Fig. 5: LCMV CL13-infected CD11b/c+ DCs cannot prime functional OT-I T cells specific for the antigens of additional infecting pathogens. | Experimental & Molecular Medicine

Fig. 5: LCMV CL13-infected CD11b/c+ DCs cannot prime functional OT-I T cells specific for the antigens of additional infecting pathogens.

From: Chronic LCMV infection regulates the effector T cell response by inducing the generation of less immunogenic dendritic cells

Fig. 5: LCMV CL13-infected CD11b/c+ DCs cannot prime functional OT-I T cells specific for the antigens of additional infecting pathogens.The alternative text for this image may have been generated using AI.

a Experimental scheme of the in vitro OT-I T cell functional assay. SPDCs were isolated from naive and LCMV-infected mice. SPDCs were pulsed with the OVA protein (20 μg/ml) for 2 hours or OVA257-264 SIINFEKL peptide (0.2 μg/ml) for 2 hours and cocultured with CellTrace-labeled OT-I T cells. b Analysis of the capacity of each SPDC to take up, process and present exogenous OVA antigen by using an H-2Kb-SIINFEKL complex-capturing antibody. The mean percentage of H-2Kb-SIINFEKL-positive cells at each timepoint is shown in the bar graph. c The proliferation and division of OT-I T cells primed with OVA-pulsed SPDCs from uninfected or CL13-infected mice were analyzed by CellTrace dye staining. The bar graph presents the percentage of cell populations in each divided number. d Analysis of the effector function of cocultured OT-I T cells. For intracellular cytokine analysis, 0.5 μg/ml OVA257-264 SIINFEKL peptide was added to cultured cells for the last 5 hours. e Experimental scheme of the in vivo OT-I T cell functional assay. A total of 5 × 105 OT-I cells were adoptively transferred by intravenous injection one day before VV-OVA infection. The mice in each group were infected with 2 × 106 PFU of VV-OVA by intraperitoneal injection. f The proliferation and division of adoptively transferred OT-I T cells were analyzed by CellTrace dye staining. The bar graph presents the percentage of the cell populations in each division number. g The cytokine-secreting potential of adoptively transferred OT-I T cells was analyzed by intracellular cytokine staining and CellTrace dye staining. The kinetics of effector cytokine expression during OT-I T cell proliferation were analyzed (top three FACS plots and bar graphs). The expression levels of IFN-γ, IL-2, and TNF-α in activated CD44hi proliferating OT-I T cells were investigated (bottom three FACS plots and bar graphs). The bar graphs show the means ± SDs (6 mice in each group). Each experiment was repeated twice. The p values in the figures indicate the following: *P < 0.05; **P < 0.01; **P < 0.01; ***P < 0.001.

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