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il 18 treatment  (InvivoGen)


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    InvivoGen il 18 treatment
    A Representative fluorescence images of HEK293T cells transfected with HaloTagged constructs of human IL18R1-Long or IL18R1-Short and stimulated with 100 ng recombinant <t>human</t> <t>IL-18</t> for 30 min. Images are shown from top to bottom: 405 nm (DAPI), 488 nm (WGA; plasma membrane stain), 561 nm (Janelia Fluor 549; HaloTag ligand), and merged channels. Scale bar, 50 µm. B IL-18-mediated NFκB activity in human HEK293T cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant IL-18. Control reaction: human IL18R1-Long + IL18RAP; test reactions: human IL18R1-Long + IL18RAP + human IL18R1-Short (500ng) or IL18BP (500ng). NanoLuc luciferase values were normalized to Firefly. Error bars represent ± standard error (SE) of the mean; N = 6 treatment replicates. Significance values shown correspond to test condition vs control, unless otherwise specified. C Relative expression of mouse ll18r1 isoforms 24h after siRNA transfection in EL4 cells, normalized to Ctcf and to the non-targeting control (2^ΔΔCq). N = 2 qPCR replicates; KD = siRNA knockdown. D Mouse 11-18-mediated Nfκb activity in EL4 cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant Il-18. NanoLuc (NL) luciferase values are normalized to Firefly luciferase (FF); NUFF ratios are normalized to untreated conditions. Error bars represent ± SE of the mean; N= 3 treatment replicates. Significance values shown correspond to isoform KD vs non-targeting control. E IL1BR1 isoform maps and schematic representation of IL18R1-Long and IL18R1-Short receptor proteins and their signaling capacities. Intracellular signaling cascade is depicted culminating in NFκB activation and signaling. P-values were calculated using unpaired two-tailed I-tests(**, p ≤ 0.001;***, p ≤ 0.001;;***, p ≤ 0.0001).
    Il 18 Treatment, supplied by InvivoGen, used in various techniques. Bioz Stars score: 94/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+il+18/Recombinant+human+IL-18/bio_rxiv__64898__2026__05__25__727421-274-1-23
    Average 94 stars, based on 23 article reviews
    il 18 treatment - by Bioz Stars, 2026-10
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    Images

    1) Product Images from "An ancient retrotransposon provides species-specific tuning of IL-18 inflammatory signaling"

    Article Title: An ancient retrotransposon provides species-specific tuning of IL-18 inflammatory signaling

