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Il 25

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IL-25 is a cytokine that plays a role in the regulation of immune responses. It is a member of the IL-17 family of cytokines. IL-25 can be detected and quantified using appropriate immunoassay techniques.

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Lab products found in correlation

Il 33, by BioLegend (7 mentions) Interleukin 7 (il 7), by BioLegend (4 mentions) Interleukin 2 (il 2), by BioLegend (3 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Il 13, by Thermo Fisher Scientific (1 mentions) Il 13 elisa, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Atlanta Biologicals (1 mentions) αifn γ xmg1, by BioLegend (1 mentions) Interleukin 3 (il 3), by Fujifilm (1 mentions) Interleukin 4 (il 4), by Fujifilm (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Il 22, by BioLegend (1 mentions) Ionomycin, by Thermo Fisher Scientific (1 mentions) Fortessa flow cytometer, by BD (1 mentions) Legendplex murine th2 panel v03 multiplex beads based assay, by BioLegend (1 mentions) Legendplex cloud based analysis software suite, by BioLegend (1 mentions) U bottom microtiter plates, by Thermo Fisher Scientific (1 mentions) Facsdiva software, by BD (1 mentions) Il 15, by BioLegend (1 mentions)

13 protocols using il 25

1

Flt3L-Induced Differentiation of BM Cells

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1×106 BM cells were incubated in complete RPMI media (Life Technologies-Thermo-Fisher, Walkersville MD), 1/5 of which from supernatants of B16 cells-expressing Flt3L (gift from Dr. Mora, Milennium Pharmaceuticals). Fresh media was added to the cultures every two-three days. On day 8, 5×105 Flt3L-BMDCs were incubated with/without IL-25 (Biolegend, San Diego CA) and IL-13 (Peprotech, Rocky Hill NJ).
Claudio E., Wang H., Kamenyeva O., Tang W., Ha H.L, & Siebenlist U. (2019). IL-25 orchestrates activation of Thelper cells via conventional dendritic cells in tissue to exacerbate chronic HDM-induced asthma pathology. Journal of immunology (Baltimore, Md. : 1950), 203(8), 2319-2327.
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2

Profiling Cytokine Secretion in ILC2s

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Bone marrow cells (5 × 105 cells per well) were plated in a round bottom 96-well plate and co-cultured with IL-2, IL-7, IL-33 at 10 ng/ml (Biolegend), and recombinant parasite-derived proteins, at 37°C, 5% CO2 for 5 days. Supernatants were collected and analysed by IL-5, IL-6 and IL-13 ELISA according to manufacturer’s instructions (Thermo Fisher). Where biotinylated constructs were used to assess binding to bone marrow ILC2s, BALB/c or ST2-deficient bone marrow cells were cultured for 5 days in IL-2, IL-7 and IL-25 at 10 ng/ml (Biolegend) to support ILC2 proliferation, prior to incubation for 20 min at 4°C with HES and recombinant proteins, followed by biotinylated recombinant proteins for a further 20 min, as indicated, and avidin-PE to detect binding.
Vacca F., Chauché C., Jamwal A., Hinchy E.C., Heieis G., Webster H., Ogunkanbi A., Sekne Z., Gregory W.F., Wear M., Perona-Wright G., Higgins M.K., Nys J.A., Cohen E.S, & McSorley H.J. (2020). A helminth-derived suppressor of ST2 blocks allergic responses. eLife, 9, e54017.
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3

Expansion and Maintenance of Cell Types

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Cell culture was performed using RPMI-1640 medium supplemented with penicillin, streptomycin, gentamicin, 2-mercaptoethanol, 2mM sodium pyruvate, glutamine, 10mM HEPES (all Life Technologies) and 10% fetal calf serum (Atlanta Biologicals). Cells were incubated at 37°C humidified atmosphere with 5% CO2. Mast cells (BMMC) were cultured from bone marrow precursors in the presence of IL-3 and SCF (both 20ng/ml, Shenandoah Biotechnology) as previously described [56 (link)]. ILC2 were expanded from lamina propria leukocytes with 20ng/ml IL-33 (Biolegend), 25ng/ml IL-25 (Biolegend), 10ng/ml IL-4 (Shenandoah Biotechnology) and 5ng/ml IL-2 (as IL-2-transfected X63 supernatant) combined with 5μg/ml αIFNγ (XMG1.2, Biolegend) for 3–7 days prior to cell sorting, and were subsequently maintained in IL-2-supplemented medium.
Burton O.T., Medina-Tamayo J., Stranks A.J., Miller S., Koleoglou K.J., Weinberg E.O, & Oettgen H.C. (2018). IgE promotes type 2 innate lymphoid cells in murine food allergy. Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, 48(3), 288-296.
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4

