The P2X(7) loss-of-function Glu496Ala polymorphism affects ex vivo cytokine release and protects against the cytotoxic effects of high ATP-levels

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Abstract

BACKGROUND: The P2X(7) receptor plays an important role in cytokine release during the inflammatory response in vivo. Polymorphisms within the P2X(7) receptor gene that lead to loss of receptor function may contribute to impaired cytokine release by immune cells. Therefore, we investigated whether a known loss-of-function polymorphism (Glu496Ala) in the P2X(7) receptor gene leads to alterations in cytokine release in response to ATP.

RESULTS: An ex vivo whole blood model was used to induce an inflammatory reaction with the pro-inflammatory stimuli LPS and PHA (phytohemagglutinin). Blood from n=9 subjects with the Glu496Ala P2X7 SNP (P2X7MUT) and n=7 'wild-type' subjects (no P2X7 SNP; P2X7WT) was used.Addition of ATP (0.9-3 mM) to LPS/PHA-stimulated whole blood induced an increase in IL-1β release in P2X7MUT subjects, whereas decreased release was observed in P2X7WT subjects. Decreased levels of IL-6 and TNF-α in response to ATP were shown in both P2X7MUT and P2X7WT subjects, which was less pronounced in P2X7MUT subjects. ATP at 3 mM also significantly decreased levels of lactate dehydrogenase (LDH) in P2X7MUT subjects compared to P2X7WT subjects.

CONCLUSIONS: The presence of the non-synonymous Glu496Ala loss-of-function polymorphism within the P2X(7) receptor gene is likely to be of importance in the release of cytokines during inflammation. Furthermore, this study suggests that carriers of the Glu496Ala loss-of-function polymorphism are protected against the cytotoxic effects of high ATP-levels.

Original languageEnglish
Pages (from-to)64
JournalBMC IMMUNOLOGY
Volume13
DOIs
Publication statusPublished - 4 Dec 2012

Keywords

  • Adenosine Triphosphate
  • Aged
  • Amino Acid Substitution
  • Cell Death
  • Cytokines
  • Cytoprotection
  • Female
  • Humans
  • L-Lactate Dehydrogenase
  • Lipopolysaccharides
  • Male
  • Middle Aged
  • Polymorphism, Single Nucleotide
  • Receptors, Purinergic P2X7
  • Tetradecanoylphorbol Acetate

Cite this

@article{2d1cab0fc3344c8686682ba992b1fae4,
title = "The P2X(7) loss-of-function Glu496Ala polymorphism affects ex vivo cytokine release and protects against the cytotoxic effects of high ATP-levels",
abstract = "BACKGROUND: The P2X(7) receptor plays an important role in cytokine release during the inflammatory response in vivo. Polymorphisms within the P2X(7) receptor gene that lead to loss of receptor function may contribute to impaired cytokine release by immune cells. Therefore, we investigated whether a known loss-of-function polymorphism (Glu496Ala) in the P2X(7) receptor gene leads to alterations in cytokine release in response to ATP.RESULTS: An ex vivo whole blood model was used to induce an inflammatory reaction with the pro-inflammatory stimuli LPS and PHA (phytohemagglutinin). Blood from n=9 subjects with the Glu496Ala P2X7 SNP (P2X7MUT) and n=7 'wild-type' subjects (no P2X7 SNP; P2X7WT) was used.Addition of ATP (0.9-3 mM) to LPS/PHA-stimulated whole blood induced an increase in IL-1β release in P2X7MUT subjects, whereas decreased release was observed in P2X7WT subjects. Decreased levels of IL-6 and TNF-α in response to ATP were shown in both P2X7MUT and P2X7WT subjects, which was less pronounced in P2X7MUT subjects. ATP at 3 mM also significantly decreased levels of lactate dehydrogenase (LDH) in P2X7MUT subjects compared to P2X7WT subjects.CONCLUSIONS: The presence of the non-synonymous Glu496Ala loss-of-function polymorphism within the P2X(7) receptor gene is likely to be of importance in the release of cytokines during inflammation. Furthermore, this study suggests that carriers of the Glu496Ala loss-of-function polymorphism are protected against the cytotoxic effects of high ATP-levels.",
keywords = "Adenosine Triphosphate, Aged, Amino Acid Substitution, Cell Death, Cytokines, Cytoprotection, Female, Humans, L-Lactate Dehydrogenase, Lipopolysaccharides, Male, Middle Aged, Polymorphism, Single Nucleotide, Receptors, Purinergic P2X7, Tetradecanoylphorbol Acetate",
author = "Anke Wesselius and Bours, {Martijn J L} and Arts, {Ilja C W} and Theunisz, {Esther H E} and Piet Geusens and Dagnelie, {Pieter C}",
year = "2012",
month = "12",
day = "4",
doi = "10.1186/1471-2172-13-64",
language = "English",
volume = "13",
pages = "64",
journal = "BMC IMMUNOLOGY",
issn = "1471-2172",
publisher = "BioMed Central",

