Skip to main content
. 2023 Mar 3;42(3):112193. doi: 10.1016/j.celrep.2023.112193

Figure 3.

Figure 3

AHR activation is necessary for IL-10-induced tolerogenic functions in tolDCs generated in vitro

(A) Short-term MLR experimental design. DC stimulation of allogeneic CD4+ T cells for 5 days induces T cell proliferation, while DC-10 stimulation induces T cell hypo-proliferation.

(B) Left, representative plots of proliferation dye dilution (left plots) and CD25 and HLA-DR expression (right plots). Right, percentages of proliferating (top) and CD25+HLA-DR+ activated (bottom) cells after co-culture of total (left graphs, n = 12) and naive (right graphs, n = 7) CD4+ T cells with DC-10s, AHRinhDC-10s, and DC-10s in the presence of AHR inhibitor, relative to the results obtained with donor-matched DC-stimulated T, set at 100% (dotted line).

(C) IFNγ production by CD4+ T cells (total and naive) stimulated with DC-10s, AHRinhDC-10s, and DC-10s in presence of AHR inhibitor, expressed as ratios to the IFNγ production of T cells stimulated by donor-matched DCs, set at 1 (dotted line) (n = 11).

(D) Long-term and secondary (II) MLR experimental design. DC stimulation of allogeneic CD4+ T cells for 10 days induces T cell proliferation, DC-10 stimulation induces T cell hypo-proliferation and generation of Tr1 cells. II MLR: upon re-stimulation with mature DCs from the same donor used in priming, DC-primed T cells proliferate, while DC-10-primed T cells are anergic and produce IL-10.

(E) Long-term MLR. Left, percentage of CD4+ cells proliferating (Ki67+; top) and of Tr1 cells (CD49b+LAG3+; bottom) upon stimulation with DC-10s, AHRinhDC-10s, and DCs (n = 7). Right, representative FC plots.

(F) II MLR. Top, percentage of proliferation of T cells primed with the indicated DCs and restimulated with mature DCs from the same donor used in priming, expressed as (% proliferating cells/% proliferating cells in the donor-matched T cells primed with DCs) (n = 7); 0% = no proliferation; 100% = proliferated as DC-stimulated. Bottom, IL-10 and IFNγ amounts in culture supernatants (n = 6). p values by Wilcoxon matched pairs test. See Figure S3.