Omics-Based Approach Reveals Complement-Mediated Inflammation in Chronic Lymphocytic Inflammation With Pontine Perivascular Enhancement Responsive to Steroids (CLIPPERS).

Morten Blaabjerg,Anne Louise Hemdrup,Bjarne W Kristensen,Morten Meyer,Hans Lassmann,Peter Garred,Martin R Larsen,Zsolt Illes,Pia Jensen,Lylia Drici,Daniel Kondziella,Klemens Ruprecht,Soren W Hansen,Romana Höftberger,Friedemann Paul,Tobias Sejbaek,Helle H Nielsen

doi:10.3389/fimmu.2018.00741

Abstract

Chronic lymphocytic inflammation with pontine perivascular enhancement responsive to steroids (CLIPPERS) is a rare syndrome with relapsing brainstem/cerebellar symptoms. To examine the pathogenic processes and investigate potential biomarkers, we analyzed combined materials of brain and cerebrospinal fluid (CSF) by comprehensive methodologies. To identify major pathways of perivascular inflammation in CLIPPERS, we first compared the CSF proteome (n = 5) to a neurodegenerative condition, Alzheimer's disease (AD, n = 5). Activation of complement was confirmed by immunohistochemistry (IHC) on CLIPPERS brain samples (n = 3) and by ELISA in the CSF. For potential biomarkers, we used biomarker arrays, and compared inflammatory and vessel-associated proteins in the CSF of CLIPPERS (n = 5) with another inflammatory relapsing CNS disease, multiple sclerosis (RMS, n = 9) and healthy subjects (HS, n = 7). Two hundred and seven proteins in the CSF discriminated CLIPPERS from AD. The complement cascade, immunoglobulins, and matrix proteins were among the most frequently represented pathways. Pathway analysis of upstream regulators suggested the importance of vascular cell adhesion protein 1 (VCAM1), IFN-γ, interleukin (IL)-1, and IL-10. Differential regulation of more than 10 complement proteins of the 3 complement pathways in the CSF pointed to the role of complement activation. IHC on brain samples confirmed the perivascular complement activation, i.e., deposition of C3bc, C3d, and the terminal C5b-9 complement complex that partially overlapped with accumulation of IgG in the vessel wall. Besides endothelial cell damage, reactivity to smooth muscle actin was lost in the walls of inflamed vessels, but the glia limitans was preserved. The semi-quantitative array indicated that increased level of IL-8/CXCL8 (p < 0.05), eotaxin/CCL11 (p < 0.01), and granulocyte colony-stimulating factor (p < 0.05) in CSF could distinguish CLIPPERS from HS. The quantitative array confirmed elevated concentration of IL-8/CXCL8 and eotaxin/CCL11 compared to HS (p < 0.05, respectively) besides increased levels of ICAM-1 (p < 0.05) and VCAM-1 (p < 0.001). The increased concentration of VCAM-1 were able to differentiate CLIPPERS from RMS (p < 0.01), and a trend of elevated levels of ICAM-1 and IL-8/CXCL8 compared to RMS was also observed (p = 0.06, respectively). Complement activation, IgG deposition, and alterations of the extracellular matrix may contribute to inflammation in CLIPPERS. VCAM1, ICAM1, and IL-8 in the CSF may differentiate CLIPPERS from RMS.

Highlights

Chronic lymphocytic inflammation with pontine perivascu lar enhancement responsive to steroids (CLIPPERS) is a rare relapsing disorder with subacute brainstem features, brain MRI displaying multiple punctate or curvilinear foci of gado linium enhancement, and a clear radiological/clinical response to steroid treatment [1,2,3]
To gather further supporting evidence that complement acti vation contributes to the pathogenesis of CLIPPERS syndrome, we investigated complement activation products in the cerebrospinal fluid (CSF) and in brain biopsy/autopsy samples of patients with CLIPPERS
We examined the proteome in the CSF of CLIPPERS patients compared to non-inflammatory brain disease Alzheimer’s disease (AD)

Summary

Introduction

Chronic lymphocytic inflammation with pontine perivascu lar enhancement responsive to steroids (CLIPPERS) is a rare relapsing disorder with subacute brainstem features, brain MRI displaying multiple punctate or curvilinear foci of gado linium enhancement, and a clear radiological/clinical response to steroid treatment [1,2,3]. A homogenous group of patients has the classical features of CLIPPERS, and these patients do not develop other conditions even after a long observation period These patients require long-term immunosuppression to prevent relapses, indicating that immune-mediated CNS inflammation is a key component [3]. Biopsies from affected areas show a predominant perivascular infiltration of CD3+ T cells, most of which are CD4+ [1]. This inflammation expands to supratentorial brain regions that appear normal on 3 T MRI [1, 2, 9]. The role of B cells in the pathogenesis may be indicated by cases successfully treated with anti-CD20 (rituximab), a B cell depletion therapy to treat antibody-mediated diseases [3, 13]

Methods

Results

Conclusion