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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with <t>anti-CD28)</t> for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).
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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with <t>anti-CD28)</t> for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).
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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with <t>anti-CD28)</t> for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).
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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with <t>anti-CD28)</t> for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).
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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with <t>anti-CD28)</t> for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).
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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with <t>anti-CD28)</t> for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).
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Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with anti-CD28) for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).

Journal: bioRxiv

Article Title: SLAMF1-peptide mediated epigenetic priming reprograms innate immune responses in sepsis

doi: 10.64898/2025.12.29.696918

Figure Lengend Snippet: Diluted whole blood samples from sepsis patients (n = 5–8) and healthy donors (n = 20) were stimulated ex vivo with defined immune agonists for 24–72 h, and cytokine levels were quantified in diluted plasma supernatants. Volcano plots display log 2 fold change versus −log 10 adjusted p values (FDR < 0.05). Statistical significance was assessed using multiple unpaired t -tests across individual cytokines. (A) Stimulation with the TLR2 ligand FSL-1 (100 ng/ml) for 24 h induced significantly attenuated cytokine production in sepsis samples compared with healthy controls, as visualized by a volcano plot and corresponding bar/dot plots. Reduced cytokine levels are shown in blue, while increased IP-10 is shown in red (n = 5). (B) Stimulation with the TLR4 ligand LPS (1 ng/ml) for 24 h resulted in significantly reduced production of IP-10 and IL-1β in sepsis samples compared with healthy controls (n = 5). (C) Stimulation with the TLR8 ligand TL8-506 (300 ng/ml) for 24 h demonstrated significantly reduced IP-10 and IL-12p70 production in sepsis samples (n = 5). (D) Stimulation of diluted whole blood with a T cell/MHC class II–dependent stimulus (Cytostim combined with anti-CD28) for 72 h revealed broadly attenuated cytokine responses in sepsis samples compared with healthy controls (n = 8).

Article Snippet: T cell responses were assessed after 72 h, under non-stimulated conditions (RPMI alone) and RPMI with a synthetic superantigen (CytoStim, 2 μl/ml) from Miltenyi Biotec (cross-binding TCR and MCH) in combination with co-stimulating anti-CD28 mAb (InVivoMAb, BioXCell, 300 ng/ml).

Techniques: Ex Vivo, Clinical Proteomics