Il 17

Manufactured by Miltenyi Biotec

IL-17 is a lab equipment product manufactured by Miltenyi Biotec. It is used for the detection and quantification of the cytokine interleukin-17 in various biological samples.

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Lab products found in correlation

Il 22, by Miltenyi Biotec (2 mentions) Staurosporine, by Selleck Chemicals (1 mentions) Il 10, by Miltenyi Biotec (1 mentions) Gm csf, by Miltenyi Biotec (1 mentions) Ifn γ, by Miltenyi Biotec (1 mentions) Tnf α, by Miltenyi Biotec (1 mentions) Cd4 rm4 5, by BD (1 mentions) Cd16 cd32, by Cytek Biosciences (1 mentions) Foxp3 fixation permeabilization kit, by BD (1 mentions) Rorγt, by BD (1 mentions) Il 10, by BioLegend (1 mentions) Interleukin 4 (il 4), by BD (1 mentions) Foxp3, by Thermo Fisher Scientific (1 mentions) Gata3, by BD (1 mentions) T bet, by BioLegend (1 mentions) Ionomycin, by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Golgistop, by BD (1 mentions) Ifn γ, by BioLegend (1 mentions) Moflo xdp, by Beckman Coulter (1 mentions)

6 protocols using il 17

Cytokine-Induced Neuronal Cell Cultures

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Neuronal cell cultures were treated with IL-17, IL-10, IFNγ, TNFα, GM-CSF (concentration: 50 ng/mL; Miltenyi), l-glutamate (concentration: 250 µM; Miltenyi) or staurosporine (concentration: 0.5 µM; Selleckchem) for 24 h respectively. Concentrations of cytokines were selected based on assumed local concentrations in the CNS of PwMS as described in previous studies (Huppert et al. 2010 (link)), (Zong et al. 2016 (link)), (Ta et al. 2019 (link)), (Nasiri et al. 2020 ), (Neniskyte et al. 2014 (link)), (Riazi et al. 2008 (link)), (Schäbitz et al. 2007 ), (Vaarmann et al. 2013 (link)), (Dikmen et al. 2020 (link)). We chose the duration of treatment according to results of preliminary tests showing first signs of neuronal integrity alterations after 24 h without further changes upon prolongation of cytokine treatment.

Meyer-Arndt L., Kerkering J., Kuehl T., Infante A.G., Paul F., Rosiewicz K.S., Siffrin V, & Alisch M. (2023). Inflammatory Cytokines Associated with Multiple Sclerosis Directly Induce Alterations of Neuronal Cytoarchitecture in Human Neurons. Journal of Neuroimmune Pharmacology, 18(1-2), 145-159.

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Murine Lung Immune Cell Profiling

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P1 and P21 male and female murine lungs were isolated as described above. Cells were blocked for 30 min with CD16/CD32 (Tonbo Biosciences). For intracellular analyses, cells were stimulated with PMA (50 ng/ml, Sigma-Aldrich), ionomycin (750 ng/mL, Sigma-Aldrich), and GolgiStop (BD Biosciences) for 5 hr, and then surface stained with fluorochrome-conjugated antibodies for 30 min: CD3 (clone: 145–2 C11, BD Biosciences), CD4 (RM4-5, BD Biosciences), and CD8a (clone: 53–6.7, Biolegend). Cells were then permeabilized with FOXP3 Fixation/Permeabilization Kit (BD Biosciences) as indicated by the manufacturer, and stained for TBET (clone: 4B10, Biolegend), GATA3 (clone: L50-823, BD Biosciences), FOXP3 (clone: FJK-16s, eBioscience), RORγt (clone: Q31-378, BD Biosciences), IFNγ (clone: XMG1.2, Biolegend), IL-4 (clone: 11B11, BD Biosciences), IL-10 (clone: JES5-16E3, Biolegend), and IL-17 (clone: TC11-18H10, Miltenyi Biotec) for 30 min. Cells were read using an LSRII flow cytometer using FACSDiva software. Flow data was analyzed using FlowJo (Tree Star Inc). Flow cytometry analysis for this project was done on instruments in the Stanford Shared FACS Facility.

Domingo-Gonzalez R., Zanini F., Che X., Liu M., Jones R.C., Swift M.A., Quake S.R., Cornfield D.N, & Alvira C.M. (2020). Diverse homeostatic and immunomodulatory roles of immune cells in the developing mouse lung at single cell resolution. eLife, 9, e56890.

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Isolation of Tregs, Th1 and Th17 cells from MOG-immunized mice

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Female 13R^+/+ or 13R^-/- Foxp3.GFP mice were immunized s.c. with 200μl IFA/PBS (v/v) solution containing 100 μg MOG peptide and 200 μg of Mycobacterium tuberculosis H37Ra (Difco) in the footpads and at the base of the limbs.
For isolation of Tregs the LN were harvested on day 10 post immunization. Cells were gated on CD4 and CD25 and sorted on the basis of GFP (Foxp3⁺) expression (97% purity) using a Beckman Coulter MoFlo XDP.
For isolation of effector Th1 and Th17 cells, SP cells were harvested on day 10 post immunization. CD4⁺ T cells were isolated from the SP by negative selection of lineage specific cells using a CD4⁺ T cell isolation kit (Miltenyi Biotech, San Diego, CA). The CD4⁺ T cells were stimulated with PMA and ionomycin for 3 hours to induce cytokine secretion optimal for isolation of bulk Th17 and Th1 cells by IL-17 and IFN-γ Secretion Assay/Cell Enrichment and Detection Kits, respectively (Miltenyi Biotech, San Diego, CA). Only cells with 98% purity are used for in vitro and in vivo suppression assays.

