IL-2 & IL-15 Reporter HEK 293 Cells (CD25, CD122, & CD132)

STAT5-SEAP reporter cells

ABOUT

Human/mouse IL-2 and human IL-15 responsive STAT5-SEAP reporter assay

HEK-Blue™ IL-2/IL-15 cells* are designed to monitor human IL-2 and IL-15-induced STAT5 stimulation or inhibition through SEAP detection. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ IL-2/IL-15 cells strongly respond to recombinant human IL-2 and human IL-15, as well as mouse IL-2. The reliable and consistent performance of HEK-Blue™ IL-2/IL-15 cells makes them suitable for release assays of therapeutic molecules that inhibit cytokine signaling, such as Ordesekimab, a monoclonal antibody targeting IL-15 (see figures).

Key features

  • Readily assessable STAT5-SEAP reporter activity
  • Convenient readout using  QUANTI-Blue™ Solution
  • High sensitivity to human (h) IL-2 and hIL-15 activity
  • Stability guaranteed for 20 passages

Applications

  • Therapeutic development
  • Drug screening
  • Release assay

 

IL-2 and IL-15 are closely related cytokines that belong to the IL-2 family and display important functions in the immune system [1]. They share the heterodimeric CD122 (IL-2Rβ)/CD132 (IL-2Rγ) receptor to deliver their signals within target cells. Their specificity of action is conferred by their α receptor chains, CD25 (IL-2Rα) and CD215 (IL-15Rα) [1].

more detailsMore details

 

InvivoGen also offers HEK-Blue™ CD122/132 cells, expressing the CD122 (IL-2Rβ) and CD132 (IL-2Rγ) subunits, showing a different detection range for mouse IL-2 and IL-15 (see table and figures).

* Note: This cell line has been renamed. It was formerly known as "HEK-Blue™ IL-2 Cells".

Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

Free cytokine
Signaling pathway in HEK-Blue™-IL-2/IL-15 cells
Signaling pathway in HEK-Blue™-IL-2/IL-15 cells
IL-2 and IL-15 detection potency
IL-2 and IL-15 detection potency

SPECIFICATIONS

Specifications

Target

IL-2, IL-15

Target species

Human, Mouse

Tested applications

Detection and quantification of IL-2 or IL-15 activity

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human embryonic kidney cells
Reporter gene
SEAP
Detection method
Colorimetric
Detection range

0.03 - 100 ng/ml (hIL-2 and mIL-2)
0.01 - 100 ng/ml (hIL-15)

Antibiotic resistance
Blasticidin
Hygromycin
Puromycin
Zeocin®
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    HEK-Blue™ IL-2/IL-15 Cells
  • Cat code: 
    hkb-il2-2
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml Puromycin (10 mg/ml)
  • 2 x 1 ml HEK-Blue™ CLR Selection (250X concentrate); a solution containing several selection antibiotics.
  • 1 ml Normocin® (50 mg/ml)
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent).

Shipping & Storage

  • Shipping method:  Dry ice

    • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

Cell line description

HEK-Blue™ IL-2/IL-15 cells were generated by stable transfection of the human embryonic kidney HEK293 cell line with the genes encoding for the human CD25 (IL-2Rα), CD122 (IL-2Rβ), and CD132 (IL-2Rγ) subunits. They also feature a stable expression of the human JAK3 and STAT5 to obtain a fully functional IL-2/IL-15 signaling pathway, as well as a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. he binding of IL-2 or IL-15 to their receptors triggers a signaling cascade leading to the activation of STAT5 and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.

HEK-Blue™ IL-2/IL-15 cells respond to human IL-2 and IL-15 and thus can be used for the screening of anti-hIL-2 and anti-hIL-15 antibodies. The addition of CD25 in HEK-Blue™ IL-2/IL-15 cells does not increase their sensitivity to human IL-2; however, it does increase their sensitivity to mouse IL-2& (see figures). HEK-Blue™ IL-2/IL-15 cells can be used to validate the functionality, toxicity, and variable dosage effects of human or mouse IL-2. However, this cell line does not respond to mouse IL-15.

 

IL-2 and IL-15 background

IL-2 and IL-15 are two closely related cytokines that belong to the IL-2 family and display important functions in the immune system [1]. They share the heterodimeric CD122 (IL-2Rβ)/CD132 (IL-2Rγ) receptor to deliver their signals within target cells. Their specificity of action is conferred by their α receptor chains, CD25 (IL-2Rα) and CD215 (IL-15Rα). The binding of IL-2 and IL-15 to their heterodimeric or heterotrimeric receptors activates a signaling cascade through tyrosine kinases of the Janus family (JAK1 and JAK3) and signal transducer and transcription activator 5 (STAT5) [1]. This results in the subsequent induction of proliferation and anti-apoptotic transcriptional program and can, if uncontrolled, potentially lead to malignant transformation [2]. IL-15 can sustain chronic inflammation, autoimmunity, and uncontrolled T cell proliferation, leading to T cell lymphoma development [1].

 

1. Sindaco P, et al., 2023. The role of interleukin-15 in the development and treatment of hematological malignancies. Front Immunol.14:1141208.

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