Multi-omics Analysis of Immune Cells Induced by Artificial Adjuvant Vector Cell (aAVC) Therapy in HPV-Positive Tumor-Bearing Mice [scRNA-seq]

Immune evasion by tumors highlights the need to activate both CTLs and NK cells against “immune-desert” type of tumor, yet the synergistic potential of dual activation in immune-desert tumors remains unclear. Here, we uncover a spatiotemporal synergy between NK cells and CTLs that drives functional antitumor effects in an immune-desert tumor model. Using a multi-omics imaging approach integrating spatial transcriptomics, scRNA-seq re-clustering, and multicolor flow cytometry, we show that vaccination rapidly reshapes lymphocyte localization and function in the tumor microenvironment (TME). Early activated IRF8+Klrg1+NK cell infiltration establishes a niche that guides subsequent antigen-primed CXCR3+CTL influx, accompanied by dendritic cell activation and inflammatory monocyte recruitment. This coordinated response relies on structural and chemotactic remodeling of the TME, facilitating NK-CTL interactions at the tumor margin. Our findings demonstrate how integrated multi-omics analyses can uncover previously inaccessible interactions and provide a framework for designing next-generation immunotherapies.