IL21R (Interleukin 21 Receptor) is a Protein Coding gene. Diseases associated with IL21R include Immunodeficiency 56 and Ige Responsiveness, Atopic. Among its related pathways are Cytokine Signaling in Immune system and IL-4 Signaling and its Primary Biological Effects in Different Immune Cell Types. The protein encoded by the IL21R (interleukin 21 receptor) gene is a cytokine receptor for interleukin 21 (IL21), which belongs to the type I cytokine receptor, and has been shown to form a heterodimeric receptor complex with a common γ chain, which is a receptor subunit also shared by the interleukin 2, 4, 7, 9 and 15 receptors. This receptor mediates the growth-promoting signaling of IL21 and is important for the proliferation and differentiation of T cells, B cells, and NK cells. Ligand binding of this receptor results in the activation of a variety of downstream signaling molecules, including JAK1, JAK3, STAT1, and STAT3. For example, by promoting the proliferation and development of Tfh and Th17 cells, balancing helper T cell subsets, inducing B cell generation and differentiation into plasma cells, and enhancing immunoglobulin production. These effects are mainly mediated by the activation of JAK/STAT, MAPK and PI3K pathways. In addition, there are studies examining the association between genetic variants in WSB1 and IL21R and Th2 immune responses during childhood A. lumbricoides infection suggest that the WSB1/IL21R pathway may provide a potential target for the treatment of Th2-mediated diseases.
Ba/F3 cell, a murine interleukin-3 dependent pro-B cell line, is a popular system for exploring both kinases and their inhibitors, because some protein kinases can render the Ba/F3 cells to be depended on the activation of the kinases instead of IL-3 supplement, while their inhibitors can antagonize the kinase-dependent growth effects.
Cell Line Name:
Ba/F3 IL21R cell line
Host Cell Line:
Stable Ba/F3 cell line expressing exogenous IL21R gene
Mostly single, round (some polymorph) cells in suspension
Split saturated culture 1:10 every 3 days; seed out at about 1-3 × 105 cells/mL
37 °C with 5% CO2
Liquid nitrogen immediately upon receiving
Approximately 20 hours
Cell Line Generation
Ba/F3-IL21R cell line was generated using lentivirus methods.
Characterization using PCR sequencing and WB
1. Prewarm culture medium (RPMI1640 + 10% FBS + 8μg/mL mouse IL-3 +1μg/mL Puromycin) in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:10 every 3 days; seed out at about 1-3 × 105 cells/mL.
1. Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage
1. Davis MR, Zhu Z, Hansen DM, Bai Q, Fang Y. The role of IL-21 in immunity and cancer. Cancer Lett. 2015 Mar 28.
2. Long D, Chen Y, Wu H, Zhao M, Lu Q. Clinical significance and immunobiology of IL-21 in autoimmunity. J Autoimmun. 2019 May.
3. Carneiro VL, da Silva HBF, Queiroz GA, Veiga RV, Oliveira PRS, Carneiro NVQ, Pires AO, da Silva RR, Sena F, Belitardo E, Nascimento R, Silva M, Marques CR, Costa RDS, Alcantra-Neves NM, Barreto ML, Cooper PJ, Figueiredo CA. WSB1 and IL21R Genetic Variants Are Involved in Th2 Immune Responses to Ascaris lumbricoides. Front Immunol. 2021 Feb 22.
F2 Building, Yizhuang Biomedical Park, No. 88, Kechuang Six Street, Beijing. P.R. China. 101111.