Caffeine modulates TNF-α production by cord blood monocytes: The role of adenosine receptors

Raul Chavez-Valdez, Marsha Wills-Karp, Rajni Ahlawat, Elizabeth A. Cristofalo, Amy Nathan, Estelle B Gauda

Research output: Contribution to journalArticle

Abstract

Caffeine, a nonspecific adenosine receptor (AR) antagonist is widely used to treat apnea of prematurity. Because adenosine modulates multiple biologic processes including inflammation, we hypothesized that AR blockade by caffeine would increase cytokine release from neonatal monocytes. Using cord blood monocytes (CBM), we investigated 1) the changes in AR mRNA profile by real time quantitative reverse-transcription polymerase-chain-reaction (qRT-PCR) and protein expression (western blot) after in vitro culture, caffeine or lipopolysaccharide (LPS) exposure, and 2) the modulation of cytokine release and cyclic adenosine monophosphate (cAMP) production by enzyme-linked immunosorbent assay (ELISA) induced by caffeine and specific AR antagonists: DPCPX(A 1R), ZM241385(A2aR), MRS1754(A2bR), and MRS1220(A3R). After 48 h in culture, A2aR and A 2bR gene expression increased 1.9 (p = 0.04) and 2.5-fold (p = 0.003), respectively. A1R protein expression directly correlated with increasing LPS concentrations (p = 0.01), with minimal expression preexposure. Only caffeine (50 μMu) and DPCPX (10 nM) decreased tumor necrosis factor-alpha (TNF-α) release from LPS activated-CBM by 20 and 25% (p = 0.01) and TNF-α gene expression by 30 and 50%, respectively, in conjunction with a ≥2-fold increase in cAMP (p <0.05). AR blockade did not modulate other measured cytokines. The induction of A1R after LPS exposure suggests an important role of this receptor in the control of inflammation in neonates. Our findings also suggest that caffeine, via A 1R blockade, increases cAMP production and inhibits pretranscriptional TNF-a production by CBM.

Original languageEnglish (US)
Pages (from-to)203-208
Number of pages6
JournalPediatric Research
Volume65
Issue number2
DOIs
StatePublished - Feb 2009

Fingerprint

Purinergic P1 Receptors
Caffeine
Fetal Blood
Monocytes
Tumor Necrosis Factor-alpha
Lipopolysaccharides
Cyclic AMP
Purinergic P1 Receptor Antagonists
Cytokines
Inflammation
Gene Expression
Apnea
Adenosine
Reverse Transcription
Proteins
Western Blotting
Enzyme-Linked Immunosorbent Assay
Polymerase Chain Reaction
Messenger RNA

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Caffeine modulates TNF-α production by cord blood monocytes : The role of adenosine receptors. / Chavez-Valdez, Raul; Wills-Karp, Marsha; Ahlawat, Rajni; Cristofalo, Elizabeth A.; Nathan, Amy; Gauda, Estelle B.

In: Pediatric Research, Vol. 65, No. 2, 02.2009, p. 203-208.

Research output: Contribution to journalArticle

@article{5beea4da5785486798a18e33cf98bbef,
title = "Caffeine modulates TNF-α production by cord blood monocytes: The role of adenosine receptors",
abstract = "Caffeine, a nonspecific adenosine receptor (AR) antagonist is widely used to treat apnea of prematurity. Because adenosine modulates multiple biologic processes including inflammation, we hypothesized that AR blockade by caffeine would increase cytokine release from neonatal monocytes. Using cord blood monocytes (CBM), we investigated 1) the changes in AR mRNA profile by real time quantitative reverse-transcription polymerase-chain-reaction (qRT-PCR) and protein expression (western blot) after in vitro culture, caffeine or lipopolysaccharide (LPS) exposure, and 2) the modulation of cytokine release and cyclic adenosine monophosphate (cAMP) production by enzyme-linked immunosorbent assay (ELISA) induced by caffeine and specific AR antagonists: DPCPX(A 1R), ZM241385(A2aR), MRS1754(A2bR), and MRS1220(A3R). After 48 h in culture, A2aR and A 2bR gene expression increased 1.9 (p = 0.04) and 2.5-fold (p = 0.003), respectively. A1R protein expression directly correlated with increasing LPS concentrations (p = 0.01), with minimal expression preexposure. Only caffeine (50 μMu) and DPCPX (10 nM) decreased tumor necrosis factor-alpha (TNF-α) release from LPS activated-CBM by 20 and 25{\%} (p = 0.01) and TNF-α gene expression by 30 and 50{\%}, respectively, in conjunction with a ≥2-fold increase in cAMP (p <0.05). AR blockade did not modulate other measured cytokines. The induction of A1R after LPS exposure suggests an important role of this receptor in the control of inflammation in neonates. Our findings also suggest that caffeine, via A 1R blockade, increases cAMP production and inhibits pretranscriptional TNF-a production by CBM.",
author = "Raul Chavez-Valdez and Marsha Wills-Karp and Rajni Ahlawat and Cristofalo, {Elizabeth A.} and Amy Nathan and Gauda, {Estelle B}",
year = "2009",
month = "2",
doi = "10.1203/PDR.0b013e31818d66b1",
language = "English (US)",
volume = "65",
pages = "203--208",
journal = "Pediatric Research",
issn = "0031-3998",
publisher = "Lippincott Williams and Wilkins",
number = "2",

