and NK cell function

Abstract Chronic Lymphocytic Leukemia (CLL) is the predominant hematologic malignancy in the U.S. BTK inhibitors (BTKi) demonstrate prolonged survival compared to standard chemotherapy, but they also show notable toxicity, including increased risk for infections. While BTKi have shown both immunosuppressive and -stimulatory effects, the overall effect on T cells (TC), including tumor-reactive TC, is not well understood. Because non-BTK TEC-family and homologous kinases have critical roles in immune cells, we tested whether differentially specific BTKi have differing immunomodulatory effects. We treated mice with ibrutinib or acalabrutinib in vivo and analyzed their immune cell repertoire. Mass cytometry revealed that acalabrutinib, but not ibrutinib, increased the number of NK cells and myeloid cells in spleens and tumors of A20-lymphoma bearing mice, while both BTKi increased TC numbers in A20 tumors. Ibrutinib inhibited the activation of CD8 TC and NK cells isolated from BTKi-treated mice and their ability to lyse tumor cells ex vivo, while acalabrutinib improved CD8 TC-mediated cytolysis. A20 tumor growth was not influenced by BTKi in vivo, but ibrutinib inhibited the ability of transferred, antigen-specific TC to induce tumor regression. To translate our findings, we tested the effect of BTKi on immunogenicity of primary human CLL/MCL cells and their ability to stimulate TC. Ibrutinib inhibited upregulation of activation markers on tumor cells to a higher extent than acalabrutinib. Similarly, only ibrutinib inhibited autologous TC activation after co-culture with tumor cells and superantigen. In conclusion, ibrutinib, but not acalabrutinib inhibits the activation and function of anti-tumor T and NK cells in vivo and in vitro.