Alterations of milk oligosaccharides in mothers with gestational diabetes mellitus impede colonization of beneficial bacteria and development of RORγt ⁺ Treg cell-mediated immune tolerance in neonates

Abstract

Gestational diabetes mellitus (GDM) is an increasing public health concern that significantly increases the risk of early childhood allergic diseases. Altered maternal milk glycobiome may strongly affect gut microbiota and enteric-specific Treg cell-mediated development of immune tolerance in GDM infants. In this study, we found that, compared with healthy Chinese mothers, mothers with GDM had significantly lower levels of total and specific human milk oligosaccharides (HMOs) in their colostrum that subsequently increased with extension of lactation. This alteration in HMO profiles significantly delayed colonization of <i>Lactobacillus</i> and <i>Bifidobacterium</i> spp. in their breast-fed infants, resulting in a distinct gut microbial structure and metabolome. Further experiments in GDM mouse models indicated that decreased contents of milk oligosaccharides, mainly 3'-sialyllactose (3'-SL), in GDM maternal mice reduced colonization of bacteria, such as <i>L. reuteri</i> and <i>L. johnsonii</i>, in the neonatal gut, which impeded development of RORγt⁺ regulatory T (Treg) cell-mediated immune tolerance. Treatment of GDM neonates with 3'-SL, <i>Lactobacillus reuteri</i> (<i>L. reuteri</i>) and <i>L. johnsonii</i> promoted the proliferation of enteric Treg cells and expression of transcription factor RORγt, which may have contributed to compromising ovalbumin (OVA)-induced allergic responses. <i>In vitro</i> experiments showed that 3'-SL, metabolites of <i>L. johnsonii</i>, and lysates of <i>L. reuteri</i> stimulated differentiation of mouse RORγt⁺ Treg cells through multiple regulatory effects on Toll-like receptor, MAPK, p53, and NOD-like receptor signaling pathways. This study provides new ideas for the development of gut microbiota and immune tolerance in GDM newborns.