Effects of Azithromycin on Blood Inflammatory Gene Expression and Cytokine Production in Sarcoidosis

Fraser, Simon D.; Thackray-Nocera, Susannah; Wright, Caroline; Flockton, Rachel; James, Sally R.; Crooks, Michael G.; Kaye, Paul M.; Hart, Simon P.

Abstract

Introduction: In sarcoidosis granulomas, monocyte-derived macrophages are activated by pro-inflammatory cytokines including TNF and IL-6. Current drug treatment for sarcoidosis aims to suppress inflammation but disabling side effects can ensue. The macrolide azithromycin may be anti-inflammatory. We aimed to determine whether treatment with azithromycin affects blood inflammatory gene expression and monocyte functions in sarcoidosis. Methods: Blood samples were collected from patients with chronic pulmonary sarcoidosis enrolled in a single arm, open label clinical trial who received oral azithromycin 250 mg once daily for 3 months. Whole blood inflammatory gene expression with or without LPS stimulation was measured using a 770-mRNA panel. Phenotypic analysis and cytokine production were conducted by flow cytometry and ELISA after 24h stimulation with growth factors and TLR ligands. mTOR activity was assessed by measuring phosphorylated S6RP. Results: Differential gene expression analysis indicated a state of heightened myeloid cell activation in sarcoidosis. Compared with controls, sarcoidosis patients showed increased LPS responses for several cytokines and chemokines. Treatment with azithromycin had minimal effect on blood gene expression overall, but supervised clustering analysis identified several chemokine genes that were upregulated. At the protein level, azithromycin treatment increased LPS-stimulated TNF and unstimulated IL-8 production. No other cytokines showed significant changes following azithromycin. Blood neutrophil counts fell during azithromycin treatment whereas mononuclear cells remained stable. Azithromycin had no detectable effects on mTOR activity or activation markers. Conclusion: Blood myeloid cells are activated in sarcoidosis, but azithromycin therapy did not suppress inflammatory gene expression or cytokine production in blood. Trial registration: EudraCT 2019-000580-24 (17 May 2019)

Citation

Fraser, S. D., Thackray-Nocera, S., Wright, C., Flockton, R., James, S. R., Crooks, M. G., Kaye, P. M., & Hart, S. P. (2024). Effects of Azithromycin on Blood Inflammatory Gene Expression and Cytokine Production in Sarcoidosis. Lung, https://doi.org/10.1007/s00408-024-00743-w

• Journal Article Type: Article
• Acceptance Date: Aug 26, 2024
• Online Publication Date: Sep 16, 2024
• Publication Date: Jan 1, 2024
• Deposit Date: Sep 17, 2024
• Publicly Available Date: Sep 20, 2024
• Journal: Lung
• Print ISSN: 0341-2040
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1007/s00408-024-00743-w
• Public URL: https://hull-repository.worktribe.com/output/4832143