Human Tregs at the materno-fetal interface show site-specific adaptation reminiscent of tumor Tregs
Judith Wienke, Laura Brouwers, Leone M. van der Burg, Michal Mokry, Rianne C. Scholman, Peter G.J. Nikkels, Bas B. van Rijn, Femke van Wijk
Judith Wienke, Laura Brouwers, Leone M. van der Burg, Michal Mokry, Rianne C. Scholman, Peter G.J. Nikkels, Bas B. van Rijn, Femke van Wijk
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Research Article Immunology Reproductive biology

Human Tregs at the materno-fetal interface show site-specific adaptation reminiscent of tumor Tregs

Abstract

Tregs are crucial for maintaining maternal immunotolerance against the semiallogeneic fetus. We investigated the elusive transcriptional profile and functional adaptation of human uterine Tregs (uTregs) during pregnancy. Uterine biopsies, from placental bed (materno-fetal interface) and incision site (control) and blood were obtained from women with uncomplicated pregnancies undergoing cesarean section. Tregs and CD4+ non-Tregs were isolated for transcriptomic profiling by Cel-Seq2. Results were validated on protein and single cell levels by flow cytometry. Placental bed uTregs showed elevated expression of Treg signature markers, including FOXP3, CTLA-4, and TIGIT. Their transcriptional profile was indicative of late-stage effector Treg differentiation and chronic activation, with increased expression of immune checkpoints GITR, TNFR2, OX-40, and 4-1BB; genes associated with suppressive capacity (HAVCR2, IL10, LAYN, and PDCD1); and transcription factors MAF, PRDM1, BATF, and VDR. uTregs mirrored non-Treg Th1 polarization and tissue residency. The particular transcriptional signature of placental bed uTregs overlapped strongly with that of tumor-infiltrating Tregs and was remarkably pronounced at the placental bed compared with uterine control site. In conclusion, human uTregs acquire a differentiated effector Treg profile similar to tumor-infiltrating Tregs, specifically at the materno-fetal interface. This introduces the concept of site-specific transcriptional adaptation of Tregs within 1 organ.

Authors

Judith Wienke, Laura Brouwers, Leone M. van der Burg, Michal Mokry, Rianne C. Scholman, Peter G.J. Nikkels, Bas B. van Rijn, Femke van Wijk

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Figure 5

uTreg and uTconv polarization at the materno-fetal interface.

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uTreg and uTconv polarization at the materno-fetal interface. (A) Gene e...
(A) Gene expression of lineage-defining transcription factors TBX21 (T-bet), GATA3 (GATA-3), and RORC (RORγt). P values from differential gene expression analysis. DE, differentially expressed Padj < 0.05. Box plots with median; box indicates 25th to 75th percentiles, and whiskers indicate minimum and maximum values (all n = 5). (B–D) Protein expression of RORγt (B) and T-bet (C and D). uTregs were gated as CD3+CD4+CD25hiFOXP3+ cells. MFI, median fluorescence intensity. Multiplicity adjusted P values of 2-way ANOVA with Tukey’s post hoc test.(n = 5) (E) Gene and protein expression of IL18R1 (IL-18R1). Gene expression: box plots with median — box indicates 25th to 75th percentiles, and whiskers indicate minimum and maximum values (n = 5). DE, differentially expressed Padj < 0.05. Protein expression: uTregs were gated as CD3+CD4+CD25hiFOXP3+ cells. Multiplicity adjusted P values of 2-way ANOVA with Tukey’s post hoc test (n = 5). (F) Heatmap showing gene expression of chemokine receptors. Expression values were mean centered and scaled per gene. DE, differentially expressed Padj < 0.05. (G–I) Protein expression of chemokine receptors downregulated (G) and upregulated (H and I) in uTregs. uTregs were gated as CD3+CD4+CD25hiFOXP3+ cells. P values of 2-way ANOVA with Tukey’s post hoc test (n = 5). (J) GSEA with published gene set of CXCR3+T-bet+ Tregs from the pancreas of prediabetic mice (38), comparing uTregs and bTregs. NES, normalized enrichment score; FDR, FDR adjusted P value.