    Journal: bioRxiv

    doi: 10.64898/2026.05.25.727421

    A Representative fluorescence images of HEK293T cells transfected with HaloTagged constructs of human IL18R1-Long or IL18R1-Short and stimulated with 100 ng recombinant human IL-18 for 30 min. Images are shown from top to bottom: 405 nm (DAPI), 488 nm (WGA; plasma membrane stain), 561 nm (Janelia Fluor 549; HaloTag ligand), and merged channels. Scale bar, 50 µm. B IL-18-mediated NFκB activity in human HEK293T cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant IL-18. Control reaction: human IL18R1-Long + IL18RAP; test reactions: human IL18R1-Long + IL18RAP + human IL18R1-Short (500ng) or IL18BP (500ng). NanoLuc luciferase values were normalized to Firefly. Error bars represent ± standard error (SE) of the mean; N = 6 treatment replicates. Significance values shown correspond to test condition vs control, unless otherwise specified. C Relative expression of mouse ll18r1 isoforms 24h after siRNA transfection in EL4 cells, normalized to Ctcf and to the non-targeting control (2^ΔΔCq). N = 2 qPCR replicates; KD = siRNA knockdown. D Mouse 11-18-mediated Nfκb activity in EL4 cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant Il-18. NanoLuc (NL) luciferase values are normalized to Firefly luciferase (FF); NUFF ratios are normalized to untreated conditions. Error bars represent ± SE of the mean; N= 3 treatment replicates. Significance values shown correspond to isoform KD vs non-targeting control. E IL1BR1 isoform maps and schematic representation of IL18R1-Long and IL18R1-Short receptor proteins and their signaling capacities. Intracellular signaling cascade is depicted culminating in NFκB activation and signaling. P-values were calculated using unpaired two-tailed I-tests(**, p ≤ 0.001;***, p ≤ 0.001;;***, p ≤ 0.0001).
    Figure Legend Snippet: A Representative fluorescence images of HEK293T cells transfected with HaloTagged constructs of human IL18R1-Long or IL18R1-Short and stimulated with 100 ng recombinant human IL-18 for 30 min. Images are shown from top to bottom: 405 nm (DAPI), 488 nm (WGA; plasma membrane stain), 561 nm (Janelia Fluor 549; HaloTag ligand), and merged channels. Scale bar, 50 µm. B IL-18-mediated NFκB activity in human HEK293T cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant IL-18. Control reaction: human IL18R1-Long + IL18RAP; test reactions: human IL18R1-Long + IL18RAP + human IL18R1-Short (500ng) or IL18BP (500ng). NanoLuc luciferase values were normalized to Firefly. Error bars represent ± standard error (SE) of the mean; N = 6 treatment replicates. Significance values shown correspond to test condition vs control, unless otherwise specified. C Relative expression of mouse ll18r1 isoforms 24h after siRNA transfection in EL4 cells, normalized to Ctcf and to the non-targeting control (2^ΔΔCq). N = 2 qPCR replicates; KD = siRNA knockdown. D Mouse 11-18-mediated Nfκb activity in EL4 cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant Il-18. NanoLuc (NL) luciferase values are normalized to Firefly luciferase (FF); NUFF ratios are normalized to untreated conditions. Error bars represent ± SE of the mean; N= 3 treatment replicates. Significance values shown correspond to isoform KD vs non-targeting control. E IL1BR1 isoform maps and schematic representation of IL18R1-Long and IL18R1-Short receptor proteins and their signaling capacities. Intracellular signaling cascade is depicted culminating in NFκB activation and signaling. P-values were calculated using unpaired two-tailed I-tests(**, p ≤ 0.001;***, p ≤ 0.001;;***, p ≤ 0.0001).

    Techniques Used: Fluorescence, Transfection, Construct, Recombinant, Clinical Proteomics, Membrane, Staining, Activity Assay, Luciferase, Reporter Assay, Control, Expressing, Knockdown, Activation Assay, Two Tailed Test

    A Genome-wide comparison of Il-18 treatment responses in mouse EL4 cells: control vs ll18r1-short KD. Orange points: genes with a stronger Il-18 treatment response in ll18r1-short KD cells than in control cells (Sh_FC - NT_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Blue points: genes with a stronger 11-18 treatment response in control cells than in II18r1-short KD cells (NT_FC - Sh_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Dark gray points indicate genes with similar responses in both conditions and light gray points indicate genes that did not reach statistical significance in either condition. Black dashed line indicates equal response; dotted lines mark ±0.5 log2FC difference thresholds. B Heatmap of top 20 differentially expressed genes by fold change difference. Rows represent individual genes clustered by expression pattern and assigned to a gene function category; columns show treatment-induced responses (log2FC) in control and ll18r1-short KD conditions. Color intensity indicates response magnitude (blue= downregulation, orange= upregulation). C Normalized counts for key Il18 response genes. Error bars represent± SE of the mean; n = 2 treatment replicates; individual data points overlaid. Asterisks indicate Wald test significance of differential responses: ** p < 0.01, * p < 0.05 (unadjusted p-values for hypothesis-driven genes).
    Figure Legend Snippet: A Genome-wide comparison of Il-18 treatment responses in mouse EL4 cells: control vs ll18r1-short KD. Orange points: genes with a stronger Il-18 treatment response in ll18r1-short KD cells than in control cells (Sh_FC - NT_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Blue points: genes with a stronger 11-18 treatment response in control cells than in II18r1-short KD cells (NT_FC - Sh_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Dark gray points indicate genes with similar responses in both conditions and light gray points indicate genes that did not reach statistical significance in either condition. Black dashed line indicates equal response; dotted lines mark ±0.5 log2FC difference thresholds. B Heatmap of top 20 differentially expressed genes by fold change difference. Rows represent individual genes clustered by expression pattern and assigned to a gene function category; columns show treatment-induced responses (log2FC) in control and ll18r1-short KD conditions. Color intensity indicates response magnitude (blue= downregulation, orange= upregulation). C Normalized counts for key Il18 response genes. Error bars represent± SE of the mean; n = 2 treatment replicates; individual data points overlaid. Asterisks indicate Wald test significance of differential responses: ** p < 0.01, * p < 0.05 (unadjusted p-values for hypothesis-driven genes).