}

TY - JOUR

T1 - The P2X(7) loss-of-function Glu496Ala polymorphism affects ex vivo cytokine release and protects against the cytotoxic effects of high ATP-levels

AU - Wesselius, Anke

AU - Bours, Martijn J L

AU - Arts, Ilja C W

AU - Theunisz, Esther H E

AU - Geusens, Piet

AU - Dagnelie, Pieter C

PY - 2012/12/4

Y1 - 2012/12/4

N2 - BACKGROUND: The P2X(7) receptor plays an important role in cytokine release during the inflammatory response in vivo. Polymorphisms within the P2X(7) receptor gene that lead to loss of receptor function may contribute to impaired cytokine release by immune cells. Therefore, we investigated whether a known loss-of-function polymorphism (Glu496Ala) in the P2X(7) receptor gene leads to alterations in cytokine release in response to ATP.RESULTS: An ex vivo whole blood model was used to induce an inflammatory reaction with the pro-inflammatory stimuli LPS and PHA (phytohemagglutinin). Blood from n=9 subjects with the Glu496Ala P2X7 SNP (P2X7MUT) and n=7 'wild-type' subjects (no P2X7 SNP; P2X7WT) was used.Addition of ATP (0.9-3 mM) to LPS/PHA-stimulated whole blood induced an increase in IL-1β release in P2X7MUT subjects, whereas decreased release was observed in P2X7WT subjects. Decreased levels of IL-6 and TNF-α in response to ATP were shown in both P2X7MUT and P2X7WT subjects, which was less pronounced in P2X7MUT subjects. ATP at 3 mM also significantly decreased levels of lactate dehydrogenase (LDH) in P2X7MUT subjects compared to P2X7WT subjects.CONCLUSIONS: The presence of the non-synonymous Glu496Ala loss-of-function polymorphism within the P2X(7) receptor gene is likely to be of importance in the release of cytokines during inflammation. Furthermore, this study suggests that carriers of the Glu496Ala loss-of-function polymorphism are protected against the cytotoxic effects of high ATP-levels.

AB - BACKGROUND: The P2X(7) receptor plays an important role in cytokine release during the inflammatory response in vivo. Polymorphisms within the P2X(7) receptor gene that lead to loss of receptor function may contribute to impaired cytokine release by immune cells. Therefore, we investigated whether a known loss-of-function polymorphism (Glu496Ala) in the P2X(7) receptor gene leads to alterations in cytokine release in response to ATP.RESULTS: An ex vivo whole blood model was used to induce an inflammatory reaction with the pro-inflammatory stimuli LPS and PHA (phytohemagglutinin). Blood from n=9 subjects with the Glu496Ala P2X7 SNP (P2X7MUT) and n=7 'wild-type' subjects (no P2X7 SNP; P2X7WT) was used.Addition of ATP (0.9-3 mM) to LPS/PHA-stimulated whole blood induced an increase in IL-1β release in P2X7MUT subjects, whereas decreased release was observed in P2X7WT subjects. Decreased levels of IL-6 and TNF-α in response to ATP were shown in both P2X7MUT and P2X7WT subjects, which was less pronounced in P2X7MUT subjects. ATP at 3 mM also significantly decreased levels of lactate dehydrogenase (LDH) in P2X7MUT subjects compared to P2X7WT subjects.CONCLUSIONS: The presence of the non-synonymous Glu496Ala loss-of-function polymorphism within the P2X(7) receptor gene is likely to be of importance in the release of cytokines during inflammation. Furthermore, this study suggests that carriers of the Glu496Ala loss-of-function polymorphism are protected against the cytotoxic effects of high ATP-levels.

KW - Adenosine Triphosphate

KW - Aged

KW - Amino Acid Substitution

KW - Cell Death

KW - Cytokines

KW - Cytoprotection

KW - Female

KW - Humans

KW - L-Lactate Dehydrogenase

KW - Lipopolysaccharides

KW - Male

KW - Middle Aged

KW - Polymorphism, Single Nucleotide

KW - Receptors, Purinergic P2X7

KW - Tetradecanoylphorbol Acetate

U2 - 10.1186/1471-2172-13-64

DO - 10.1186/1471-2172-13-64

M3 - Article

C2 - 23210974

VL - 13

SP - 64

JO - BMC IMMUNOLOGY

JF - BMC IMMUNOLOGY

SN - 1471-2172

ER -