Abstract: Objective: In this study, we investigated the therapeutic effects and underlying mechanisms of Laetiporus sulphureus in a mouse model of food allergy (FA). Methods: Mice were randomly divided into the following four groups: control, FA, L. sulphureus prevention and L. sulphureus treatment. The FA model was established via combined transdermal calcipotriol ointment mixed with ovalbumin (OVA) and oral OVA challenges. The prevention group received daily oral gavage of a suspension of L. sulphureus (2×10⁹ CFU/day) from the initiation of the experiment, whereas the treatment group was administered the same dosage of the bacterial suspension for 4 weeks following successful model establishment. Systemic allergic responses were assessed by monitoring body temperature and clinical symptoms post-challenge and serum IgE and mMCP-1 levels were quantified using ELISA. In addition, histopathological changes were evaluated using hematoxylin and eosin and toluidine blue staining, and Western blotting was performed to analyze the TLR4/NF-κB/MyD88 signaling pathway in lung tissues to assess its potential role in the response of L. sulphureus to OVA-induced FA. Results: Compared with the control mice, those in the FA group were characterized by hypothermia, elevated levels of IgE and mMCP-1, intestinal mucosal damage, mast cell infiltration, and pulmonary vascular dilation, along with the upregulated expression of TLR4, phosphorylated NF-κB, and MyD88. Intervention with L. sulphureus was found to contribute to a significant mitigation of the aforementioned adverse effects, reducing hypothermia, lowering IgE/mMCP-1 levels, attenuating tissue damage, and downregulating activation of the TLR4/NF-κB/MyD88 pathway. Conclusion: Laetiporus sulphureus can contribute to alleviating OVA-induced FA in mice by modulating the TLR4/NF-κB/MyD88 pathway, thereby indicating the potential utility of this bacterium as a therapeutic agent for the treatment of food allergies.