    Techniques Used: Genome Wide, Comparison, Control, Expressing

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    Article Snippet: .. Fifty ng/ml recombinant human IL2 (PeproTech, Rocky Hill, NJ, USA) was added to the cell cultures together with 50 ng/ml recombinant human IL-12 (PeproTech, Rocky Hill, NJ, USA) and 50 ng/ml recombinant human IL-18 (Invivogen, San Diego, CA, USA) for 3 days [6]. ..

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    Article Snippet: .. Cells were activated using a mix of 10 ng/mL recombinant human IL-12 (rcyc-hil12, iNVIVOGEN) and 10 ng/mL recombinant human IL-18 (rcyec-hil18, iNVIVOGEN). .. After four hours, golgistop (BD Biosciences, 554724) was added (1:2000) to the cells.

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    Article Snippet: .. All co-cultures were supplemented with recombinant human IL-2 (100 U x ml -1 , PeproTech) and further stimulated with indicated combinations of recombinant human IL-1β (50 ng x ml, BioLegend), recombinant human IL-18 (5 ng x ml, Invivogen), and recombinant human IL-23 (50 ng x ml -1 , BioLegend), as well as anti-human IFN γ (1 μg x ml -1 , clone: NIB42, Thermo Fisher Scientific Inc.), ..

    Blocking Assay:

    Article Title: Aberrant naive CD4+ T Cell differentiation in systemic juvenile idiopathic arthritis is committed to B cell help
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    A Representative fluorescence images of HEK293T cells transfected with HaloTagged constructs of human IL18R1-Long or IL18R1-Short and stimulated with 100 ng recombinant <t>human</t> <t>IL-18</t> for 30 min. Images are shown from top to bottom: 405 nm (DAPI), 488 nm (WGA; plasma membrane stain), 561 nm (Janelia Fluor 549; HaloTag ligand), and merged channels. Scale bar, 50 µm. B IL-18-mediated NFκB activity in human HEK293T cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant IL-18. Control reaction: human IL18R1-Long + IL18RAP; test reactions: human IL18R1-Long + IL18RAP + human IL18R1-Short (500ng) or IL18BP (500ng). NanoLuc luciferase values were normalized to Firefly. Error bars represent ± standard error (SE) of the mean; N = 6 treatment replicates. Significance values shown correspond to test condition vs control, unless otherwise specified. C Relative expression of mouse ll18r1 isoforms 24h after siRNA transfection in EL4 cells, normalized to Ctcf and to the non-targeting control (2^ΔΔCq). N = 2 qPCR replicates; KD = siRNA knockdown. D Mouse 11-18-mediated Nfκb activity in EL4 cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant Il-18. NanoLuc (NL) luciferase values are normalized to Firefly luciferase (FF); NUFF ratios are normalized to untreated conditions. Error bars represent ± SE of the mean; N= 3 treatment replicates. Significance values shown correspond to isoform KD vs non-targeting control. E IL1BR1 isoform maps and schematic representation of IL18R1-Long and IL18R1-Short receptor proteins and their signaling capacities. Intracellular signaling cascade is depicted culminating in NFκB activation and signaling. P-values were calculated using unpaired two-tailed I-tests(**, p ≤ 0.001;***, p ≤ 0.001;;***, p ≤ 0.0001).
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    Image Search Results


