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Application: Screen and characterize modulators of IL-15-triggered signaling.
Background: IL-15 is a proinflammatory cytokine mainly produced in dendric cells (DC) and monocytes. It remains associated with IL-15Rα (IL-15 receptor alpha subunit) on the surface of DC and monocytes and binds to the CD122 (IL-12/IL-15 receptor β)/CD132 (γ subunit) heterodimeric receptor present at the surface of various target immune cells, including CD8+ memory T cells, resulting in their proliferation and differentiation. IL-15 is a cytokine of interest in cancer therapy and in immunology. It is produced in cells as a stable heterodimer composed of the IL-15 polypeptide single chain and IL-15Rα, which functions not as receptor but a binding partner that allows transport to the cell membrane. Although there is no indication that unassociated IL-15 exists in vivo, soluble recombinant IL-15 and its analogs have been examined for their therapeutic potential.
Description: The Human IL-15 Responsive Luciferase Reporter Ba/F3 Cell Line is a murine Ba/F3 cell line engineered to express both human CD122 (hIL2RB, NM_000878.5) and CD132 (hIL2RG, NM_000206.3) separated by a self-cleaving P2A peptide. The construct was delivered by lentiviral transduction of STAT5 Luciferase Reporter Ba/F3 cells (#79772), which express a firefly luciferase reporter driven by STAT5 (Signal transducer and activator of transcription 5) response elements located upstream of the minimal TATA promoter. After activation by IL-15, the endogenous transcription factor STAT5 binds to the response elements, inducing transcription of the luciferase reporter. This cell line has been validated to respond to human IL-15 and IL-2. Functional validation demonstrates that IL-15-induced luciferase activity can be inhibited by anti-IL15 neutralizing antibody Ordesekimab. Figure 1: Illustration of the mechanism of Human IL-15 Luciferase Reporter Ba/F3 Cell Line. IL-2 (interleukin 2) and IL-15 (interleukin 15) both bind to and signal through a complex composed of IL-2/IL-15 receptor beta chain (CD122) and the common gamma-chain (gamma-C, CD132). Activation of the receptor activates downstream JAK (janus kinase) tyrosine kinases, which phosphorylate the transcription factor STAT5. STAT5 phosphorylation triggers the formation of a homodimer and translocation to the nucleus, where it can activate the transcription of the Firefly luciferase reporter driven by STAT5 response elements present in the promoter.
Genbank: hIL2RB (NM_000878.5), hIL2RG (NM_000206.3)
Host Cell Line: Ba/F3, mouse IL-3-dependent pro-B cell line, suspension.
Mycoplasma Testing: The cell line has been screened to confirm the absence of Mycoplasma species.
Storage Stability: Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage. Contact technical support at support@bpsbioscience.com if the cells are not frozen in dry ice upon arrival.
Supplied As: Each vial contains >1 x 106 cells in 1 ml of Cell Freezing Medium (BPS Bioscience #79796)
Target: IL-15
Uniprot: P14784 (hIL2RB), P31785 (hIL2RG)
Biosafety Level: BSL-1
References: Bahatt RS., et. al., 2021 Cancer Immunol Res. 9(2): 156-169. Lui G., et al., 2023 Cells 12(12):1611.
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