Cytokine Modulation of Parasite Infection

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Where indicated, 5 µg of IL-25 (#587306; BioLegend), or sterile PBS as a vehicle control, was administered by intraperitoneal (i.p.) injection every other day between Days 3 and 11 following T. muris infection, and 5 µg of IL-22 (#576206; BioLegend), or PBS, was administered by i.p. injection on Days 0, 2, and 4 following C. rodentium infection.
Israelson H., Vedsted-Jakobsen A., Zhu L., Gagnaire A., von Münchow A., Polakovicova N., Valente A.H., Raza A., Andersen-Civil A.I., Olsen J.E., Myhill L.J., Geldhof P, & Williams A.R. (2024). Diet composition drives tissue-specific intensity of murine enteric infections. mBio, 15(2), e02603-23.
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5

Cytokine Profiling of Activated Immune Cells

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Bulk purified immune cells were incubated in DMEM with high glucose supplemented with 10% FCS, 10 mM Hepes, 1 mM sodium pyruvate, non-essential amino acids, 80 μM 2-Mercaptoethanol, 2 mM Glutamine, 100 U/ml Penicillin and 100 μg/ml Streptomycin (all from Gibco) in 96-well U-bottom microtiter plates (Nunc) for 8 h at 37°C and 5% CO2. If indicated, the culture was supplemented with a cocktail of IL-2, IL-7, IL-25, IL-33 (Biolegend, 100 ng/ml, each) or phorbol 12-myristate 13-acetate (PMA, 1 μg/ml) and ionomycin (Invitrogen, 0.5 μg/ml).
Cytokine concentration in culture supernatant or serum samples were determined by using the LEGENDplex murine Th Cytokine Panel V03 and LEGENDplex murine TH2 Panel V03 multiplex beads-based assay (Biolegend) or the BD Cytometric Bead Array Mouse IL-5 Enhanced Sensitivity Flex Set (BD Biosciences) according to the manufacture’s protocol. Samples were recorded on a custom configuration Fortessa flow cytometer and the FACS Diva software (BD Biosciences) and the flow cytometry data files were analyzed using the Legendplex cloud-based analysis software suite (Biolegend).
Jarick K.J., Topczewska P.M., Jakob M.O., Yano H., Arifuzzaman M., Gao X., Boulekou S., Stokic-Trtica V., Leclere P., Preusser A., Rompe Z.A., Stamm A., Tsou A.M., Chu C., Heinrich F.R., Guerra G.M., Durek P., Ivanov A., Beule D., Helfrich S., Duerr C.U., Kühl A.A., Stehle C., Romagnani C., Mashreghi M.F., Diefenbach A., Artis D, & Klose C.S. (2022). Non-redundant functions of group 2 innate lymphoid cells. Nature, 611(7937), 794-800.
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6

Mature Adipocyte Macrophage Co-culture

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Confluent 3T3-L1 (ATCC) cells seeded in six-well plates were induced into mature adipocytes with 0.5 mM isobutylmethylxanthine (Sigma-Aldrich), 1 μM dexamethasone (Sigma-Aldrich) and 10 μg/ml insulin (Sigma-Aldrich) in dulbecco's modified eagle medium containing 10% fetal bovine serum for 2 days and then treated with dulbecco's modified eagle medium containing 10 μg/ml insulin and 10% fetal bovine serum for 7 days. Then, 3 × 105 RAW264.7 cells/well were seeded into Transwell chambers (Corning, New York, NY, USA) on top of the mature adipocytes for 2 days. Different cytokines, such as IL-5 (50 ng/ml, Biolegend), IL-15 (25 ng/ml, Biolegend), IL-25 (50 ng/ml, Biolegend), IFNγ (50 ng/ml, Biolegend) and lipopolysaccharide (LPS; 10 ng/ml, Sigma), were administered for treatment of the macrophages.
Feng J., Li L., Ou Z., Li Q., Gong B., Zhao Z., Qi W., Zhou T., Zhong J., Cai W., Yang X., Zhao A., Gao G, & Yang Z. (2017). IL-25 stimulates M2 macrophage polarization and thereby promotes mitochondrial respiratory capacity and lipolysis in adipose tissues against obesity. Cellular and Molecular Immunology, 15(5), 493-505.
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7

