Recombinant human IL-4 protein - Bioactive cytokine

Human IL-4 protein - E. coli -expressed, tag-free, with HSA

Recombinant human IL-4 is a high-quality and biologically active cytokine, validated using proprietary IL-4/IL-13 reporter cells. This member of the IL-2/γc superfamily is produced in E. coli and thoroughly purified to remove endotoxins.

Recombinant human IL-4 can be used together with HEK-Blue™ IL-4/IL-13 cells for the screening of inhibitory molecules, such as Dupilumab, a therapeutic monoclonal antibody targeting the IL-4Rα subunit of the IL-4 receptor (see figures).

IL-4 signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ IL-4/IL-13 cells
• Endotoxin < 0.1 EU/µg
• 0.2 µm sterile-filtered

Applications

• Standard for IL-4 detection and quantification assays
• Screening and release assays for antibodies blocking IL-4 signaling
• Screening and release assays for engineered IL-4

Interleukin 4 (IL-4) shares a common receptor subunit, IL-4Rα, with IL-13. These two cytokines play an important role in regulating inflammation and immune responses. They contribute to Th2-mediated diseases, including asthma and atopic dermatitis.

 More details

InvivoGen’s products are for research use only, and not for clinical or veterinary use.

Figures

SDS PAGE of the recombinant human (h)IL-4 cytokine. 1.5 μg of hIL-4 was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. A band was detected at ~15 kDa.

Dose-response in HEK-Blue™ IL-4/IL-13 cells to recombinant IL-4 cytokine. Cells were stimulated with increasing concentrations of recombinant human IL-4. After overnight incubation, the NF-κB-induced SEAP activity was determined using QUANTI-Blue™, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Dose-dependent inhibition of HEK-Blue™ IL-4/IL-13 cells response using Dupilumab biosimilar.  HEK-Blue™ IL-4/IL13 cells were incubated with increasing concentrations of Anti-hIL-4R-hIgG4 (S228P) (0.1 ng/ml - 10 µg/ml) for 1 h before the addition of either recombinant human IL-4 (1 ng/ml) or recombinant human IL-13 (10 ng/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage of activity (%) (mean ± SEM).

 

Specifications

Source: E. coli

Species: Human

Alternative names: B-cell stimulatory factor 1, Lymphocyte stimulatory factor 1

Carrier: HSA (human serum albumin)

Tag: Tag-free

Accession number: P05112

Molecular weight: ~ 15 kDa (SDS-PAGE)

Solubility: 100 μg/ml in water

Formulation: Phosphate buffer saline (pH 7.4), 5% saccharose, 2% HSA

Sterility: 0.2 µm filtration

Form: Lyophilized

Reconstitution buffer: Endotoxin-free water (provided)

Purity: ≥98% (SDS-PAGE and HPLC)

Endotoxin level: ≤ 0.1 EU/μg (measurement by kinetic chromogenic LAL assay)

Tested applications: Cellular assays

Quality control: Each lot is functionally tested and validated

Contents

• 10 μg of recombinant human IL-4 provided as a white lyophilized powder
• 1.5 ml endotoxin-free water

 Recombinant human IL-4 is shipped at room temperature.

Upon receipt, the product should be stored at -20°C.

 Avoid repeated freeze-thaw cycles.

Details

IL-4 background

IL-4, also known as B-cell stimulatory factor 1 or Lymphocyte stimulatory factor 1, is a cytokine that belongs to the IL-2/γc superfamily. IL-4 is produced as a secreted monomeric molecule by T cells and innate cells, including mast cells, basophils, or eosinophils [1].

IL-4 binds two types of heterodimeric receptors and exerts pleiotropic actions on multiple cell lineages. Type I receptors are composed of the IL-4Rα and the common γ chains. Type II receptors are composed of the IL-4Rα and IL-13Rα1 chains. The binding of IL-4 to its receptors triggers a signaling cascade leading to the activation of STAT6. Subsequent gene expression drives the differentiation of Th2 and Th9 helper subsets while suppressing the development of Th1 and Th17 cells.in B cells, IL-4 promotes proliferation and immunoglobulin class switching [1, 2]. In non-hematopoietic cells, the functions of IL-4 span form mucus hypersecretion in airway epithelial cells, extracellular matrix protein deposition by fibroblasts, and growth of endothelial cells [1].

Relevance for therapeutics development

IL-4 is a key cytokine for controlling infections by extracellular parasites. However, along with IL-13, which shares the IL-4Rα/IL-13Rα1 receptor, it also contributes to harmful allergic responses [1]. As the prevalence of chronic allergic diseases such as asthma and atopic dermatitis is increasing worldwide, there has been a keen interest in the therapeutic blocking of IL-4 and IL-13 signaling.

Dupilumab is a fully human monoclonal antibody (mAb) that targets the IL-4Rα subunit [3]. It acts like a receptor antagonist and inhibits signaling of both IL-4 and IL-13 [3]. Dupilimab was FDA-approved in 2017 for treating asthma, atopic dermatitis, and chronic sinusitis [3, 4]. Due to its favorable safety profile and established clinical use, Dupilumab holds significant promise for amending treatment options for many dermatologic conditions [5].

The development of engineered IL-4 cytokines has also been explored. For example, IL-4 "superkines" featuring higher affinity for type I or type II receptors have been generated to target cells expressing lower second chain (γc or IL-13Rα1) numbers. To go further, engineered super-IL-4 fused to Pseudomonas toxin has been tested for targeted toxin delivery in tumor cells expressing IL-4 receptors [6].

References:

1. Keegan, A.D., et al., 2021. Recent advances in understanding the role of IL-4 signaling. Fac Rev. 10: 71.
2. Adkis, M., et al., 2016. Interleukins (from IL-1 to IL-38), interferons, transforming growth factor β, and TNF-α: Receptors, functions, and roles in diseases. J Allergy Clin Immunol. 138(4):984-1010.
3. Harb, H. & Chatila, T.A., 2020. Mechanisms of Dupilumab. Clin Exp Allergy. 50(1):5-14. 
4. U.S. Food and Drug Administration (FDA) (Report). 2018. New Drug Therapy Approvals 2018.
5. Olbrich, H., et al., 2023. Dupilumab in Inflammatory Skin Diseases: A Systematic Review. Biomolecules.13(4):634.
6. Leonard, W.J & Lin, J.X, 2023. Strategies to therapeutically modulate cytokine action. Nat Rev Drug Discov. 22(10):827-854.