Airway smooth muscle CXCR3 ligand production: Regulation by JAK-STAT1 and intracellular Ca<sup>2+</sup>

Tan, X; Alrashdan, YA; Alkhouri, H; Oliver, BGG; Armour, CL; Hughes, JM

Airway smooth muscle CXCR3 ligand production: Regulation by JAK-STAT1 and intracellular Ca<sup>2+</sup>

Tan, X Alrashdan, YA Alkhouri, H Oliver, BGG

Armour, CL Hughes, JM

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Publication Type:: Journal Article
Citation:: American Journal of Physiology - Lung Cellular and Molecular Physiology, 2013, 304 (11)
Issue Date:: 2013-06-11

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Field	Value	Language
dc.contributor.author	Tan, X	en_US
dc.contributor.author	Alrashdan, YA	en_US
dc.contributor.author	Alkhouri, H	en_US
dc.contributor.author	Oliver, BGG https://orcid.org/0000-0002-7122-9262	en_US
dc.contributor.author	Armour, CL	en_US
dc.contributor.author	Hughes, JM	en_US
dc.date.issued	2013-06-11	en_US
dc.identifier.citation	American Journal of Physiology - Lung Cellular and Molecular Physiology, 2013, 304 (11)	en_US
dc.identifier.issn	1040-0605	en_US
dc.identifier.uri	http://hdl.handle.net/10453/29766
dc.description.abstract	In asthma, airway smooth muscle (ASM) chemokine (C-X-C motif) receptor 3 (CXCR3) ligand production may attract mast cells or T lymphocytes to the ASM, where they can modulate ASM functions. In ASM cells (ASMCs) from people with or without asthma, we aimed to investigate JAK-STAT1, JNK, and Ca2+ involvement in chemokine (C-X-C motif) ligand (CXCL)10 and CXCL11 production stimulated by interferon-γ, IL-1β, and TNF-α combined (cytomix). Confluent, growth-arrested ASMC were treated with inhibitors for pan-JAK (pyridone-6), JAK2 (AG-490), JNK (SP-600125), or the sarco(endo)plasmic reticulum Ca2+ATPase (SERCA) pump (thapsigargin), Ca2+ chelator (BAPTA-AM), or vehicle before and during cytomix stimulation for up to 24 h. Signaling protein activation as well as CXCL10/CXCL11 mRNA and protein production were examined using immunoblot analysis, real-time PCR, and ELISA, respectively. Cytomix-induced STAT1 activation was lower and CXCR3 ligand mRNA production was more sensitive to pyridone-6 and AG-490 in asthmatic than nonasthmatic ASMCs, but CXCL10/CXCL11 release was inhibited by the same proportion. Neither agent caused additional inhibition of release when used in combination with the JNK inhibitor SP-600125. Conversely, p65 NF-κB activation was higher in asthmatic than nonasthmatic ASMCs. BAPTA-AM abolished early CXCL10/CXCL11 mRNA production, whereas thapsigargin reduced it in asthmatic cells and inhibited CXCL10/CXCL11 release by both ASMC types. Despite these inhibitory effects, neither Ca2+ agent affected early activation of STAT1, JNK, or p65 NF-κB. In conclusion, intracellular Ca2+ regulated CXCL10/CXCL11 production but not early activation of the signaling molecules involved. In asthma, reduced ASM STAT1-JNK activation, increased NF-κB activation, and altered Ca2+ handling may contribute to rapid CXCR3 ligand production and enhanced inflammatory cell recruitment. © 2013 the American Physiological Society.	en_US
dc.relation.ispartof	American Journal of Physiology - Lung Cellular and Molecular Physiology	en_US
dc.relation.isbasedon	10.1152/ajplung.00356.2012	en_US
dc.subject.classification	Respiratory System	en_US
dc.subject.mesh	Lung	en_US
dc.subject.mesh	Myocytes, Smooth Muscle	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Lung Neoplasms	en_US
dc.subject.mesh	Asthma	en_US
dc.subject.mesh	Lung Diseases	en_US
dc.subject.mesh	Calcium	en_US
dc.subject.mesh	Tumor Necrosis Factor-alpha	en_US
dc.subject.mesh	NF-kappa B	en_US
dc.subject.mesh	Chemokines, CXC	en_US
dc.subject.mesh	Adolescent	en_US
dc.subject.mesh	Adult	en_US
dc.subject.mesh	Aged	en_US
dc.subject.mesh	Middle Aged	en_US
dc.subject.mesh	Female	en_US
dc.subject.mesh	Male	en_US
dc.subject.mesh	STAT1 Transcription Factor	en_US
dc.subject.mesh	Interleukin-1beta	en_US
dc.subject.mesh	Chemokine CXCL10	en_US
dc.subject.mesh	Receptors, CXCR3	en_US
dc.subject.mesh	Chemokine CXCL11	en_US
dc.subject.mesh	Interferon-gamma	en_US
dc.title	Airway smooth muscle CXCR3 ligand production: Regulation by JAK-STAT1 and intracellular Ca<sup>2+</sup>	en_US
dc.type	Journal Article
utslib.citation.volume	11	en_US
utslib.citation.volume	304	en_US
utslib.for	1116 Medical Physiology	en_US
utslib.for	0606 Physiology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
utslib.copyright.status	open_access
pubs.issue	11	en_US
pubs.publication-status	Published	en_US
pubs.volume	304	en_US

Abstract:

In asthma, airway smooth muscle (ASM) chemokine (C-X-C motif) receptor 3 (CXCR3) ligand production may attract mast cells or T lymphocytes to the ASM, where they can modulate ASM functions. In ASM cells (ASMCs) from people with or without asthma, we aimed to investigate JAK-STAT1, JNK, and Ca2+ involvement in chemokine (C-X-C motif) ligand (CXCL)10 and CXCL11 production stimulated by interferon-γ, IL-1β, and TNF-α combined (cytomix). Confluent, growth-arrested ASMC were treated with inhibitors for pan-JAK (pyridone-6), JAK2 (AG-490), JNK (SP-600125), or the sarco(endo)plasmic reticulum Ca2+ATPase (SERCA) pump (thapsigargin), Ca2+ chelator (BAPTA-AM), or vehicle before and during cytomix stimulation for up to 24 h. Signaling protein activation as well as CXCL10/CXCL11 mRNA and protein production were examined using immunoblot analysis, real-time PCR, and ELISA, respectively. Cytomix-induced STAT1 activation was lower and CXCR3 ligand mRNA production was more sensitive to pyridone-6 and AG-490 in asthmatic than nonasthmatic ASMCs, but CXCL10/CXCL11 release was inhibited by the same proportion. Neither agent caused additional inhibition of release when used in combination with the JNK inhibitor SP-600125. Conversely, p65 NF-κB activation was higher in asthmatic than nonasthmatic ASMCs. BAPTA-AM abolished early CXCL10/CXCL11 mRNA production, whereas thapsigargin reduced it in asthmatic cells and inhibited CXCL10/CXCL11 release by both ASMC types. Despite these inhibitory effects, neither Ca2+ agent affected early activation of STAT1, JNK, or p65 NF-κB. In conclusion, intracellular Ca2+ regulated CXCL10/CXCL11 production but not early activation of the signaling molecules involved. In asthma, reduced ASM STAT1-JNK activation, increased NF-κB activation, and altered Ca2+ handling may contribute to rapid CXCR3 ligand production and enhanced inflammatory cell recruitment. © 2013 the American Physiological Society.

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http://hdl.handle.net/10453/29766