nonreactive ot-i siinfkel (257- –264) peptide  (GenScript corporation)

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: 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