Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Takuya Matsushita Last modified date:2019.06.28

Lecturer / Neurology / Kyushu University Hospital


Papers
1. , James F. Howard, Kimiaki Utsugisawa, Michael Benatar, Hiroyuki Murai, Richard J. Barohn, Isabel Illa, Saiju Jacob, John Vissing, Ted M. Burns, John T. Kissel, Srikanth Muppidi, Richard J. Nowak, Fanny O'Brien, Jing Jing Wang, Renato Mantegazza, Claudio Gabriel Mazia, Miguel Wilken, Carolina Ortea, Juliet Saba, Marcelo Rugiero, Mariela Bettini, Gonzalo Vidal, Alejandra Dalila Garcia, Phillipa Lamont, Wai Kuen Leong, Heidi Boterhoven, Beverly Fyfe, Leslie Roberts, Mahi Jasinarachchi, Natasha Willlems, Julia Wanschitz, Wolfgang Löscher, Jan De Bleecker, Guy Van den Abeele, Kathy de Koning, Katrien De Mey, Rudy Mercelis, Linda Wagemaekers, Delphine Mahieu, Philip Van Damme, Charlotte Smetcoren, Olivier Stevens, Sarah Verjans, Ann D'Hondt, Petra Tilkin, Alzira Alves de Siqueira Carvalho, Katsuhisa Masaki, Dai Matsuse, Takuya Matsushita, Taira Uehara, Safety and efficacy of eculizumab in anti-acetylcholine receptor antibody-positive refractory generalised myasthenia gravis (REGAIN)
a phase 3, randomised, double-blind, placebo-controlled, multicentre study, The Lancet Neurology, 10.1016/S1474-4422(17)30369-1, 16, 12, 976-986, 2017.12, Background Complement is likely to have a role in refractory generalised myasthenia gravis, but no approved therapies specifically target this system. Results from a phase 2 study suggested that eculizumab, a terminal complement inhibitor, produced clinically meaningful improvements in patients with anti-acetylcholine receptor antibody-positive refractory generalised myasthenia gravis. We further assessed the efficacy and safety of eculizumab in this patient population in a phase 3 trial. Methods We did a phase 3, randomised, double-blind, placebo-controlled, multicentre study (REGAIN) in 76 hospitals and specialised clinics in 17 countries across North America, Latin America, Europe, and Asia. Eligible patients were aged at least 18 years, with a Myasthenia Gravis-Activities of Daily Living (MG-ADL) score of 6 or more, Myasthenia Gravis Foundation of America (MGFA) class II–IV disease, vaccination against Neisseria meningitides, and previous treatment with at least two immunosuppressive therapies or one immunosuppressive therapy and chronic intravenous immunoglobulin or plasma exchange for 12 months without symptom control. Patients with a history of thymoma or thymic neoplasms, thymectomy within 12 months before screening, or use of intravenous immunoglobulin or plasma exchange within 4 weeks before randomisation, or rituximab within 6 months before screening, were excluded. We randomly assigned participants (1:1) to either intravenous eculizumab or intravenous matched placebo for 26 weeks. Dosing for eculizumab was 900 mg on day 1 and at weeks 1, 2, and 3; 1200 mg at week 4; and 1200 mg given every second week thereafter as maintenance dosing. Randomisation was done centrally with an interactive voice or web-response system with patients stratified to one of four groups based on MGFA disease classification. Where possible, patients were maintained on existing myasthenia gravis therapies and rescue medication was allowed at the study physician's discretion. Patients, investigators, staff, and outcome assessors were masked to treatment assignment. The primary efficacy endpoint was the change from baseline to week 26 in MG-ADL total score measured by worst-rank ANCOVA. The efficacy population set was defined as all patients randomly assigned to treatment groups who received at least one dose of study drug, had a valid baseline MG-ADL assessment, and at least one post-baseline MG-ADL assessment. The safety analyses included all randomly assigned patients who received eculizumab or placebo. This trial is registered with ClinicalTrials.gov, number NCT01997229. Findings Between April 30, 2014, and Feb 19, 2016, we randomly assigned and treated 125 patients, 62 with eculizumab and 63 with placebo. The primary analysis showed no significant difference between eculizumab and placebo (least-squares mean rank 56·6 [SEM 4·5] vs 68·3 [4·5]; rank-based treatment difference −11·7, 95% CI −24·3 to 0·96; p=0·0698). No deaths or cases of meningococcal infection occurred during the study. The most common adverse events in both groups were headache and upper respiratory tract infection (ten [16%] for both events in the eculizumab group and 12 [19%] for both in the placebo group). Myasthenia gravis exacerbations were reported by six (10%) patients in the eculizumab group and 15 (24%) in the placebo group. Six (10%) patients in the eculizumab group and 12 (19%) in the placebo group required rescue therapy. Interpretation The change in the MG-ADL score was not statistically significant between eculizumab and placebo, as measured by the worst-rank analysis. Eculizumab was well tolerated. The use of a worst-rank analytical approach proved to be an important limitation of this study since the secondary and sensitivity analyses results were inconsistent with the primary endpoint result; further research into the role of complement is needed. Funding Alexion Pharmaceuticals..
2. Takayuki Fujii, Ryo Yamasaki, Kyoko Iinuma, Daisuke Tsuchimoto, Yoshinori Hayashi, Ban yu Saitoh, Takuya Matsushita, Mizuho A. Kido, Shinichi Aishima, Hiroshi Nakanishi, Yusaku Nakabeppu, Jun ichi Kira, A Novel Autoantibody against Plexin D1 in Patients with Neuropathic Pain, Annals of Neurology, 10.1002/ana.25279, 84, 2, 208-224, 2018.08, Objective: To identify novel autoantibodies for neuropathic pain (NeP). Methods: We screened autoantibodies that selectively bind to mouse unmyelinated C-fiber type dorsal root ganglion (DRG) neurons using tissue-based indirect immunofluorescence assays (IFA) with sera from 110 NeP patients with various inflammatory and allergic neurologic diseases or other neuropathies, and 50 controls without NeP including 20 healthy subjects and 30 patients with neurodegenerative diseases or systemic inflammatory diseases. IgG purified from IFA-positive patients' sera was subjected to Western blotting (WB) and immunoprecipitation (IP) using mouse DRG lysates. Immunoprecipitates were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify target autoantigens. Results: Antiunmyelinated C-fiber type DRG neuron antibodies were more frequent in patients with NeP than non-NeP subjects (10% vs 0%; p < 0.05). These autoantibodies were all from the IgG2 subclass and colocalized mostly with isolectin B4- and P2X3-positive pain-conducting small neurons but not with S100β-positive myelinated neurons. WB revealed a common immunoreactive band (approximately 220kDa). IP and LC-MS/MS studies identified plexin D1 as a target autoantigen. Immunoadsorption tests with recombinant human plexin D1 in IFA revealed that all 11 anti–small DRG neuron antibody-positive patients had anti–plexin D1 antibodies. Application of anti–plexin D1 antibody-positive patient sera to cultured DRG neurons increased membrane permeability, leading to cellular swelling. NeP patients with anti–plexin D1 antibodies commonly developed burning pain and current perception threshold abnormalities for C-fibers. Main comorbidities were atopy and collagen-vascular disease. Immunotherapies ameliorated NeP in 7 treated cases. Interpretation: Anti–plexin D1 antibodies are a novel biomarker for immunotherapy-responsive NeP..
