ScopeFood allergy has become a world recognized public health problem due to its versatility and lack of efficacious methods for its treatment. Probiotics supplement is a potential way to prevent food allergy.Methods and resultsIn this study, potential strains are screen out by peripheral blood mononuclear cells (PBMCs), and their abilities of alleviating food allergy are examined using a mouse model induced by ovalbumin (OVA). The results show that six strains increase ratio of interferon‐γ (IFN‐γ)/interleukin (IL)‐4 secreted by PBMCs with good abilities in intestinal adhesion and gastrointestinal tolerance. Oral administration of Bifidobacterium animalis KV9 (KV9) and Lactobacillus vaginalis FN3 (FN3) attenuates allergic responses in allergy mice, including allergic symptoms, mast cells aggregation and activity, serum OVA‐special‐immunoglobulins E (OVA‐sIgE) production. KV9 and FN3 upregulate the production of IFN‐γ/IL‐4 in splenocytes, increase the genes and proteins expression of toll‐like receptor 4 (TLR4), myeloid differentiation primary response 88 (Myd88) and interferon regulatory factor (IRF)‐1 in allergic mice spleen, and decrease the IRF‐4.ConclusionThe study demonstrates that KV9 and FN3 possess anti‐allergic activities via activation of TLR4 pathway and modulating the expression of IRF‐1 and IRF‐4 which leads to T helper type 1 (Th1)/T helper type 2 (Th2) cell immunology balance.