Background: Non-celiac wheat sensitivity (NCWS) is a poorly understood gluten-related disorder (GRD) and its prominent symptoms can be ameliorated by gluten avoidance. This study aimed to determine the effectiveness of a probiotic mixture in hydrolyzing gliadin peptides (toxic components of gluten) and suppressing gliadin-induced inflammatory responses in Caco-2 cells. Methods: Wheat dough was fermented with a probiotic mix for 0, 2, 4, and 6 h. The effect of the probiotic mix on gliadin degradation was monitored by SDS-PAGE. The expression levels of IL-6, IL-17A, INF-γ, IL-10, and TGF-β were evaluated using ELISA and qRT-PCR methods. Results: According to our findings, fermenting wheat dough with a mix ofB. longum,L. acidophilus, andL. plantarumfor 6 h was effective in gliadin degradation. This process also reduced levels of IL-6 (p= 0.004), IL-17A (p= 0.004), and IFN-γ (p= 0.01) mRNA, as well as decreased IL-6 (p= 0.006) and IFN-γ (p= 0.0009) protein secretion. 4 h fermentation led to a significant decrease in IL-17A (p= 0.001) and IFN-γ (p= 0.003) mRNA, as well as reduced levels of IL-6 (p= 0.002) and IFN-γ (p< 0.0001) protein secretion. This process was also observed to increase the expression levels of IL-10 (p< 0.0001) and TGF-β (p< 0.0001) mRNA. Conclusions: 4 h fermentation of wheat flour with the proposed probiotic mix might be a good strategy to develop an affordable gluten-free wheat dough for NCWS and probably other GRD patients.