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Measuring NAADP-Evoked Ca2+ Release in Permeabilized T Cells
Kai J Winterberg1, Andreas Bauche1, Ralf Fliegert1
1The Calcium Signalling Group, Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Abstract:
Nicotinic acid adenine dinucleotide phosphate (NAADP) is a highly potent Ca2+ releasing second messenger with a distinct role during early T cell activation. While studies have proposed NAADP signaling in T cells as a target for treating autoimmune diseases, there are currently no pharmacologic inhibitors available for clinical trial. This is in part due to the fact, that investigating NAADP-evoked Ca2+ signals in T cells currently requires laborious techniques such as microinjections. We have combined recent advances on NAADP signaling to develop a protocol for measuring NAADP-evoked Ca2+ release from permeabilized T cells, that can be used to further characterize this pathway in T cells and validate new pharmacologic agents.
Insights
Nicotinic acid adenine dinucleotide phosphate (NAADP) is crucial for T cell activation. Researchers developed a new method to study NAADP signaling, aiding the search for autoimmune disease treatments.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent calcium (Ca2+) releasing second messenger.
- NAADP signaling plays a critical role in early T cell activation.
- Targeting NAADP signaling in T cells is a potential strategy for autoimmune disease treatment, but lacks pharmacologic inhibitors.
Purpose of the Study:
- To develop a simplified protocol for measuring NAADP-evoked Ca2+ release in T cells.
- To facilitate the characterization of NAADP signaling pathways in T cells.
- To enable the validation of novel pharmacologic agents targeting NAADP signaling.
Main Methods:
- Utilized recent advancements in NAADP signaling research.
- Developed a protocol for measuring Ca2+ release from permeabilized T cells.
- Focused on quantifying NAADP-evoked Ca2+ signals.
Main Results:
- Successfully established a protocol for measuring NAADP-evoked Ca2+ release.
- The new method simplifies the investigation of NAADP signaling in T cells.
- The protocol is suitable for validating potential pharmacologic agents.
Conclusions:
- The developed protocol offers a more accessible method to study NAADP signaling in T cells.
- This advancement will aid in understanding T cell activation and autoimmune diseases.
- Facilitates the development and testing of new drugs targeting NAADP pathways.
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