S1pr is involved in immune regulation, and Nfatc1 and Pak2 support the phenotype of Foxp3+ Treg cells and maintain the function of suppressive Treg cells.30,31 The maintenance of immune tolerance by Treg cells is critical to prevent allergic and autoimmune diseases.60 Thus, our results suggested that supplementation of 3’-SL, L. reuteri, and L. johnsonii synergistically promoted the proliferation of intestinal RORγt+ Treg cells through multiple pathways, which in turn contributed to establishment of immune tolerance to OVA stimulation in offspring. Here, NFATC1 is linked to autoimmune disease.