    A Representative fluorescence images of HEK293T cells transfected with HaloTagged constructs of human IL18R1-Long or IL18R1-Short and stimulated with 100 ng recombinant human IL-18 for 30 min. Images are shown from top to bottom: 405 nm (DAPI), 488 nm (WGA; plasma membrane stain), 561 nm (Janelia Fluor 549; HaloTag ligand), and merged channels. Scale bar, 50 µm. B IL-18-mediated NFκB activity in human HEK293T cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant IL-18. Control reaction: human IL18R1-Long + IL18RAP; test reactions: human IL18R1-Long + IL18RAP + human IL18R1-Short (500ng) or IL18BP (500ng). NanoLuc luciferase values were normalized to Firefly. Error bars represent ± standard error (SE) of the mean; N = 6 treatment replicates. Significance values shown correspond to test condition vs control, unless otherwise specified. C Relative expression of mouse ll18r1 isoforms 24h after siRNA transfection in EL4 cells, normalized to Ctcf and to the non-targeting control (2^ΔΔCq). N = 2 qPCR replicates; KD = siRNA knockdown. D Mouse 11-18-mediated Nfκb activity in EL4 cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant Il-18. NanoLuc (NL) luciferase values are normalized to Firefly luciferase (FF); NUFF ratios are normalized to untreated conditions. Error bars represent ± SE of the mean; N= 3 treatment replicates. Significance values shown correspond to isoform KD vs non-targeting control. E IL1BR1 isoform maps and schematic representation of IL18R1-Long and IL18R1-Short receptor proteins and their signaling capacities. Intracellular signaling cascade is depicted culminating in NFκB activation and signaling. P-values were calculated using unpaired two-tailed I-tests(**, p ≤ 0.001;***, p ≤ 0.001;;***, p ≤ 0.0001).

    Journal: bioRxiv

    Article Title: An ancient retrotransposon provides species-specific tuning of IL-18 inflammatory signaling

    doi: 10.64898/2026.05.25.727421

    Figure Lengend Snippet: A Representative fluorescence images of HEK293T cells transfected with HaloTagged constructs of human IL18R1-Long or IL18R1-Short and stimulated with 100 ng recombinant human IL-18 for 30 min. Images are shown from top to bottom: 405 nm (DAPI), 488 nm (WGA; plasma membrane stain), 561 nm (Janelia Fluor 549; HaloTag ligand), and merged channels. Scale bar, 50 µm. B IL-18-mediated NFκB activity in human HEK293T cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant IL-18. Control reaction: human IL18R1-Long + IL18RAP; test reactions: human IL18R1-Long + IL18RAP + human IL18R1-Short (500ng) or IL18BP (500ng). NanoLuc luciferase values were normalized to Firefly. Error bars represent ± standard error (SE) of the mean; N = 6 treatment replicates. Significance values shown correspond to test condition vs control, unless otherwise specified. C Relative expression of mouse ll18r1 isoforms 24h after siRNA transfection in EL4 cells, normalized to Ctcf and to the non-targeting control (2^ΔΔCq). N = 2 qPCR replicates; KD = siRNA knockdown. D Mouse 11-18-mediated Nfκb activity in EL4 cells measured by dual-luciferase reporter assay across increasing concentrations of recombinant Il-18. NanoLuc (NL) luciferase values are normalized to Firefly luciferase (FF); NUFF ratios are normalized to untreated conditions. Error bars represent ± SE of the mean; N= 3 treatment replicates. Significance values shown correspond to isoform KD vs non-targeting control. E IL1BR1 isoform maps and schematic representation of IL18R1-Long and IL18R1-Short receptor proteins and their signaling capacities. Intracellular signaling cascade is depicted culminating in NFκB activation and signaling. P-values were calculated using unpaired two-tailed I-tests(**, p ≤ 0.001;***, p ≤ 0.001;;***, p ≤ 0.0001).

