636959
research-article2016
MSJ0010.1177/1352458516636959Multiple Sclerosis JournalA Zinger, SL Latham
MULTIPLE SCLEROSIS MSJ JOURNAL
Short Report
Plasma levels of endothelial and B-cell-derived microparticles are restored by fingolimod treatment in multiple sclerosis patients
Multiple Sclerosis Journal 1–5 DOI: 10.1177/ 1352458516636959 © The Author(s), 2016. Reprints and permissions: http://www.sagepub.co.uk/ journalsPermissions.nav
Anna Zinger, Sharissa L Latham, Valery Combes, Scott Byrne, Michael H Barnett, Simon Hawke and Georges E Grau
Abstract Background: No molecular marker can monitor disease progression and treatment efficacy in multiple sclerosis (MS). Circulating microparticles represent a potential snapshot of disease activity at the blood brain barrier. Objectives and methods: To profile plasma microparticles by flow cytometry in MS and determine how fingolimod could impact endothelial microparticles production. Results: In non-treated MS patients compared to healthy and fingolimod-treated patients, endothelial microparticles were higher, while B-cell-microparticle numbers were lower. Fingolimod dramatically reduced tumour necrosis factor (TNF)-induced endothelial microparticle release in vitro. Conclusion: Fingolimod restored dysregulated endothelial and B-cell-microparticle numbers, which could serve as a biomarker in MS.
Keywords: Multiple sclerosis, immunology, plasma microparticles, vascular endothelium, B cells, fingolimod, biological marker Date received: 3 September 2015; revised: 27 January 2016; accepted: 10 February 2016
Introduction Numerous effective treatments are currently available in multiple sclerosis (MS), but their long-term efficacy remains unknown. There is no definitive way of determining whether the disease has been suppressed at the molecular level and there is a pressing need to identify biomarkers that indicate effective disease suppression. Fingolimod, the first oral treatment for MS, is a relatively new therapy and the mechanism of action has not been fully elucidated. While we know that the primary effect is likely to be on lymphocyte egress from peripheral lymphoid tissue, the class of molecule (S1P1R agonists) may have broader effects. Microparticles (MP) are plasma membrane-derived vesicles released under physiological and pathological conditions. Their production is increased in inflammation,1 as occurs in MS.2
MPs may have a pathophysiological role in rheumatoid arthritis, cancer and Alzheimer’s disease. They may also play a role in neuroprotection, synaptic activity and regeneration in central nervous system (CNS).3 Increased MP levels have been detected in the cerebrospinal fluid (CSF) and plasma of MS patients.4,5 While endothelial MPs are detected at lower numbers in MS than in vascular disorders, they are detectable in plasma and thus might provide a profile of events occurring at the CNS vasculature. A number of studies have suggested an active role of MP in MS progression,5–7 but here we explore the role of MPs in monitoring MS treatment, focussing on fingolimod.