3. yuri nakamura, Laura Gaetano, Takuya Matsushita, Altermatt Anna, Till Sprenger, Ernst Wilhelm Radue, Jens Wuerfel, Lorena Bauer, Michael Amann, Koji Shinoda, Noriko Isobe, Ryo Yamasaki, Takahiko Saida, Ludwig Kappos, Jun-Ichi Kira, A comparison of brain magnetic resonance imaging lesions in multiple sclerosis by race with reference to disability progression, Journal of Neuroinflammation, 10.1186/s12974-018-1295-1, 15, 1, 2018.09, Background: We compared the magnetic resonance imaging (MRI) features between Japanese and Caucasian patients with multiple sclerosis (MS), and identified the relationships between MRI features and disability. Methods: From the baseline data of phase II fingolimod trials, 95 Japanese and 246 Caucasian relapsing-remitting MS patients were enrolled. The number, volume, and distribution of brain MRI lesions were evaluated using T2-weighted (T2W) images. Cross-sectional total normalized brain volume (NBV), normalized cortical gray matter volume, normalized deep gray matter volume (NDGMV), normalized white matter volume (NWMV), and normalized thalamic volume were measured. Results: Japanese patients had significantly lower Expanded Disability Status Scale (EDSS) scores than Caucasian patients (mean 2.0 vs. 2.3, p = 0.008), despite a similar disease duration. Japanese patients showed a trend towards fewer T2W-lesions (median 50 vs. 65, p = 0.08) and significantly lower frequencies of cerebellar and parietal lobe lesions (p = 0.02 for both) than Caucasian patients. There were no differences in T2W-lesion volume between races, whereas Japanese patients had a significantly larger T2W-lesion volume per lesion compared with Caucasian patients (median 140 mm3 vs. 85 mm3, p < 0.0001). T2W-lesion volumes were positively correlated with EDSS scores in Japanese patients (p < 0.0001). In both races, NBV, normalized cortical gray matter volume, NDGMV, and thalamic volume were negatively correlated with disease duration and EDSS scores (p < 0.01 for all). NWMV was negatively correlated with disease duration and EDSS scores only in Caucasian patients (p = 0.03 and p = 0.004, respectively). NBV, NDGMV, NWMV, and thalamic volume were consistently smaller in Japanese compared with Caucasian patients throughout the entire examined disease duration (p = 0.046, p = 0.01, p = 0.005, and p = 0.04, respectively). Japanese patients had a significantly faster reduction in NDGMV (p = 0.001), particularly for thalamic volume (p = 0.001), with disease duration compared with Caucasian patients. Conclusions: Gray matter atrophy is a common denominator for disability in Japanese and Caucasian patients. Additional contributory factors for disability include T2W-lesion volume in Japanese patients and white matter atrophy in Caucasian patients. Less frequent parietal and cerebellar involvement with fewer T2W-lesions may underlie milder disability in Japanese patients..
4. Yuko Kobayakawa, Katsuhisa Masaki, Ryo Yamasaki, Wataru Shiraishi, Shotaro Hayashida, Shintaro Hayashi, Koichi Okamoto, Takuya Matsushita, Jun-Ichi Kira, Downregulation of neuronal and dendritic Connexin36-made electrical synapses without glutamatergic axon terminals in spinal anterior horn cells from the early stage of amyotrophic lateral sclerosis, Frontiers in Neuroscience, 10.3389/fnins.2018.00894, 12, NOV, 2018.11, Connexin36 (Cx36) forms gap junctions between neurons, which are called electrical synapses, enabling adjacent neurons to communicate directly. The participation of chemical synapses in neurodegeneration in amyotrophic lateral sclerosis (ALS) has long been indicated, but it remains unclear whether electrical synapses are involved in the pathogenesis of ALS. We performed extensive immunopathological analyses using mutant superoxide dismutase 1 (SOD1G93A) transgenic mice and their littermates to investigate whether Cx36-made electrical synapses are affected in motor neuron diseases. We found that in the lamina IX of the lumbar spinal cord from wild type mice, about half of the Cx36 puncta existed independently of chemical synapse markers, while the rest coexisted with chemical synapse markers, such as vesicular glutamate transporter 1 (VGLUT1), which is a glutamatergic axon terminal marker, and/or glutamate decarboxylase 65 (GAD65), which is a GABAergic axon terminal marker. Cx36 single or Cx36/GAD65 double positive puncta, but not VGLUT1-containing puncta, were preferentially decreased on neuronal and dendritic surfaces of the anterior horn cells in the early stage of SOD1G93A ALS mice. Moreover, in five human autopsied sporadic ALS cases with bulbar or upper limb onset, Cx36 immunoreactivity was diminished in the proximal dendrites and neuropils of well-preserved large motor neurons in the lumbar anterior horns. These findings suggest that downregulation of neuronal and dendritic Cx36 in the spinal anterior horns commonly occurs from the early stage of hereditary and sporadic ALS. Cx36-made electrical synapses without glutamatergic signaling appear to be more vulnerable than other chemical synapses and electrical synapses with glutamatergic signaling in the early stage of motor neuron degeneration, suggesting involvement of Cx36-made electrical synapses in the pathogenesis of human ALS..
5. Yu Hashimoto, Hidenori Ogata, Ryo Yamasaki, Takakazu Sasaguri, Senri Ko, kenichiro yamashita, Zhang Xu, Takuya Matsushita, Takahisa Tateishi, Shin'Ichi Akiyama, Shoichi Maruyama, Akifumi Yamamoto, Jun-Ichi Kira, Chronic inflammatory demyelinating polyneuropathy with concurrent membranous nephropathy
An anti-paranode and podocyte protein antibody study and literature survey, Frontiers in Neurology, 10.3389/fneur.2018.00997, 9, NOV, 2018.11, Background: Several case reports have described the concurrence of chronic inflammatory demyelinating polyneuropathy (CIDP) and membranous nephropathy (MN). The presence of autoantibodies against podocyte antigens phospholipase A2 receptor (PLA2R) and thrombospondin type 1 domain containing 7A (THSD7A) in MN suggests an autoimmune mechanism. Some CIDP patients also harbor autoantibodies against paranodal proteins such as neurofascin 155 (NF155) and contactin-1 (CNTN1). We investigated the relationship between CIDP and MN by assaying autoantibodies against paranodal and podocyte antigens in a CIDP patient with MN, and by a literature survey on the clinical features of CIDP with MN. Methods: Anti-CNTN1 and NF155 antibodies were measured by flow cytometry using HEK293 cell lines stably expressing human CNTN1 or NF155. Binding capacity of antibodies was validated by immunostaining mouse teased sciatic nerve fibers. Anti-PLA2R antibodies were measured by enzyme-linked sorbent assay and anti-THSD7A antibodies by indirect immunofluorescence assay. Clinical features between 14 CIDP with MN cases including two with anti-CNTN1 antibodies and 20 anti-CNTN1 antibody-positive CIDP cases were compared. Results: A patient whose ages was in the late 70 s complained of progressive weakness and superficial and deep sensory impairment in four extremities over 6 months. Nerve conduction studies showed prominent demyelination patterns. The patient presented with nephrotic syndrome. Renal biopsy disclosed basement membrane thickening with local subepithelial projections and glomerular deposits of IgG4, compatible with MN. Autoantibody assays revealed the presence of IgG4 and IgG1 anti-CNTN1 antibodies, but an absence of anti-NF155, anti-PLA2R, and anti-THSD7A antibodies. The patient's serum stained paranodes of teased sciatic nerves. CIDP with MN and anti-CNTN1 antibody-positive CIDP commonly showed male preponderance, relatively higher age of onset, acute to subacute onset in 35-50% of cases, distal dominant sensorimotor neuropathy, proprioceptive impairment leading to sensory ataxia, and very high cerebrospinal fluid protein levels. However, 11 of 13 CIDP patients with MN had a favorable response to mono- or combined immunotherapies whereas anti-CNTN1 antibody-positive CIDP was frequently refractory to corticosteroids and intravenous immunoglobulin administration. Conclusion: CIDP with MN and anti-CNTN1 antibody-positive CIDP show considerable overlap but are not identical. CIDP with MN is probably heterogeneous and some cases harbor anti-CNTN1 antibodies..
6. kenichiro yamashita, Taira Uehara, Pukovisa Prawiroharjo, Koji Yamashita, Osamu Togao, Hiwatashi Akio, Yoshihide Taniwaki, Hidetsuna Utsunomiya, Takuya Matsushita, Ryo Yamasaki, Jun-Ichi Kira, Functional connectivity change between posterior cingulate cortex and ventral attention network relates to the impairment of orientation for time in Alzheimer’s disease patients, Brain Imaging and Behavior, 10.1007/s11682-018-9860-x, 13, 1, 154-161, 2019.02, Alzheimer’s disease (AD) patients exhibit various cognitive dysfunctions, including impairment of orientation for time (OT). The brain regions underlying OT impairment remain to be elucidated. A previous single-photon emission computed tomography study has indicated hypoperfusion of the posterior cingulate cortex (PCC) in relation to deterioration of OT. In this study, we investigated whole brain functional connectivity changes of PCC using resting-state functional magnetic resonance imaging. Voxel-based functional connectivity with PCC was analyzed in OT-poor or OT-good AD patients, classified according to the mean OT scores of the Mini-Mental State Examination subscale. The connectivities of dorsal frontal lobe, and lateral parietal and lateral temporal lobes with PCC in the right hemisphere were reduced in the OT-poor AD group compared with the OT-good AD group. A subtraction connectivity map of OT score differences (OT-good minus OT-poor) revealed the right middle temporal gyrus near the temporo-parietal junction as a significantly connected region with PCC. These results suggest that the right posterior part of the middle temporal gyrus may play an important role in OT in conjunction with PCC, and that disconnection between PCC and the right ventral attention network may cause OT disturbance in AD patients..
7. Noriko Isobe, Toshikazu Baba, Yuri Nakamura, Koji Shinoda, mitsuru watanabe, Takuya Matsushita, Jun-Ichi Kira, Case of central nervous system inflammatory disease in late pregnancy, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12503, 10, S1, 54-58, 2019.03, Background: During pregnancy, dynamic changes occur not only in coagulation, but also in immune responses. We report a case of a patient who developed an inflammatory disease in the central nervous system during late pregnancy and was challenging to diagnose. Case presentation: A 27-year-old woman experienced dysarthria during pregnancy at approximately 33 gestational weeks. From the 38th week of pregnancy, she developed general fatigue, clumsiness of the left hand and numbness of the right side of the body. After delivery of her child at 41 weeks’ gestation, she was moved to the neurology ward for further investigation. She showed limb ataxia of the left extremities and sensory disturbance on the right side of the body, including the face. Magnetic resonance imaging detected multiple gadolinium-enhanced lesions at the brainstem and cerebrum. High-dose intravenous methylprednisolone pulse therapy was effective, but her symptoms still remained, including palatal myoclonus. Diagnosing this patient was challenging, but clinically isolated syndrome or multiple sclerosis and neuro-Behҫet's disease were the main candidates. Conclusions: We present a case of a pregnant woman who developed brain inflammatory lesions with unknown etiology. Longitudinal follow up is mandatory for diagnosis with careful tapering of oral prednisolone..
8. Yiwen Cui, Katsuhisa Masaki, Xu Zhang, Ryo Yamasaki, Takayuki Fujii, Hidenori Ogata, Shotaro Hayashida, Hiroo Yamaguchi, Fuminori Hyodo, Hinako Eto, Sachiko Koyama, Kyoko Iinuma, Tomomi Yonekawa, Takuya Matsushita, Mari Yoshida, Kazunori Yamada, Mitsuhiro Kawano, Marie Malissen, Bernard Malissen, Jun-Ichi Kira, A novel model for treatment of hypertrophic pachymeningitis, Annals of Clinical and Translational Neurology, 10.1002/acn3.715, 6, 3, 431-444, 2019.03, Objective: Immunoglobulin (Ig)G4-related disease is a major cause of hypertrophic pachymeningitis (HP), presenting as a progressive thickening of the dura mater. HP lacks an animal model to determine its underlying mechanisms. We developed a suitable animal model for the treatment of HP. Methods: We longitudinally evaluated dura in mice with a mutation (Y136F) in the linker for activation of T cells (LAT), which induced type 2 T helper (Th2) cell proliferation and IgG1 (IgG4 human equivalent) overexpression. Mice were therapeutically administered daily oral irbesartan from 3 to 6 weeks of age. Human IgG4-related, anti-neutrophil cytoplasmic antibody-related, and idiopathic HP dura were also immunohistochemically examined. Results: LATY136F mice showing dural gadolinium enhancement on magnetic resonance imaging had massive infiltration of B220+ B cells, IgG1+ cells, CD138+ plasma cells, CD3+ T cells, F4/80+ macrophages, and polymorphonuclear leukocytes in the dura at 3 weeks of age, followed by marked fibrotic thickening. In dural lesions, transforming growth factor (TGF)-β1 was produced preferentially in B cells and macrophages while TGF-β receptor I (TGF-βRI) was markedly upregulated on fibroblasts. Quantitative western blotting revealed significant upregulation of TGF-β1, TGF-βRI, and phosphorylated SMAD2/SMAD3 in dura of LATY136F mice aged 13 weeks. A similar upregulation of TGF-βRI, SMAD2/SMAD3, and phosphorylated SMAD2/SMAD3 was present in autopsied dura of all three types of human HP. Irbesartan abolished dural inflammatory cell infiltration and fibrotic thickening in all treated LATY136F mice with reduced TGF-β1 and nonphosphorylated and phosphorylated SMAD2/SMAD3. Interpretation: TGF-β1/SMAD2/SMAD3 pathway is critical in HP and is a potential novel therapeutic target..
9. Xu Zhang, Takayuki Fujii, Hidenori Ogata, Ryo Yamasaki, Katsuhisa Masaki, Yiwen Cui, Takuya Matsushita, Noriko Isobe, Jun-Ichi Kira, Cerebrospinal fluid cytokine/chemokine/growth factor profiles in idiopathic hypertrophic pachymeningitis, Journal of Neuroimmunology, 10.1016/j.jneuroim.2019.01.010, 330, 38-43, 2019.05, Hypertrophic pachymeningitis (HP) is a rare neurologic disease causing inflammatory fibrous thickening of the brain and spinal dura mater. We investigated the cerebrospinal fluid cytokine profile of HP by measuring 28 cytokines/chemokines/growth factors with a multiplexed fluorescent immunoassay in 8 patients with HP (6 idiopathic, 1 IgG4-related, 1 anti-neutrophil cytoplasmic antibody-related), and 11 with other non-inflammatory neurologic diseases (OND). Interleukin (IL)-4, IL-5, IL-9, IL-10, TNF-α, and CXCL8/IL-8 levels were significantly higher in idiopathic HP (IHP) than OND. Cluster analyses disclosed two major clusters: one mainly consisted of IHP and the other of OND, suggesting a unique cytokine profile in IHP..
