Synergistic Effects of Azithromycin and STING Agonist Promote IFN-I Production by Enhancing the Activation of STING-TBK1 Signaling

Overview

Journal J Exp Pharmacol

Publisher Dove Medical Press

Date 2023 Nov 7

PMID 37933302

Authors

Kanoktip Petcharat

Narongsuk Munkong

Rungthip Thongboontho

Widsanusan Chartarrayawadee

Arthid Thim-Uam

Affiliations

Soon will be listed here.

Abstract

Background: Azithromycin (AZM) is a macrolide antibiotic that exhibits anti-inflammatory and anti-viral infection properties by enhancing type-I interferon (IFN-I) responses. The stimulator of interferon genes (STING) can directly induce IFN-I production. However, elevated IFN-I induces auto-immune phenotypes such as systemic lupus erythematosus (SLE). The effects of AZM and STING on the production of IFN-I are unclear.

Objective: Therefore, this study aims to evaluate the role of AZM and STING on IFN-I responses in macrophages.

Methods: RAW 264.7 macrophages were treated with AZM with and without a STING-agonist (DMXAA), and the maturation of macrophages was determined using flow cytometry. Gene expression and pro-inflammatory cytokines were analyzed using qPCR and ELISA, respectively. Moreover, protein expression was investigated using Western blot assays and immunofluorescence.

Results: Our results show that AZM significantly induced M1 phenotypes, promoting surface molecule expansion of CD80 and MHC-II and production of IL-6 and TNF-α cytokines on DMXAA-stimulated macrophages. Furthermore, we found that AZM-increased mRNA levels of interferon-stimulated genes (ISGs) could be due to the high expression of STNG-TBK1 signaling in the presence of DMXAA.

Conclusion: Our data suggest that AZM enhancement of IFN-I responses was STING dependent in DMXAA-stimulated macrophages. These data underline a novel approach to AZM action-mediated STING-TBK1 signaling for regulating IFN-I responses and may further augment the scientific basis and potential use of AZM in clinical applications.

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References

Zahid A, Ismail H, Li B, Jin T . Molecular and Structural Basis of DNA Sensors in Antiviral Innate Immunity. Front Immunol. 2020; 11:613039. PMC: 7734173. DOI: 10.3389/fimmu.2020.613039. View

Gielen V, Johnston S, Edwards M . Azithromycin induces anti-viral responses in bronchial epithelial cells. Eur Respir J. 2010; 36(3):646-54. DOI: 10.1183/09031936.00095809. View

Menzel M, Akbarshahi H, Tufvesson E, Persson C, Bjermer L, Uller L . Azithromycin augments rhinovirus-induced IFNβ via cytosolic MDA5 in experimental models of asthma exacerbation. Oncotarget. 2017; 8(19):31601-31611. PMC: 5458233. DOI: 10.18632/oncotarget.16364. View

Zhu X, Guo Q, Zou J, Wang B, Zhang Z, Wei R . MiR-19a-3p Suppresses M1 Macrophage Polarization by Inhibiting STAT1/IRF1 Pathway. Front Pharmacol. 2021; 12:614044. PMC: 8129022. DOI: 10.3389/fphar.2021.614044. View

Nujic K, Banjanac M, Munic V, Polancec D, Erakovic Haber V . Impairment of lysosomal functions by azithromycin and chloroquine contributes to anti-inflammatory phenotype. Cell Immunol. 2012; 279(1):78-86. DOI: 10.1016/j.cellimm.2012.09.007. View

Luo K, Li N, Ye W, Gao H, Luo X, Cheng B . Activation of Stimulation of Interferon Genes (STING) Signal and Cancer Immunotherapy. Molecules. 2022; 27(14). PMC: 9325158. DOI: 10.3390/molecules27144638. View

Kournoutou G, Dinos G . Azithromycin through the Lens of the COVID-19 Treatment. Antibiotics (Basel). 2022; 11(8). PMC: 9404997. DOI: 10.3390/antibiotics11081063. View

Singh R, Vidhyasagar V, Yang S, Arna A, Yadav M, Aggarwal A . DDX41 is required for cGAS-STING activation against DNA virus infection. Cell Rep. 2022; 39(8):110856. PMC: 9205463. DOI: 10.1016/j.celrep.2022.110856. View

Aghai Z, Kode A, Saslow J, Nakhla T, Farhath S, Stahl G . Azithromycin suppresses activation of nuclear factor-kappa B and synthesis of pro-inflammatory cytokines in tracheal aspirate cells from premature infants. Pediatr Res. 2007; 62(4):483-8. DOI: 10.1203/PDR.0b013e318142582d. View

10.