Correspondence to: A Zinger Vascular Immunology Unit, Discipline of Pathology, Sydney Medical School, The University of Sydney, Medical Foundation Building (K25), Room 208, 92–94 Parramatta Rd, Camperdown, NSW 2050, Australia. [email protected] Anna Zinger Sharissa L Latham Valery Combes Georges E Grau Vascular Immunology Unit, Discipline of Pathology, Sydney Medical School, The University of Sydney, Camperdown, NSW, Australia Scott Byrne Cellular Photoimmunology Group, Discipline of Infectious Diseases and Immunology, Sydney Medical School, The Charles Perkins Centre Hub at The University of Sydney, Camperdown, NSW, Australia Michael H Barnett Brain and Mind Centre, The University of Sydney, Camperdown, NSW, Australia Simon Hawke Vascular Immunology Unit, Discipline of Pathology, Sydney Medical School, The University of Sydney, Camperdown, NSW, Australia/Brain and Mind Centre, The University of Sydney, Camperdown, NSW, Australia/Central West Neurology & Neurosurgery, Orange, NSW, Australia
Patients and methods MS patients were recruited from MS Clinics in Orange NSW and at the Brain and Mind Centre, University of
http://msj.sagepub.com 1
Downloaded from msj.sagepub.com at Gazi University on March 7, 2016
Multiple Sclerosis Journal Table 1. Demographic and clinical characteristics of subjects participating in the study. Groups
Healthy subjects
Non-treated MS patients
Fingolimod-treated MS patients
N Sex: female, N (%) Age at first symptom (years; mean ± SD) Age at sampling (years; mean ± SD) Active disease during the 6 months prior to sampling, N (%)a Previous treatments, N (%) Duration of fingolimod treatment prior to sampling (months; mean ± SD) PBMC (×106/mL; mean ± SD)b,c CD3+ lymphocytes (×106/mL; mean ± SD)d,e CD19+ lymphocytes (×106/mL; mean ± SD)f,g CD14+ monocytes (×106/mL; mean ± SD)
19 12 (63) n/a 38.23 ± 11.9 n/a
15 11 (73) 31.06 ± 9.05 37.41 ± 9.08 12 (80)
19 13 (68) 31.47 ± 8.28 38.23 ± 6.99 4 (21)
n/a n/a
6 (40) n/a
9 (47) 6.69 ± 5.25
2.02 ± 0.7 1.07 ± 0.31
2.22 ± 0.6 1.28 ± 0.3
0.95 ± 0.54 0.16 ± 0.05
0.14 ± 0.07
0.13 ± 0.09
0.01 ± 0.01
0.12 ± 0.06
0.11 ± 0.06
0.09 ± 0.05
MS: multiple sclerosis; PBMC: peripheral blood mononuclear cells; SD: standard deviation. Comparisons were performed with Kruskal–Wallis analysis of variance with Dunn multiple comparison test or t test, except for sex, active disease and previous treatment, which were compared using chi-square with Fisher exact test. ap
research-article2016
MSJ0010.1177/1352458516636959Multiple Sclerosis JournalA Zinger, SL Latham
MULTIPLE SCLEROSIS MSJ JOURNAL
Short Report
Plasma levels of endothelial and B-cell-derived microparticles are restored by fingolimod treatment in multiple sclerosis patients
Multiple Sclerosis Journal 1–5 DOI: 10.1177/ 1352458516636959 © The Author(s), 2016. Reprints and permissions: http://www.sagepub.co.uk/ journalsPermissions.nav
Anna Zinger, Sharissa L Latham, Valery Combes, Scott Byrne, Michael H Barnett, Simon Hawke and Georges E Grau
Abstract Background: No molecular marker can monitor disease progression and treatment efficacy in multiple sclerosis (MS). Circulating microparticles represent a potential snapshot of disease activity at the blood brain barrier. Objectives and methods: To profile plasma microparticles by flow cytometry in MS and determine how fingolimod could impact endothelial microparticles production. Results: In non-treated MS patients compared to healthy and fingolimod-treated patients, endothelial microparticles were higher, while B-cell-microparticle numbers were lower. Fingolimod dramatically reduced tumour necrosis factor (TNF)-induced endothelial microparticle release in vitro. Conclusion: Fingolimod restored dysregulated endothelial and B-cell-microparticle numbers, which could serve as a biomarker in MS.
