Journal: Frontiers in Immunology

Article Title: Carbohydrate fatty acid monosulphate ester adjuvant enhances the immunogenicity of influenza antigens via TLR4/2-dependent mechanisms

doi: 10.3389/fimmu.2026.1787181

Figure Lengend Snippet: CMS activates DCs through TLR4 and TLR2-dependent mechanisms. (A) Heat map representation depicting the expression of different TLRs comparing unstimulated DCs vs CMS-stimulated DCs. (B) THP-1 human monocytic cell line stably expressing a secreted embryonic alkaline phosphatase (SEAP) reporter inducible by NF-kB was treated for 24 hrs with CMS at the indicated concentrations. The activation of NF-κB was assessed by measuring the activity of SEAP in the supernatant using QUANTI−Blue™ Solution. Results of THP1 MD2-CD14-TLR4, THP1 and THP1 KO-TLR4 are shown (mean ± SD, n=4). (C, E) Effect of CLI095 (TLR4 pathway inhibitor), or TL2-C29 (TLR2 pathway inhibitor), compared to DMSO (solvent control) on the expression of CMS-induced DC maturation markers. Scatter plots representing the mean ± SD (n = 8 donors) values of CD80, CD86, and HLA-DR are presented. (D, F) Inhibitory effect of CLI095 or TL2-C29 on the secretion (mean ± SD, n = 8) of IL-8, IL-6, and IL-12p70 (all in pg/ml) by CMS-stimulated DCs. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 as analyzed by one-way ANOVA with Tukey’s multiple comparisons post-test (C-F) . CA, cells alone; DMSO, dimethyl sulfoxide; SD, standard deviation.

Article Snippet: THP1-Dual, THP1-Dual MD2-CD14-TLR4, and THP1-Dual KO-TLR4 reporter cells (Invivogen) were resuspended at a concentration of 100,000 cells per well in a 96-well U-bottom plate in 200 μl RPMI 1640 Medium, GlutaMAX Supplement (Gibco), supplemented with 25mM HEPES, 10% FBS, 100 IU/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Expressing, Stable Transfection, Activation Assay, Activity Assay, Solvent, Control, Standard Deviation

Journal:

Article Title: Helicobacter pylori Activates Toll-Like Receptor 4 Expression in Gastrointestinal Epithelial Cells

doi: 10.1128/IAI.71.6.3496-3502.2003

Figure Lengend Snippet: Monoclonal anti-TLR4 antibodies do not block H. pylori-induced IL-8 secretion from gastric epithelial cells. (A) The presence of TLR4 monoclonal antibody (HTA 125 clone; 100 μg/ml for 1 h at room temperature) prevented E. coli-derived LPS-induced IL-8 secretion from THP-1 cells. Medium, unstimulated cells without TLR4 antibody; LPS, macrophages stimulated with LPS (100 ng/ml for 4 h); TLR4, THP-1 cells incubated with the monoclonal antibody alone; TLR4+LPS, THP-1 cells challenged with E. coli LPS following incubation with the anti-TLR4 monoclonal antibody. Concentrations of IL-8 in culture supernatants were measured by immunoassay (P < 0.05 by ANOVA). (B) AGS cells were incubated with antibodies and their isotype controls (1 h) followed by infection with H. pylori for 4 h at 37°C. No difference was observed between H. pylori-infected cells and infected cells treated with TLR4 antibodies and isotype controls. Data are presented as means ± SEM of three separate experiments. ***, P < 0.001 by ANOVA.

Article Snippet: A THP-1 whole-cell lysate (Santa Cruz Biotechnology) served as the positive control for TLR4 expression ( 5 ).

Techniques: Blocking Assay, Derivative Assay, Incubation, Infection

Journal:

Article Title: Helicobacter pylori Activates Toll-Like Receptor 4 Expression in Gastrointestinal Epithelial Cells

doi: 10.1128/IAI.71.6.3496-3502.2003

Figure Lengend Snippet: H. pylori LPS induces IL-8 secretion from THP-1 cells but not from AGS cells. (A) AGS and THP-1 cells were either infected with H. pylori (MOI, 100:1) or treated with purified H. pylori- or E. coli-derived LPS (100 ng/ml) for 4 h at 37°C. Levels of IL-8 in cell-free tissue culture medium supernatants were measured by immunoassay. Data are presented as means ± SEM of three separate experiments. ***, P < 0.001 by ANOVA. (B) mCD14 staining of THP-1 and AGS cells measured by flow cytometry. The black line depicts the negative control (R-PE conjugate mouse IgG2a), and the grey line shows R-PE-conjugated monoclonal anti-human mCD14 staining. THP-1 cells (lower graph), but not AGS cells (upper graph), expressed CD14.

Article Snippet: A THP-1 whole-cell lysate (Santa Cruz Biotechnology) served as the positive control for TLR4 expression ( 5 ).

