Journal: European Journal of Immunology
Article Title: Type I IFN signaling facilitates the development of IL‐10‐producing effector CD8 + T cells during murine influenza virus infection
doi: 10.1002/eji.201646548
Figure Lengend Snippet: Blimp‐1 and IRF4 are required for the development of IL‐10‐producing effector CD8 + T cells induced by type I IFNs plus IL‐2 and IL‐27 . CD8 + T cells were cultured under indicated conditions (no cytokines, IFN‐α, IL‐2 plus IL‐27 or IFN‐α plus IL‐2 and IL‐27). (A) Prdm1 (Blimp‐1 gene) expression in CD8 + T was determined by normalizing the data to HPRT expression through real‐time RT‐PCR. (B) Prdm1 expression in CD8 + T cells was determined by normalizing the data to HPRT expression through real‐time RT‐PCR in total of three experiments. (C) Production of IL‐10 by control or Blimp‐1 cKO CD8 + T cells was determined by flow cytometry. (D) IL‐10 protein production in the supernatants following restimulation of control or Blimp‐1 cKO CD8 + T cells cultured with indicated conditions was determined by ELISA. (E) IRF4 expression in CD8 + T cells cultured with indicated conditions was determined by flow cytometry. (F) Production of IL‐10 by control or IRF4 cKO CD8 + T was determined by flow cytometry. (G) IL‐10 protein production in the supernatants following restimulation of WT or IRF4 cKO CD8 + T was determined by ELISA. (H) Production of IL‐10 and IFN‐γ by lung CD8 + T cells from WT or IRF4 cKO mice was determined by flow cytometry following in vitro antigenic stimulation with influenza‐infected WT BMDCs. (I) Normalized percentages of IL‐10 + cells in influenza‐specific lung CD8 + T cells (IFN‐γ + ) from infected control or IRF4 cKO mice were determined by flow cytometry. (J) Model of type‐I‐IFN‐dependent IL‐10 production by CD8 + T cells. Data (A, C, D, E, F, and G) are from a single experiment representative of two to three independent experiments with one to two mice per experiment. Data (B) are pooled from total of three experiments with one to two mice per experiment. Data (H) in density plots are from a single experiment representative of two independent experiments with two to three mice per experiment. Data (I) in graphs are shown as mean + SEM and are representative of two separate experiments with two to three mice per experiment. Statistics were determined by paired (B) or unpaired (I) two‐tailed Student's t ‐test ( ∗ p < 0.05).
Article Snippet: WT C57/BL6 mice were purchased from the Jackson Laboratory; Blimp‐1 control ( Prdm1 fl /fl , where Prdm is PR domain 1), CD4‐Cre transgenic, CD8‐cre transgenic, IRF4 control ( Irf4 fl /fl ), and CD45.1 congenic mice were originally from the Jackson Laboratory and bred inhouse.
Techniques: Cell Culture, Gene Expression, Expressing, Quantitative RT-PCR, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, In Vitro, Infection, Two Tailed Test
