Author Of 3 Presentations
P0056 - CSF inflammatory profile of primary progressive multiple sclerosis (ID 1657)
Abstract
Background
Background: So far, no specific biomarkers that help to stratify primary progressive multiple sclerosis (PPMS) from relapsing remitting multiple sclerosis (RRMS) patients are still unknown. In the diagnosis of PPMS, among with clinical and imaging assessment, the analysis of cerebrospinal fluid with regard to evidence of oligoclonal bands and/or elevated IgG index is helpful.
Objectives
Objectives: To evaluate the levels of 69 pro and anti-inflammatory cytokines and chemochines as well as Nf-L in the CSF of PPMS at the diagnosis.
Methods
Methods: Levels of 69 inflammatory mediators and of NF-L has been evaluated in the CSF obtained at the diagnosis from 16 patients with PPMS, 80 patients with RRMS and 12 patients with central nervous system non-inflammatory neurological disorders by mean of immune-assay multiplex techniques based on the Luminex technology and Human NF-light enzyme-linked immunosorbentassays. Clinical assessment including EDSS and white and grey matter (WM and GM) lesion volume and numbers were collected by using 3-T MRI analysis.
Results
Results: No significant differences were noticed in IgG index, CSF lymphocyte count, NF-L levels, WM and GM lesion volume and number were not significantly different between PPMS and RRMS. PPMS patients had higher EDSS when compared to RRMS group (median[IQR] 3 [3-4] vs 2[1-2.375]) and older age (mean54.5±9.6y vs 36.8 ± 11.9, p<0.001). Among selected molecules that appeared increased in both PPMS and RRMS groups respect to controls, a multivariate logistic regression analysis showed that at diagnosis altered levels of some B-cell related cytokines such as IL-10 (OR=0.28, CI95%[0.09-0.96]) and CXCL12 (OR=3.97, CI95%[1.34-11.7]) and the monocyte-related osteopontin (OR=2.24, CI95%[1.01-4.99]) were predictive for a primary progressive course of the disease instead of a relapsing one. Kegg pathway analysis confirmed that most of molecules characterizing PPMS CSF profile are involved in chronic immune inflammatory diseases.
Conclusions
Conclusions: At the diagnosis, CSF of PPMS patients appeared characterized by high levels of inflammatory mediators. A detailed CSF profiling obtained at the diagnosis could help to differentiate progressive forms of MS, providing new insights into its pathogenesis and useful tools in clinical practice.
P0957 - Does peripheral immune profile reflect intrathecal inflammation in multiple sclerosis patients at time of diagnosis? (ID 1728)
Abstract
Background
Intrathecal inflammation, possibly compartmentalized in meninges, perivascular cuffs and cerebrospinal fluid (CSF), represents one of the main features of Multiple Sclerosis (MS). Recent studies have shown that a specific panel of CSF molecules present at diagnosis may be able to stratify naïve MS patients in two subgroups according to the high (SMhigh) or low (MSlow) level of CSF inflammation and cortical damage and to predict their disease outcome.
Objectives
Verify whether the inflammatory CSF intrathecal profile may correlate with the protein and cell inflammatory profile of the paired blood at diagnosis.
Methods
Immunoassay protein analysis of 79 inflammatory molecules was evaluated in paired CSF and serum obtained at diagnosis by 71 patients with MS and 36 subjects with other neurodegenerative diseases. Each patient was annually subjected to detailed clinical/radiological examination by 3 T-MRI. By using flow-cytometry we also analysed the specific blood immunophenotypes in paired CSF and blood samples of a subgroup of 15 MS patients.
