Journal: Molecular Medicine

Article Title: IL17F + naïve and IFNγ + memory CD8 T cells drive hepatic dysfunction in the cecal ligation and puncture model of sepsis

doi: 10.1186/s10020-025-01411-2

Figure Lengend Snippet: Effects of Immune Education, CLP, and Specific Cytokine Blockade on T cell Cytokine Production. A-C Immune Educated or Control IFNγ-eYFP reporter mice on a C57Bl/6 background were sacrificed prior to or 24 h. following CLP. IFNγ + T cells were assessed. Percent IFNγ + CD4 (Left) and CD8 (Right) T cells in the ( A ) spleen and ( B ) liver. C Histograms of IFNγ + (Red) and negative (Gray) splenic CD4 T cells in Educated mice following CLP. D-E Immune Educated or Control IL17A EGFP /IL17F mCherry reporter mice on a C57Bl/6 background were sacrificed prior to or 24 h. following CLP. IL17A + and IL17F + T cells were assessed. Percent IL17A + CD4 (Left), IL17A + CD8 (Middle Left), IL17F + CD4 (Middle Right) and IL17F + CD8 (Right) T cells in the ( D ) inguinal lymph node (ILN) and ( E ) liver. F Histograms of IL17A + (Orange) vs IL17F + (Green) ILN CD4 T cells in Educated mice following CLP. A-F * = p < 0.05 for comparisons between two groups, † = p < 0.05 for interaction effect, # = p < 0.05 for CLP effect, ^ = p < 0.05 for Education effect. N = 3–4/group. G Hepatic leukocytes were isolated from Educated or control C57Bl/6 mice prior to or 24 h. following CLP alone or with specific cytokine blockade and intracellular cytokine production was assessed following in vitro PMA/Ionomycin stimulation. Percent IFNγ + (Left) and IL17A + (Right) CD8 T cells. Analyzed by 2-way ANOVA. * = p < 0.05 for comparisons between two groups, # = p < 0.05 for specific cytokine effect, p values for interactions, effect of education or effect of blockade displayed. N = 3–5/group. Gating: FSC/SSC, Singlets, Live, CD90/CD4 or CD90/CD8, eYFP, eGFP, mCherry, IFNγ, IL17A, or TNF

Article Snippet: IFNγ (XMG1.2, dose: 0.5 mg), IL12p40 (C17.8, dose: 1 mg), TNF (XT3.11, 1 mg), IL17A (17F3, dose: 0.5 mg) and IL17F (MM17F8F5.1A9, dose: 0.5 mg) blocking antibodies were all obtained from BioXCell (Lebanon, NH), diluted from stock concentration in sterile PBS and administered through intraperitoneal injection at in vivo blocking doses as previously described in the literature. (Jordan et al. ; Taylor et al. ; Naik et al. ; Lemaire et al. ).

Techniques: Control, Isolation, In Vitro

Journal: Molecular Medicine

Article Title: IL17F + naïve and IFNγ + memory CD8 T cells drive hepatic dysfunction in the cecal ligation and puncture model of sepsis

doi: 10.1186/s10020-025-01411-2

Figure Lengend Snippet: Effects of Immune Education, CLP, and Specific Cytokine Blockade on Innate and T Cell Immune Responses. Hepatic leukocytes were isolated from Educated or control C57Bl/6 mice prior to or 24 h. following CLP alone or with specific cytokine blockade; innate immune cell numbers and cytokine production were assessed. Total number of hepatic neutrophils ( A ) or MoDCs ( B ), and percent TNF + ( C ), IL1β + ( D ) or IL12p40 + MoDCs ( E ) and total number of hepatic CD8 T cells ( F ) in control (Blue) or Educated (Red) mice at baseline, following CLP or following CLP with IFNγ, IL12p40, TNF, or IL17A/F combined blockade. CLP with IL17A or IL17F blockade shown to the right. Analyzed by 2-way ANOVA. IL17A and IL17F analyzed separately. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 for comparisons between two groups, # = p < 0.05 for cytokine effect compared to CLP. N = 3–12/group. Cytokine production analyzed in a subset of samples. Gating: Neutrophils = FSC/SSC, Singlets, Live Cells, CD11b + /CD11c − , Ly6G + /Ly6C + , MoDCs = FSC/SSC, Singlets, Live, CD11b + /CD11c − , Ly6G − , MHCII + , Macrophages = FSC/SSC, Singlets, Live, CD11b + /CD11c − , Ly6G − , MHCII − , Ly6C Hi /CD64 + , CD8 + T cells = FSC/SSC, Singlets, Live, CD90 + /CD8 +

Article Snippet: IFNγ (XMG1.2, dose: 0.5 mg), IL12p40 (C17.8, dose: 1 mg), TNF (XT3.11, 1 mg), IL17A (17F3, dose: 0.5 mg) and IL17F (MM17F8F5.1A9, dose: 0.5 mg) blocking antibodies were all obtained from BioXCell (Lebanon, NH), diluted from stock concentration in sterile PBS and administered through intraperitoneal injection at in vivo blocking doses as previously described in the literature. (Jordan et al. ; Taylor et al. ; Naik et al. ; Lemaire et al. ).