    Article Snippet: For IL-18 treatment conditions, cells were exposed during the final 30 min of this incubation to either 0 or 100 ng/mL human IL-18 (InvivoGen; rcyec-hil18) in medium containing 200 nM JFX-549.

    Techniques: Fluorescence, Transfection, Construct, Recombinant, Clinical Proteomics, Membrane, Staining, Activity Assay, Luciferase, Reporter Assay, Control, Expressing, Knockdown, Activation Assay, Two Tailed Test

    A Genome-wide comparison of Il-18 treatment responses in mouse EL4 cells: control vs ll18r1-short KD. Orange points: genes with a stronger Il-18 treatment response in ll18r1-short KD cells than in control cells (Sh_FC - NT_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Blue points: genes with a stronger 11-18 treatment response in control cells than in II18r1-short KD cells (NT_FC - Sh_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Dark gray points indicate genes with similar responses in both conditions and light gray points indicate genes that did not reach statistical significance in either condition. Black dashed line indicates equal response; dotted lines mark ±0.5 log2FC difference thresholds. B Heatmap of top 20 differentially expressed genes by fold change difference. Rows represent individual genes clustered by expression pattern and assigned to a gene function category; columns show treatment-induced responses (log2FC) in control and ll18r1-short KD conditions. Color intensity indicates response magnitude (blue= downregulation, orange= upregulation). C Normalized counts for key Il18 response genes. Error bars represent± SE of the mean; n = 2 treatment replicates; individual data points overlaid. Asterisks indicate Wald test significance of differential responses: ** p < 0.01, * p < 0.05 (unadjusted p-values for hypothesis-driven genes).

    Journal: bioRxiv

    Article Title: An ancient retrotransposon provides species-specific tuning of IL-18 inflammatory signaling

    doi: 10.64898/2026.05.25.727421

    Figure Lengend Snippet: A Genome-wide comparison of Il-18 treatment responses in mouse EL4 cells: control vs ll18r1-short KD. Orange points: genes with a stronger Il-18 treatment response in ll18r1-short KD cells than in control cells (Sh_FC - NT_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Blue points: genes with a stronger 11-18 treatment response in control cells than in II18r1-short KD cells (NT_FC - Sh_FC > 0.5 log2FC, padj ≤ 0.05 in both conditions). Dark gray points indicate genes with similar responses in both conditions and light gray points indicate genes that did not reach statistical significance in either condition. Black dashed line indicates equal response; dotted lines mark ±0.5 log2FC difference thresholds. B Heatmap of top 20 differentially expressed genes by fold change difference. Rows represent individual genes clustered by expression pattern and assigned to a gene function category; columns show treatment-induced responses (log2FC) in control and ll18r1-short KD conditions. Color intensity indicates response magnitude (blue= downregulation, orange= upregulation). C Normalized counts for key Il18 response genes. Error bars represent± SE of the mean; n = 2 treatment replicates; individual data points overlaid. Asterisks indicate Wald test significance of differential responses: ** p < 0.01, * p < 0.05 (unadjusted p-values for hypothesis-driven genes).

    Article Snippet: For IL-18 treatment conditions, cells were exposed during the final 30 min of this incubation to either 0 or 100 ng/mL human IL-18 (InvivoGen; rcyec-hil18) in medium containing 200 nM JFX-549.