Results
We found significant (p<0,01) correlations between number of spinal lesions both at diagnosis (T0) and after 2 years and the expressions of CXCL16, (r=0.54), CXCL5, (r=0.46), sTNFR1, (r= 0.39), TWEAK (r=0.43) and finally IL12p70 (r=0.39), as well as with number of blood total CD19+ B cells (r= -0.93; p<0.05) in MS patients but not in controls. However, no correlation between the protein inflammatory profile of paired CSF/serum samples has been revealed, with the only exception of interleukin 10 (IL-10), which was found elevated in both CSF (p<0.01) and serum (p<0.05) of MShigh compared to MSlow patients. Significant correlation was measured between IL-10 serum levels and the number of B cells in the blood (r=0.67; p<0.01), in particular with a subset of switched memory CD19+CD27+IgD- B cells and in particular in the MShigh group. In addition, positive correlation (p<0.05) was found between blood exhausted CD19+CD27-IgD- B cells and number white matter lesions at diagnosis (r=0,55), volume of white matter lesions (r=0.69), global cortical thickness (r=0.71), number of cortical lesions (r=0.79), volume of cortical lesions (r=0.80) and global cortical thickness (r=0.82). On the contrary, the percentage of some memory CD19+CD27-IgG+B cells correlates (p<0.05) with the cortical thickness (r=0.70) and age at diagnosis (r= -0.67). When we analysed possible correlation between blood and CSF immunophenotypes at diagnosis, we found significant correlation (r=0.74; p<0.05) between blood naïve B cells (CD19+CD27-IgD+) and CSF memory CD19+CD27-IgG+B cells.
Conclusions
Serum inflammatory profile at diagnosis only partially reflects intrathecal CSF inflammation. Only IL-10 appeared to represent a useful link between CSF and serum inflammation at diagnosis and may help to identify a subgroup pf MS patients with high involvement of B lymphocytes in early MS pathogenesis.
P0969 - Imbalance of TNF and TNF receptors in the cerebrospinal fluid of naïve multiple sclerosis patients (ID 1602)
Abstract
Background
Background: An imbalance in TNF signaling in the inflammatory milieu generated in the cortical subarachnoid space of the multiple sclerosis (MS) brain is thought to lead to increased cortical pathology.
Objectives
Objectives: To investigate the changes in TNF and its soluble receptors in CSF at the early stages of MS and how they associate with clinical outcome.
Methods
Methods: Protein levels of TNF, sTNFR1 and sTNFR2 were assayed in CSF collected from 122 naïve MS patients and 34 subjects with other neurological conditions at diagnosis. Potential correlations with other molecules of TNF family and other cytokines/chemokines were evaluated by using BioPlex System immunoassay and Ingenuity pathway analysis. TNF and TNFRs levels in CSF were correlated with clinical and imaging parameters at diagnosis (T0) and after 2 years of follow-up (T2).
Results
Results: Significant increased levels of TNF (fold change:7.739; p<0.001), sTNFR1 (fold change:1.693; p<0.001) and sTNFR2 (fold change:2.189; p<0.001) were detected in CSF of MS patients compared to the control group at T0, and were found especially high in patients with enhanced CSF inflammation. Increased TNF levels in CSF were significantly (p<0.01) associated with increased EDSS change (r=0.43), relapses (r=0.48) and new white matter lesions (r=0.49) at T2. Elevated CSF levels of sTNFR1 were associated with higher cortical lesion volume (r=0.41) at T0, as well as with new cortical lesions (r=0.56) and signs of disease activity (r= 0.61) at T2, whilst no correlation could be found between sTNFR2 levels inCSF and clinical or MRI features. By performing combined correlation and pathway analysis, CSF TNF signalling (encompassing elevated levels of BAFF, IFN-G, IL-1beta, IL-10, IL-8, IL-16, CCL21, haptoglobin and fibrinogen) was found predominantly related to interplay between innate and adaptive immune cell. CSF sTNFR1 pathway (encompassing high levels of CXCL13, TWEAK, LIGHT, IL35, osteopontin, pentraxin-3, sCD163 and chitinase-3-L1) was mainly related to dendritic cell maturation and acute immune response pathway. Finally, CSF sTNFR2 pathway (encompassing high CSF levels of IFN-B, IFN-L2, sIL-6Ra) was linked to Th cell differentiation and regulatory cytokine pathway.
Conclusions
Conclusions: CSF dysregulation of TNF and TNFRs pathways can be early identified in MS patients at time of diagnosis and associates with a specific clinical/MRI profile. This, in turn, could have a key role in predicting the disease outcome.