Journal: Life Science Alliance

Article Title: Dectin-1 epigenetic reprogramming rescues senescent-like Treg function in allergic asthma

doi: 10.26508/lsa.202503552

Figure Lengend Snippet: FACS-sorted Tregs from patients with asthma and healthy controls (n = 25 per group for all panels) were evaluated for suppressive capacity, intracellular cytokine expression, and secreted cytokine production. (A, B) Suppressive capacity. Representative flow cytometry plots of responder T-cell proliferation and quantitative analysis of Treg-mediated suppression. Tregs from asthma patients exhibited reduced suppressive efficiency compared with controls. (C) Intracellular cytokine expression. Frequencies of Tregs expressing anti-inflammatory cytokines (IL-10, TGF-β) and pro-inflammatory cytokines (IFN-γ, IL-17). Asthma-derived Tregs showed decreased IL-10 + and TGF-β + populations and increased IFN-γ + and IL-17 + populations relative to controls. (D) Secreted cytokine production. Concentrations of IL-10 and active TGF-β1 in 72-h culture supernatants from highly purified Tregs. Asthma Tregs secreted lower levels of both cytokines compared with healthy controls. Data are presented as the mean ± SD, with each dot representing an individual sample. Statistical significance was determined using Welch’s unpaired t test or one-way ANOVA followed by Tukey’s post hoc test, as appropriate. For panels (B, C), P -values were Bonferroni-corrected for multiple comparisons. Significant differences are indicated by asterisks (**** P < 0.001).

Article Snippet: Human IL-10, TGF-β1, IFN-γ, and IL-17 ELISA kits (R&D Systems) were used for the detection of cytokines.

Techniques: Expressing, Flow Cytometry, Derivative Assay, Purification

Journal: Life Science Alliance

Article Title: Dectin-1 epigenetic reprogramming rescues senescent-like Treg function in allergic asthma

doi: 10.26508/lsa.202503552

Figure Lengend Snippet: (A, B) Purity and batch consistency analysis of KQS-1. HPLC analysis of KQS-1 production batches. (A) Quantitative analysis of purity across three independent production batches (n = 5 replicates per batch). Individual data points (black dots) represent single measurements, and the vertical distribution shows batch uniformity. The red dashed line denotes the minimum purity requirement of 95%. Statistical analysis by one-way ANOVA indicates no significant variation between batches (ns, P > 0.05), confirming high-level production consistency. (B) Representative HPLC chromatogram overlay of three batches showing consistent peak profiles and identical retention times, confirming the chemical stability and absence of batch-to-batch variation in the manufacturing process. (C, D, E) Impact of PBS vehicle on the function and viability of asthma patient–derived Tregs. Assessment of PBS treatment on FACS-sorted CD4 + CD25 + CD127 − Tregs isolated from asthma patients (n = 25 patients per group). (C) Treg suppressive efficiency (%) measured by the inhibition of responder T-cell proliferation. No significant difference was observed between the untreated control and the PBS vehicle group. (D) Frequencies (%) of intracellular anti-inflammatory (IL-10, TGF-β) and pro-inflammatory (IFN-γ, IL-17) cytokines determined by flow cytometry after 48 h. Comparisons were adjusted using the Bonferroni correction; no significant alterations were detected across all markers. (E) Treg survival rate (%) after 48 h of culture in the presence or absence of PBS. In all panels, box-and-whisker plots represent the median, interquartile range (IQR), and range, whereas individual black dots denote data from independent patient samples. Statistical significance was evaluated using Welch’s unpaired t test. ns indicates P > 0.05, confirming that PBS serves as a neutral vehicle for KQS-1.

Article Snippet: Human IL-10, TGF-β1, IFN-γ, and IL-17 ELISA kits (R&D Systems) were used for the detection of cytokines.

Techniques: Derivative Assay, Isolation, Inhibition, Control, Flow Cytometry, Whisker Assay

Journal: Life Science Alliance

Article Title: Dectin-1 epigenetic reprogramming rescues senescent-like Treg function in allergic asthma

doi: 10.26508/lsa.202503552

Figure Lengend Snippet: (A) Suppression assays demonstrating that KQS-1–treated Tregs exhibit enhanced inhibition of responder T-cell proliferation compared with vehicle-treated controls. (B, C) Cytokine profiling after 48-h KQS-1 exposure. ELISA analysis of culture supernatants shows a shift toward an anti-inflammatory phenotype, characterized by increased IL-10 and TGF-β1 production and reduced IFN-γ and IL-17 levels. (D) Treg survival analysis in patient-derived cells. Representative and quantitative assessment of Annexin V + /PI − apoptotic Tregs after 48-h treatment with KQS-1 or vehicle control. Paired samples are connected by lines. Data are presented as the mean ± SD. (A, B, C, D) Sample sizes: n = 8 (A, B, C) and n = 12 (D). Statistical significance was determined using unpaired or paired two-tailed t tests, as appropriate, with Bonferroni correction for multiple comparisons where applicable. ** P < 0.01, *** P < 0.001.