10. Ban yu Saitoh, Ryo Yamasaki, Hiwatashi Akio, Takuya Matsushita, Shintaro Hayashi, Yoshihiro Mitsunaga, Yasuhiro Maeda, Noriko Isobe, Kunihiro Yoshida, Shu ichi Ikeda, Jun-Ichi Kira, Discriminative clinical and neuroimaging features of motor-predominant hereditary diffuse leukoencephalopathy with axonal spheroids and primary progressive multiple sclerosis
A preliminary cross-sectional study, Multiple Sclerosis and Related Disorders, 10.1016/j.msard.2019.03.008, 31, 22-31, 2019.06, Background: Hereditary diffuse leukoencephalopathy with axonal spheroids (HDLS) is a rare autosomal-dominant white matter disease, typically characterized by juvenile cognitive decline and frontoparietal white matter lesions. A portion of HDLS patients exhibit preferential motor dysfunctions as their initial symptoms, mimicking multiple sclerosis (MS). However, there is no study comparing this phenotype of HDLS and primary progressive multiple sclerosis (PPMS), which greatly resemble each other. This is the first preliminary study to clarify the clinical and neuroimaging features of motor-predominant HDLS, and compare it with PPMS, using cases whose colony stimulating factor 1 receptor (CSF1R) were sequenced. Methods: Clinical and radiological data from Japanese patients at the Department of Neurology, Kyushu University Hospital, Fukuoka, Japan, were evaluated retrospectively and cross-sectionally. Twenty-nine brain and 18 spinal cord magnetic resonance imaging (MRI) scans from four motor-predominant HDLS patients with CSF1R mutations and 15 PPMS patients without CSF1R mutations, were evaluated using an HDLS MRI scoring system. Results: Two patients with HDLS were initially diagnosed with MS and received immunotherapy. Clinically, motor-predominant HDLS and PPMS patients resembled each other in onset age and disability. However, motor-predominant HDLS patients had a significantly higher frequency of frontal release signs, lower positivity rates of oligoclonal IgG bands (OCB), and lower IgG index values. Total HDLS MRI scores, total white matter lesions (WMLs), and brain atrophy were similar between the diseases. However, motor-predominant HDLS patients had more marked atrophy of the corpus callosum (CC) body, more WMLs in the deep and subcortical regions of the frontoparietal lobes, fewer WMLs in the occipitotemporal periventricular regions, and more restricted diffusivity lesions on MRI than PPMS patients. There was a stronger association between disease duration and CC index in HDLS, suggesting more rapid progression compared with PPMS. Conclusions: Motor-predominant HDLS has characteristic frequent frontal release signs, normal findings for OCB and the IgG index, severe CC body atrophy, abundant deep and subcortical WMLs in the frontoparietal lobes, subtle occipitotemporal lobe periventricular WMLs, and more restricted diffusivity lesions on MRI. Although the present study was limited by the small number of HDLS cases, we propose that immunotherapy should be avoided in such cases..
11. Guangrui Li, Ryo Yamasaki, Mei Fang, Katsuhisa Masaki, Hirofumi Ochi, Takuya Matsushita, Jun-Ichi Kira, Novel disease-modifying anti-rheumatic drug iguratimod suppresses chronic experimental autoimmune encephalomyelitis by down-regulating activation of macrophages/microglia through an NF-κB pathway, Scientific Reports, 10.1038/s41598-018-20390-5, 8, 1, 2018.12, We aimed to elucidate the effects of iguratimod, a widely used anti-rheumatic drug with no severe side effects, on chronic experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Iguratimod was orally administered to mice immunised with myelin oligodendrocyte glycoprotein peptide 35-55. Preventive administration of iguratimod from the time of immunisation was found to markedly reduce the clinical severity of acute and chronic EAE. Pathologically, iguratimod treatment significantly reduced demyelination and infiltration of CD3 + T, F4/80 + , and CD169 + cells into the spinal cord, and suppressed macrophage/microglia activation in the parenchyma at the acute and chronic stages compared with vehicle treatment. Therapeutic administration of iguratimod after the onset of clinical symptoms significantly ameliorated the clinical severity of chronic EAE and reduced demyelination, T helper (Th)1/Th17 cell infiltration, macrophage/microglia activation, and nuclear factor (NF)-κB p65 and cyclooxygenase-2 expression in the spinal cord. In vitro, iguratimod treatment inhibited nuclear translocation of NF-κB p65 and down-regulated pro-inflammatory responses in macrophages and microglia. Our results suggest that iguratimod ameliorates acute and chronic EAE by suppressing inflammatory cell infiltration and immune cell activation, partly through inhibition of NF-κB p65, supporting the therapeutic potential of this drug for not only acute, but also chronic MS..
12. Koji Shinoda, Takuya Matsushita, yuri nakamura, Katsuhisa Masaki, Ryo Yamasaki, Jun-Ichi Kira, HLA genotype and cortical lesions
Response to the letter from Spencer et al., Multiple Sclerosis, 10.1177/1352458517734072, 24, 6, 819-820, 2018.05.
13. Guzailiayi Maimaitijiang, Koji Shinoda, yuri nakamura, Katsuhisa Masaki, Takuya Matsushita, Noriko Isobe, Ryo Yamasaki, Yasunobu Yoshikai, Jun-Ichi Kira, Association of decreased percentage of Vδ2+Vγ9+ γδ T cells with disease severity in multiple sclerosis, Frontiers in Immunology, 10.3389/fimmu.2018.00748, 9, APR, 2018.04, We recently reported that deletion-type copy number variations of the T cell receptor (TCR) γ, α, and δ genes greatly enhanced susceptibility to multiple sclerosis (MS). However, the effect of abnormal TCR γδ gene rearrangement on MS pathogenesis remains unknown. In the present study, we aimed to clarify γδ TCR repertoire alterations and their relationship to clinical and immunological parameters in MS patients by comprehensive flow cytometric immunophenotyping. Peripheral blood mononuclear cells obtained from 30 untreated MS patients in remission and 23 age- and sex-matched healthy controls (HCs) were stained for surface markers and intracellular cytokines after stimulation with phorbol 12-myristate 13-acetate and ionomycin, and analyzed by flow cytometry. MS patients showed significantly decreased percentages of Vδ2+ and Vδ2+Vγ9+ cells in γδ T cells (pcorr = 0.0297 and pcorr = 0.0288, respectively) and elevated Vδ1/Vδ2 ratios compared with HCs (p = 0.0033). The percentages of interferon (IFN)-γ+Vδ2+ and interleukin (IL)-17A+IFN-γ+Vδ2+ cells in γδ T cells, as well as IFN-γ+ cells in Vδ2+ γδ T cells, were significantly lower in MS patients than in HCs (pcorr < 0.0009, pcorr = 0.0135, and pcorr = 0.0054, respectively). The percentages of Vδ2+ and Vδ2+Vγ9+ cells in γδ T cells were negatively correlated with both the Expanded Disability Status Scale score (r = -0.5006, p = 0.0048; and r = -0.5040, p = 0.0045, respectively) and Multiple Sclerosis Severity Score (r = -0.4682, p = 0.0091; and r = -0.4706, p = 0.0087, respectively), but not with age at disease onset, disease duration, or annualized relapse rate. In HCs, the percentages of Vδ2+ and Vδ2+Vγ9+ cells of total CD3+ T cells had strong positive correlations with the percentage of CD25+CD127low/- cells in CD4+ T cells (r = 0.7826, p < 0.0001; and r = 0.7848, p < 0.0001, respectively), whereas such correlations were totally absent in MS patients. These findings suggest that decreased Vδ2+Vγ9+ γδ T cells are associated with disability in MS. Therefore, the Vδ1/Vδ2 ratio might be a candidate biomarker for predicting disease severity in MS..
14. Atsushi Fujita, Hidenori Ogata, Ryo Yamasaki, Takuya Matsushita, Jun-Ichi Kira, Parallel fluctuation of anti-neurofascin 155 antibody levels with clinico-electrophysiological findings in patients with chronic inflammatory demyelinating polyradiculoneuropathy, Journal of the Neurological Sciences, 10.1016/j.jns.2017.11.035, 384, 107-112, 2018.01, Background The long-term clinical course and closely related biomarkers in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) with anti-neurofascin 155 (NF155) antibodies remain to be elucidated. Methods We retrospectively studied the longitudinal clinical courses of three Japanese male anti-NF155 antibody-positive CIDP patients. Anti-NF155 antibody levels were measured by flow cytometry using HEK293 cell lines stably expressing human NF155. Results All three patients presented with chronic progressive sensorimotor disturbance, with ages at onset of 16, 26, and 34 years old, and they were followed for 58, 31, and 38 months, respectively, from the onset. All patients had postural tremor and generalized decreased deep tendon reflexes. Peak cerebrospinal fluid protein levels were > 400 mg/dl, and nerve conduction studies (NCS) showed severe demyelination patterns. Combined immunotherapies including intravenous immunoglobulin, plasma exchange, corticosteroids, and other immunosuppressants ameliorated clinical severity and NCS abnormalities, with improvements of > 10 kg in grip strength and at least 20% in F-wave latencies. However, their symptoms exacerbated after the immunotherapies were tapered. Anti-NF155 antibody levels varied in parallel with the clinical and electrophysiological changes, or preceded them. Conclusion The patients’ clinical courses suggest that anti-NF155 antibody levels and NCS findings could be disease activity markers in anti-NF155 antibody-positive CIDP..
15. Yu Hashimoto, Koji Shinoda, Eizo Tanaka, Taira Uehara, Takuya Matsushita, Ryo Yamasaki, Jun-ichi Kira, Re-emergence of a tumefactive demyelinating lesion after initiation of fingolimod therapy, JOURNAL OF THE NEUROLOGICAL SCIENCES, 10.1016/j.jns.2017.06.002, 379, 167-168, 2017.08.
16. Shotaro Hayashida, Katsuhisa Masaki, Tomomi Yonekawa, Satoshi O Suzuki, Hiwatashi Akio, Takuya Matsushita, Mitsuru Watanabe, Ryo Yamasaki, Toshihiko Suenaga, Toru Iwaki, Hiroyuki Murai, Jun-ichi Kira, Early and extensive spinal white matter involvement in neuromyelitis optica, BRAIN PATHOLOGY, 10.1111/bpa.12386, 27, 3, 249-265, 2017.05.
17. Koji Shinoda, Takuya Matsushita, Yuri Nakamura, Katsuhisa Masaki, Ryo Yamasaki, hiroo yamaguchi, Osamu Togao, Akio Hiwatashi, Jun-Ichi Kira, HLA-DRB1*04:05 allele is associated with intracortical lesions on three-dimensional double inversion recovery images in Japanese patients with multiple sclerosis, Multiple Sclerosis, 10.1177/1352458517707067, 1352458517707067, 2017.05, BACKGROUND: Cortical lesions (CLs) frequently observed in Caucasian patients with multiple sclerosis (MS) contribute to disability. However, it remains unclear whether CLs are associated with clinical features and genetic risk factors, such as HLA-DRB1*15:01 and -DRB1*04:05 in Asian MS patients.
OBJECTIVE: To elucidate the frequency of CLs and their association with HLA-DRB1 and DPB1 alleles in Japanese MS patients.
METHODS: Three-dimensional double inversion recovery imaging and clinical information were retrospectively obtained from 92 Japanese MS patients.
RESULTS: CLs of any type, intracortical lesions (ICLs), and leukocortical lesions (LCLs) were detected in 39.1%, 26.1%, and 28.3% of patients, respectively. MS patients with ICLs had a significantly higher frequency of secondary progression and greater Expanded Disability Status Scale (EDSS) scores than those without ICLs. Similar trends were observed with CLs and LCLs. The number of all three lesion types positively correlated with EDSS scores. The frequency and number of ICLs were significantly higher in HLA-DRB1*15:01 carriers than in HLA-DRB1*15:01 non-carriers, but significantly lower in HLA-DRB1*04:05 carriers than in HLA-DRB1*04:05 non-carriers. Multivariate logistic regression analysis revealed a negative association of HLA-DRB1*04:05 with ICLs.
CONCLUSION: ICLs are associated with greater disease severity in Japanese MS patients and are partly suppressed by the HLA-DRB1*04:05 allele..
18. Ryo Yamasaki, Hiroo Yamaguchi, Takuya Matsushita, Takayuki Fujii, Hiwatashi Akio, Jun-ichi Kira, Early strong intrathecal inflammation in cerebellar type multiple system atrophy by cerebrospinal fluid cytokine/chemokine profiles: a case control study, JOURNAL OF NEUROINFLAMMATION, 10.1186/s12974-017-0863-0, 14, 2017.04.
19. Mitsuru Watanabe, Masaki Katsuhisa, Ryo Yamasaki, Jun Kawanoukuchi, Hideyuki Takeuchi, Takuya Matsushita, Akio Suzumura, Jun-ichi Kira, Th1 cells downregulate connexin 43 gap junctions in astrocytes via microglial activation, SCIENTIFIC REPORTS, 10.1038/srep38387, 6, 2016.12.
20. K. Shinoda, T. Iwata, yuri nakamura, K. Masaki, Takuya Matsushita, Ryo Yamasaki, Jun-Ichi Kira, Minocycline-induced human herpesvirus 6 encephalomyelitis with drastically disseminated contrast-enhanced lesions, European Journal of Neurology, 10.1111/ene.13190, 23, 12, e76-e77, 2016.12.
21. Ryo Yamasaki, Takayuki Fujii, Wang Bing, Masaki Katsuhisa, Kido MA, Mari Yoshida, Takuya Matsushita, Jun-ichi Kira, Allergic Inflammation Leads to Neuropathic Pain via Glial Cell Activation, JOURNAL OF NEUROSCIENCE, 10.1523/JNEUROSCI.1981-16.2016, 36, 47, 11929-11945, 2016.11.
22. Yuri Nakamura, Kei-ichiro Takase, Takuya Matsushita, Yoshimura Satoshi, Ryo Yamasaki, Hiroyuki Murai, Kazufumi Kikuchi, Jun-ichi Kira, Recurrent Hemorrhagic Venous Infarctions Caused by Thrombosis of a Pontine Developmental Venous Anomaly and Protein S Mutation, JOURNAL OF STROKE & CEREBROVASCULAR DISEASES, 10.1016/j.jstrokecerebrovasdis.2016.08.040, 25, 11, 2016.11.
23. Zi Ye Song, yuri nakamura, Ryo Yamasaki, Yuji Kawano, Koji Shinoda, Maimaitijiang Guzailiayi, Katsuhisa Masaki, hiroo yamaguchi, Takuya Matsushita, Jun-Ichi Kira, Peripheral blood T cell subset characteristics of multiple sclerosis in remission phase correlate with annualized relapse rates, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12321, 7, 4, 346-352, 2016.11, Objective: A large number of disease-modifying drugs are available for multiple sclerosis (MS); however, there is no established biomarker to predict long-term disease severity and future relapses in MS. We aimed to clarify the alterations in peripheral blood T cell subsets that are associated with MS relapse and disease severity, according to cytokine production profiles in the remission phase. Methods: Blood samples collected from 29 relapsing–remitting MS patients in the remission phase and 21 healthy controls (HC) were analyzed for various cytokine-producing T cell subsets by flow cytometry. Results: MS patients in the remission phase had significantly higher percentages of interleukin (IL)-17+CD4+ T cells, IL-4+CD4+ T cells, IL-9+CD4+ T cells, interferon-γ+CD8+ T cells and IL-4+CD8+ T cells than HC (P = 0.047, P = 0.007, P = 0.026, P = 0.015 and P = 0.007, respectively). In MS, the percentages of IL-9+CD4+ T cells, IL-9+CD8+ T cells and IFN-γ+IL-17+CD8+ T cells showed a significant positive correlation with annualized relapse rates (ARR) (P = 0.011, r = 0.47, P = 0.007, r = 0.49 and P = 0.044, r = 0.38, respectively). Conclusions: In the remission phase of MS, both anti-inflammatory cytokine-producing T cells and pro-inflammatory cytokine-producing T cells are increased; however, only the percentages of pro-inflammatory cytokine-producing T cells, such as IL-9-producing CD4+ T cells, IL-9-producing CD8+ T cells and IFN-γ- and IL-17-producing CD8+ T cells, correlate with ARR. These pro-inflammatory cytokine-producing T cells in the remission phase might be candidate biomarkers for future relapses in MS patients..
24. Jack Antel, Maria Ban, Sergio Baranzini, Lisa Barcellos, Nadia Barizzone, Ashley Beecham, Tone Berge, Bernardinelli Luisa, David Booth, Bos Steffan, Drothea Buck, Mariusz Butkiewicz, Elisabeth G Celius, Manuel Comabella, Alastair Compston, Katarina Dedham, Chris Cotsapas, Sandra Alfonso, Takuya Matsushita, NR1H3 p.Arg415Gln Is Not Associated to Multiple Sclerosis Risk, NEURON, 10.1016/j.neuron.2016.09.052, 92, 2, 333-335, 2016.10.
25. Yuri Nakamura, Takuya Matsushita, Shinya Sato, Masaaki Niino, Toshiyuki Fukazawa, Yoshimura Satoshi, Shin Hisahara, Noriko Isobe, Shun Shimohama, Mitsuru Watanabe, Kazuto Yoshida, Hideki Houzen, Yusei Miyazaki, Ryo Yamasaki, Seiji Kikuchi, Jun-ichi Kira, Latitude and HLA-DRB1*04:05 independently influence disease severity in Japanese multiple sclerosis: a cross-sectional study, JOURNAL OF NEUROINFLAMMATION, 10.1186/s12974-016-0695-3, 13, 2016.09.
26. Ryo Yamasaki, Takuya Matsushita, Toshiyuki Fukazawa, Kazumasa Yokoyama, Kazuo Fujihara, Meiko Ogino, Takanori Yokota, Katsuichi Miyamoto, Masaaki Niino, Kyouichi Nomura, Ryo Tomioka, Masami Tanaka, Kawachi Izumi, Takashi Ohashi, Takashi Kanda, Ken-ichi Kaida, Makoto Matsui, Yuji Nakatsuji, Hirofumi Ochi, Efficacy of intravenous methylprednisolone pulse therapy in patients with multiple sclerosis and neuromyelitis optica, MULTIPLE SCLEROSIS JOURNAL, 10.1177/1352458515617248, 22, 10, 1337-1348, 2016.09.
27. Satoshi Nagata, Koji Shinoda, Taira Uehara, Takuya Matsushita, Ryo Yamasaki, Jun-ichi Kira, Clear detection of lower medullary lesions by three-dimensional double inversion recovery imaging in a patient with neuromyelitis optica spectrum disorder, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12330, 7, 355-356, 2016.07.
28. Kazumasa Saigoh, Yoshimura Satoshi, Tomomi Izumikawa, Shinji Miyata, Yasuharu Tabara, Takuya Matsushita, Tetsuro Miki, Katsuichi Miyamoto, Makito Hirano, Hiroshi Kitagawa, Jun-ichi Kira, Susumu Kusunoki, Chondroitin sulfate beta-1,4-N-acetylgalactosaminyltransferase-1 (ChGn-1) polymorphism: Association with progression of multiple sclerosis, NEUROSCIENCE RESEARCH, 10.1016/j.neures.2016.01.002, 108, 55-59, 2016.07.
29. Hidenori Ogata, Dai Matsuse, Ryo Yamasaki, Nobutoshi Kawamura, Takuya Matsushita, Tomomi Yonekawa, Makoto Hirotani, Hiroyuki Murai, Jun-ichi Kira, A nationwide survey of combined central and peripheral demyelination in Japan, JOURNAL OF NEUROLOGY NEUROSURGERY AND PSYCHIATRY, 10.1136/jnnp-2014-309831, 87, 1, 29-36, 2016.01.
30. Sudarshini Ramanathan, Shinya Sato, Takuya Matsushita, Katsuhisa Masaki, Ryo Yamasaki, Russell C. Dale, Jun-Ichi Kira, Fabienne Brilot, Antibodies to myelin oligodendrocyte glycoprotein are uncommon in Japanese opticospinal multiple sclerosis, Multiple Sclerosis, 10.1177/1352458515586089, 22, 1, 127-128, 2016.01.
31. Sudarshini Ramanathan, Shinya Sato, Takuya Matsushita, Katsuhisa Masaki, Ryo Yamasaki, Russell C. Dale, Jun-Ichi Kira, Fabienne Brilot, Antibodies to myelin oligodendrocyte glycoprotein are uncommon in Japanese opticospinal multiple sclerosis, Multiple Sclerosis, 10.1177/1352458515586089, 22, 1, 127-128, 2016.01.
32. Pouya Khankhanian, Wendy Cozen, Daniel S. Himmelstein, Lohith Madireddy, Lennox Din, Anke van den Berg, Takuya Matsushita, Sally L. Glaser, Jayaji M. Moré, Karin E. Smedby, Sergio E. Baranzini, Thomas M. Mack, Antoine Lizée, Silvia de Sanjosé, Pierre Antoine Gourraud, Alexandra Nieters, Stephen L. Hauser, Pierluigi Cocco, Marc Maynadié, Lenka Foretová, Anthony Staines, Manon Delahaye-Sourdeix, Dalin Li, Smita Bhatia, Mads Melbye, Kenan Onel, Ruth Jarrett, James D. McKay, Jorge R. Oksenberg, Henrik Hjalgrim, Meta-analysis of genome-wide association studies reveals genetic overlap between hodgkin lymphoma and multiple sclerosis, International Journal of Epidemiology, 10.1093/IJE/DYV364, 45, 3, 728-740, 2016.01, Background: Based on epidemiological commonalities, multiple sclerosis (MS) and Hodgkin lymphoma (HL), two clinically distinct conditions, have long been suspected to be aetiologically related. MS and HL occur in roughly the same age groups, both are associated with Epstein-Barr virus infection and ultraviolet (UV) light exposure, and they cluster mutually in families (though not in individuals). We speculated if in addition to sharing environmental risk factors, MS and HL were also genetically related. Using data from genome-wide association studies (GWAS) of 1816 HL patients, 9772MS patients and 25 255 controls, we therefore investigated the genetic overlap between the two diseases. Methods: From among a common denominator of 404 K single nucleotide polymorphisms (SNPs) studied, we identified SNPs and human leukocyte antigen (HLA) alleles independently associated with both diseases. Next, we assessed the cumulative genome- wide effect of MS-associated SNPs on HL and of HL-associated SNPs on MS. To provide an interpretational frame of reference, we used data from published GWAS to create a genetic network of diseases within which we analysed proximity of HL and MS to autoimmune diseases and haematological and non-haematological malignancies. Results: SNP analyses revealed genome-wide overlap between HL and MS, most prominently in the HLA region. Polygenic HL risk scores explained 4.44% of HL risk (Nagelkerke R2), but also 2.36% of MS risk. Conversely, polygenic MS risk scores explained 8.08% of MS risk and 1.94% of HL risk. In the genetic disease network, HL was closer to autoimmune diseases than to solid cancers. Conclusions: HL displays considerable genetic overlap with MS and other autoimmune diseases..
33. Shotaro Hayashida, Katsuhisa Masaki, Takuya Matsushita, mitsuru watanabe, Ryo Yamasaki, Hiroyuki Murai, Jun-Ichi Kira, Holocord involvement with sparing of the peripheral white matter rim in longitudinally extensive spinal cord lesions of neuromyelitis optica, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12274, 6, 78-79, 2015.12.
34. Shinya Sato, Ken Yamamoto, Takuya Matsushita, Noriko Isobe, Yuji Kawano, Kyoko Iinuma, Masaaki Niino, Toshiyuki Fukazawa, yuri nakamura, mitsuru watanabe, Tomomi Yonekawa, Katsuhisa Masaki, Satoshi Yoshimura, Hiroyuki Murai, Ryo Yamasaki, Jun-Ichi Kira, Copy number variations in multiple sclerosis and neuromyelitis optica, Annals of Neurology, 10.1002/ana.24511, 78, 5, 762-774, 2015.11, Objective To clarify the potential association of copy number variations (CNVs) with multiple sclerosis (MS) and neuromyelitis optica (NMO) in Japanese cases. Methods Genome-wide association analyses of CNVs among 277 MS patients, 135 NMO/NMO spectrum disorder (NMOSD) patients, and 288 healthy individuals as a discovery cohort, and among 296 MS patients, 76 NMO/NMOSD patients, and 790 healthy individuals as a replication cohort were performed using high-density single nucleotide polymorphism microarrays. Results A series of discovery and replication studies revealed that most identified CNVs were 5 to 50kb deletions at particular T cell receptor (TCR) gamma and alpha loci regions. Among these CNVs, a TCR gamma locus deletion was found in 16.40% of MS patients (p = 2.44E-40, odds ratio [OR] = 52.6), and deletion at the TCR alpha locus was found in 17.28% of MS patients (p = 1.70E-31, OR = 13.0) and 13.27% of NMO/NMOSD patients (p = 5.79E-20, OR = 54.6). These CNVs were observed in peripheral blood T-cell subsets only, suggesting the CNVs were somatically acquired. NMO/NMOSD patients carrying the CNV tended to be seronegative for anti-aquaporin-4 antibody or had significantly lower titers than those without CNV. Interpretation Deletion-type CNVs at specific TCR loci regions contribute to MS and NMO susceptibility..
35. Hidenori Ogata, Ryo Yamasaki, Hiwatashi Akio, Nobuyuki Oka, Nobutoshi Kawamura, Dai Matsuse, Motoi Kuwahara, Hidekazu Suzuki, Susumu Kusunoki, Yuichi Fujimoto, Koji Ikezoe, Hitaru Kishida, Fumiaki Tanaka, Takuya Matsushita, Hiroyuki Murai, Jun-Ichi Kira, Characterization of IgG4 anti-neurofascin 155 antibody-positive polyneuropathy, Annals of Clinical and Translational Neurology, 10.1002/acn3.248, 2, 10, 960-971, 2015.10, Objective: To investigate anti-neurofascin 155 (NF155) antibody-positive chronic inflammatory demyelinating polyneuropathy (CIDP). Methods: Sera from 50 consecutive CIDP patients diagnosed in our clinic, 32 patients with multiple sclerosis, 40 patients with other neuropathies including 26 with Guillain–Barré syndrome (GBS)/Fisher syndrome, and 30 healthy controls were measured for anti-NF antibodies by flow cytometry using HEK293 cell lines stably expressing human NF155 or NF186. Four additional CIDP patients with anti-NF155 antibodies referred from other clinics were enrolled for clinical characterization. Results: The positivity rate for anti-NF155 antibodies in CIDP patients was 18% (9/50), who all showed a predominance of IgG4 subclass. No other subjects were positive, except one GBS patient harboring IgG1 anti-NF155 antibodies. No anti-NF155 antibody carriers had anti-NF186 antibodies. Anti-NF155 antibody-positive CIDP patients had a significantly younger onset age, higher frequency of drop foot, gait disturbance, tremor and distal acquired demyelinating symmetric phenotype, greater cervical root diameter on magnetic resonance imaging neurography, higher cerebrospinal fluid protein levels, and longer distal and F-wave latencies than anti-NF155 antibody-negative patients. Marked symmetric hypertrophy of cervical and lumbosacral roots/plexuses was present in all anti-NF155 antibody-positive CIDP patients examined by neurography. Biopsied sural nerves from two patients with anti-NF155 antibodies demonstrated subperineurial edema and occasional paranodal demyelination, but no vasculitis, inflammatory cell infiltrates, or onion bulbs. Among anti-NF155 antibody-positive patients, treatment responders more frequently had daily oral corticosteroids and/or immunosuppressants in addition to intravenous immunoglobulins than nonresponders did. Interpretation: Anti-NF155 antibodies occur in a subset of CIDP patients with distal-dominant involvement and symmetric nerve hypertrophy..
36. Takuya Matsushita, Pouya Khankhanian, Lohith Madireddy, Antoine Lizée, Lennox Din, Jayaji M Moré, Pierre-Antoine Gourraud, Stephen L Hauser, Sergio E Baranzini, Jorge R Oksenberg, Genetic contribution to multiple sclerosis
risk among Ashkenazi Jews, BMC medical genetics, 10.1186/s12881-015-0201-2, 16, 1, 55-64, 2015.07, Background: Multiple sclerosis (MS) is an autoimmune disease of the central nervous system, with a strong genetic component. Over 100 genetic loci have been implicated in susceptibility to MS in European populations, the most prominent being the 15:01 allele of the HLA-DRB1 gene. The prevalence of MS is high in European populations including those of Ashkenazi origin, and low in African and Asian populations including those of Jewish origin.
Methods: Here we identified and extracted a total of 213 Ashkenazi MS cases and 546 ethnically matched healthy control individuals from two previous genome-wide case-control association analyses, and 72 trios (affected proband and two unaffected parents) from a previous genome-wide transmission disequilibrium association study, using genetic data to define Ashkenazi. We compared the pattern of genetic risk between Ashkenazi and non-Ashkenazi Europeans. We also sought to identify novel Ashkenazi-specific risk loci by performing association tests on the subset of Ashkenazi cases, controls, probands, and parents from each study.
Results: The HLA-DRB1*15:01 allele and the non-HLA risk alleles were present at relatively low frequencies among Ashkenazi and explained a smaller fraction of the population-level risk when compared to non-Ashkenazi Europeans. Alternative HLA susceptibility alleles were identified in an Ashkenazi-only association study, including HLA-A*68:02 and one or both genes in the HLA-B*38:01-HLA-C*12:03 haplotype. The genome-wide screen in Ashkenazi did not reveal any loci associated with MS risk.
Conclusion: These results suggest that genetic susceptibility to MS in Ashkenazi Jews has not been as well established as that of non-Ashkenazi Europeans. This implies value in studying large well-characterized Ashkenazi populations to accelerate gene discovery in complex genetic diseases.
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37. Noriko Isobe, Lohith Madireddy, Pouya Khankhanian, Takuya Matsushita, Stacy J. Caillier, Jayaji M. Moré, Pierre Antoine Gourraud, Jacob L. McCauley, Ashley H. Beecham, Laura Piccio, Joseph Herbert, Omar Khan, Jeffrey Cohen, Lael Stone, Adam Santaniello, Bruce A.C. Cree, Suna Onengut-Gumuscu, Stephen S. Rich, Stephen L. Hauser, Stephen Sawcer, Jorge R. Oksenberg, An ImmunoChip study of multiple sclerosis risk in African Americans, Brain, 10.1093/brain/awv078, 138, 6, 1518-1530, 2015.06, The aims of this study were: (i) to determine to what degree multiple sclerosis-associated loci discovered in European populations also influence susceptibility in African Americans; (ii) to assess the extent to which the unique linkage disequilibrium patterns in African Americans can contribute to localizing the functionally relevant regions or genes; and (iii) to search for novel African American multiple sclerosis-associated loci. Using the ImmunoChip custom array we genotyped 803 African American cases with multiple sclerosis and 1516 African American control subjects at 130 135 autosomal single nucleotide polymorphisms. We conducted association analysis with rigorous adjustments for population stratification and admixture. Of the 110 non-major histocompatibility complex multiple sclerosis-associated variants identified in Europeans, 96 passed stringent quality control in our African American data set and of these, >70% (69) showed over-representation of the same allele amongst cases, including 21 with nominally significant evidence for association (one-tailed test P<0.05). At a further eight loci we found nominally significant association with an alternate correlated risk-tagging single nucleotide polymorphism from the same region. Outside the regions known to be associated in Europeans, we found seven potentially associated novel candidate multiple sclerosis variants (P<10-4), one of which (rs2702180) also showed nominally significant evidence for association (one-tailed test P = 0.034) in an independent second cohort of 620 African American cases and 1565 control subjects. However, none of these novel associations reached genome-wide significance (combined P = 6.3 × 10-5). Our data demonstrate substantial overlap between African American and European multiple sclerosis variants, indicating common genetic contributions to multiple sclerosis risk..
