Research Article: Utility of CSF Cytokine/Chemokines as Markers of Active Intrathecal Inflammation: Comparison of Demyelinating, Anti-NMDAR and Enteroviral Encephalitis

Date Published: August 30, 2016

Publisher: Public Library of Science

Author(s): Kavitha Kothur, Louise Wienholt, Shekeeb S. Mohammad, Esther M. Tantsis, Sekhar Pillai, Philip N. Britton, Cheryl A. Jones, Rajeshwar R. Angiti, Elizabeth H. Barnes, Timothy Schlub, Sushil Bandodkar, Fabienne Brilot, Russell C. Dale, Edgar Meinl.


Despite the discovery of CSF and serum diagnostic autoantibodies in autoimmune encephalitis, there are still very limited CSF biomarkers for diagnostic and monitoring purposes in children with inflammatory or autoimmune brain disease. The cause of encephalitis is unknown in up to a third of encephalitis cohorts, and it is important to differentiate infective from autoimmune encephalitis given the therapeutic implications.

To study CSF cytokines and chemokines as diagnostic biomarkers of active neuroinflammation, and assess their role in differentiating demyelinating, autoimmune, and viral encephalitis.

We measured and compared 32 cytokine/chemokines using multiplex immunoassay and APRIL and BAFF using ELISA in CSF collected prior to commencing treatment from paediatric patients with confirmed acute disseminated encephalomyelitis (ADEM, n = 16), anti-NMDAR encephalitis (anti-NMDAR E, n = 11), and enteroviral encephalitis (EVE, n = 16). We generated normative data using CSF from 20 non-inflammatory neurological controls. The sensitivity of CSF cytokine/chemokines to diagnose encephalitis cases was calculated using 95th centile of control values as cut off. We correlated CSF cytokine/chemokines with disease severity and follow up outcome based on modified Rankin scale. One-way hierarchical correlational cluster analysis of molecules was performed in different encephalitis and outcome groups.

In descending order, CSF TNF-α, IL-10, IFN-α, IL-6, CXCL13 and CXCL10 had the best sensitivity (>79.1%) when all encephalitis patients were included. The combination of IL-6 and IFN-α was most predictive of inflammation on multiple logistic regression with area under the ROC curve 0.99 (CI 0.97–1.00). There were no differences in CSF cytokine concentrations between EVE and anti-NMDAR E, whereas ADEM showed more pronounced elevation of Th17 related (IL-17, IL-21) and Th2 (IL-4, CCL17) related cytokine/chemokines. Unlike EVE, heat map analysis showed similar clustering of cytokine/chemokine molecules in immune mediated encephalitis (ADEM and anti-NMDAR E). Th1 and B cell (CXCL13 and CXCL10) molecules clustered together in patients with severe encephalopathy at admission and worse disability at follow up in all encephalitis. There was no correlation between CSF neopterin and IFN-γ or IFN-α.

A combination panel of cytokine/chemokines consisting of CSF TNF-α, IL-10, IFN-α, IL-6, CXCL13 and CXCL10 measured using multiplex immunoassay may be used to diagnose and monitor intrathecal inflammation in the brain. Given their association with worse outcome, certain key chemokines (CXCL13, CXCL10) could represent potential therapeutic targets in encephalitis.

Partial Text

Paediatric neuroinflammatory disorders represent a heterogeneous group of disorders including encephalitis, demyelinating disorders and other immune-mediated autoinflammatory CNS disorders [1]. It is important to differentiate neuroinflammatory disorders from other neurological conditions, as the former are potentially treatable. There is a need to develop CSF biomarkers in neuroinflammatory disorders of the brain to determine if persistent neurological symptoms are due to active ongoing CNS inflammation or alternatively to residual brain injury [2, 3]. Currently available investigations do not have high sensitivity in detecting neuroinflammation, for example MRI is frequently normal in autoimmune encephalitis. The currently available CSF markers including pleocytosis, neopterin, oligoclonal bands lack sensitivity and specificity, and cell neuronal surface antibodies are found only in a small proportion of neuroinflammatory conditions [4–8]. The etiology of 37–63% patients with encephalitis remains elusive in encephalitis cohort studies despite extensive investigations, and it can be difficult to differentiate infective from autoimmune encephalitis [4, 9–11].

As part of the ethically approved protocol, we wrote to all parents and gained written consent to use the stored CSF for this study. This study was approved by HREC of Sydney Children’s Hospital network (LNRSSA/14/SCHN/283).

The aim of the present study was to describe the CSF cytokine/chemokine concentrations in patients in well-defined encephalitis groups and discuss their utility as biomarkers of inflammation. This is based on the hypothesis that CSF cytokines may correspond to the intrathecal activation of immunoactive cells, and the evaluation of these cytokines might be a good indicator of the type of immune activation, the severity of inflammation and disease activity in neuroinflammatory disorders of the brain. In our cohort, among currently available investigations, the sensitivity of pleocytosis, oligoclonal bands and neopterin were similar to previous studies [4, 7, 9, 18, 19]. CSF neopterin performed the best of these commonly used CSF biomarkers, although is not used commonly in neurology practice outside of HIV neurology [20]. These findings highlight the need to develop more diagnostic markers, which can guide clinicians in diagnosing, monitoring and treating neuroinflammation.

To conclude, we presented cytokine/chemokine profile from relatively well-defined encephalitis groups. Our study suggests that combining several biomarkers will enhance our ability to diagnose and monitor CNS inflammation. Despite similarities between CSF cytokine/chemokine concentrations in EVE and anti NMDAR E, we identified differential cytokine/chemokine interactions on cluster analysis, which may play a role in autoimmune pathogenesis (ADEM and anti NMDAR E). Some of these molecules (CXCL13 and CXCL10) may represent potential therapeutic targets in view of their association with severity of encephalopathy at admission and worse disability at follow up in all encephalitis cases.