Journal: eLife

Article Title: IL-2/JES6-1 mAb complexes dramatically increase sensitivity to LPS through IFN-γ production by CD25 + Foxp3 - T cells

doi: 10.7554/eLife.62432

Figure Lengend Snippet: Purified CD8 + and CD4 + T cells from OT-I/Ly5.1 and OT-II/Ly5.1 mice (2 and 4 × 10 6 /mouse respectively), were adoptively transferred (AT) to C56BL/6 mice. One day later, mice were i.p. injected with PBS (Control), with both OT-I and OT-II specific peptides plus polyI:C (75 µg/mice) or with the latter plus αIFN-γ mAb (250 µg). Expansion of AT T cells was determined 3 d post stimulation by flow cytometry. Dot plots showing one representative mouse ( A ) out of two mice analysed by flow cytometry and a bar graph showing mean ± SD ( B ) are presented. ( C ) Three groups of C56BL/6 mice described above and one control group (no AT) were challenged with LPS (50% of MNLD). ( D ) IFN-γ -/- and normal C56BL/6 mice (IFN-γ +/+ ) were treated with IL-2/JES6 and challenged with LPS (50% of MNLD) as shown in . IFN-γ -/- C56BL/6 mice challenged with the same dose of LPS were used as the control. ( E ) Data pooled from three independent experiments (n = 13–16 technical replicates) described in D showing body temperature of mice 8 h after LPS challenge. ( F ) Scheme showing the proposed mechanism of how IL-2/JES6 induces LPS hyperreactivity. Experiments A-D were done at least twice with similar results; n = 2–6 technical replicates. Data were analysed using unpaired two-tailed Student’s t-test. Significant differences to control are shown (* p ≤ 0.05; *** p ≤ 0.001). Figure 6—source data 1. Source data for , panels B-E.

Article Snippet: C57BL/6 mice were injected i.p. with PBS, OT-I plus OT-II peptides (10 and 50 μg/mouse, respectively; MBL International, Woburn, Massachusetts, USA; or Genscript, Piscataway, New Jersey, USA, respectively) plus polyI:C (75 μg/mouse; Sigma-Aldrich, St. Louis, Missouri, USA) or with the latter plus αIFN-γ mAb (250 μg/mouse; XMG1.2; BioXcell, Lebanon, New Hampshire, USA).

Techniques: Purification, Injection, Control, Flow Cytometry, Two Tailed Test

Journal: Nature communications

Article Title: p38 MAPK-inhibited dendritic cells induce superior antitumor immune responses and overcome regulatory T cell-mediated immunosuppression

doi: 10.1038/ncomms5229

Figure Lengend Snippet: Antitumor activity of DC immunization and Treg depletion using anti-CD25 antibody. (a–c) Comparison of antitumor activity against pre-established tumors. C57BL/6 mice (5 per group) were inoculated subcutaneously with B16-OVA (a,b) or B16 (c) tumor cells (3 × 105) and 3 days later were immunized with 1 × 106 OT-I peptide-loaded (a), OT-II peptide-loaded (b) or TRP2 peptide-loaded (c) mDCs or mSBDCs twice at a weekly interval. Arrows represent the timing of DC-therapy. Anti-CD25 antibodies (50 μg/mouse) were administered intraperitoneally into some groups of mice 3 days before immunization. Error bars represent s.d. Representative results from one of three repeated experiments are shown. (d) IFN-γ ICS of splenocytes from tumor-bearing mice 10 days after second immunization was performed after restimulation with the immunizing peptides overnight in vitro. Numbers represent percentages of IFN-γ secreting lymphocytes gated on CD4+ or CD8+ T cells from one representative experiment of three performed. (e) In vivo CTL assay. CFSElo (left population in gates, non-target) and CFSEhi (right population in gates, target) cells were pulsed with control peptide and OT-I peptide, respectively, and were injected into mice 7 days after OT-I peptide-pulsed DC immunization. Splenocytes were collected 6 hours later for detection of CFSE-labeled cells. Numbers represents percentages of specific killing. Results are representative of one of three independent experiments. (f) Pooled data (n = 3) for in vivo CTL assay. Splenocytes were collected 36 hours later for detection of CFSE-labeled cells in OT-II and TRP2 peptide-loaded DC immunization. Error bars represent s.d. P values were calculated with Student’s t-test.

Article Snippet: MHC class II–restricted TRP1 (SGHNCGTCRPGWRGAACNQKILTVR), H2-K b –restricted TRP2 (SVYDFFVWL), H2-K b –restricted OT-I (SIINFEKL), and MHC class II–restricted OT-II (ISQAVHAAHAEINEAGR) peptides were purchased from GenScript.

Techniques: Activity Assay, In Vitro, In Vivo, CTL Assay, Injection, Labeling