Journal: bioRxiv

Article Title: Metabolic and Thermal Cues Shape IL-6 Responses and Disease Tolerance Mechanisms in Severe Malaria

doi: 10.64898/2026.04.14.718305

Figure Lengend Snippet: Estimated marginal means (EMMs) of IL‐6 secretion from PBn-parasite co‐cultures under factorial combinations of temperature (37°C, 40°C), pipecolic acid (PA; present/absent), lysophosphatidylcholine (LPC; replete/depleted), and parasite exposure (parasite vs no parasite). Panels A and B show the effects of LPC depletion and repletion, respectively. Panel C shows the effect of parasite exposure independent of metabolic conditions. IL‐6 production is highest under febrile temperature (40°C) when PA is present, and LPC is available. Parasite exposure modestly increases IL‐6 across conditions, but metabolic context remains the dominant determinant of IL‐6 output.

Article Snippet: IL-6 concentrations in co-culture supernatants were quantified using the AccuBind IL-6 ELISA kit (Monobind Inc., USA).

Techniques:

Journal: bioRxiv

Article Title: Metabolic and Thermal Cues Shape IL-6 Responses and Disease Tolerance Mechanisms in Severe Malaria

doi: 10.64898/2026.04.14.718305

Figure Lengend Snippet: Interaction plot showing estimated marginal means (EMMs) of IL‐6 secretion across factorial combinations of temperature (37°C, 40°C), pipecolic acid (PA; absent/present), and lysophosphatidylcholine (LPC; depleted/replete). Lines represent model‐adjusted means. IL‐6 production increases under febrile temperature (40°C), particularly when PA is present, and LPC is replete. Panel A: LPC‐depleted conditions. Panel B: LPC‐replete conditions.

Article Snippet: IL-6 concentrations in co-culture supernatants were quantified using the AccuBind IL-6 ELISA kit (Monobind Inc., USA).

Techniques:

Journal: Shock (Augusta, Ga.)

Article Title: The Protective Effect of a Short Peptide Derived from Cold-inducible RNA-binding Protein in Renal Ischemia-Reperfusion Injury

doi: 10.1097/SHK.0000000000000988

Figure Lengend Snippet: Serum was collected 24 h after RIR and used to measure (A) TNF-α, (B) IL-6, (C) IL-1β using ELISA. Data are expressed as mean ± SEM (sham: n = 4 mice; RIR + vehicle: n = 5 mice; RIR + vehicle: n = 5 mice) and compared by one-way analysis of variance and Student-Newman-Keuls method. *P < 0.05 versus sham; #P < 0.05 versus vehicle. RIR, renal ischemia-reperfusion; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Serum TNF-α, IL-6, and IL-1β were determined with an ELISA kit, specific to mouse TNF-α, IL-6, and IL-1β (BD Biosciences, San Diego, CA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: The Journal of Experimental Medicine

Article Title: Helicobacter pylori metabolites exacerbate gastritis through C-type lectin receptors

doi: 10.1084/jem.20200815

Figure Lengend Snippet: αCAG induces innate and acquired immune responses as potent as TDM. (A) Reporter cells expressing Mincle + FcRγ were stimulated with αCG, αCAG C14:0, or TDM (0.003, 0.03, and 0.3 nmol/well) for 20 h and analyzed for GFP expression. (B) Binding of Mincle-Ig to plate-coated αCG, αCAG C14:0, or TDM (0.001, 0.01 and 0.1 nmol/well). Bound proteins were detected with anti-human IgG-HRP. (C) WT and Clec4e −/− BMDCs were stimulated with αCAG C14:0 (0.001, 0.01, 0.1, and 1 nmol/well) or LPS for 1 d and cytokine production was quantified. (D) WT and Clec4e −/− BMDCs were stimulated with αCAG C14:0 for 1 d and analyzed for the surface expression of CD80 and CD40. (E) IFN-γ–primed BMDCs were stimulated with αCAG C14:0 for 1 d and analyzed for the expression of intracellular NOS2. SSC, side scatter. (F) Reporter cells expressing human Mincle + FcRγ were stimulated with αCG, αCAG C14:0, or TDM (0.003, 0.03, and 0.3 nmol/well) and analyzed for GFP expression. (G) hMoDCs were stimulated with αCG, αCAG C14:0 or TDM (0.04, 0.1, and 0.3 nmol/well) for 1 d, and IL-8 production was quantified. (H) BMDCs were stimulated with 0.1 nmol/well of αCAG C14:0, and co-cultured with OT-II CD4 + T cells in the presence of OVA 323–339 peptide for 3 d. Cytokine concentrations were measured by ELISA. (I) Mice were immunized with OVA + αCAG C14:0 and challenged with heat-aggregated OVA at 7 d after immunization. At day 7 after OVA challenge, B cell–depleted inguinal LN cells were stimulated with indicated amounts of OVA for 4 d, and IFN-γ production was quantified. The results of the statistical analysis show the difference between WT and Clec4e −/− mice. Data are presented as the mean ± SD of triplicate (B, C, G, and H) or duplicate (A, F, and I) assays and representative of three (A, B, F, and H), five (C and D), two (E and I), or four (G) independent experiments. An unpaired two-tailed Student’s t test was used for the statistical analyses. *, P < 0.05; **, P < 0.01.

