Background: There is an urgent need for risk assessment and mechanism exploration of fish allergens to provide a theoretical basis and methodological reference for food safety. PV is a major marine food allergen that can cause allergic reactions in fish allergic patients. This study aims to break through the previous model of evaluating the sensitivity of food allergens on single effector cells but to construct a coupling cell co-culture system for the process of dendritic cell maturation and T cell differentiation during sensitization. Methods: A new type of highly effective allergen sensitization evaluation model was proposed based on the effect of PV-induced differentiation of dendritic cells and T cells on the sensitization process. Results: Flow cytometry, ELISA, proteomics and transcriptomics approaches revealed that PV could promote the maturation of dendritic cells, mainly by inducing BMDC to express MHC Ⅱ, CD86, and other phenotypic molecules, while promoting cytokines/chemokines IL-6, IFN-γ, IL-23, and IL-12p70, whereas inhibiting TNF-α expression. It could strengthen the synthesis process of biological protein of DC cells and improve the expression level of genes such as the migration ability of DC cells in vivo. Finally, PV could induce T cells to shift to Th2 differentiation by inducing the increase of IL-4 level, which leads to allergic behavior. Conclusions: Therefore, the new allergen sensitization evaluation coupled with the cell co-culture system, in combination with the changes of the above markers could be used for quick and efficient determination of the potential sensitization possibility and sensitization intensity of fish allergens.