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Human il 18

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Sourced in Australia, United States

Human IL-18 is a recombinant human protein. It is a member of the interleukin-1 family and is involved in the regulation of immune and inflammatory responses.

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Lab products found in correlation

Mouse il 18, by R&D Systems (3 mentions) Human ifn β, by R&D Systems (2 mentions) Mouse ifn β, by R&D Systems (2 mentions) Tlrl msu, by InvivoGen (2 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions) Human il 12, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Merck Group (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Human il 10, by BioLegend (1 mentions) Rneasy kit, by Qiagen (1 mentions) Human il 1β, by Thermo Fisher Scientific (1 mentions) Mouse il 1β, by Thermo Fisher Scientific (1 mentions) Ccl 86, by American Type Culture Collection (1 mentions) Cxcl1 kc, by R&D Systems (1 mentions) Cxcl2 mip 2, by R&D Systems (1 mentions) Cxcl8, by BD (1 mentions) Mouse il 1β, by BD (1 mentions) Il 18, by InvivoGen (1 mentions) Il 18, by R&D Systems (1 mentions) Il 33, by Thermo Fisher Scientific (1 mentions)

10 protocols using human il 18

1

Cytokine and LDH Quantification in PBMCs

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Commercial ELISA kits were used to determine the amounts of human IL-1β (Mabtech), human IL-18 (R&D Systems), human IL-1α (Biolegend), and human IL-10 (Biolegend) in the culture supernatants of PBMCs and CD14+ monocytes. A colorimetric lactate dehydrogenase (LDH) assay kit (Abcam) was used to determine the LDH activity in culture supernatants of PBMCs and CD14+ monocytes.
Jung B.G., Dean K., Wadle C., Samten B., Tripathi D., Wallace RJ J.r., Brown-Elliott B.A., Tucker T., Idell S., Philley J.V, & Vankayalapati R. (2022). Decreased Interleukin-1 Family Cytokine Production in Patients with Nontuberculous Mycobacterial Lung Disease. Microbiology Spectrum, 10(6), e03110-22.
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2

Cytokine Secretion Assay in Mouse RPE Cells

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Secreted human and mouse interferon-β and IL-18 in the media were detected using ELISA kits (mouse IFN-β, R&D Systems Cat# 42400-1; mouse IL-18, R&D Systems Cat# 7625; human IFN-β, R&D Systems Cat# 41410; human IL-18, R&D systems Cat# DY318-05) according to the manufacturer′s instructions. Primary mouse cells were cultured as above. WT, Gsdmd−/−, Casp11−/−, or Mb21d1−/− mouse RPE cells were seeded at a density of 250,000 cells/well in a 12-well plate. When confluency reached 60–70%, cells were transfected with 20 pmol of in vitro transcribed Alu RNA or mock using Lipofectamine 2000 reagent (Life Technologies, Carlsbad, CA) following the manufacturer’s protocol. Media was collected to detect secreted cytokine content at 8 to 24 h post-transfection. For examination of the induction of IL-18 secretion by monosodium urate (MSU) crystals (Invivogen Cat# tlrl-msu), mouse RPE cells were primed with LPS (500 ng/ml) for 6 h and exposed to MSU (250 μg/ml) for 16 h, and media was collected to detect secreted cytokine.
Kerur N., Fukuda S., Banerjee D., Kim Y., Fu D., Apicella I., Varshney A., Yasuma R., Fowler B.J., Baghdasaryan E., Marion K.M., Huang X., Yasuma T., Hirano Y., Serbulea V., Ambati M., Ambati V.L., Kajiwara Y., Ambati K., Bastos-Carvalho A., Ogura Y., Terasaki H., Oshika T., Kim K.B., Hinton D.R., Leitinger N., Cambier J.C., Buxbaum J.D., Kenney M.C., Jazwinski S.M., Nagai H., Hara I., West A.P., Fitzgerald K.A., Sadda S.R., Gelfand B.D, & Ambati J. (2017). cGAS drives non-canonical inflammasome activation in age-related macular degeneration. Nature medicine, 24(1), 50-61.
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3

