Cladribine modifies functional properties of microglia. (6th July 2020)

• Record Type: Journal Article
• Authors: Jørgensen, L. Ø.; Hyrlov, K. H.; Elkjaer, M. L.; Weber, A. B.; Pedersen, A. E.; Svenningsen, Å. Fex; Illes, Z.
• Is Part Of: Clinical and experimental immunology. Volume 201:Number 3(2020)
• Page Start: 328
• Page End: 340
• Publication Date: 2020-07-06
• Subjects: cladribine -- deoxycytidine kinase -- microglia -- migration -- multiple sclerosis -- phagocytosis -- viability; Immunopathology -- Periodicals; 616.079
• DOI: 10.1111/cei.13473
• Languages: English
• ISSNs: 0009-9104

Abstract

Summary: Cladribine (CdA), an oral prodrug approved for the treatment of relapsing multiple sclerosis, selectively depletes lymphocytes. CdA passes the blood–brain barrier, suggesting a potential effect on central nervous system (CNS) resident cells. We examined if CdA modifies the phenotype and function of naive and activated primary mouse microglia, when applied in the concentrations 0·1–1 μM that putatively overlap human cerebrospinal fluid (CSF) concentrations. Primary microglia cultures without stimulation or in the presence of proinflammatory lipopolysaccharide (LPS) or anti‐inflammatory interleukin (IL)‐4 were treated with different concentrations of CdA for 24 h. Viability was assessed by MTT [3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide] assay. Phagocytotic ability and morphology were examined by flow cytometry and random migration using IncuCyte Zoom and TrackMate. Change in gene expression was examined by quantitative polymerase chain reaction (qPCR) and protein secretion by Meso Scale Discovery. We found that LPS and IL‐4 up‐regulated deoxycytidine kinase (DCK) expression. Only activated microglia were affected by CdA, and this was unrelated to viability. CdA 0·1–1 μM significantly reduced granularity, phagocytotic ability and random migration of activated microglia. CdA 10 μM increased the IL‐4‐induced gene expression of arginase 1 (Arg1) and LPS‐induced expression of IL‐1β, tumor necrosis factor (TNF), inducible nitric oxide synthase (iNOS) and Arg1, but protein secretion remained unaffected. CdA 10 μM potentiated the increased expression of anti‐inflammatory TNF receptor 2 (TNF‐R2) but not TNF‐R1 induced by LPS. This suggests that microglia acquire a less activated phenotype when treated with 0·1–1 μM CdA that putatively overlaps human CSF concentrations. This may be related to the up‐regulated gene expression of DCK upon activation, and suggests a potential alternative mechanism of CdA with direct effect on CNS resident cells. Abstract : Oral cladribine approved for the treatment of relapsing multiple sclerosis is a synthetic deoxyadenosine analogue that selectively depletes lymphocytes, but also crosses the blood brain barrier. We examined, if cladribine modifies the phenotype and function of naïve and activated primary mouse microglia, when applied in the concentrations that putatively overlaps human CSF concentrations. We found that microglia acquire a less activated phenotype when co‐treated with cladribine in such concentrations in vitro, suggesting a potential alternative mechanism with direct effect on CNS resident cells.