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