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Human IL-1 Reporter Cells

Signaling pathway in HEK-Blue™ IL-1β cells

HEK-Blue™ IL-1β cells were engineered from the human embryonic kidney HEK293 cell line to detect bioactive human interleukin-1 β (IL-1β) in various biological samples (cell culture supernatant and serum) by monitoring the activation of the NF-κB and AP-1 pathways. These cells are useful to monitor IL-1β secretion in inflammasome activation studies. Additionally, HEK-Blue™ IL-1β detect bioactive human IL-1α. HEK-Blue™ IL-1β cells can also be used for screening antibodies or small molecule inhibitors targeting the IL-1 pathway.

Both IL‑1α and IL-1β are secreted pro‑inflammatory cytokine that play a critical role in immune responses and inflammation [1].

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Cell line description

HEK-Blue™ IL-1β cells endogenously express the human (h) IL-1 receptor, which binds both IL-1α and IL-1β. They were transfected with a NF-κB/AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-1α or IL-1β to its receptor triggers a signaling cascade leading to NF-κB/AP-1 activation 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-1β cells respond to hIL-1α and hIL-1β (see figures). They also respond to a lesser extent to murine (m) IL-1α and mIL-1β (see figures). The specificity of the HEK-Blue™ IL-1β cells for the detection of IL-1α or IL-1β can be confirmed using a neutralizing antibody against human IL-1α or IL-1β, such as anti-hIL-1α-IgG and anti-hIL-1β-IgG. Of note, these cells do no respond to hTNF-α or mTNF-α (see figures).

Key features

Fully functional IL-1β signaling pathway
Readily assessable NF-κB/AP-1-inducible SEAP reporter activity
Strong response to human (h) IL-1α and hIL-1β
Low response to murine (m) IL-1α and mIL-1β
No response to hTNF-α and mTNF-α

Applications

Detection and quantification of hIL-1β activity, and to a lesser extent mIL-1β activity
Detection of hIL-1β in inflammasome studies
Screening of anti-IL-1α or -IL-1β antibodies and anti-IL-1 receptor antibodies
Screening of small molecule inhibitors of the IL-1 pathway

HEK-Blue™ IL-1β and HEK-Blue™ IL-1R cells

HEK-Blue™ IL-1β cells are more sensitive to human IL-1 isoforms than murine isoforms. We recommend using this cell line to test supernatant from human inflammasome cellular assays.
HEK-Blue™ IL-1R cells are stably transfected to additionally express the murine IL-1R, conferring a higher sensitivity to mIL-1α and mIL-1β, when compared to HEK-Blue™ IL-1β cells.

Download our Practical guide on Inflammasomes

Note: This cell line has been replaced with another clone which is KO for TLR3, TLR5, and TNFR1. Thus, it has a new cat. code: hkb-il1bv2.

Reference:

1. Dinarello C., 2018. Overview of the IL-1 family in innate inflammation and acquired immunity. Immunol Rev. 281(1): 8–27.

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