anti human il 17a monoclonal antibody  (R&D Systems)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Aiolos and Eos drive distinct human TH17 functional states

doi: 10.1007/s00018-026-06089-1

Figure Lengend Snippet: Identification of Distinct Human TH17 Cell Subsets and Generation of Stable TH17 Clones from PBMC for Functional Characterization. A Schematic representation of the workflow to generate T H 17-IL22 + /IFNg + and T H 17-IL-10 + clones used to perform bulk ATAC-seq and RNA-seq data sets. In brief, peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood using density gradient centrifugation. The samples were enriched for CD4 + CCR6 + CXCR3- TH17 cells, referred to as “bulk TH17 cells.” Viable IL-17-producing cells were isolated by flow cytometry following a 3-hour stimulation with PMA and ionomycin using a IL-17 capture assay. The single TH17 cell clones were sorted into 384-well plates and expanded with allogeneic γ-irradiated feeder cells and phytohemagglutinin in complete medium containing IL-2. After approximately ten days, clones were transferred to 96-well plates for expansion, and following 2–3 weeks, their cytokine profiles were analyzed. T cell clones were then evaluated at two stages: day 0 (resting state) and day 5 (activated state). On day 5, they were stimulated for 48 hours with anti-CD3 and CD28, followed by an additional 3 days in uncoated plates. On both evaluation days, cells underwent further stimulation — 5 hours for protein analysis and 2 hours for RNA and chromatin-accessibility (ATAC-seq) analysis. Only TH17 clones exhibiting a stable cytokine profile after two rounds of resting and reactivation were selected for RNA-seq and ATAC-seq analysis. B Intracellular staining of IL-17 and IFNγ (top) and IL-22 and IL-10 (bottom) in a T H 17-IL10 + clone (right) and a T H 17-IL22 + /IFNg + clone (left) in the resting state (Day 0) and 5 days post-activation (Day 5). Numbers in quadrants indicate percent cells. C Frequency of IL-17+, IL-10+, IFNγ+, and IL-22+ cells among 6 independent TH17-IL-22 + /IFNγ + (left) and TH17-IL-10 + (right) clones at Day 0 and Day 5. Each symbol represents an individual T cell clone ( n = 6); data are shown as mean ± s.e.m. * P < 0.05, ** P < 0.01 (one-way ANOVA). TH17 clones were selected for RNA and ATAC-seq analysis based on the following criteria: ≥50% IL-17A+ cells at Day 0, ≥15% IL-22+ cells at Day 0 and Day 5, ≥15% IFNγ+ cells at Day 0 and Day 5 for TH17-IL-22 + /IFNγ + clones; ≥50% IL-17A+ cells at Day 0, ≥15% IL-10+ cells at Day 5 for TH17-IL-10 + clones

Article Snippet: PE IL-17 Secretion Assay- detection kit , Miltenyi , 130-094-536.

Techniques: Clone Assay, Functional Assay, RNA Sequencing, Isolation, Gradient Centrifugation, Flow Cytometry, Irradiation, Staining, Activation Assay

Journal: bioRxiv

Article Title: Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

doi: 10.64898/2025.12.19.695390

Figure Lengend Snippet: (A) Akata and Mutu I cells were treated with increasing concentrations of IL-17 (0 to 1,000 ng/ml) for 48 h, and BZLF1 (Zta) mRNA was quantified by RT-qPCR. Expression was normalized to GAPDH and is shown as log 2 (fold change; 2⁻ΔΔCᴛ) relative to untreated cells. (B) Dose-response curves for Zta induction in Akata and Mutu I cells. Fold changes from panel A were fit by nonlinear regression (four-parameter logistic), yielding EC₅₀ values of ∼100 ng/ml (Akata) and ∼140 ng/ml (Mutu I). (C) Akata and Mutu I cells were treated with IL-17 (500 ng/ml), and mRNA levels of BZLF1 (Zta), BMRF1, and BLLF1 (VCA) were quantified by RT-qPCR at the indicated times (16 to 72 h). Expression was normalized to GAPDH and is shown as log 2 (fold change; 2⁻ΔΔCᴛ) relative to time-matched controls. (D) EBV-negative 293 and DG75 cells were transfected with a luciferase reporter driven by the BZLF1 promoter (Zp) and then treated with vehicle or IL-17 (500 ng/ml) for 24 h. Relative luciferase activity is shown normalized to control. For all RT-qPCR panels in this figure, bars/points represent means ± SEM from ≥ 2 biological replicates; each biological replicate was assayed in technical triplicate. Statistical significance was assessed by t test versus the corresponding control. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: For neutralization assays, IL-17A was mixed with neutralizing anti-human IL-17A monoclonal antibody (R&D Systems, Cat# MAB31711-100) at the indicated concentrations and preincubated for 1 h at 37°C to allow immune-complex formation prior to addition to cells.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Luciferase, Activity Assay, Control

Journal: bioRxiv

Article Title: Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

doi: 10.64898/2025.12.19.695390

Figure Lengend Snippet: (A) Akata and Mutu I cells were treated with vehicle or IL-17 (500 ng/ml) for 48 h, and lysates were analyzed by immunoblotting with antibodies against Zta, EAD, and p18 (VCA). GAPDH served as a loading control. (B) RT-qPCR analysis of BZLF1, BMRF1, and BLLF1 (VCA) transcripts in Akata and Mutu I cells treated as in panel A. Expression was normalized to GAPDH and shown as fold change relative to control. (C) Akata and Mutu I cells stably expressing a lytic reporter (GFP under the BMRF1 promoter) were treated with vehicle or IL-17 (500 ng/ml) for 48 h. GFP-positive cells were quantified from fluorescence images using ImageJ and expressed as the percentage of GFP-positive cells. Scale bar: 200 μm. (D) DNase-resistant cell-free EBV DNA in culture supernatants from Akata and Mutu I cells treated with vehicle or IL-17 (500 ng/ml) for 72 h was quantified by qPCR. EBV genome copies were measured using three primer pairs targeting distinct noncoding/non-transcribed regions of the viral genome (Orilyt-H1, Rp, and Cp). For all RT-qPCR and qPCR panels in this figure (B and D), data represent means ± SEM from ≥ 2 biological replicates; each biological replicate was assayed in technical triplicate. Statistical significance was assessed by t test versus the corresponding control. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: For neutralization assays, IL-17A was mixed with neutralizing anti-human IL-17A monoclonal antibody (R&D Systems, Cat# MAB31711-100) at the indicated concentrations and preincubated for 1 h at 37°C to allow immune-complex formation prior to addition to cells.

