Figure 1. Defining the immune landscape of term decidua.
(A) Experimental design: CD45+ leukocytes were isolated from maternal blood and decidua from term cesarean deliveries (n = 4). Single-cell suspensions of decidual leukocytes and matched PBMCs were subjected to gene expression profiling using 10× 3′ single-cell gene expression protocol (scRNA-seq).
Additionally, sorted decidual and peripheral T cells were surface stained with DNA oligo-tagged antibodies and hashing antibodies and subjected to 5′ gene expression and TCR on the 10x platform (n = 5). Finally, the frequency, phenotype, and function of various immune cells were assayed using flow cytometry.
(B) Uniform Manifold Approximation and Projection (UMAP) of 12,698 CD45+ decidual leukocytes from four donors.
(C) Violin plots showing log-transformed normalized expression levels of major cell-type determining markers identified using Seurat.
(D) Stacked bar graph comparing frequencies of major immune cell types in term decidua (n = 62) using multiparameter flow cytometry. Error bars represent the mean and standard error of the mean (SEM).
(E) Changes in frequencies of major immune cell subsets in the decidua based on scRNA-seq obtained from first-trimester (T1, weeks 6–12) decidua9 integrated with week 37 (T3) data generated in this study.
(F) Feature plots comparing expression of transcription factors, surface markers, and cytokines associated with immune activation at T3 relative to T1.