Fig 2: hUC-MSCs suppress the activation and pro-inflammatory cytokine secretion of hepatic T cells through CHI3L1 in Con A-induced hepatitis.A Representative plots and quantification (B) of flow cytometry analysis for CD3+ T cells in liver MNCs 24 h after Con A administration in indicated groups. C–F Representative plots and quantification of flow cytometry analysis for CD25 (C–D) and CD69 (E–F) expression in CD3+ T cells in liver MNCs 24 h after Con A administration in indicated groups. G–J Representative plots and quantification of flow cytometry analysis for IFN-? (G–H) and TNF-a (I–J) -producing CD3+ T cells in liver MNCs 24 h after Con A administration in indicated groups. K Serum concentrations of IL6, IFN-?, and TNF-a 24 h after Con A administration in indicated groups were detected by CBA kit. Analyses in (G–J) were conducted with liver MNCs stimulated with PMA (50 ng/mL), ionomycin (500 ng/mL) and BFA (10 µg/mL) for 6 h after isolation. Data in (B), (D), (F), (H), (J), and (K) are shown as mean ± SD (n = 5 for each group) with indicated significance (*p < 0.05, **p < 0.01).

Fig 3: Murine ATOs Generate Functional CD8SP and CD4SP Mature T Cells and Foxp3+ CD4+ Treg-like Cells(A) Representative kinetic analysis of T cell maturation from LSK cells isolated from C57BL/6 WT mice at the indicated time points gated on CD45+ TCR?d- Lin-. CD4 and CD8 mature single-positive cells (CD4SP and CD8SP respectively) are gated on TCRß+ CD3+ cells (bottom row).(B) Frequencies of TCRß+ CD3+ cells (left) are shown as percentage of total CD45+ TCR?d- Lin- cells; CD8SP cells (CD8+ CD4- CD3+ TCRß+) (middle) and CD4SP cells (CD8- CD4+ CD3+ TCRß+) (right) are shown as percentage of CD45+ TCRß+ CD3+ cells in M-ATOs and in the thymus. Error bar denotes ±SD (M-ATO, n = 6 independent experiments; thymus, n = 3).(C) Expression of the maturation marker CD62L on CD8SP and CD4SP cells in week 7 M-ATOs. Summary of data on CD62L expression shown as percentage of CD8SP and CD4SP (n = 12 independent M-ATOSs). Frequencies of CD62L+ and naive CD4SP (CD4SP CD62L+) are shown as percentage of CD8SP and CD4SP, respectively (n = 12 independent M-ATOs).(D) Polyfunctional cytokine production by thymic or M-ATO-derived CD62L+ CD8SP (top) and CD62L+ CD4SP (bottom) after treatment with PMA + ionomycin for 6 h. Error bar denotes ± SEM (thymus SP8 and SP4, n = 2; M-ATO SP8, n = 5; M-ATO SP4, n = 2).(E) Proliferation (as measured by dilution of CellTrace Violet (CTV) and activation (upregulation of CD25) of thymic (left) and M-ATO-derived (right) CD62L+ CD8SP and CD62L+ CD4SP after 3 days of treatment with IL2 ± anti-CD3/CD28 stimulation. Data are representative of three independent experiments.(F) Representative immunofluorescence analysis (n = 3) for CD8 (green) and mDLL4 (red) expression (upper) and CD4 (green) and mDLL4 (red) (lower) in week 4 M-ATOs. Nuclei were stained with DAPI. Scale bars, 50 µm.(G) GFP expression in CD4SP cells from thymocytes harvested from the Foxp3-GFP reporter mouse (left), generated in week 7 M-ATOs from LSK cells isolated from Foxp3-GFP (middle) or C57BL/6 WT mice (right). Frequencies of Foxp3+ CD25+ Treg-like T cells are shown as percentage of total CD4SP T cells in thymus (n = 4) or in M-ATOs over time (n = 15 independent M-ATOs).

Fig 4: Blood and pancreatic tissue samples of CP patients contain increased numbers of Treg cells.T cells were isolated from EDTA blood samples of patients (CP) and blood doners (BD) and were analyzed by flow cytometry. a The T cell activation marker CD25 (p = 0.0317, BD n = 10/CP n = 7) was significantly elevated in CP patients compared to BD, whereas the elevation of the surface expression of CD69 (BD n = 10/CP n = 7) did not reach significance. T cell differentiation was analyzed by intracellular transcription factor staining of FOXP3, GATA3 and TBET. b FOXP3+CD25+ Treg cells were significantly elevated in the blood of CP patients (p = 0.0345, BD n = 10/CP n = 7), c in the same manner as GATA3+ Th2 cells (p = 0.0026, BD n = 10/CP n = 7). d Interestingly also TBET+ Th1 cells were significantly increased (p = 0.0478, BD n = 10/CP n = 7). e Representative immunofluorescence labeling images of CD4 and FOXP3 illustrate a high number of CD4+FOXP3+ Treg cells in human CP tissue. f Summary illustration showing how regulatory T cells control organ fibrosis during chronic pancreatitis by suppressing Th2/ILC2-mediated alternative activation of macrophages via IL-4/IL-13. Alternatively activated macrophages in turn induce activation of pancreatic stellate cells (PSC) which contribute to extracellular matrix production and fibrosis. All data were presented as means ± SEM, statistically significant differences were tested by unpaired two-tailed students t-test for independent samples and significance levels of p < 0.05 are marked by an asterisk (a–d). Source data are provided as a Source Data file.

Fig 5: EGFR Expression on Th2 Cells Is Induced by TCR Activation and Maintained by Cytokines(A–E) WT H. polygyrus-infected mice were either treated with pyrantel embonate or left untreated; mLN were collected 14 days after treatment.(A) EGFR expression on activated (CD69+, red bars) or memory (CD25–CD45RBlowCD44hi, blue bars) CD4+ T cells was analyzed by flow cytometry.(B) Activated CD4+ T cells were flow cytometry-sorted from infected mice into ST2+ EGFR+ (red) and ST2+ EGFR– (orange) populations. Memory CD4+ T cells (CD25–CD45RBlowCD44hi) were flow cytometry-sorted from dewormed mice.(C) After sorting, activated EGFR+ and memory cells were stimulated with rIL-33 or media only, and expression of IL-13 was determined by flow cytometry analysis.(D–E) Activated EGFR– and memory cells were cultured overnight with rTSLP or anti-CD3 antibodies, and expression of EGFR on CD4+ T cells was determined by flow cytometry analysis (D) or were re-stimulated with rIL-33 and expression of IL-13 determined by intra-cellular cytokine staining and flow cytometry analysis (E).(F–G) Egfrfl/flxCd4-cre (EGFR?CD4) mice were infected with H. polygyrus and received either flow cytometry-sorted activated CD4 T cells (red) (F) or flow cytometry-sorted naive or memory T cells (blue) (G) from OVA-challenged mice. Worm burden and egg counts in feces were determined 2 weeks post transfer.Data in (A)–(E) are representative of at least two independent experiments (mean ± SEM); results for individual mice are shown as dots. See also Figure S5.