Immovable cultural heritage—archaeological sites, historic buildings, and culturally significant infrastructure—has traditionally been approached through conservation, material preservation, and identity-based perspectives. However, the evolution of heritage theory and the emergence of systemic paradigms such as One Health call for its reinterpretation as an active component within interconnected human, animal, and environmental systems. Although One Health recognizes the interdependence of these domains, no operational framework currently assesses the functional contribution of immovable cultural heritage. This study develops a formal methodological framework that operationalizes immovable cultural heritage as a functional element within the One Health system. The framework integrates environmental, animal, and human health domains through structured indicators, mathematical formalization, and internal validation procedures. It explicitly incorporates the coexistence of tangible and intangible heritage dimensions, acknowledging their embedded socio-ecological relationships. The plausibility and coherence of the framework is validated against established scientific literature, environmental assessment models, and foundational One Health principles. Results demonstrate that the proposed approach enables systematic, reproducible, and domain-complete assessment of immovable cultural heritage within the One Health paradigm, overcoming methodological fragmentation and supporting integration with sustainability analysis, environmental governance, resilience planning, and long-term socio-ecological stability.