Article Snippet: Human IL-10, TGF-β1, IFN-γ, and IL-17 ELISA kits (R&D Systems) were used for the detection of cytokines.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Derivative Assay, Control, Two Tailed Test

Journal: PLOS One

Article Title: Clinical proof of concept for small molecule mediated inhibition of IL-17 in psoriasis

doi: 10.1371/journal.pone.0341049

Figure Lengend Snippet: (A) Inhibition of IL-17 bioactivity as secreted by human TH17 cells and assayed by a reporter cell assay. Dotted lines represent positive and negative assay controls. 1 representative of 7 experiments with different donors is shown. The error bars represent the SEM. (B, C) Evaluation of DC-806 in rat CIA. Rats were randomized on Day 11 and dosed as indicated. Daily measurements of ankle thickness (B) and terminal measurement of footpad weights (C) were used as efficacy readouts. (D) Preclinical evaluation of DC-806 serum levels. PK sampling for exposure determination was performed on Days 11, 16, at 1 and 12 hours post dose. The dotted lines represent the uncorrected IC 50 for rat IL-17AA and rat IL-17AF as listed in . The dashed line represents the LLQ of the quantification assay. All error bars represent the SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Abbreviations : anti-F, anti-IL-17F; BID, twice daily; CIA, collagen-induced arthritis; Dex, dexamethasone; IC 50 , half-maximal inhibitory concentration; IL-17, interleukin-17; LLQ, lower limit of quantification; PK, pharmacokinetic; QD, once daily; SEM, standard error of mean; TH17, T-helper 17.

Article Snippet: Serum IL-17A level was measured using Quantikine ® High Sensitivity (HS) Human IL-17 Immunoassay kit #HS170 (R&D Systems), which mostly detects IL-17AA with approximately 3% cross-reactivity to IL-17AF [ ].

Techniques: Inhibition, Sampling, Concentration Assay

Journal: PLOS One

Article Title: Clinical proof of concept for small molecule mediated inhibition of IL-17 in psoriasis

doi: 10.1371/journal.pone.0341049

Figure Lengend Snippet: Graph detailing the group mean ± SD DC-806 plasma concentration by 200 mg BID (green circle) and 800 mg BID (blue diamond) groups on a semilogarithmic scale through Study Day 29. PK in psoriasis patients was comparable to healthy volunteer PK, with consistent trough levels over 28 days after achieving steady state around Day 3. The dotted line represents IC 50 for recombinant human IL-17AA, as obtained using HEK-blue IL-17 cell-based assay . PK Concentration Analysis Set. Abbreviations : BID, twice daily; HEK: human embryonic kidney; IC 50 , half-maximal inhibitory concentration; IL-17, interleukin-17; PK, pharmacokinetic; SD, standard deviation.

Article Snippet: Serum IL-17A level was measured using Quantikine ® High Sensitivity (HS) Human IL-17 Immunoassay kit #HS170 (R&D Systems), which mostly detects IL-17AA with approximately 3% cross-reactivity to IL-17AF [ ].

Techniques: Clinical Proteomics, Concentration Assay, Recombinant, Cell Based Assay, Standard Deviation

Journal: PLOS One

Article Title: Clinical proof of concept for small molecule mediated inhibition of IL-17 in psoriasis

doi: 10.1371/journal.pone.0341049

Figure Lengend Snippet: (A–C) DC-806 demonstrated biological effects as evidenced by dose-dependent responses of biomarkers of interest compared to placebo. Blue diamonds represent the 800 mg BID group; Green circles represent the 200 mg BID group; Black triangles represent the placebo group. (A) Change in mean ± SEM serum IL-17AA levels from time of first dose through 28 days post first dose. The dotted line represents low limit of quantification of the assay. (B) Change in mean ± SEM serum BD-2 levels from time of first dose through 28 days thereafter. *Except on Day 28 when n = 10; **Except on Day 28 when n = 9. (C) Change in mean ± SEM plasma IL-19 levels from time of first dose through 28 days post first dose. The dotted line represents normalization value of 21 pg/mL. PD Biomarker Analysis Set. Abbreviations: BD-2, beta defensin-2; BID, twice daily; BL, baseline; IL-17, interleukin-17; PASI, psoriasis area and severity index; PD, pharmacodynamic; SEM, standard error of mean.