Techniques: Isolation, Control

Journal: Molecular Medicine

Article Title: IL17F + naïve and IFNγ + memory CD8 T cells drive hepatic dysfunction in the cecal ligation and puncture model of sepsis

doi: 10.1186/s10020-025-01411-2

Figure Lengend Snippet: Effects of Immune Education, CLP, and Specific Cytokine Blockade on CLP-Induced Hepatic Dysfunction. Immune Educated (Red) or Control (Blue) C57Bl/6 mice were sacrificed prior to or 24 h. following CLP alone or with cytokine blockade. A Hepatic tissue was assessed by RT-PCR for log 10 transformed relative transcription levels of SCLO1a1, SLC10, and ABCC2. B Serum Alanine aminotransferase (ALT). C Log 10 transformed peritoneal bacterial colony forming units. D Probability of Survival in the days following CLP with blockade as noted. Data as mean ± SD. Analyzed by 2-way ANOVA. * = p < 0.05 for comparisons between two groups. P Int = Significant Interaction between Blockade and Education, P Block = Significant Effect of Treatment (CLP + antibody blockade), P Edu = Significant Effect of Control vs Immune Education. For A , primary 2-way ANOVA comparison between Baseline, CLP, CLP + αIFNγ, CLP + αIL12p40, CLP + αTNF, and CLP + αIL17A/F. IL17A and IL17F blockade compared to each other separately from other comparisons. Survival analyzed by Log-rank Test. N = 4–22/group

Article Snippet: IFNγ (XMG1.2, dose: 0.5 mg), IL12p40 (C17.8, dose: 1 mg), TNF (XT3.11, 1 mg), IL17A (17F3, dose: 0.5 mg) and IL17F (MM17F8F5.1A9, dose: 0.5 mg) blocking antibodies were all obtained from BioXCell (Lebanon, NH), diluted from stock concentration in sterile PBS and administered through intraperitoneal injection at in vivo blocking doses as previously described in the literature. (Jordan et al. ; Taylor et al. ; Naik et al. ; Lemaire et al. ).

Techniques: Control, Reverse Transcription Polymerase Chain Reaction, Transformation Assay, Blocking Assay, Comparison

Journal: Molecular Medicine

Article Title: IL17F + naïve and IFNγ + memory CD8 T cells drive hepatic dysfunction in the cecal ligation and puncture model of sepsis

doi: 10.1186/s10020-025-01411-2

Figure Lengend Snippet: Effects of CLP on Pet Shop and SPF J:ARC(S) Swiss Outbred Mice. Pet Store or Control J:ARC(S) Swiss Outbred mice were sacrificed 24 h. following CLP. A Serum levels of IFNγ, IL12p40, and IL17. B Total number of hepatic neutrophils, MoDCs, or cDCs. C RT-PCR log 10 transformed relative transcription levels of SCLO1a1 from hepatic tissue, serum alanine aminotransferase (ALT), and log 10 transformed peritoneal bacterial colony forming units. Analyzed by 2-way ANOVA. P Int = Significant Interaction between Blockade and Pet Store/Laboratory, P Block = Significant Effect of Treatment (CLP + antibody blockade), P group = Significant Effect of Pet Store/Laboratory Strain. 2-way ANOVA comparison between Baseline, CLP, CLP + αIFNγ, and CLP + αIL17F. * = p < 0.05 for comparisons between two groups. N = 4–7/group. Gating: Neutrophils = FSC/SSC, Singlets, Live, CD11b + /CD11c − , Ly6G + /Ly6C + , MoDCs = FSC/SSC, Singlets, Live, CD11b + /CD11c − , Ly6G − , MHCII + , cDCs = FSC/SSC, Singlets, Live, CD11c + , MHCII +

Article Snippet: IFNγ (XMG1.2, dose: 0.5 mg), IL12p40 (C17.8, dose: 1 mg), TNF (XT3.11, 1 mg), IL17A (17F3, dose: 0.5 mg) and IL17F (MM17F8F5.1A9, dose: 0.5 mg) blocking antibodies were all obtained from BioXCell (Lebanon, NH), diluted from stock concentration in sterile PBS and administered through intraperitoneal injection at in vivo blocking doses as previously described in the literature. (Jordan et al. ; Taylor et al. ; Naik et al. ; Lemaire et al. ).

Techniques: Control, Reverse Transcription Polymerase Chain Reaction, Transformation Assay, Blocking Assay, Comparison