Culturing Purified Intestinal ILC2s

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Sort-purified intestinal ILC2s were cultured in DMEM with high glucose supplemented with 10% FBS, 10 mM Hepes, 1 mM sodium pyruvate, 1x MEM non-essential amino acid solution, 80 μM 2-Mercaptoethanol, 2 mM L-Glutamine, 100 U/ml Penicillin and 100 μg/ml Streptomycin in 96-well microtiter plates at 37°C and 5% CO2 for 3 days, in the presence of IL-7, IL-25 and IL-33 (20 ng/ml each, all from Biolegend) as indicated.
Flamar A.L., Klose C.S., Moeller J.B., Mahlakõiv T., Bessman N.J., Zhang W., Moriyama S., Stokic-Trtica V., Rankin L.C., Putzel G.G., Rodewald H.R., He Z., Chen L., Lira S.A., Karsenty G, & Artis D. (2020). Interleukin-33 induces the enzyme tryptophan hydroxylase 1 to promote inflammatory group 2 innate lymphoid cell-mediated immunity. Immunity, 52(4), 606-619.e6.
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8

Cytokine Profiling in Mouse and Human

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Mouse IL-13, AREG, and human IL-25 ELISA kits were purchased from R&D Systems and used according to the manufacturer’s instructions. Mouse legend plex was used for IL-33, IL-25, TSLP, and interferon-β were purchased from BioLegend.
Krishnamoorthy N., Walker K.H., Brüggemann T.R., Tavares L.P., Smith E.W., Nijmeh J., Bai Y., Ai X., Cagnina R.E., Duvall M.G., Lehoczky J.A, & Levy B.D. (2023). The Maresin 1–LGR6 axis decreases respiratory syncytial virus-induced lung inflammation. Proceedings of the National Academy of Sciences of the United States of America, 120(2), e2206480120.
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9

Isolation and Culture of Mouse Lung Macrophages

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Mouse pulmonary macrophages were isolated from lung tissue of C57BL/6J mice aged around 6-week-old, the mice were euthanized and their lung tissues were removed for enzymatic digestion and processed into single cell suspension. After centrifugation, the cell sediment was removed, and the cells were resuspended in RPMI-1640 medium (Hyclone) with 10% FBS (Gibco), 100 IU/mL penicillin, and 100 mg/mL streptomycin and cell cultures were maintained at 37 °C and 5% CO2 for 1 h. The medium was discarded and washed with PBS to remove the suspended cells. The culture was replaced with complete medium overnight, and Giemsa staining confirmed that more than 95% of the adherent cells were lung macrophages. Based on the above steps, approximately 1*10^6 macrophages can be extracted from each mouse lung tissue, which can be used for inoculation with a six well plate. Escherichia coli–derived LPS (1 µg/mL; Sigma-Aldrich), IL-25 (50 ng/mL, Biolegend), Stattic (5 µM, MedChemExpress) were used when indicated.
Chang C., Chen G., Wu W., Chen D., Chen S., Gao J., Feng Y, & Zhen G. (2023). Exogenous IL-25 ameliorates airway neutrophilia via suppressing macrophage M1 polarization and the expression of IL-12 and IL-23 in asthma. Respiratory Research, 24, 260.
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10

IL-25 and IL-33 Intraperitoneal Injection

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Mice were given intraperitoneal injections of 500 ng IL-25 or IL-33 (BioLegend) on days 0–3. On day 4, tissues were collected for analysis.
Yu Y., Wang C., Clare S., Wang J., Lee S.C., Brandt C., Burke S., Lu L., He D., Jenkins N.A., Copeland N.G., Dougan G, & Liu P. (2015). The transcription factor Bcl11b is specifically expressed in group 2 innate lymphoid cells and is essential for their development. The Journal of Experimental Medicine, 212(6), 865-874.
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