    Techniques: Genome Wide, Comparison, Control, Expressing

    a Gene set enrichment analysis of Vγ9Vδ2 γδ T cells compared with non-Vγ9Vδ2 γδ T cells using published MAIT cell signatures ( GSE200173 ). NES, normalized enrichment score. b Proportion of IFN-γ + cells among non-Vδ2 and Vδ2 γδ T cells after IL-12 and IL-18 stimulation (50 ng/mL each, 18 h), with PMA/ionomycin as a positive control ( n = 12 biologically independent donors). c Violin plots showing ADT expression levels of CD26 and CD161 across Vγ9Vδ2 γδ T cell clusters (C1 and C2/C3). Horizontal lines indicate the median expression level. d Representative flow cytometry plots of Vδ2 γδ T-cell clusters, categorized into CD26 + CD161 + (C1) and non-CD26 + CD161 + (C2/C3) cells. e Representative flow cytometry histograms and cumulative data presenting the PLZF expression in the Vδ2 γδ T-cell population ( n = 23 biologically independent donors). Violin plots showing expressions of MAIT gene set module score ( f ) and KLRB1 ( g ) in Vγ9Vδ2 clusters (C1–C3). Horizontal lines indicate the median expression level. h Proportions of IFN-γ + and TNF + cells among CD26 + CD161 + (C1) and non-CD26 + CD161 + (C2/C3) cells after IL-12 and IL-18 stimulation (50 ng/mL each, 18 h), with PMA/ionomycin as a positive control ( n = 12 biologically independent donors). i Expression of IL-18Rα (CD218a) in the Vδ2 γδ T-cell population ( n = 12 biologically independent donors). j Representative flow cytometry plots and proportion of IL-17A + cells among CD26 + CD161 + (C1), non-CD26 + CD161 + (C2/C3), and MAIT cells as a positive control from peripheral blood (squares) and liver sinusoidal (circles) mononuclear cells after PMA/ionomycin stimulation (10 ng/mL; 500 ng/mL, 6 h) ( n = 8 biologically independent donors). Data are presented as mean ± s.d. Statistics: Gene set enrichment analysis with false discovery rate correction ( a ); two-sided Wilcoxon signed-rank tests ( b , e , h , i , j ); Source data are provided as a file.

    Journal: Nature Communications

    Article Title: Single-cell landscape of human liver sinusoidal γδ T cells in the context of latent human cytomegalovirus infection

    doi: 10.1038/s41467-026-77255-z

    Figure Lengend Snippet: a Gene set enrichment analysis of Vγ9Vδ2 γδ T cells compared with non-Vγ9Vδ2 γδ T cells using published MAIT cell signatures ( GSE200173 ). NES, normalized enrichment score. b Proportion of IFN-γ + cells among non-Vδ2 and Vδ2 γδ T cells after IL-12 and IL-18 stimulation (50 ng/mL each, 18 h), with PMA/ionomycin as a positive control ( n = 12 biologically independent donors). c Violin plots showing ADT expression levels of CD26 and CD161 across Vγ9Vδ2 γδ T cell clusters (C1 and C2/C3). Horizontal lines indicate the median expression level. d Representative flow cytometry plots of Vδ2 γδ T-cell clusters, categorized into CD26 + CD161 + (C1) and non-CD26 + CD161 + (C2/C3) cells. e Representative flow cytometry histograms and cumulative data presenting the PLZF expression in the Vδ2 γδ T-cell population ( n = 23 biologically independent donors). Violin plots showing expressions of MAIT gene set module score ( f ) and KLRB1 ( g ) in Vγ9Vδ2 clusters (C1–C3). Horizontal lines indicate the median expression level. h Proportions of IFN-γ + and TNF + cells among CD26 + CD161 + (C1) and non-CD26 + CD161 + (C2/C3) cells after IL-12 and IL-18 stimulation (50 ng/mL each, 18 h), with PMA/ionomycin as a positive control ( n = 12 biologically independent donors). i Expression of IL-18Rα (CD218a) in the Vδ2 γδ T-cell population ( n = 12 biologically independent donors). j Representative flow cytometry plots and proportion of IL-17A + cells among CD26 + CD161 + (C1), non-CD26 + CD161 + (C2/C3), and MAIT cells as a positive control from peripheral blood (squares) and liver sinusoidal (circles) mononuclear cells after PMA/ionomycin stimulation (10 ng/mL; 500 ng/mL, 6 h) ( n = 8 biologically independent donors). Data are presented as mean ± s.d. Statistics: Gene set enrichment analysis with false discovery rate correction ( a ); two-sided Wilcoxon signed-rank tests ( b , e , h , i , j ); Source data are provided as a file.

