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HEK-Blue CD40L Cells - CD40L Reporter Cells
➥ Study CD40L-CD40 interactions
➥ Screen for molecules that interfere with CD40L-CD40 cross-talk
Background
CD40 ligand (CD40L) is a member of the tumor necrosis factor (TNF)
family of cell surface interaction molecules. It is mainly expressed in
CD4+-T cells and interacts with CD40 on antigen-presenting cells to
regulate both humoral and cellular immune responses. CD40L-CD40
interactions are thought to play an important role in the pathogenesis of
certain diseases, including AIDS, Alzheimer’s disease and rheumatoid
arthritis.
Description
HEK-Blue CD40L cells can serve to measure the bioactivity of CD40L
through the secretion of embryonic alkaline phosphatase (SEAP) upon
NF-kB activation. These cells were generated by stable transfection of
HEK293 cells with the human CD40 gene and an NF-kB-inducible SEAP
construct. The SEAP construct consists of the SEAP reporter gene under
the control of the IFN-b minimal promoter fused to five NF-kB binding
sites. Binding of CD40L to its receptor CD40 triggers a signaling cascade
leading to the activation of NF-kB and the subsequent production of
SEAP (figure 1). CD40L-CD40 interaction can be monitored by assessing
the levels of SEAP using a SEAP detection assay, such as QUANTI-Blue
(figure 2). HEK293 cells express endogenously the receptors for the
cytokines IL-1b and TNF-a which share a common signaling pathway with
CD40L. Consequently, HEK-Blue CD40L cells also respond to IL-1b and
TNF-a. IL-1b- and TNF-a-mediated SEAP production can be blocked
using neutralizing antibodies, such as anti-hIL-1b-IgA and anti-hTNF-a-IgA
antibodies respectively (see page 104).
• Detection range for CD40L: 5 ng - 1 μg/ml
HEK-Blue CD40L cells are resistant to the selective antibiotics blasticidin
and Zeocin .
Contents and Storage
HEK-Blue CD40L cells are grown in DMEM medium, 2mM L-glutamine,
10% FBS supplemented with 100 μg/ml Zeocin and 30 μg/ml blasticidin.
Each vial contains 5-7 x 10 6
cells and is supplied with 10 mg Zeocin ,
1 mg blasticidin, 50 mg Normocin and 1 pouch of QUANTI-Blue . Cells
are shipped on dry ice.
HEK-Blue CD40L cells are also available in a kit, see below: Figure 2: Stimulation of HEK-Blue CD40L cells by human CD40L was assessed by
measuring the levels of SEAP using QUANTI-Blue .
HEK-Blue CD40L Kit
• HEK-Blue CD40L Cells 5-7 x 10 6 cells
• Anti-hCD40L-IgA antibody 100 μg
• QUANTI-Blue 1 pouch (100 ml)
• Normocin 1 ml (50 mg/ml)
• Blasticidin 100 μl (10 mg/ml)
• Zeocin 100 μl (100 mg/ml)
Figure 1: CD40L-induced NF-kB signaling pathway.
PRODUCT QUANTITY CAT. CODE
HEK-Blue CD40L Cells 5-7 x 10 6 cells hkb-cd40
HEK-Blue CD40L Kit 1 kit hkb-cd40-kit
www.invivogen.com/cytokine-sensor-cells 115
CYTOKINE SIGNALING 4