Both IFN and non-IFN pathways induce sIL-15 complexes upon CD40 stimulation.

<p>A) Serum was isolated from Wt mice 0, 1, and 2 days after treatment with anti-CD40 mAb (200 μg i.p.). Levels of sIL-15 complexes in serum were measured using ELISA. n = 3–4 mice/group. Similar results were observed in at least 5 experiments. B,C) WT and IFNAR-/- mice were treated with anti-CD40 Ab (50 μg i.p.). Serum was isolated 24 hours later and analyzed for levels of B) sIL-15 complexes and C) IFN-α via ELISA. Graph shows average cytokine levels from 3 experiments. n = 12 mice/group. Error bar represents standard error of the mean. * indicates p<0.05.</p

Viral infection transiently induces sIL-15 complexes <i>in vivo</i> independent of IFN signaling.

<p>A) Serum levels of sIL-15 complexes in Wt mice, at indicated times post VSV infection (1x10⁶ PFU/ mouse, i.v.) (n = 2–5 mice/group), one representative experiment of five total is shown. B,C) Cell surface IL-15 expression in myeloid cells one day after VSV infection or Poly I:C injection (150 μg/mouse, i.p.) as determined by immunofluorescence staining and flow cytometry. Histograms depict representative staining with control Ig (black histogram) and anti-IL-15 Ab (grey histograms) in macrophages (lineage-CD11b+CD11c+F480+). Graph shows average MFI of IL-15 expression in DCs (lineage-CD11b+/-CD11c+F480), macrophages, and monocyte (lineage-CD11b+CD11c-F480-) of indicated mice. n = 3 mice/group, one representative of three experiments shown. D) Levels of sIL-15 complexes in serum from WT and IFNAR-/- mice one day post VSV infection. n = 3–4 mice/group; data is representative of three experiments. * indicates p<0.05.</p

Type I IFN signaling is required for increasing sIL-15 complexes after total body irradiation.

<p>Serum was isolated 24 hours after treating WT mice with varying levels of total body irradiation. Levels of sIL-15 complexes in serum were measured using ELISA. n = 3–4 mice/group. Induction of sIL-15 complexes after TBI has been observed in at least 8 experiments. (B) IFN-α levels were measured in serum isolated from WT mice 24hours after treatment with 1000 RADs of total body irradiation. One of two experiments is shown. (C) Serum was isolated from WT and IFNAR1-/- mice 24 hours after treatment with TBI (1000 RADs). Levels of sIL-15 complexes in serum were measured using ELISA. n = 3–4 mice/group; data is representative of 4 experiments. * indicates p<0.05.</p

IFN-α increases sIL-15 complexes <i>in vitro</i> and <i>in vivo</i>.

<p>(A) A plasmid encoding IFN-α, (pIFNα) or an empty vector control plasmid (pORF) were injected i.v. by hydrodynamic injection. Serum was isolated A) 48 hrs post injection and B) after various times post plasmid injection. Levels of sIL-15 complexes in serum were measured using ELISA. n = 2–3 mice/group. Data is representative of three experiments. C) Culture supernatants were collected from WT and IFNAR1-/- BMDC treated with rIFN-α (300 U/mL) or Poly I:C (50 μg/mL) for 24 hr. Levels of sIL-15 complexes in culture supernatants were measured using ELISA. Data is representative of three experiments. D) Levels of sIL-15 complexes in serum from Wt and IFNAR1-/- mice 24 hours after treatment with Poly I:C (150 μg, i.p.) were measured using ELISA. n = 3 mice/group. Data is representative of three experiments. E) Cell-surface ADAM17 expression in WT and IFNAR1-/- BMDCs was detected by indirect immunofluorescence staining and flow cytometric analysis after either no treatment or stimulation with rIFN-α (300U/mL) for 24 hr. F) Non-transduced and TAT-Cre transduced BMDCs generated from ADAM17^fl/fl mice were left untreated (open bars) or stimulated with rIFN-α (300U/mL) (filled bars). Culture supernatants were collected 24hrs later and analyzed for sIL-15 complexes. n = 1–3 wells/group. Data is representative of two experiments, * indicates p<0.05.</p

Supplementary Data from Targeted Inhibition of Inducible Nitric Oxide Synthase Inhibits Growth of Human Melanoma <i>In vivo</i> and Synergizes with Chemotherapy

Supplementary Data from Targeted Inhibition of Inducible Nitric Oxide Synthase Inhibits Growth of Human Melanoma In vivo and Synergizes with Chemotherap

Movie 1 from Multifaceted Role of BTLA in the Control of CD8⁺ T-cell Fate after Antigen Encounter

Movie 1 from Multifaceted Role of BTLA in the Control of CD8+ T-cell Fate after Antigen Encounte

Supplementary Table 1 from Multifaceted Role of BTLA in the Control of CD8⁺ T-cell Fate after Antigen Encounter

Sequence of the primers used to amplify mouse BTLA</p

Supplementary Figure 3 from Multifaceted Role of BTLA in the Control of CD8⁺ T-cell Fate after Antigen Encounter

Expression of BTLA in human CD8+ T cells according to differentiation status.</p

Supplementary Figure 4 from Multifaceted Role of BTLA in the Control of CD8⁺ T-cell Fate after Antigen Encounter

Greatly attenuated recall response in OT-1 BTLA KO T cells in response to immunization with OVA peptide SIINFEKL.</p

Supplementary Methods from The Glutaminase Inhibitor CB-839 (Telaglenastat) Enhances the Antimelanoma Activity of T-Cell–Mediated Immunotherapies

Supplementary Methods</p