Article Snippet: The ELISA kits for TNF, IL-6, and IFN-γ were from BD Biosciences.

Techniques: Expressing, Binding Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: The Journal of Experimental Medicine

Article Title: Helicobacter pylori metabolites exacerbate gastritis through C-type lectin receptors

doi: 10.1084/jem.20200815

Figure Lengend Snippet: Pathogenic role of Mincle in H. pylori –induced gastritis. (A and B) WT or Clec4e −/− mice were orally administered with 3 × 10 8 CFU of H. pylori SS1 three times in 2 d. At 6 wk after infection, cells from gastric LNs (A) or Peyer’s patches (B) were stimulated with indicated concentrations of H. pylori lysates (Ag) in the presence of BMDCs for 4 d. Uninfected WT mice were used as control. The concentrations of IFN-γ (left panel) and IL-17 (right panel) in the supernatant were determined by ELISA. (C) The number of bacteria recovered from the stomachs of WT and Clec4e −/− mice at 8 wk after H. pylori infection. Bacterial numbers were determined by counting the number of colonies on H. pylori selective agar plates. (D) H&E (HE) staining and immunohistochemical staining of anti-CD3 and F4/80 of stomach sections from uninfected and H. pylori –infected mice after 8 wk. Scale bar, 100 µm. (E) The numbers of Ly6G + CD11b + or F4/80 + CD11b + cells in gastric MNC from mice at 6 wk after infection. (F) Heat map of differentially expressed genes based on RNA-sequencing analysis using RNA extracted from the stomachs of H. pylori –infected WT or Clec4e −/− mice ( n = 3 in each group) after 8 wk. (G–I) H. pylori –infected mice were injected with anti-Mincle mAb or rat IgG as a control Ab (cont. Ab). After 8 wk, single-cell suspensions of mesenteric LN (G), spleen (H), or splenic CD4 + T cells (I; in the presence of BMDCs) were stimulated with H. pylori lysates (2, 20, and 200 µg/ml) for 4 d. Uninfected WT mice were used as control. The concentrations of IFN-γ in the supernatant were determined by ELISA. (J) The frequency of Ly6G + CD11b + cells in gastric MNCs was analyzed at 6 wk after infection with anti-Mincle mAb or cont. Ab treatment. Bars indicate the average number. (K) Bacterial CFU in the stomach of infected mice was determined by counting the number of colonies on H. pylori selective agar plates. Littermates (A–F) or C57BL/6J mice obtained from CLEA Japan (G–K) were used as WT mice. Data are presented as the mean ± SD of triplicate assays (A, B, and G–I) from three (WT vs. Clec4e −/− ) or two (Control Ab vs. Anti-Mincle) independent experiments (at least six infected mice in each group) with similar results. An unpaired two-tailed Student’s t test was used for the statistical analyses. *, P < 0.05; **, P < 0.01.

Article Snippet: The ELISA kits for TNF, IL-6, and IFN-γ were from BD Biosciences.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemical staining, RNA Sequencing Assay, Injection, Two Tailed Test

Journal: The Journal of Experimental Medicine

Article Title: Helicobacter pylori metabolites exacerbate gastritis through C-type lectin receptors