Barik S., Ellis J.S., Cascio J.A., Miller M.M., Ukah T.K., Cattin-Roy A.N, & Zaghouani H. (2017). IL-4/IL-13 Heteroreceptor Influences Th17 Cell Conversion and Sensitivity to Treg Suppression to Restrain Experimental Allergic Encephalomyelitis. Journal of immunology (Baltimore, Md. : 1950), 199(7), 2236-2248.

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Proliferative signaling in human airway smooth muscle cells

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Primary aSMCs were isolated from human bronchial biopsies. Additional detail on the method is provided in an online data supplement. Human aSMCs were seeded into 24-well plates (10 000 aSMCs/well) and allowed to adhere during 6 hour before serum starvation during 24 hours. When indicated, human aSMCs were treated with the Rac inhibitor, EHT1864 (10⁻⁵ M; Tocris Bioscience), P21-activated kinases (Pak) inhibitor IPA3 (10⁻⁵ M; Tocris Bioscience), Akt inhibitor VIII (10⁻⁵ M; Calbiochem), MEK inhibitor PD98059 (10⁻⁵ M; ThermoFisher), JAK inhibitor ruxolitinib (10⁻⁵ M; InvivoGen) added 30 min before stimulation with bFGF (25 ng/mL; Miltenyi Biotech), PDGF-bb (25 ng/mL; Miltenyi Biotech), interleukins (IL)-13 (10 ng/mL; Miltenyi Biotech), IL-33 (10 ng/mL; Miltenyi Biotech), IL-17 (20 ng/mL; Miltenyi Biotech), IL-9 (10 ng/mL; Miltenyi Biotech) or TSLP (10 ng/mL; Miltenyi Biotech) for 48 hours. Cells were stained with EdU for 12 hours at 10⁻⁵ M according to the manufacturer’s indications (EdU Staining Proliferation Kit iFluor 488, ID: ab219801, Abcam). Proliferation was quantified by the ratio of EdU-positive cells over the total number of cells. Proliferative signalling pathways were analysed by immunoblotting detailed in an online data supplement.

Dilasser F., Rose L., Hassoun D., Klein M., Rousselle M., Brosseau C., Guignabert C., Taillé C., Dombret M.C., Di Candia L., Heddebaut N., Bouchaud G., Pretolani M., Magnan A., Loirand G, & Sauzeau V. (2021). Essential role of smooth muscle Rac1 in severe asthma-associated airway remodelling. Thorax, 76(4), 326-334.

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Colonic Biopsy Cytokine Stimulation

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Colonic biopsies (n=8) for ex vivo stimulations were collected in unaffected areas obtained after surgical resection of colorectal tumors (at least 10 cm distant to the tumor). Written informed consents were obtained under institutional review board protocol # DC-2008-402.
Biopsies were transported in ice-cold RPMI and processed within a maximum of 2h. Biopsies were controlled for weight and placed into 4-well Petri dishes filled with in 500μL serum-free medium (RPMI 1640, Gibco^™) supplemented with BSA (0.01%), 200μg/mL Penicillin/Streptomycin (Gibco ^™; #15140–122) and 0.25μg/mL Fungizone (Gibco^™; #15290–026). Biopsies were cultured during 6 h at 37°C in a 95% O₂ / 5% CO₂ atmosphere on a low-speed rocking platform in the presence of cytokines (IL-22; 100ng/mL [Miltenyi, # 130-096-295], IL-17; 100ng/mL [Miltenyi, #130-093-959], IL-22BP; 200ng/mL [Miltenyi, #8498-BP-025]). Supernatants were collected and stored at −80°C until use.

Jha D., Al-Taie Z., Krek A., Eshghi S.T., Fantou A., Laurent T., Tankelevich M., Cao X., Meringer H., Livanos A.E., Tokuyama M., Cossarini F., Bourreille A., Josien R., Hou R., Canales-Herrerias P., Ungaro R.C., Kayal M., Marion J., Polydorides A.D., Ko H.M., D’souza D., Merand R., Kim-Schulze S., Hackney J.A., Nguyen A., McBride J.M., Yuan G.C., Colombel J.F., Martin J.C., Argmann C., Suárez-Fariñas M., Petralia F, & Mehandru S. (2023). Myeloid cell influx into the colonic epithelium is associated with disease severity and non-response to anti-Tumor Necrosis Factor Therapy in patients with Ulcerative Colitis. bioRxiv.

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Cytokine Stimulation of Keratinocytes

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KCs were cultivated in 24-well plates and at 70% to 80% confluence were stimulated for 24 or 72 hours with 10 ng/mL tumor necrosis factor (TNF)-a, 5 ng/mL IL-1b, or 20 ng/mL IL-6, IL-8, IL-17, IL-22, epidermal growth factor (Miltenyi), 50 ng/mL IL-19, IL-23 (Miltenyi), or 50 ng/mL IL-20 (Roche).

Saalbach A., Tremel J., Herbert D., Schwede K., Wandel E., Schirmer C., Anderegg U., Beck-Sickinger A.G., Heiker J.T., Schultz S., Magin T, & Simon J.C. (2016). Anti-Inflammatory Action of Keratinocyte-Derived Vaspin: Relevance for the Pathogenesis of Psoriasis. The American journal of pathology, 186(3).

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