Keywords: Multiple sclerosis, immunology, plasma microparticles, vascular endothelium, B cells, fingolimod, biological marker Date received: 3 September 2015; revised: 27 January 2016; accepted: 10 February 2016
Introduction Numerous effective treatments are currently available in multiple sclerosis (MS), but their long-term efficacy remains unknown. There is no definitive way of determining whether the disease has been suppressed at the molecular level and there is a pressing need to identify biomarkers that indicate effective disease suppression. Fingolimod, the first oral treatment for MS, is a relatively new therapy and the mechanism of action has not been fully elucidated. While we know that the primary effect is likely to be on lymphocyte egress from peripheral lymphoid tissue, the class of molecule (S1P1R agonists) may have broader effects. Microparticles (MP) are plasma membrane-derived vesicles released under physiological and pathological conditions. Their production is increased in inflammation,1 as occurs in MS.2
MPs may have a pathophysiological role in rheumatoid arthritis, cancer and Alzheimer’s disease. They may also play a role in neuroprotection, synaptic activity and regeneration in central nervous system (CNS).3 Increased MP levels have been detected in the cerebrospinal fluid (CSF) and plasma of MS patients.4,5 While endothelial MPs are detected at lower numbers in MS than in vascular disorders, they are detectable in plasma and thus might provide a profile of events occurring at the CNS vasculature. A number of studies have suggested an active role of MP in MS progression,5–7 but here we explore the role of MPs in monitoring MS treatment, focussing on fingolimod.
Correspondence to: A Zinger Vascular Immunology Unit, Discipline of Pathology, Sydney Medical School, The University of Sydney, Medical Foundation Building (K25), Room 208, 92–94 Parramatta Rd, Camperdown, NSW 2050, Australia. [email protected] Anna Zinger Sharissa L Latham Valery Combes Georges E Grau Vascular Immunology Unit, Discipline of Pathology, Sydney Medical School, The University of Sydney, Camperdown, NSW, Australia Scott Byrne Cellular Photoimmunology Group, Discipline of Infectious Diseases and Immunology, Sydney Medical School, The Charles Perkins Centre Hub at The University of Sydney, Camperdown, NSW, Australia Michael H Barnett Brain and Mind Centre, The University of Sydney, Camperdown, NSW, Australia Simon Hawke Vascular Immunology Unit, Discipline of Pathology, Sydney Medical School, The University of Sydney, Camperdown, NSW, Australia/Brain and Mind Centre, The University of Sydney, Camperdown, NSW, Australia/Central West Neurology & Neurosurgery, Orange, NSW, Australia
Patients and methods MS patients were recruited from MS Clinics in Orange NSW and at the Brain and Mind Centre, University of
http://msj.sagepub.com 1
Downloaded from msj.sagepub.com at Gazi University on March 7, 2016
Multiple Sclerosis Journal Table 1. Demographic and clinical characteristics of subjects participating in the study. Groups
Healthy subjects
Non-treated MS patients
Fingolimod-treated MS patients
N Sex: female, N (%) Age at first symptom (years; mean ± SD) Age at sampling (years; mean ± SD) Active disease during the 6 months prior to sampling, N (%)a Previous treatments, N (%) Duration of fingolimod treatment prior to sampling (months; mean ± SD) PBMC (×106/mL; mean ± SD)b,c CD3+ lymphocytes (×106/mL; mean ± SD)d,e CD19+ lymphocytes (×106/mL; mean ± SD)f,g CD14+ monocytes (×106/mL; mean ± SD)
19 12 (63) n/a 38.23 ± 11.9 n/a
15 11 (73) 31.06 ± 9.05 37.41 ± 9.08 12 (80)
19 13 (68) 31.47 ± 8.28 38.23 ± 6.99 4 (21)
n/a n/a
6 (40) n/a
9 (47) 6.69 ± 5.25
2.02 ± 0.7 1.07 ± 0.31
2.22 ± 0.6 1.28 ± 0.3
0.95 ± 0.54 0.16 ± 0.05
0.14 ± 0.07
0.13 ± 0.09
0.01 ± 0.01
0.12 ± 0.06
0.11 ± 0.06
0.09 ± 0.05
MS: multiple sclerosis; PBMC: peripheral blood mononuclear cells; SD: standard deviation. Comparisons were performed with Kruskal–Wallis analysis of variance with Dunn multiple comparison test or t test, except for sex, active disease and previous treatment, which were compared using chi-square with Fisher exact test. ap