    Article Snippet: Cultures were incubated at 37 °C for 6 h. After the first hour of stimulation, brefeldin A (cat #555029; BD Biosciences) and monensin (cat #554724; BD Biosciences) were added, and their presence was maintained for the remaining 5 h. For intracellular cytokine staining (ICS) following IL-12 and IL-18 stimulation, cells were treated with recombinant human IL-12 (50 ng/mL; cat #200-12; PeproTech) and IL-18 (50 ng/mL; cat #9124-IL; R&D Systems) at 37 °C for 18 h. Brefeldin A and monensin were added during the final 5 h of incubation.

    Techniques: Analysis, Positive Control, Expressing, Flow Cytometry

    ADAM17 blockade restores HIV-1–specific NK cell ADCC. Healthy donor peripheral blood mononuclear cells were incubated in the presence of IL-18, alone or in combination with an ADAM17-blocking antibody (aADAM17), or with the ADAM17 inhibitor TAPI-1. Following overnight culture, NK cells were isolated and co-cultured with antibody-coated HIV-1 − ( B-E ) or HIV-1 + ( F ) CEM.NKR.CCR5 cell lines. ( A ) Levels of CD16 on NK cells were measured by flow cytometry ( n = 25). ( B - F ) The increase in the percentage of Annexin V⁺ CEM.NKR.CCR5 cells ( B ; n = 12; F ; n = 6) were quantified by flow cytometry following co-culture. The statistics for total, non-responders (triangle), and responders (square) were tabulated. ( C ) Correlation between CD16 expression and % ADCC is shown, and the Spearman correlation coefficient was calculated. ( D , E ) The fold changes of CD16 MFI on NK cells from untreated to IL-18-treated ( D ) and from IL-18-treated to IL-18 and TAPI-1-treated conditions ( E ) were calculated for responders and non-responders. (** p < 0.01, *** p < 0.005, **** p < 0.001, Wilcoxon tests)

    Journal: Virology Journal

    Article Title: ADAM17 blockade restores natural killer cell antibody-dependent cytotoxicity against human immunodeficiency virus-1

    doi: 10.1186/s12985-026-03223-0

    Figure Lengend Snippet: ADAM17 blockade restores HIV-1–specific NK cell ADCC. Healthy donor peripheral blood mononuclear cells were incubated in the presence of IL-18, alone or in combination with an ADAM17-blocking antibody (aADAM17), or with the ADAM17 inhibitor TAPI-1. Following overnight culture, NK cells were isolated and co-cultured with antibody-coated HIV-1 − ( B-E ) or HIV-1 + ( F ) CEM.NKR.CCR5 cell lines. ( A ) Levels of CD16 on NK cells were measured by flow cytometry ( n = 25). ( B - F ) The increase in the percentage of Annexin V⁺ CEM.NKR.CCR5 cells ( B ; n = 12; F ; n = 6) were quantified by flow cytometry following co-culture. The statistics for total, non-responders (triangle), and responders (square) were tabulated. ( C ) Correlation between CD16 expression and % ADCC is shown, and the Spearman correlation coefficient was calculated. ( D , E ) The fold changes of CD16 MFI on NK cells from untreated to IL-18-treated ( D ) and from IL-18-treated to IL-18 and TAPI-1-treated conditions ( E ) were calculated for responders and non-responders. (** p < 0.01, *** p < 0.005, **** p < 0.001, Wilcoxon tests)

    Article Snippet: Human PBMCs were cultured in RPMI medium (Corning Inc., Corning, NY) with 10% fetal bovine serum (ThermoFisher Scientific) (R10), with and without 1 ng/mL recombinant human IL-15 (Miltenyi Biotec, Gaithersburg, MD), 10 ng/mL recombinant human IL-18 (R&D Systems, Minneapolis, MN), 1 μg/mL anti-TACE antibody (Millipore Sigma, Burlington, MA; clone D1[A12]), and 2.5 μM ADAM17 inhibitor TAPI-1 (Selleckchem) [ , , ] overnight.

    Techniques: Incubation, Blocking Assay, Isolation, Cell Culture, Flow Cytometry, Co-culture Assay, Expressing