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The IRAK-ERK-p67phox-Nox-2 axis mediates TLR4, 2-induced ROS production for IL-1β transcription and processing in monocytes

Cellular & Molecular Immunology volume 13, pages 745–763 (2016)Cite this article

A Correction to this article was published on 11 September 2019

Abstract

In monocytic cells, Toll-like receptor 4 (TLR4)- and TLR2-induced reactive oxygen species (ROS) cause oxidative stress and inflammatory response; however, the mechanism is not well understood. The present study investigated the role of interleukin-1 receptor-associated kinase (IRAK), extracellular signal-regulated kinase (ERK), p67phox and Nox-2 in TLR4- and TLR2-induced ROS generation during interleukin-1 beta (IL-1β) transcription, processing, and secretion. An IRAK1/4 inhibitor, U0126, PD98059, an NADPH oxidase inhibitor (diphenyleneiodonium (DPI)), and a free radical scavenger (N-acetyl cysteine (NAC))-attenuated TLR4 (lipopolysaccharide (LPS))- and TLR2 (Pam3csk4)-induced ROS generation and IL-1β production in THP-1 and primary human monocytes. An IRAK1/4 inhibitor and siRNA-attenuated LPS- and Pam3csk4-induced ERK-IRAK1 association and ERK phosphorylation and activity. LPS and Pam3csk4 also induced IRAK1/4-, ERK- and ROS-dependent activation of activator protein-1 (AP-1), IL-1β transcription, and IL-1β processing because significant inhibition in AP-1 activity, IL-1β transcription, Pro- and mature IL-β expression, and caspase-1 activity was observed with PD98059, U0126, DPI, NAC, an IRAK1/4 inhibitor, tanshinone IIa, and IRAK1 siRNA treatment. IRAK-dependent ERK-p67phox interaction, p67phox translocation, and p67phox–Nox-2 interaction were observed. Nox-2 siRNA significantly reduced secreted IL-1β, IL-1β transcript, pro- and mature IL-1β expression, and caspase-1 activity indicating a role for Nox-2 in LPS- and Pam3csk4-induced IL-1β production, transcription, and processing. In the present study, we demonstrate that the TLR4- and TLR2-induced IRAK-ERK pathway cross-talks with p67phox-Nox-2 for ROS generation, thus regulating IL-1β transcription and processing in monocytic cells.

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Acknowledgements

The authors gratefully acknowledge the technical help provided by Mr. C.P. Pandey, Mrs. M. Chaturvedi, and Mr. A.L. Vishwakarma. Funding from THUNDER BSC0102 CSIR Network project to Manoj Kumar Barthwal is gratefully acknowledged. Fellowships from University Grants Commission to Ankita Singh and Council of Scientific and Industrial Research to Vishal Singh and Rajiv L Tiwari are acknowledged. CDRI communication number: 9008.

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  1. Dr. MK Barthwal, Pharmacology Division, CSIR-Central Drug Research Institute, B.S 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031, Uttar Pradesh, India. E-mail: manojbarthwal@cdri.res.in

Authors and Affiliations

  1. Pharmacology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Sitapur Road, Jankipuram Extension, Lucknow, 226031, Uttar Pradesh, India

    Ankita Singh, Vishal Singh, Rajiv L. Tiwari, Madhu Dikshit & Manoj K. Barthwal

  2. Department of Transfusion Medicine and Blood Bank, King George’s Medical University, Lucknow, Uttar Pradesh, India

    Tulika Chandra

  3. Department of Pathology, King George’s Medical University, Lucknow, Uttar Pradesh, India

    Ashutosh Kumar

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Singh, A., Singh, V., Tiwari, R. et al. The IRAK-ERK-p67phox-Nox-2 axis mediates TLR4, 2-induced ROS production for IL-1β transcription and processing in monocytes. Cell Mol Immunol 13, 745–763 (2016). https://doi.org/10.1038/cmi.2015.62

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