Journal: Cell reports
Article Title: Aryl hydrocarbon receptor activity downstream of IL-10 signaling is required to promote regulatory functions in human dendritic cells.
doi: 10.1016/j.celrep.2023.112193
Figure Lengend Snippet: 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) IFNg 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 IFNg 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 IFNg amounts in culture supernatants (n = 6). p values by Wilcoxon matched pairs test. See Figure S3.
Article Snippet: Coating Ab Clone Brand Concentration Purified Rat Anti-Human and Viral IL-10 JES3-9D7 BD Biosciences 1 mg/mL Purified Mouse Anti-Human IFNg NIB42 BD Biosciences 1 mg/mL Detection Ab Clone Brand Concentration Biotin Anti-Human and Viral IL-10 JES3-12G8 BD Biosciences 1 mg/mL Biotin Anti-Human IFNg 4S.B3 BD Biosciences 1 mg/mL Standard Curve Brand Concentration (max:min) Recombinant Human IL-10 protein – CellGenix 2000 : 15,625 pg/mL Recombinant Human IFNg – R&D Systems 2000 : 15,625 pg/mL Quantification of signal was performed using Streptavidin covalently coupled to horseradish peroxidase (strep-POD) and Tetramethylbenzidine (TMB) substrate, and optical density (OD) was read at 640nm on a Multiskan GO (Thermo Fisher Scientific) and analyzed by SkanIt 4.1.
Techniques: Activation Assay, Generated, In Vitro, Expressing, Co-Culture Assay
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