Probiotic-based oral vaccine mucosal delivery system enabling genetically encoded dual-antigen arrays

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Oral vaccines provide a non-invasive approach for cancer immunotherapy but face challenges in gastrointestinal stability, antigen presentation, and mucosal delivery. Here, we present an engineered probiotic-based oral vaccine system, BacOR-Fn-T+phiX174, featuring genetically encoded dual-antigen ferritin arrays and inducible bacterial lysis. Upon oral administration and arabinose induction, the probiotic strain lyses in situ, releasing OVA/TRP2-decorated ferritin nanoparticles that efficiently traverse the intestinal barrier via M-cell targeting and activate mucosal dendritic cells. This platform robustly stimulates CD8+ and CD4+ T-cell responses, enhances B-cell and macrophage activation, reduces regulatory T cells, and provides therapeutic efficacy against melanoma in both lung metastasis and subcutaneous tumor models. It also establishes durable immunological memory without disrupting systemic or mucosal homeostasis. This work offers a programmable bacterial chassis for precise antigen array presentation and controlled delivery, representing a promising strategy for next-generation, needle-free cancer vaccines. Minimally invasive oral vaccine delivery is a desired approach. Here, the authors report on an engineered probiotic-based oral vaccine system that enables controlled mucosal delivery of ferritin-displayed dual antigens, eliciting robust mucosal and systemic antitumor immune responses.

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