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R&D Systems il 18

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 <t>and</t> <t>IL-18</t> 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.

Il 18, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/recombinant+human+il+18/Recombinant+Human+IL-18%2FIL-1F4+Protein/pmc13623833-380-62-67
Average 94 stars, based on 31 article reviews

il 18 - by Bioz Stars, 2026-10

94/100 stars

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1) Product Images from "Single-cell landscape of human liver sinusoidal γδ T cells in the context of latent human cytomegalovirus infection"

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

Journal: Nature Communications

doi: 10.1038/s41467-026-77255-z

Figure Legend 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.

Techniques Used: Analysis, Positive Control, Expressing, Flow Cytometry

Recombinant:

Article Title: A novel method for generating chimeric antigen receptor-transduced primary human natural killer cells by the use of multiple cytokines and anti-CD2 and anti-NKp46 stimulatory antibodies.
Article Snippet: PBMCs containing 60,000 NK (CD56+, CD3-) cells were seeded in 8 mL of CellGenix GMP SCGM (CellGenix GmbH, Freiburg, Germany) supplemented with 10 % FBS. .. The following cytokines were added at the following concentrations: recombinant human IL-2 (27 ng/mL) (Peprotech, Cranbury, NJ, USA), recombinant human IL-12 (10 ng/mL) (R&D Systems, MN, USA), recombinant human IL-18 (10 ng/mL) (R&D Systems, MN, USA), recombinant human IL-21 (10 ng/mL) (R&D Systems, MN, USA). .. The PBMCs were cultured with dissolvable hydrogel microspheres harboring anti-CD2 and anti-NKp46 antibodies (Cloudz Human NK Cell Expansion Kit, Bio-Techne, MN, USA) in a T-25 flask at 37◦C and 5 % CO2 in an incubator according to the manufacture’s instruction with slight modifications.

Article Title: A Feeder Cell-Free System for Chimeric Antigen Receptor Gene Transduction Into Natural Killer Cells
Article Snippet: Recombinant human IL-12 (R&D Systems, catalog number: 10018-IL) 9. .. Recombinant human IL-18 (R&D Systems, catalog number: 9124-IL) 10. .. Recombinant human IL-21 (R&D Systems, catalog number: 8879-IL) 11.

Article Title: ADAM17 blockade restores natural killer cell antibody-dependent cytotoxicity against human immunodeficiency virus-1
Article Snippet: To minimize the background cell death, dead cells were removed prior to antibody-coating using the Dead Cell Removal kit (Miltenyi Biotec). .. 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. ..

Article Title: Selective agonists of KIR and NKG2A to evade missing self response of natural killer cells.
Article Snippet: NK92 cells were cultured in MyeloCult H5100 medium (STEMCELL Technologies, ST05150) supplemented with 500 IU/mL Recombinant Human IL-2 (Nipro, 87–890; or Clinigen, Proleukin) and 1% (v/v) Streptomycin/Penicillin. .. PBMC-derived NK (PBMC-NK) cells were cultured in MyeloCult H5100 medium supplemented with 1000 IU/mL Recombinant Human IL-2, 100 ng/mL Recombinant Human IL-18 (R&D Systems, 9124-IL) and 1% (v/v) Streptomycin/Penicillin. .. PBMC-derived T (PBMC-T) cells were cultured in AIM V Medium (Thermo Fisher Scientific, 12055083) supplemented with 10% (v/v) FBS and 100 IU/mL Recombinant Human IL-2.

Article Title: IL-18 potentiates platelet activation and thrombosis through IL-18Rα-dependent MAPKs and PI3K/Akt signaling.
Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.