doi: 10.1084/jem.20200815

Figure Lengend Snippet: Cellular immunity during H. pylori infection. (A) Gating strategy for analyzing gastric MNCs by flow cytometry. Numbers indicate the percentages of cells in each gate. PI, propidium iodide; SSC, side scatter; FSC, forward scatter. (B) Gastric MNCs from uninfected and H. pylori –infected WT or Clec4e −/− mice were stained with anti-Ly6G, F4/80, CD11b, TCRβ and B220 at 6 wk after infection. Numbers indicate the percentages of cells in each gate. (C) Metagenome analysis of gastric mucosal swabs from uninfected WT mice, H. pylori –infected WT and Clec4e −/− mice ( n = 3 in each group) after 12 wk. (D) H. pylori –infected WT mice were injected with anti-Mincle mAb or rat IgG as a control Ab (cont. Ab). At 8 wk after infection, Peyer’s patch cells were collected and cultured for 4 d in the absence of exogenous H. pylori lysates. Uninfected WT mice were used as control. The concentrations of IFN-γ and IL-17 in the supernatants were determined by ELISA. (E) H&E (HE) staining and immunohistochemical staining with anti-CD3 of stomach sections from uninfected and H. pylori –infected mice treated with anti-Mincle mAb or control Ab. Scale bar, 100 µm. (F) Mouse invariant NKT hybridoma cells (DN32.D3) expressing CD1d were incubated with indicated amounts of αCG, αCAG, αCPG, or α-GalCer for 1 d. Each lipid was dissolved in DMSO. The expressions of CD69 were analyzed by flow cytometry. (G) The number of bacteria recovered from the stomachs of WT and Traj18 −/− mice at 8 wk after H. pylori infection. Bacterial numbers were determined by counting the number of colonies on H. pylori selective agar plates. (H) H&E staining of stomach sections from H. pylori –infected WT and Traj18 −/− mice after 8 wk. Scale bar, 100 µm. Data are presented as the mean ± SD of triplicate assays (D and F) from two independent experiments (at least six infected mice in each group) with similar results. An unpaired two-tailed Student’s t test was used for the statistical analyses. *, P < 0.05; **, P < 0.01.

Article Snippet: The ELISA kits for TNF, IL-6, and IFN-γ were from BD Biosciences.

Techniques: Infection, Flow Cytometry, Staining, Injection, Cell Culture, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Expressing, Incubation, Two Tailed Test

Journal: Journal of Innate Immunity

Article Title: SR-A, MARCO and TLRs Differentially Recognise Selected Surface Proteins from Neisseria meningitidis : an Example of Fine Specificity in Microbial Ligand Recognition by Innate Immune Receptors

doi: 10.1159/000155227

Figure Lengend Snippet: Polyanionic ligands do not stimulate TLR-mediated cytokine induction. Bg-PMϕ from WT and MyD88–/– mice were stimulated with 1 µg/ml of each polyanionic ligand and the induction of IL-6 (a) and TNF-α (b) was measured. Cells were separately stimulated with E. coli LPS (100 ng/ml), which served as a positive control for cytokine induction.

Article Snippet: The ELISA kits for murine tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were from BD Biosciences Pharmingen (San Diego, Calif., USA).

Techniques: Positive Control

Journal: Journal of Innate Immunity

Article Title: SR-A, MARCO and TLRs Differentially Recognise Selected Surface Proteins from Neisseria meningitidis : an Example of Fine Specificity in Microbial Ligand Recognition by Innate Immune Receptors

doi: 10.1159/000155227

Figure Lengend Snippet: Selected N. meningitidis surface proteins stimulate TLR-mediated cytokine induction. Bg-PMϕ from WT and MyD88–/– mice were stimulated with 1 µg/ml of the purified recombinant N. meningitidis surface proteins and the induction of IL-6 (a) and TNF-α (b) was measured. Cells were separately stimulated with E. coli LPS (100 ng/ml), which served as a positive control for cytokine induction.

Article Snippet: The ELISA kits for murine tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were from BD Biosciences Pharmingen (San Diego, Calif., USA).

Techniques: Purification, Recombinant, Positive Control

Journal: Journal of Innate Immunity

Article Title: SR-A, MARCO and TLRs Differentially Recognise Selected Surface Proteins from Neisseria meningitidis : an Example of Fine Specificity in Microbial Ligand Recognition by Innate Immune Receptors

doi: 10.1159/000155227

Figure Lengend Snippet: Cytokine induction in SR-A–/–, MARCO–/– and double knock-out Mϕ. RPMϕ from WT, SR-A–/–, MARCO–/– and SR-A-MARCO–/– double knock-out animals were stimulated with 1 µg/ml of the N. meningitidis proteins that were showntobeTLR agonists in figure ​figure44 and the induction of IL-6 (a) and TNF-α (b) was measured. Each strain of Mϕ was also stimulated with E. coli LPS (100 ng/ml), which served as a positive control for cytokine induction.

Article Snippet: The ELISA kits for murine tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were from BD Biosciences Pharmingen (San Diego, Calif., USA).

Techniques: Knock-Out, Positive Control

Journal: Journal of Innate Immunity

Article Title: SR-A, MARCO and TLRs Differentially Recognise Selected Surface Proteins from Neisseria meningitidis : an Example of Fine Specificity in Microbial Ligand Recognition by Innate Immune Receptors

doi: 10.1159/000155227

Figure Lengend Snippet: Summary of receptor interaction with N. meningitidis proteins

Article Snippet: The ELISA kits for murine tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were from BD Biosciences Pharmingen (San Diego, Calif., USA).

Techniques: