Additional file 1: Figure S1. of Early strong intrathecal inflammation in cerebellar type multiple system atrophy by cerebrospinal fluid cytokine/chemokine profiles: a case control study

A schematic drawing indicating measurement of each part of the hindbrain. 1. Vertical diameter of vermis. 2. Anteroposterior diameter of vermis. 3. Anteroposterior diameter of pontine base. 4. Anteroposterior diameter of medulla oblongata. Figure S2. Clustering of correlations between each CSF cytokine level in SCA patients using the same order of cytokines as in MSA-C. Color codes indicate R values of correlations calculated using Pearson’s correlation coefficient. * p < 0.05, ** p < 0.01. CSF: cerebrospinal fluid, SCA: spinocerebellar ataxia, MSA-C: multiple system atrophy cerebellar-type. Figure S3. Heatmap analysis of correlations between cytokine levels and disease duration in SCA patients. R values of Pearson’s correlation coefficient analysis are divided into quintiles, and p-values calculated using one-way ANOVA are indicated as a heatmap. There were no significant correlations between cytokines and disease duration in SCA patients. SCA: spinocerebellar ataxia. Figure S4. Heatmap analysis of correlations between cytokine levels and MRI measurement in SCA. R-values of Pearson’s correlation coefficient analysis are divided into quintiles, and p-values calculated using one-way ANOVA are indicated as a heatmap. Among the 27 cytokines studied, only PDGF, IL-12(p70), GM-CSF, and MIP-1α showed significant negative correlations with MRI measurements. GM-CSF: granulocyte-macrophage colony-stimulating factor, IL: interleukin, MIP: macrophage inflammatory protein, MRI: magnetic resonance imaging, PDGF: platelet-derived growth factor, SCA: spinocerebellar ataxia. Table S1. Detection rates of cytokines/chemokines and growth factors in cerebrospinal fluid. Table S2. Summary of dysregulated cerebrospinal fluid cytokines (PDF 188 kb

Comparisons of the phenotype frequencies of the <i>HLA-DRB1</i> alleles.

<p>*^aCompared with HCs, p^corr = 0.0196, OR  = 2.217, 95% CI  = 1.389–3.539.</p><p>*^bCompared with HCs, p^corr = 0.0056, OR  = 3.660, 95% CI  = 1.802–7.431.</p><p>*^cCompared with HCs, p^corr = 0.0084, OR  = 0.279, 95% CI  = 0.135–0.575.</p><p>*^dCompared with HCs, p^corr = 0.0392, OR  = 2.624, 95% CI 1.432–4.809.</p><p>p^uncorr was corrected by multiplying the value by 28 to calculate p^corr.</p><p>CI, confidence interval; CSF, cerebrospinal fluid; HCs, healthy controls; MS, multiple sclerosis; OR, odds ratio; p^corr, corrected p value.</p

Proportions of patients with CSF IgG abnormality by year of birth.

<p>Among the MS patients, the proportion of patients with CSF IgG abnormality did not change significantly with advancing year of birth. CSF, cerebrospinal fluid; MS, multiple sclerosis.</p

Comparisons of the frequencies of antibodies against common infectious agents.

<p>*p = 0.0119, compared with IgG abnormality (−) MS patients.</p><p>**p = 0.0451, compared with HCs.</p><p>***p = 0.0474, compared with HCs.</p><p>*^,**^,***Significant difference between the linked values (p<0.05).</p><p>The age of the patients during examination did not differ significantly among HCs and MS patients, regardless of the presence or absence of IgG abnormality (mean ± SD in years: 37.21±12.54 for MS; 36.19±11.36 for IgG abnormality-positive MS; 39.00±14.39 for IgG abnormality-negative MS; and 38.93±12.11 for HCs).</p><p>CSF, cerebrospinal fluid; EBV, Epstein-Barr virus; HCs, healthy controls; MS, multiple sclerosis; p^corr, corrected p value; VZV, varicella zoster virus.</p

Correlation of cytokine and chemokine levels in patients with RRMS and NMO/NMOSD in the relapse phase.

<p>Among the cytokines and chemokines analyzed, distances of each pair of cytokines/chemokines, based on Spearman's correlation coefficient, of RRMS and NMO/NMOSD in the relapse phase were shown as a heatmap. NMO = neuromyelitis optica; NMOSD = neuromyelitis optica spectrum disorder; RRMS = relapsing remitting multiple sclerosis.</p

Comparisons of the demographic features of MS patients according to the CSF IgG abnormality status.

a<p>Values represent the mean ± SD.</p>b<p>Brain MRI lesions meeting the Barkhof criteria <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0095367#pone.0095367-Barkhof1" target="_blank">[34]</a>.</p>c<p>Opticospinal form of MS with short spinal cord lesions extending less than three vertebral segments.</p><p>CSF, cerebrospinal fluid; EDSS, Kurtzke's Expanded Disability Status Scale; MS, multiple sclerosis.</p

Proportions of patients with <i>Helicobacter pylori</i> infection by year of birth.

<p>Among the MS patients, the proportion of patients with <i>H. pylori</i> decreased markedly in those born after 1965. MS, multiple sclerosis.</p

Demographic features of patients.

<p>NA = not applicable; NMO = neuromyelitis optica; NMOSD = neuromyelitis optica spectrum disorder; OND = other non-inflammatory neurological diseases; PPMS = primary progressive multiple sclerosis; RRMS = relapsing remitting multiple sclerosis; CSF = cerebrospinal fluid; SD = standard deviation; EDSS = Expanded Disability Status Scale.</p>*, #<p>p<0.05.</p

Correlations of cytokines/chemokines in RRMS and NMO/NMOSD in the relapse phase.

<p>NMO = neuromyelitis optica; NMOSD = neuromyelitis optica spectrum disorder; OND = other non-inflammatory neurological diseases; RRMS = relapsing remitting multiple sclerosis. rho = Spearman's correlation coefficient, <i>^corrp</i> = corrected p value by Benjamini-Hochberg method.</p

Genetic and Infectious Profiles of Japanese Multiple Sclerosis Patients

<div><h3>Background</h3><p>Nationwide surveys conducted in Japan over the past thirty years have revealed a four-fold increase in the estimated number of multiple sclerosis (MS) patients, a decrease in the age at onset, and successive increases in patients with conventional MS, which shows an involvement of multiple sites in the central nervous system, including the cerebrum and cerebellum. We aimed to clarify whether genetic and infectious backgrounds correlate to distinct disease phenotypes of MS in Japanese patients.</p> <h3>Methodology/Principal Findings</h3><p>We analyzed <em>HLA-DRB1</em> and <em>-DPB1</em> alleles, and IgG antibodies specific for <em>Helicobacter pylori</em>, <em>Chlamydia pneumoniae</em>, varicella zoster virus, and Epstein-Barr virus nuclear antigen (EBNA) in 145 MS patients and 367 healthy controls (HCs). Frequencies of <em>DRB1*0405</em> and <em>DPB1*0301</em> were significantly higher, and <em>DRB1*0901</em> and <em>DPB1*0401</em> significantly lower, in MS patients as compared with HCs. MS patients with <em>DRB1*0405</em> had a significantly earlier age of onset and lower Progression Index than patients without this allele. The proportion and absolute number of patients with <em>DRB1*0405</em> successively increased with advancing year of birth. In MS patients without <em>DRB1*0405</em>, the frequency of the <em>DRB1*1501</em> allele was significantly higher, while the <em>DRB1*0901</em> allele was significantly lower, compared with HCs. Furthermore, <em>DRB1*0405</em>-negative MS patients were significantly more likely to be positive for EBNA antibodies compared with HCs.</p> <h3>Conclusions</h3><p>Our study suggests that MS patients harboring <em>DRB1*0405</em>, a genetic risk factor for MS in the Japanese population, have a younger age at onset and a relatively benign disease course, while <em>DRB1*0405</em>-negative MS patients have features similar to Western-type MS in terms of association with Epstein-Barr virus infection and <em>DRB1*1501</em>. The recent increase of MS in young Japanese people may be caused, in part, by an increase in <em>DRB1*0405</em>-positive MS patients.</p> </div