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Rise and Fall of Kir2.2 Current by TLR4 Signaling in Human Monocytes: PKC-Dependent Trafficking and PI3K-Mediated

Kyung Soo Kim1, Ji Hyun Jang1, Haiyue Lin1

  • 1Department of Physiology, Seoul National University College of Medicine, Seoul 110-799, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 110-799, Republic of Korea;

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|September 2, 2015
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Toll-like receptor 4 (TLR4) stimulation by lipopolysaccharides (LPS) induces inwardly rectifying potassium (Kir2.2) channels in monocytes. Protein kinase C (PKC) mediates Kir2.2 trafficking, influencing calcium influx and cytokine release.

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Area of Science:

  • Immunology
  • Cell Biology
  • Ion Channel Physiology

Background:

  • Lipopolysaccharides (LPS) are potent stimulators of Toll-like receptor 4 (TLR4) in immune cells.
  • The precise effects of LPS on ion channel activity in immune cells, particularly potassium channels, remain largely unexplored.
  • Understanding these interactions is crucial for elucidating immune cell activation and function.

Purpose of the Study:

  • To investigate the impact of LPS on inwardly rectifying potassium (Kir) channel currents in human monocytes.
  • To identify the specific Kir channel subtypes involved and the molecular mechanisms regulating their activity.
  • To determine the functional consequences of LPS-induced Kir channel modulation on immune cell responses.

Main Methods:

  • Electrophysiological recordings (patch-clamp) were used to measure Kir channel currents in LPS-treated THP-1 cells and human monocytes.
  • Quantitative PCR and Western blotting assessed Kir2.1 and Kir2.2 mRNA and protein expression.
  • Small interfering RNA (siRNA) knockdown, pharmacological inhibitors (PKC, PI3K), and immunofluorescence microscopy were employed to dissect signaling pathways and protein localization.

Main Results:

  • LPS treatment induced a novel inwardly rectifying potassium current (IKir,LPS) in monocytes, primarily mediated by Kir2.2.
  • LPS promoted the plasma membrane translocation of Kir2.2 in a protein kinase C (PKC)-dependent manner.
  • The decay of IKir,LPS was linked to the conversion of phosphatidylinositol (4,5)-bisphosphate (PIP2) to PIP3, modulated by PI3K signaling.

Conclusions:

  • TLR4 stimulation by LPS induces Kir2.2 channel activity via PKC-dependent membrane trafficking in monocytes.
  • The dynamic regulation of Kir2.2 by PIP2/PIP3 metabolism influences LPS-driven immune responses, including calcium influx and cytokine release.
  • These findings reveal a novel role for Kir2.2 in TLR4-mediated monocyte activation and suggest potential therapeutic targets.