hUC-MSCs Attenuate Acute Graft-Versus-Host Disease through Chi3l1 Repression of Th17 Differentiation

Figure 7

hUC-MSC-secreted Chi3l1 represses CD4 differentiation to Th17 cells by inhibiting STAT3 activation. (a) Reactome pathway showing that Chi3l1 is associated with interleukin-6 family signaling. (b) Gene Ontology showing that Chi3l1 is associated with lymphocyte differentiation. (c) STRING interaction network displaying that Chi3l1 interacts with STAT3. (d–f) CD4⁺ T cells () obtained from the spleen of C57BL/6J mice were differentiated into Th17 cells in 96-well plates in the presence or absence of hUC-MSCs, sh-NC-MSCs, sh-Chi3l1-MSCs, or sh-Chi3l1-MSCs () plus Stattic (20 μM) at a CD4⁺ T/hUC-MSC ratio of 100 : 1 for 72 hours. (e) The proportion of Th17 cells was analyzed using flow cytometry. (d) The percentage of Th17-positive cells was detected by flow cytometry (one-way ANOVA and Tukey’s multiple comparison test, , three independent experiments). (f) Protein expression level of p-STAT3 measured by western blot. STAT3: signal transducer and activator of transcription 3; sh-NC: sh-NC-MSCs; sh-Chi3l1: sh-Chi3l1-MSCs. Stattic: p-STAT3 inhibitor.

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