Innate-adaptive IFN cross-talk reprograms myeloid and T cells for antitumor immunity with a second-generation recombinant MVA

Resistance to immune checkpoint blockade (ICB) remains a major obstacle in cancer immunotherapy. We rationally engineered a second-generation recombinant Modified Vaccinia virus Ankara (MQ833) by deleting viral immune-evasion genes (E3L, E5R, WR199) and incorporating Flt3L, OX40L, and matrix-anchored IL12. Intratumoral MQ833 elicited robust tumor regressions across multiple models, including ICB-resistant and MHC-I-deficient tumors. Single-cell RNA sequencing revealed extensive remodeling of the tumor microenvironment, characterized by neutrophil and monocyte recruitment and activation, M2 macrophage depletion, M1 polarization, and effector T-cell differentiation and proliferation. Conditional Ifnar1 knockout mice demonstrated that MQ833 efficacy requires type I interferon signaling in neutrophils, macrophages/monocytes, and T cells. Moreover, Nos2 deficiency impaired therapeutic efficacy, confirming iNOS myeloid cells as key effectors. Together, these findings show that MQ833 activates innate-adaptive IFN cross-talk to reprogram myeloid and T cells, defining a rationally designed viral immunotherapy capable of overcoming ICB resistance. GRAPHIC ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=179 SRC="FIGDIR/small/509429v2_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@496f37org.highwire.dtl.DTLVardef@124bd78org.highwire.dtl.DTLVardef@1134528org.highwire.dtl.DTLVardef@216da6_HPS_FORMAT_FIGEXP M_FIG C_FIG