Article Snippet: Serum IL-17A level was measured using Quantikine ® High Sensitivity (HS) Human IL-17 Immunoassay kit #HS170 (R&D Systems), which mostly detects IL-17AA with approximately 3% cross-reactivity to IL-17AF [ ].

Techniques: Clinical Proteomics, Biomarker Discovery

Journal: PNAS Nexus

Article Title: Soluble CD14 promotes Th17 expansion and differentiation through gamma-aminobutyric acid and expands infidel innate lymphoid cells

doi: 10.1093/pnasnexus/pgaf406

Figure Lengend Snippet: The supernatant of sCD14-treated PBMCs promotes Th17 phenotype. A) The schematic plot of the experimental design. B) Volcano plot of differentially expressed genes in isolated T cells stimulated with anti-CD3/CD28, with and without supernatant from sCD14-treated PBMCs. C) Heat map comparing differentially expressed genes associated with Th17 cells in isolated T cells stimulated with anti-CD3/CD28 plus supernatant from unstimulated autologous PBMCs, and treated with the supernatant from sCD14-treated PBMCs. D) Abundance (TPM) of various genes associated with Th17 cells in isolated T cells, with or without treatment with the supernatant from sCD14-treated PBMCs. E) Bar plot showing the up- and down-regulated transcriptional factors in isolated T cells after treatment with the supernatant from sCD14-treated PBMCs. F) Fold change in IL-17A gene expression in isolated T cells after treatment with the supernatant from sCD14-treated PBMCs. G) IL-17A levels in culture supernatants of isolated T cells treated with the supernatant from sCD14-treated PBMCs, as quantified by ELISA. H) Isolated T cells (1 × 10 5 ) were stimulated in the presence of the supernatant from sCD14-treated PBMCs, and IL-17A-secreting cells were visualized and enumerated by ELISpot. I) Box plot showing the expression of CTLA-4 gene in T cells stimulated with anti-CD3/CD28, in the presence and absence of the supernatant from sCD14-treated PBMCs. P values were calculated using two-tailed Mann–Whitney U test (D, F, I) and Wilcoxon matched-pairs signed-rank tests (G, H); * P < 0.05, **<0.01, ****<0.0001. Each dot represents sample from a study subject.

Article Snippet: The concentrations of IL-17A and sCD14 were measured by DuoSet ELISA kit (R&D systems, Cat# DY317 and Cat# DY383-05, respectively) following the manufacturer's protocols.

Techniques: Isolation, Gene Expression, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: PNAS Nexus

Article Title: Soluble CD14 promotes Th17 expansion and differentiation through gamma-aminobutyric acid and expands infidel innate lymphoid cells

doi: 10.1093/pnasnexus/pgaf406

Figure Lengend Snippet: Increased GABA in the supernatant of sCD14-treated monocytes promotes IL-17A secretion. A) Volcano plot illustrating the magnitude and significance of differences in cytokine/chemokine concentrations in the supernatant of isolated monocytes from HCs ( n = 10) with and without overnight treatment with sCD14. B) Normalized and calculated concentrations of shown cytokines in the supernatant of isolated monocytes from HCs with and without overnight treatment with sCD14. C) Metabolic pathway enrichment analysis plot showing significantly altered pathways in the supernatant of monocytes isolated from 12 HCs (six males, six females) after treatment with sCD14, with box plots showing the metabolites altered in each pathway. D) Metabolic pathway enrichment analysis plot showing significantly altered pathways in the supernatant of monocytes isolated from male HCs ( n = 6) and E) female HCs ( n = 6) after treatment with sCD14, with box plots showing the metabolites altered in each pathway. F) Culture supernatant concentrations of IL-17A in isolated T cells treated with the supernatant from sCD14-treated PBMCs, GABA, hydroxyproline, arginine, and Th17-polarizing conditions, as quantified by ELISA. G) Fold change in IL17A gene expression in isolated T cells after treatment with the supernatant from sCD14-treated PBMCs and GABA. P values were calculated using two-tailed Mann–Whitney U test (C, D, E), Kruskal–Wallis analysis with Dunn's multiple comparisons test (G); * P < 0.05, **<0.01, ***<0.001. Each dot represents sample from a human study subject.

Article Snippet: The concentrations of IL-17A and sCD14 were measured by DuoSet ELISA kit (R&D systems, Cat# DY317 and Cat# DY383-05, respectively) following the manufacturer's protocols.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Gene Expression, Two Tailed Test, MANN-WHITNEY