Techniques: Infection, Purification, Derivative Assay, Staining, Flow Cytometry, Negative Control

Journal: Translational Cancer Research

Article Title: Pan-cancer analysis identifies FAM111B as a biomarker for immune suppression microenvironment in low-grade gliomas

doi: 10.21037/tcr-2025-1762

Figure Lengend Snippet: FAM111B promotes the malignant progression of LGG through immunosuppression. (A) The protein expression of AKT, p-AKT, P53 and CD276 in FAM111B knockdown or overexpression LN229 and A172 cells were detected by Western blot analysis. (B) Representative bioluminescence images of FAM111B overexpression LN229 and control cells derived xenografts (n=6). (C,D) IL-10 cytokines concentration of supernatant in THP-1 cells and LN229 or A172 co-culture system, detected by ELISA. (E) Immunofluorescence for CD163 and iNOS expression to detect macrophages in glioma tissue from control and FAM111B overexpression mice. Two-tailed Student’s test, *, P<0.05; **, P<0.01; ***, P<0.001. DAPI, 4',6-diamidino-2-phenylindole; ELISA, enzyme linked immunosorbent assay; FAM111B, FAM111 trypsin-like peptidase B; IL-10, interleukin-10; iNOS, inducible nitric oxide synthase; LGG, lower-grade glioma; NC, negative control; OE-FAM111B, overexpression of FAM111B; siFAM111B, small interfering RNA targeting FAM111B; THP-1, tumor-derived human promonocytic cell line-1; WT, wild-type.

Article Snippet: Subsequently, supernatant from THP-1 cells was collected, and the levels of interleukin-10 (IL-10) were assessed using an ELISA kit (Elabscience) to evaluate the polarization degree of macrophages.

Techniques: Expressing, Knockdown, Over Expression, Western Blot, Control, Derivative Assay, Concentration Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Two Tailed Test, Negative Control, Small Interfering RNA

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Schematic diagram showing the strategy for the drug screen. B ) THP-1 ASC-GFP monocytic cells without any treatment. C ) THP-1 ASC-GFP monocytic cells treated with LPS (1μg/ml) for 2 hours. D ) THP-1 ASC-GFP monocytic cells treated with LPS (1μg/ml) for 4 hours and Nigericin (10 μM) for 1 hour.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques:

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) Vehicle-treated or drug-treated, 20 μM, 16h ( B-P ) THP-1 ASC-GFP monocytic cells were treated with LPS (1μg/ml) for 4 hours. THP-1 ASC-GFP cells were pretreated with selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Incubation

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (10 μM) for 1 hour. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei. B ) RAW-ASC cells pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Nigericin (5 μM) for 1 hour. Cells were fixed and permeabilized and stained with anti-ASC antibody and Alexa-labeled secondary antibody. ASC-puncta were visualized using fluorescent microscopy, with DAPI used to stain nuclei.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Incubation, Microscopy, Staining, Labeling

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A, B ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in THP-1 ASC-GFP cells pretreated with vehicle or selected drugs for 16h. C, D ) Western blot analysis of basal or LPS-induced expression of various inflammasome components in RAW- ASC cells pretreated with vehicle or selected drugs for 16h. E ) Age-matched (10-week-old) male WTC57BL6J mice fed with chow diet were i.p injected with saline or saquinavir. After 2 hours, mice were primed for inflammasome assembly by an I.P. injection of LPS (5μg/mouse). After 4h of LPS injection, the NLRP3 inflammasome assembly was induced by I.P. injection of ATP (0.5 mL of 30 mM, pH 7.0). The peritoneal cavity was lavaged with 5 mL sterile PBS, and IL-1β levels in peritoneal lavage were determined by ELISA (N = 6, mean ± SD for all groups, ∗∗p < 0.01 by two-tailed t-test). Mouse plasma was used in multiplex analysis to determine levels of F ) TNF-α, G ) IL-1β, H ) IL-17A, I ) IL-33, and J ) IL-15 (N=3, mean ± SD for all groups, ∗p < 0.05 by two-tailed t-test).

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Western Blot, Expressing, Injection, Saline, Sterility, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Clinical Proteomics, Multiplex Assay

Journal: bioRxiv

Article Title: FDA-approved drug library screen identifies antidepressants, antimicrobials, anti-COPD, and anti-CVD agents as blockers of NLRP3 inflammasome and sepsis in a sex-dependent manner

doi: 10.64898/2026.03.05.709979

Figure Lengend Snippet: A ) THP-1 ASC-GFP macrophages pretreated with vehicle or selected drugs for 16h, followed by washing with PBS and incubation with LPS (1μg/ml) for 2 hours, followed by incubation with Mitosox Red dye and fluorescent microscopy. DAPI staining is used to visualize nuclei.

Article Snippet: THP-1-ASC-GFP cells (Invivogen #thp-ascgfp) were maintained in RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml NormocinTM,1% Pen/Strep.

Techniques: Incubation, Microscopy, Staining