P2Y6 receptors are involved in mediating the effect of inactivated avian influenza virus H5N1 on IL-6 & CXCL8 mRNA expression in respiratory epithelium.

Abstract

One of the key pathophysiologies of H5N1 infection is excessive proinflammatory cytokine response (cytokine storm) characterized by increases in IFN-β, TNF-α, IL-6, CXCL10, CCL4, CCL2 and CCL5 in the respiratory tract. H5N1-induced cytokine release can occur via an infection-independent mechanism, however, detail of the cellular signaling involved is poorly understood. To elucidate this mechanism, the effect of inactivated (β-propiolactone-treated) H5N1 on the cytokine and chemokine mRNA expression in 16HBE14o- human respiratory epithelial cells was investigated. We found that the inactivated-H5N1 increased mRNA for IL-6 and CXCL8 but not TNF-α, CCL5 or CXCL10. This effect of the inactivated-H5N1 was inhibited by sialic acid receptor inhibitor (α-2,3 sialidase), adenosine diphosphatase (apyrase), P2Y receptor (P2YR) inhibitor (suramin), P2Y6R antagonist (MRS2578), phospholipase C inhibitor (U73122), protein kinase C inhibitors (BIM and Gö6976) and cell-permeant Ca2+ chelator (BAPTA-AM). Inhibitors of MAPK signaling, including of ERK1/2 (PD98059), p38 MAPK (SB203580) and JNK (SP600125) significantly suppressed the inactivated-H5N1-induced mRNA expression of CXCL8. On the other hand, the inactivated-H5N1-induced mRNA expression of IL-6 was inhibited by SB203580, but not PD98059 or SP600125, whereas SN-50, an inhibitor of NF-κB, inhibited the effect of virus on mRNA expression of both of IL-6 and CXCL8. Taken together, our data suggest that, without infection, inactivated-H5N1 induces mRNA expression of IL-6 and CXCL8 by a mechanism, or mechanisms, requiring interaction between viral hemagglutinin and α-2,3 sialic acid receptors at the cell membrane of host cells, and involves activation of P2Y6 purinergic receptors.

• Authors: Nawiya Huipao, Suparerk Borwornpinyo, Suwimon Wiboon-Ut, Craig R Campbell, Il-Ha Lee, Siriphun Hiranyachattada, Chonlaphat Sukasem, Arunee Thitithanyanont, Chumpol Pholpramool, David I Cook, Anuwat Dinudom
• Journal: PloS one (PLoS One) Vol. 12 Issue 5 Pg. e0176974 ( 2017) ISSN: 1932-6203 [Electronic] United States
• PMID: 28494003 (Publication Type: Journal Article)

Chemical References

• CXCL8 protein, human
• Interleukin-6
• Interleukin-8
• RNA, Messenger
• Receptors, Purinergic P2
• purinoceptor P2Y6

Topics

• Animals
• Cell Line
• Chickens
• Gene Expression Regulation
• Humans
• Influenza A Virus, H5N1 Subtype (physiology)
• Influenza, Human (genetics, metabolism, virology)
• Interleukin-6 (genetics)
• Interleukin-8 (genetics)
• RNA, Messenger (genetics)
• Receptors, Purinergic P2 (metabolism)
• Respiratory Mucosa (metabolism, virology)
• Signal Transduction