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A novel assay to detect nucleotide receptor P2X7 genetic polymorphisms influencing numerous innate immune functions

Department of Medicine, University of Wisconsin Medical School, Madison, Wisconsin, USA, ldenling{at}wisc.edu, Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison Wisconsin, USA

Kathleen Schell

Comprehensive Cancer Center, University of Wisconsin Medical School, Madison, Wisconsin, USA

Giuditta Angelini

Department of Anesthesiology, University of Wisconsin Medical School, Madison, Wisconsin, USA

Dawn Green

Department of Anesthesiology, University of Wisconsin Medical School, Madison, Wisconsin, USA

Arturo Guadarrama

Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison Wisconsin, USA

Usha Prabhu

Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison Wisconsin, USA

Douglas B. Coursin

Department of Medicine, University of Wisconsin Medical School, Madison, Wisconsin, USA, Department of Anesthesiology, University of Wisconsin Medical School, Madison, Wisconsin, USA

Kirk Hogan

Department of Anesthesiology, University of Wisconsin Medical School, Madison, Wisconsin, USA

Paul J. Bertics

Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison Wisconsin, USA, Comprehensive Cancer Center, University of Wisconsin Medical School, Madison, Wisconsin, USA

The importance of accessory signaling pathways amplifying endotoxin responses has recently been highlighted by genetic studies describing LPS-hyporesponsive individuals despite carrying the common allele for TLR4. The nucleotide receptor P2X₇ modulates the production of numerous LPS-stimulated inflammatory mediators. We have recently described the largest phenotypic screen known for genetic polymorphisms associated with the nucleotide receptor P2X₇, a global regulator of leukocyte function. This required the development of a novel monocyte pore assay with numerous advantages over previous methods and with the potential to facilitate rapid (< 3 h), multiplex analysis of clinical samples. This paper addresses aspects pertinent to the development of the monocyte pore assay, briefly summarizes our results suggestingthat P2X₇ alleles modulate LPSstimulated cytokine production, and discusses a model wherein P2X₇ may serve as an amplification loop of innate immunity.

Key Words: Nucleotide receptor P2X7 • genetic polymorphisms • innate immune functions • monocyte pore assay

Journal of Endotoxin Research, Vol. 10, No. 2, 137-142 (2004)
DOI: 10.1177/09680519040100020101


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