}

TY - JOUR

T1 - Caffeine modulates TNF-α production by cord blood monocytes

T2 - The role of adenosine receptors

AU - Chavez-Valdez, Raul

AU - Wills-Karp, Marsha

AU - Ahlawat, Rajni

AU - Cristofalo, Elizabeth A.

AU - Nathan, Amy

AU - Gauda, Estelle B

PY - 2009/2

Y1 - 2009/2

N2 - Caffeine, a nonspecific adenosine receptor (AR) antagonist is widely used to treat apnea of prematurity. Because adenosine modulates multiple biologic processes including inflammation, we hypothesized that AR blockade by caffeine would increase cytokine release from neonatal monocytes. Using cord blood monocytes (CBM), we investigated 1) the changes in AR mRNA profile by real time quantitative reverse-transcription polymerase-chain-reaction (qRT-PCR) and protein expression (western blot) after in vitro culture, caffeine or lipopolysaccharide (LPS) exposure, and 2) the modulation of cytokine release and cyclic adenosine monophosphate (cAMP) production by enzyme-linked immunosorbent assay (ELISA) induced by caffeine and specific AR antagonists: DPCPX(A 1R), ZM241385(A2aR), MRS1754(A2bR), and MRS1220(A3R). After 48 h in culture, A2aR and A 2bR gene expression increased 1.9 (p = 0.04) and 2.5-fold (p = 0.003), respectively. A1R protein expression directly correlated with increasing LPS concentrations (p = 0.01), with minimal expression preexposure. Only caffeine (50 μMu) and DPCPX (10 nM) decreased tumor necrosis factor-alpha (TNF-α) release from LPS activated-CBM by 20 and 25% (p = 0.01) and TNF-α gene expression by 30 and 50%, respectively, in conjunction with a ≥2-fold increase in cAMP (p <0.05). AR blockade did not modulate other measured cytokines. The induction of A1R after LPS exposure suggests an important role of this receptor in the control of inflammation in neonates. Our findings also suggest that caffeine, via A 1R blockade, increases cAMP production and inhibits pretranscriptional TNF-a production by CBM.

AB - Caffeine, a nonspecific adenosine receptor (AR) antagonist is widely used to treat apnea of prematurity. Because adenosine modulates multiple biologic processes including inflammation, we hypothesized that AR blockade by caffeine would increase cytokine release from neonatal monocytes. Using cord blood monocytes (CBM), we investigated 1) the changes in AR mRNA profile by real time quantitative reverse-transcription polymerase-chain-reaction (qRT-PCR) and protein expression (western blot) after in vitro culture, caffeine or lipopolysaccharide (LPS) exposure, and 2) the modulation of cytokine release and cyclic adenosine monophosphate (cAMP) production by enzyme-linked immunosorbent assay (ELISA) induced by caffeine and specific AR antagonists: DPCPX(A 1R), ZM241385(A2aR), MRS1754(A2bR), and MRS1220(A3R). After 48 h in culture, A2aR and A 2bR gene expression increased 1.9 (p = 0.04) and 2.5-fold (p = 0.003), respectively. A1R protein expression directly correlated with increasing LPS concentrations (p = 0.01), with minimal expression preexposure. Only caffeine (50 μMu) and DPCPX (10 nM) decreased tumor necrosis factor-alpha (TNF-α) release from LPS activated-CBM by 20 and 25% (p = 0.01) and TNF-α gene expression by 30 and 50%, respectively, in conjunction with a ≥2-fold increase in cAMP (p <0.05). AR blockade did not modulate other measured cytokines. The induction of A1R after LPS exposure suggests an important role of this receptor in the control of inflammation in neonates. Our findings also suggest that caffeine, via A 1R blockade, increases cAMP production and inhibits pretranscriptional TNF-a production by CBM.

UR - http://www.scopus.com/inward/record.url?scp=60449103179&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=60449103179&partnerID=8YFLogxK

U2 - 10.1203/PDR.0b013e31818d66b1

DO - 10.1203/PDR.0b013e31818d66b1

M3 - Article

C2 - 19047957

AN - SCOPUS:60449103179

VL - 65

SP - 203

EP - 208

JO - Pediatric Research

JF - Pediatric Research

SN - 0031-3998

IS - 2

ER -