Single-cell and proteomic profiling unveil aberrant immune phenotypes in gestational diabetes following assisted reproductive technology

Pregnant women undergoing assisted reproductive technology (ART) have an increased risk of gestational diabetes mellitus (GDM), potentially linked to altered immune regulation, but the underlying circulating immune features remain unclear. Here we show an integrated single-cell transcriptomic, immune receptor repertoire, and plasma proteomic analysis of peripheral blood from 32 women with GDM and 31 normal pregnancies after ART. GDM is associated with increased proportions of CD8+ T cells and elevated plasma levels of CD6, CXCL5, MMP10, and 4E-BP1. Cytotoxic CD8+ T cell subsets display enhanced effector and cytotoxic activity, while B cells, monocytes, and natural killer cells exhibit activated phenotypes. Transcription factors from the FOS and JUN families and KLF6 are implicated in immune activation, accompanied by distinct T cell and B cell receptor repertoire features. The hypothesis-generating multi-omic results highlight potential therapeutic targets and offered insights for future research and management ART-associated pregnancy complications. Multimodal immune profiling of ART pregnancies reveals systemic immune hyperactivation in gestational diabetes, characterized by activated CD8⁺ T cells, altered immune repertoires, and inflammatory signaling driven by FOS/JUN and KLF6.