Article Title: Pharmacological Characterization of Zasocitinib (TAK-279): An Oral, Highly Selective, and Potent Allosteric TYK2 Inhibitor.
Article Snippet: Buffers and antibodies for flow cytometric analysis of signal transducer and activator of transcription (STAT) phosphorylation were obtained from BD Biosciences (San Diego, CA), BioLegend (San Diego, CA), and eBioscience (San Diego, CA), Miltenyi Biotec (Westphalia, Germany), R&D Systems (Minneapolis, MN), Rockland Immunochemicals (Limerick, PA), and Thermo Fisher Scientific as follows: BD Phosflow Lyse/Fix buffer (catalog number 558049), BD Phosflow Perm Buffer III (catalog number 558050), BD Brilliant Stain Buffer (catalog number 566349), BD BSA stain buffer (catalog number 554657), Phycoerythrin-conjugated mouse antihuman CD42a antibody (catalog number 558819), FITC-conjugated mouse antihuman CD3 antibody (catalog number 561807), and Alexa Fluor 647econjugated mouse antihuman Stat5 pY694 antibody (catalog number 562076) from BD Biosciences; phycoerythrin-conjugated mouse anti-STAT4 (pY693) (catalog number 941204), Alexa Fluor 647econjugated mouse antihuman CD161 antibody (catalog number 339910), Alexa Fluor 647econjugated mouse antihuman CD61 antibody (catalog number 336408), Brilliant Violet 421econjugated mouse antihuman STAT3 Phospho (Tyr705) antibody (catalog number 651010), Brilliant Violet 605econjugated mouse antihuman CD62P antibody (catalog number 304920), Alexa Fluor 647econjugated mouse antihuman STAT4 Phospho (Tyr693) antibody (catalog number 941204), APCCy7econjugated mouse antihuman CD41 antibody (catalog number 303716), and Fc receptor blocking solution (catalog number 422302) from BioLegend; Fc receptor binding inhibitor polyclonal antibody (catalog number 14-9161-71) from eBioscience; AutoMACS running buffer (catalog number 130-091-221) from Miltenyi Biotec; reconstitution buffer (catalog number RB05 or RB02), Rockland Immunochemicals BSA 30% solution (lot number 49664), BD BSA stain buffer (catalog number BSA-30), and FACS buffer comprising PBS þ 1% BSA from R&D Systems; and Hanks’ Balanced Salt Solution (catalog number 14025092), FITC-conjugated mouse antihuman STAT3 Phospho (Tyr705) antibody (catalog number 11-9033-42), and recombinant human ILs from Thermo Fisher Scientific. .. Type 1 IFN and thrombopoietin were obtained as follows: recombinant human IL-2 (catalog number PHC00267, Life Technologies, Carlsbad, CA), recombinant human IL-12 (catalog number 200-12, PeproTech US, Cranbury, NJ), recombinant human IL-18 (catalog number 9124-IL-050, R&D Systems), HumanKine recombinant human IL-23 (catalog number HZ-1254, Proteintech, Rosemont, IL), recombinant human type 1 IFN (catalog number 11200-1, PBL Assay Science, Piscataway, NJ), and recombinant human thrombopoietin (catalog number 300-18, Thermo Fisher Scientific). .. All flow cytometric analyses were carried out using a BD FACSCanto II or BD Fortessa X-20 machine or a BD Fortessa without X-20 (BD Biosciences).

Article Title: Selective agonists of KIR and NKG2A to evade missing self response of natural killer cells
Article Snippet: NK92 cells were cultured in MyeloCult H5100 medium (STEMCELL Technologies, ST-05150) supplemented with 500 IU/mL Recombinant Human IL-2 (Nipro, 87–890; or Clinigen, Proleukin) and 1% (v/v) Streptomycin/Penicillin. .. PBMC-derived NK (PBMC-NK) cells were cultured in MyeloCult H5100 medium supplemented with 1000 IU/mL Recombinant Human IL-2, 100 ng/mL Recombinant Human IL-18 (R&D Systems, 9124-IL) and 1% (v/v) Streptomycin/Penicillin. .. PBMC-derived T (PBMC-T) cells were cultured in AIM V Medium (Thermo Fisher Scientific, 12055083) supplemented with 10% (v/v) FBS and 100 IU/mL Recombinant Human IL-2.

Cell Culture:

other:

Article Title: Heterodimeric FC cytokines and uses thereof
Article Snippet: The vectors were DNA sequenced (MC Lab, South San Francisco, CA) to confirm identity.

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).

Techniques: Genome Wide, Comparison, Control, Expressing

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