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Calcium pyrophosphate dihydrate crystals
NLRP3 Inflammasome Inducer - Calcium Pyrophosphate Dihydrate crystals
Calcium pyrophosphate dihydrate (CPPD) is the aetiological agent of the joint acute inflammatory disease pseudogout. Notably, research has demonstrated that CPPD crystals induce the release of interleukin (IL)-1β in response to the activation of the NLRP3 (also known as NALP3 or cryopyrin) inflammasome . The NLRP3 inflammasome is an intracellular multi-protein complex that plays a central role in innate immunity [2, 3].
Mode of action:
Similar to uric acid, the causative agent of gout, CPPD crystals can act as endogenous danger signals that stimulate the innate immune system to produce inflammatory cytokines such as IL-1β . CPPD crystals were shown to activate caspase-1, which is required for IL-1β maturation, through the NRLP3 (NALP3) inflammasome . The NLRP3 inflammasome is a caspase-1-activating complex comprising the NLR protein NLRP3 and the adaptor ASC . Engagement of the NRLP3 inflammasome is supported by the finding that macrophages from mice deficient in various components of the inflammasome do not respond to the injection of CPPD crystals .
Key features of CPPD crystals:
- Potent inducer of the NLRP3 inflammasome
- Each lot is functionally tested
Read our review on the NLRP3 inflammasome.
Download our Practical guide on Inflammasomes.
1. Shi Y. et al., 2003. Molecular identification of a danger signal that alerts the immune system to dying cells. Nature 425(6957):516-21
2. Martinon F. et al., 2006. Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature. 440(7081):237-41.
3. Martinon F. & Tschopp J., 2004. Inflammatory caspases: linking an intracellular innate immune system to autoinflammatory diseases. Cell. 117(5):561-74.
IL-1β production in THP1-HMGB1-Lucia™ cells. Cells were primed with 1 μg/ml LPS‑EK for 3h and then incubated with inflammasome inducers; 8 μM Nigericin, 0.5 μg/ml complexed Poly(dA:dT), 200 μg/ml MSU Crystals, 100 μg/ml CPPD Crystals, or 1 mg/ml Zymosan. After 24h, supernatants were incubated with HEK-Blue™ IL-1β cells overnight and SEAP activity was assessed using QUANTI-Blue™ Solution. The optical density (OD) was read at 630 nm. Data are shown as a fold increase (mean ± SEM) over non-induced cells.
Working concentration: 1 - 50 µg/ml
CAS number: 7790-76-3
Molecular weight: 254.1 g/mol
Structure: Triclinic crystals characterized by X-ray diffraction
Solubility: InsolubleBack to the top
- 5 mg calcium pyrophosphate dihydrate (CPPD) crystals
CPPD crystals are shipped at room temperature.
Upon receipt, store at 4°C.Back to the top
THP-1/HEK-Blue™ IL-1β Assay
CPPD crystals can be used to induce the NLRP3 inflammasome in cellular assays, such as InvivoGen’s THP1/HEK-Blue™ IL-1β assay. This assay uses the secretion of IL-1β by THP1-Null2 cells as an indicator of NLRP3 inflammasome induction. The production of IL-1β by these cells is measured using HEK-Blue™ IL-1β cells.
1- Production of IL-1β by THP-1 cells: Typically, THP1-Null2 cells are primed with lipopolysaccharide (LPS). This treatment induces the production of NLRP3 and pro-IL-1β, the immature form of IL-1β. Subsequent stimulation with inflammasome inducers, such as CPPD crystals, leads to NRLP3 and caspase-1 activation resulting in IL-1β maturation and secretion.
2- Detection of IL-1β by HEK-Blue™ IL-1β cells: THP1-Null2 cell supernatants containing IL-1β can be added to HEK-Blue™ IL-1β cells. This activates NF-κB and leads to the subsequent production of secreted embryonic alkaline phosphatase (SEAP). The presence of SEAP in HEK-Blue™ IL-1β supernatants is assessed using QUANTI-Blue™ Solution, a SEAP detection medium.Back to the top