NK cell proliferation and inhibition in the presence of TGFβ1 was determined as previously described.33 (link) Enriched NK cells from human PBMC (Human NK Cell Enrichment Kit, STEMCELL Technologies, 19055) were used. NK cells were activated by recombinant human IL-2 (10 IU/mL; R&D Systems, 202-IL/CF) for 3 days in the presence or absence of various concentrations of TGFβ1 (R&D systems). NK cell proliferation was observed by staining with Ki-67-BUV395 antibody (BD Bioscience, 564071) using flow cytometry. TGFβ1 was added at the time of setup with or without SAR439459 or isotype control (IgG4) for 3 days. NK cytotoxicity assays were performed using K562 cells (ATCC® CCL-243™) as targets. NK and K562 cells were cocultured (effector-to-target ratio 5:1) in RPMI-1640 media (GIBCO, 11835–030) supplemented with 10% FBS (GIBCO, 10082–147), and delivery of granzyme B was quantitated by intracellular FACS analysis using GranToxiLux™ assay kit as described (OncoImmunin, Inc, GTL702-8), for 2 hours. In some experiments, culture supernatants were collected for cytokine, GZB, perforin analysis by MSD (Meso Scale Diagnostics) or Luminex (ThermoFisher Scientific).
202 il cf
Manufactured by R&D Systems
202-IL/CF is a laboratory instrument designed for the analysis and detection of interleukin (IL) and other cytokine-related factors (CF) in biological samples. The core function of this product is to provide researchers with a reliable and efficient tool for conducting immunoassays and quantifying the levels of specific proteins in their experiments.
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Lab products found in correlation
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ccl 243, by American Type Culture Collection (1 mentions)
Human nk cell enrichment kit, by STEMCELL (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Recombinant human il 1β, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Il 1β, by BD (1 mentions)
Il 23, by BD (1 mentions)
Interleukin 6 (il 6), by BD (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Tnf α, by BD (1 mentions)
2 protocols using 202 il cf
1
Quantifying NK Cell Proliferation and Cytotoxicity under TGFβ1 Influence
2
Naïve CD4+ T Cell Activation Assay
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Purified naïve CD4+ T cells (5 × 105) were cultured in 1 ml of complete medium containing human IL-2 (2 ng/ml, 202-IL/CF, R&D Systems) in 48-well plates and stimulated with plate-bound anti-CD3 (2 μg/ml, 555329, BD Pharmingen) and soluble anti-CD28 monoclonal antibodies (2 μg/ml, 555726, BD Pharmingen) for 5 d. The exogenous cytokines used were IL-1β (20 ng/ml), IL-6 (100 ng/ml), IL-23 (100 ng/ml), and TNF-α (50 ng/ml). Recombinant human IL-1β, IL-6, and TNF-α were purchased from Peprotech.
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