Etidronate down-regulates Toll-like receptor 2 ligand-induced chemokine production by inhibiting MyD88 expression and NF-κB activation

Abstract

Objective

Pretreatment of J774.1 cells with etidronate, a non-nitrogen-containing bisphosphonate (non-NBP) used as an antibone resorptive drug, was previously reported to inhibit Toll-like receptor (TLR) 2 agonist-induced proinflammatory cytokine production. The present study aimed to examine the effects of etidronate on chemokine production by human monocytic U937 cells incubated with Pam₃Cys-Ser-(Lys)₄ (Pam₃CSK₄, a TLR2 ligand) and lipid A (a TLR4 ligand).

Methods

U937 cells were pretreated with or without etidronate, and then incubated with or without Pam₃CSK₄ or lipid A. Levels of secreted human interleukin (IL)-8 and monocyte chemoattractant protein-1 (MCP-1) in culture supernatants and activation of nuclear factor-κB (NF-κB) p65 were measured by enzyme-linked immunosorbent assay (ELISA). Cytotoxicity was determined by measuring lactate dehydrogenase (LDH) activity in supernatants. Expression of intracellular adhesion molecule (ICAM)-1 and MyD88 was analyzed by flow cytometry and Western blot analysis, respectively.

Results

Etidronate down-regulated IL-8 and MCP-1 production and NF-κB p65 activation induced by Pam₃CSK_4, but not lipid A, in U937 cells. Etidronate also inhibited MyD88 expression in U937 cells incubated with Pam₃CSK₄.

Conclusion

Etidronate down-regulates IL-8 and MCP-1 production in U937 cells by inhibiting both the expression of MyD88 and activation of NF-κB p65 in the TLR2, but not TLR4, pathway.

Keywords:

• Non-NBP
• etidronate
• TLR
• chemokines
• myeloid differentiation factor 88 (MyD88)
• NF-κB

Correction Statement

This article has been republished with minor changes. These changes do not impact the academic content of the article.

Disclosure statement

The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article.

Additional information

Funding

This work was supported by a Grant-in-Aid for Scientific Research from the Ohu University School of Dentistry, including the President’s discretionary expense.