Project description:IL-15-responsive CD122+ macrophages (CD122+Macs) develop in the uterus during normal pregnancy. We aimed to understand the signals driving macrophages to adopt this fate. We aimed to determine how CD122+Macs and conventional uterine macrophages differ transcriptionally.
Project description:Gene expression profile was compared between CD8+CD122+ T cells and CD8+CD122- T cells. mRNA taken from CD8+CD122+ cells or CD8+CD122- cells collected by cell sorting from C57BL/6 mice spleen was amplified and analyzed by using gene chip of Agilent.
Project description:This study uses samples from the Pandion Phase 1a IL-2 mutein (PT-101) trial to investigate the mechanism of action of IL-2 mutein therapy. IL-2 mutein is engineered for increased affinity for CD25 and decreased affinity for CD122, selectively activating regulatory T cells (Tregs) while limiting stimulation of CD122-expressing effector populations (NK cells, ILCs, and CD122+ T cells). This dataset addresses whether the highest tested dose (10mg) induces an undesirable, off-target transcriptional signature in CD122+ non-myeloid cells. Single-cell RNA-seq (10x Genomics Chromium) was performed on FACS-sorted CD122+ cells from PBMC of 4 subjects in the 10mg dose cohort, at baseline (pre-treatment) and Day 8 post-dose, to assess treatment-induced changes in cell cluster composition and gene expression at single-cell resolution.
Project description:Prior to pregnancy, hormonal changes lead to cellular adaptations in the endometrium allowing for embryo implantation. Critical for successful pregnancy establishment, innate immune cells constitute a significant proportion of uterine cells prior to arrival of the embryo and throughout the first trimester in humans and animal models. Abnormal uterine immune cell function during implantation is believed to play a role in multiple adverse pregnancy outcomes. Current work in humans has focused on uterine immune cells present after pregnancy establishment, and limited in vitro models exist to explore unique functions of these cells. With single-cell RNA-sequencing (scRNAseq), we comprehensively compared the human uterine immune landscape of the endometrium during the window of implantation and the decidua during the first trimester of pregnancy. We uncovered global and cell-type-specific gene signatures for each timepoint. Immune cells in the endometrium prior to implantation expressed genes associated with immune metabolism, division, and activation. In contrast, we observed widespread interferon signaling during the first trimester of pregnancy. We also provide evidence of specific inflammatory pathways enriched in pre- and post-implantation macrophages and natural killer (NK) cells in the uterine lining. Using our novel implantation-on-a-chip (IOC) to model human implantation ex vivo, we demonstrate for the first time that uterine macrophages strongly promote invasion of extravillous trophoblasts (EVTs), a process essential for pregnancy establishment. Pre- and post-implantation uterine macrophages promoted EVT invasion to a similar degree as pre- and post-implantation NK cells on the IOC. This work provides a foundation for further investigation of the individual roles of uterine immune cell subtypes present prior to embryo implantation and during early pregnancy, which will be critical for our understanding of pregnancy complications associated with abnormal trophoblast invasion and placentation.