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: 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