Techniques: Western Blot, Control, Quantitative RT-PCR, Expressing, Stable Transfection, Fluorescence

Journal: bioRxiv

Article Title: Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

doi: 10.64898/2025.12.19.695390

Figure Lengend Snippet: Akata and Mutu I cells were treated with vehicle, IL-17, IL-17 preincubated with an anti-IL-17 neutralizing antibody, or heat-inactivated IL-17 as indicated. After 48 h, RT-qPCR was performed for EBV lytic transcripts (BZLF1, BRLF1, and BMRF1). Expression was normalized to GAPDH and shown as fold change relative to vehicle-treated cells. Bars represent means ± SEM from ≥ 2 biological replicates; each biological replicate was assayed in technical triplicate. Statistical significance was assessed by t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: For neutralization assays, IL-17A was mixed with neutralizing anti-human IL-17A monoclonal antibody (R&D Systems, Cat# MAB31711-100) at the indicated concentrations and preincubated for 1 h at 37°C to allow immune-complex formation prior to addition to cells.

Techniques: Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

doi: 10.64898/2025.12.19.695390

Figure Lengend Snippet: (A) Akata and Mutu I cells were treated with vehicle or IL-17 (500 ng/ml) for 48 h and subjected to DNBSEQ RNA-seq (paired-end 100 bp). Representative coverage across the EBV genome is shown using Integrative Genomics Viewer (IGV), together with a heat map of viral gene expression. (B) Volcano plots of host gene expression changes in Akata and Mutu I (IL-17 versus control). Dotted lines indicate thresholds (|log₂ fold change| ≥ log₂1.5; -log₁₀FDR ≥ 1.3). Selected top altered genes are labeled. (C and D) Hallmark GSEA-Preranked identified 13 pathways significantly enriched (FDR q < 0.05) in both Akata and Mutu I cells after IL-17 treatment. Normalized enrichment scores (NES) for shared pathways are shown. (E) Pooled Gene Ontology Biological Process over-representation analysis of IL-17-responsive genes from Akata and Mutu I highlights terms related to B-cell receptor signaling and B-cell activation. (F) RNA-seq-based TPM values for IL-17 receptor subunits (IL17RA, IL17RC) and the adaptor TRAF3IP2 (ACT1) in Akata and Mutu I cells treated with vehicle or IL-17.

Article Snippet: For neutralization assays, IL-17A was mixed with neutralizing anti-human IL-17A monoclonal antibody (R&D Systems, Cat# MAB31711-100) at the indicated concentrations and preincubated for 1 h at 37°C to allow immune-complex formation prior to addition to cells.

Techniques: RNA Sequencing, Gene Expression, Control, Labeling, Activation Assay

Journal: bioRxiv

Article Title: Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

doi: 10.64898/2025.12.19.695390

Figure Lengend Snippet: Rael (latency I), SNK6 (latency II), Mutu III (latency III), and Raji (latency III) EBV+ B-cell lines were treated with vehicle or IL-17 (500 ng/ml) for 48 h. Whole-cell lysates were analyzed by immunoblotting for lytic proteins (Zta, EAD, and VCA as available for each line). GAPDH served as a loading control. IL-17 induced lytic protein expression in Rael but not in SNK6, Mutu III or Raji.

Article Snippet: For neutralization assays, IL-17A was mixed with neutralizing anti-human IL-17A monoclonal antibody (R&D Systems, Cat# MAB31711-100) at the indicated concentrations and preincubated for 1 h at 37°C to allow immune-complex formation prior to addition to cells.

Techniques: Western Blot, Control, Expressing

Journal: bioRxiv

Article Title: Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

doi: 10.64898/2025.12.19.695390

Figure Lengend Snippet: IL-17 produced by Th17 cells binds IL-17RA/RC on EBV+ B cells and signals via Act1 (TRAF3IP2) and TRAF6 to activate TAK1, IKK/NF-κB, and MAPKs. In a parallel pathway, B-cell receptor (BCR) engagement activates SYK/BTK-PLCγ2/PKCβ and the CARD11-BCL10-MALT1 (CBM) complex, which also recruits TRAF6 and stimulates the same TAK1-IKK/MAPK nodes. Either pathway alone can therefore drive Zp/Rp (BZLF1/BRLF1) transactivation, leading to EBV lytic gene expression and virion production. Convergence on shared distal effectors predicts that inhibitors of TRAF6-TAK1-IKK/MAPKs will blunt lytic reactivation induced by both IL-17 and BCR, while perturbation of pathway-specific nodes will differentially affect the two inputs. Created in BioRender. Dochi, H. (2025) https://BioRender.com/xhy86xa .

Article Snippet: For neutralization assays, IL-17A was mixed with neutralizing anti-human IL-17A monoclonal antibody (R&D Systems, Cat# MAB31711-100) at the indicated concentrations and preincubated for 1 h at 37°C to allow immune-complex formation prior to addition to cells.

Techniques: Produced, Gene Expression