Corrales L, Glickman L, McWhirter S, Kanne D, Sivick K, Katibah G . Direct Activation of STING in the Tumor Microenvironment Leads to Potent and Systemic Tumor Regression and Immunity. Cell Rep. 2015; 11(7):1018-30. PMC: 4440852. DOI: 10.1016/j.celrep.2015.04.031. View

11.

Banjanac M, Munic Kos V, Nujic K, Vrancic M, Belamaric D, Crnkovic S . Anti-inflammatory mechanism of action of azithromycin in LPS-stimulated J774A.1 cells. Pharmacol Res. 2012; 66(4):357-62. DOI: 10.1016/j.phrs.2012.06.011. View

12.

Ahn J, Barber G . STING signaling and host defense against microbial infection. Exp Mol Med. 2019; 51(12):1-10. PMC: 6906460. DOI: 10.1038/s12276-019-0333-0. View

13.

Ceron S, North B, Taylor S, Leib D . The STING agonist 5,6-dimethylxanthenone-4-acetic acid (DMXAA) stimulates an antiviral state and protects mice against herpes simplex virus-induced neurological disease. Virology. 2019; 529:23-28. PMC: 6382592. DOI: 10.1016/j.virol.2019.01.006. View

14.

Karimi Y, Giles E, Vahedi F, Chew M, Nham T, Loukov D . IFN- signalling regulates RAW 264.7 macrophage activation, cytokine production, and killing activity. Innate Immun. 2019; 26(3):172-182. PMC: 7144030. DOI: 10.1177/1753425919878839. View

15.

Cigana C, Nicolis E, Pasetto M, Assael B, Melotti P . Anti-inflammatory effects of azithromycin in cystic fibrosis airway epithelial cells. Biochem Biophys Res Commun. 2006; 350(4):977-82. DOI: 10.1016/j.bbrc.2006.09.132. View

16.

Vieira R, Nascimento M, Noronha I, Vasconcelos J, Benvenuti L, Barber G . STING Signaling Drives Production of Innate Cytokines, Generation of CD8 T Cells and Enhanced Protection Against Infection. Front Immunol. 2022; 12:775346. PMC: 8795786. DOI: 10.3389/fimmu.2021.775346. View

17.

Ashley C, Abendroth A, McSharry B, Slobedman B . Interferon-Independent Upregulation of Interferon-Stimulated Genes during Human Cytomegalovirus Infection is Dependent on IRF3 Expression. Viruses. 2019; 11(3). PMC: 6466086. DOI: 10.3390/v11030246. View

18.

Li M, Ferretti M, Ying B, Descamps H, Lee E, Dittmar M . Pharmacological activation of STING blocks SARS-CoV-2 infection. Sci Immunol. 2021; 6(59). PMC: 10021026. DOI: 10.1126/sciimmunol.abi9007. View

19.

Northcott M, Jones S, Koelmeyer R, Bonin J, Vincent F, Kandane-Rathnayake R . Type 1 interferon status in systemic lupus erythematosus: a longitudinal analysis. Lupus Sci Med. 2022; 9(1). PMC: 8867321. DOI: 10.1136/lupus-2021-000625. View

20.

Toriyama K, Takano N, Kokuba H, Kazama H, Moriya S, Hiramoto M . Azithromycin enhances the cytotoxicity of DNA-damaging drugs via lysosomal membrane permeabilization in lung cancer cells. Cancer Sci. 2021; 112(8):3324-3337. PMC: 8353917. DOI: 10.1111/cas.14992. View