Cytokine-Stimulated NK Cell Profiling

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Cytokine stimulation assays were performed in 96 well plates. 1 × 105 NKs were plated in RPMI media (as above). Human IL-18 (50 ng/ml) (R&D systems) and 20 IU/ml IL-2 (NIH) were added to NK cells and plates incubated at 37 °C. NK cells were then spun down at 1400 rpm for 4 min and cell pellets frozen in RLT buffer (Qiagen RNEasy kit) + 10 µl/ml 2-Mercaptoethanol (BME) and stored at −80°C.
Todd K.L., Lai J., Sek K., Huang Y.K., Newman D.M., Derrick E.B., Koay H.F., Nguyen D., Hoang T.X., Petley E.V., Chan C.W., Munoz I., House I.G., Lee J.N., Kim J.S., Li J., Tong J., N. de Menezes M., Scheffler C.M., Yap K.M., Chen A.X., Dunbar P.A., Haugen B., Parish I.A., Johnstone R.W., Darcy P.K, & Beavis P.A. (2023). A2AR eGFP reporter mouse enables elucidation of A2AR expression dynamics during anti-tumor immune responses. Nature Communications, 14, 6990.
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4

Cytokine Quantification in Cell Culture and Sera

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Cell culture supernatants and mouse sera were assayed for human IL-1β (eBioscience), human IL-18 (R&D Systems), and mouse IL-1β (eBioscience), respectively. Colon organ culture supernatants were assayed for mouse IL-18 (R&D Systems).
Chung I.C., Yuan S.N., OuYang C.N., Lin H.C., Huang K.Y., Chen Y.J., Chung A.K., Chu C.L., Ojcius D.M., Chang Y.S, & Chen L.C. (2018). Src-family kinase-Cbl axis negatively regulates NLRP3 inflammasome activation. Cell Death & Disease, 9(11), 1109.
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5

Cytokine Production and Proliferation Assay

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For cytokine production analyses, thawed and rested PBMCs were stimulated with PMA (5 ng/mL; Sigma) plus Ionomycin (500 nmol/L; Sigma), or with human IL12 (10 ng/mL; Pepro-tech) plus human IL18 (50 ng/mL; R&D Systems). Alternatively, PBMCs were exposed to the target cell lines K562 (ATCC CCL-243) or to anti–CD20-coated Raji (ATCC CCL-86) that had been preincubated with 10 mg/mL of the antibody (30 minutes, 4°C) and washed. Stimulation and coincubations were done for 5 hours at 37°C with Brefeldin A present during the last 4 hours. For proliferation assays, sorted cells were carboxyfluorescein succinimidyl ester (CFSE)-labeled and cultured in IL15 at indicated dosages for 7 days.
Dubois S., Conlon K.C., Müller J.R., Hsu-Albert J., Beltran N., Bryant B.R, & Waldmann T.A. (2017). IL15 Infusion of Cancer Patients Expands the Subpopulation of Cytotoxic CD56bright NK Cells and Increases NK-Cell Cytokine Release Capabilities. Cancer immunology research, 5(10), 929-938.
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6

Cytokine Production Analyses of PBMCs

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For cytokine production analyses, thawed and rested PBMCs were stimulated with PMA (5 ng/mL, Sigma) plus ionomycin (500 nmol/L, Sigma), or with human IL-12 (10 ng/mL, PeproTech) plus human IL-18 (50 ng/mL, R&D Systems). Alternatively, PBMCs were exposed to the target cell lines K562 (ATCC CCL-243), MHC class I chain-related protein A (C1R-MICA) MHC class I related stress-inducible surface glycoprotein (kindly provided by Dr Veronika Groh-Spies, Fred Hutchinson Cancer Research Center) or to anti-CD20-coated Raji (ATCC CCL-86) cells that had been preincubated with 10 µg/mL of the antibody (30 min at 4°C) and washed. Stimulation and coincubations were done for 5 hours at 37°C with brefeldin A present during the last 4 hours.
Dubois S.P., Miljkovic M.D., Fleisher T.A., Pittaluga S., Hsu-Albert J., Bryant B.R., Petrus M.N., Perera L.P., Müller J.R., Shih J.H., Waldmann T.A, & Conlon K.C. (2021). Short-course IL-15 given as a continuous infusion led to a massive expansion of effective NK cells: implications for combination therapy with antitumor antibodies. Journal for Immunotherapy of Cancer, 9(4), e002193.
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7

Cytokine Secretion Assay in Mouse RPE Cells

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Secreted human and mouse interferon-β and IL-18 in the media were detected using ELISA kits (mouse IFN-β, R&D Systems Cat# 42400-1; mouse IL-18, R&D Systems Cat# 7625; human IFN-β, R&D Systems Cat# 41410; human IL-18, R&D systems Cat# DY318-05) according to the manufacturer′s instructions. Primary mouse cells were cultured as above. WT, Gsdmd−/−, Casp11−/−, or Mb21d1−/− mouse RPE cells were seeded at a density of 250,000 cells/well in a 12-well plate. When confluency reached 60–70%, cells were transfected with 20 pmol of in vitro transcribed Alu RNA or mock using Lipofectamine 2000 reagent (Life Technologies, Carlsbad, CA) following the manufacturer’s protocol. Media was collected to detect secreted cytokine content at 8 to 24 h post-transfection. For examination of the induction of IL-18 secretion by monosodium urate (MSU) crystals (Invivogen Cat# tlrl-msu), mouse RPE cells were primed with LPS (500 ng/ml) for 6 h and exposed to MSU (250 μg/ml) for 16 h, and media was collected to detect secreted cytokine.
Kerur N., Fukuda S., Banerjee D., Kim Y., Fu D., Apicella I., Varshney A., Yasuma R., Fowler B.J., Baghdasaryan E., Marion K.M., Huang X., Yasuma T., Hirano Y., Serbulea V., Ambati M., Ambati V.L., Kajiwara Y., Ambati K., Bastos-Carvalho A., Ogura Y., Terasaki H., Oshika T., Kim K.B., Hinton D.R., Leitinger N., Cambier J.C., Buxbaum J.D., Kenney M.C., Jazwinski S.M., Nagai H., Hara I., West A.P., Fitzgerald K.A., Sadda S.R., Gelfand B.D, & Ambati J. (2017). cGAS drives non-canonical inflammasome activation in age-related macular degeneration. Nature medicine, 24(1), 50-61.
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8

Quantification of Inflammatory Cytokines

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Mouse IL-1β (BD Biosciences, North Ryde, NSW, Australia), IL-18 (R&D Systems, In Vitro Technologies Pty Ltd., VIC, Australia), Cxcl1/KC (R&D Systems), Cxcl2/MIP2 (R&D Systems), human IL-18 (R&D Systems) and CXCL8 (BD Biosciences) were quantified by ELISA, as per the manufacturers’ instructions. Production of bioactive IL-18 by AGS cells was also measured using the HEK-Blue™ IL-18 reporter cell line system (InvivoGen, San Diego, CA, USA).
Tran L.S., Ying L., D’Costa K., Wray-McCann G., Kerr G., Le L., Allison C.C., Ferrand J., Chaudhry H., Emery J., De Paoli A., Colon N., Creed S., Kaparakis-Liaskos M., Como J., Dowling J.K., Johanesen P.A., Kufer T.A., Pedersen J.S., Mansell A., Philpott D.J., Elgass K.D., Abud H.E., Nachbur U., Croker B.A., Masters S.L, & Ferrero R.L. (2023). NOD1 mediates interleukin-18 processing in epithelial cells responding to Helicobacter pylori infection in mice. Nature Communications, 14, 3804.
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9

Eosinophil Activation by IL-18 and IL-33

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Blood samples were obtained from individuals with a physician diagnosis of asthma and/or allergic rhinoconjunctivitis. Granulocytes were isolated from peripheral blood by density gradient centrifugation, followed by negative immunomagnetic selection of EOS (6) . EOS were treated with human IL-18 (R&D Systems) and/or IL-33 (Peprotech). RNA was isolated from EOS, and qPCR analysis was conducted using TaqMan primer probe sets with quantification of IL13, IL18, and IFNG genes relative to the endogenous control gene HPRT1 using the ΔCt method.
Murphy R.C., Lai Y., Liu M., Al-Shaikhly T., Altman M.C., Altemeier W.A., Frevert C.W., Debley J.S., Piliponsky A.M., Ziegler S.F., Gharib S.A, & Hallstrand T.S. (2023). Distinct Epithelial-Innate Immune Cell Transcriptional Circuits Underlie Airway Hyperresponsiveness in Asthma. American journal of respiratory and critical care medicine, 207(12).
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10

Cytokine Secretion Inhibition in PBMC Stimulation

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A total of 1.0 × 10 6 PBMCs from healthy donors were stimulated with 1.0 × 10 6 copies of EBV in RPMI 1640 supplemented with 10% FBS for 24 h. To inhibit cytokine secretion, monensin was added for the last 4 h of the 24 h culture at a volume indicated in the instructions provided with the Cell Fixation/Permeabilization Kit with BD GolgiStop. Human IL-18 (R&D systems, MN, USA), anti-Human IL-18 monoclonal antibody (2.5 µg/ml), and mouse IgG1 (2.5 µg/ml) (MBL, Tokyo, Japan) were added to each culture well at the beginning of the culturing.
Ishikawa Y., Yamada M., Wada N., Takahashi E, & Imadome K.I. (2022). Mucosal-associated invariant T cells are activated in an interleukin-18-dependent manner in Epstein-Barr virus-associated T/natural killer cell lymphoproliferative diseases. Clinical and experimental immunology, 207(2).
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