38. Takuya Matsushita, Lohith Madireddy, T Sprenger, Pouya Khankhanian, S Magon, Y Naegelin, E Caverzasi, R.L.P. Lindberg, L Kappos, Stephen L Hauser, Jorge R Oksenberg, R Henry, D Palletier, Sergio E Baranzini, Genetic associations with brain cortical thickness in multiple sclerosis., Genes, brain, and behavior, 10.1111/gbb.12190, 14, 2, 217-227, 2015.05, Multiple sclerosis (MS) is characterized by temporal and spatial dissemination of demyelinating lesions in the central nervous system. Associated neurodegenerative changes contributing to disability have been recognized even at early disease stages. Recent studies show the importance of gray matter damage for the accrual of clin- ical disability rather than white matter where demyelina- tion is easily visualized by magnetic resonance imaging (MRI). The susceptibility to MS is influenced by genetic risk, but genetic factors associated with the disability are not known. We used MRI data to determine cortical thick- ness in 557 MS cases and 75 controls and in another cohort of 219 cases. We identified nine areas showing different thickness between cases and controls (regions of interest, ROI) (eight of them were negatively corre- lated with Kurtzke’s expanded disability status scale, EDSS) and conducted genome-wide association studies (GWAS) in 464 and 211 cases available from the two data sets. No marker exceeded genome-wide significance in the discovery cohort. We next combined nominal statis- tical evidence of association with physical evidence of interaction from a curated human protein interaction net- work, and searched for subnetworks enriched with nom- inally associated genes and for commonalities between the two data sets. This network-based pathway analysis of GWAS detected gene sets involved in glutamate sig- naling, neural development and an adjustment of intra- cellular calcium concentration. We report here for the first time gene sets associated with cortical thinning of MS. These genes are potentially correlated with disability of MS..
39. Zi Ye Song, Ryo Yamasaki, Yuji Kawano, Shinya Sato, Katsuhisa Masaki, Satoshi Yoshimura, Dai Matsuse, Hiroyuki Murai, Takuya Matsushita, Jun-Ichi Kira, Peripheral blood T cell dynamics predict relapse in multiple sclerosis patients on fingolimod, PLoS One, 10.1371/journal.pone.0124923, 10, 4, 2015.04, Background: Fingolimod efficiently reduces multiple sclerosis (MS) relapse by inhibiting lymphocyte egress from lymph nodes through down-modulation of sphingosine 1-phosphate (S1P) receptors. We aimed to clarify the alterations in peripheral blood T cell subsets associated with MS relapse on fingolimod. Methods/Principal Findings: Blood samples successively collected from 23 relapsing-remitting MS patients before and during fingolimod therapy (0.5 mg/day) for 12 months and 18 healthy controls (HCs) were analysed for T cell subsets by flow cytometry. In MS patients, the percentages of central memory T (CCR7+CD45RO+) cells (TCM) and naïve T (CCR7+CD45RO-) cells decreased significantly, while those of effector memory T (CCR7-CD45RA-) and suppressor precursor T (CD28-) cells increased in both CD4+T and CD8+T cells from 2 weeks to 12 months during fingolimod therapy. The percentages of regulatory T (CD4+CD25highCD127low) cells in CD4+T cells and CCR7-CD45RA+T cells in CD8+T cells also increased significantly. Eight relapsed patients demonstrated greater percentages of CD4+TCM than 15 non-relapsed patients at 3 and 6 months (p=0.0051 and p=0.0088, respectively). The IL17-, IL9-, and IL4-producing CD4+T cell percentages were significantly higher at pre-treatment in MS patients compared with HCs (p<0.01 for all), while the IL17-producing CD4+T cell percentages tended to show a transient increase at 2 weeks of fingolimod therapy (pcorr=0.0834). Conclusions: The CD4+TCM percentages at 2 weeks to 12 months during fingolimod therapy are related to relapse..
40. Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Yuji Kawano, Katsuhisa Masaki, Satoshi Yoshimura, Hiroyuki Murai, Ryo Yamasaki, Jun-Ichi Kira, Distinct features of immunoglobulin G2 aquaporin-4 antibody carriers with neuromyelitis optica, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12179, 6, 2, 154-158, 2015.01, Objective Neuromyelitis optica (NMO) is an inflammatory disease in which NMO-immunoglobulin G (IgG) targeting aquaporin-4 (AQP4) is specifically detected. Although the main subclass of AQP4 antibody was reported to be in the IgG1 subclass, other subclasses have also been described, including IgG2 AQP4 antibody, as a second common subclass. NMO patients were analyzed to clarify the clinical features of NMO patients with IgG2 AQP4 antibody. Methods Serum samples from 58 NMO patients, who met the revised 2006 criteria for NMO, were analyzed for AQP4 antibody subclass expression using an established flow cytometric assay, and clinical features were compared according to the main AQP4 antibody subclasses. Results A total of 50 patients (86.2%) had IgG1 AQP4 antibodies, while eight (13.8%) expressed IgG2 AQP4 antibody as the main subclass. Those eight individuals exhibited younger age of onset (P = 0.0089), lower AQP4 antibody titers (P = 0.0024) and a more common fulfillment of Barkhof's criteria (P = 0.0466) than patients with IgG1 AQP4 antibody expression. Conclusions Results from the present study suggest that the characteristics of individuals with IgG2 AQP4 antibody as a main subclass are more similar to multiple sclerosis and somewhat distinct from NMO patients with IgG1 AQP4 antibody..
41. Masaaki Niino, Shinya Sato, Toshiyuki Fukazawa, Satoshi Yoshimura, Shin Hisahara, Takuya Matsushita, Noriko Isobe, Kazuto Yoshida, Hideki Houzen, Yusei Miyazaki, Shun Shimohama, Seiji Kikuchi, Jun-Ichi Kira, Latitude and HLA-DRB1 alleles independently affect the emergence of cerebrospinal fluid IgG abnormality in multiple sclerosis, Multiple Sclerosis, 10.1177/1352458514560924, 21, 9, 1112-1120, 2015.01, Background: It is unclear whether the prevalence of oligoclonal IgG bands (OCBs) in multiple sclerosis (MS) is different between northern and southern regions of Asia. Objective: This study aimed to compare the prevalence of OCBs and positive cerebrospinal fluid (CSF) findings between northern and southern regions of Japan and to investigate the association of these CSF findings with HLA-DRB1 alleles. Methods: The study included 180 MS patients from Hokkaido (northern Japan) and 184 patients from Kyushu (southern Japan). The IgG index was defined as increased if it was >0.658. Presence of CSF OCBs and/or increased IgG index was defined as positive CSF findings. Results: Positive CSF findings and OCB positivity were significantly higher in MS patients from Hokkaido than in those from Kyushu (p <0.0001 for both). Logistic regression analysis revealed that after adjusting for covariates that can be related to abnormal CSF IgG production, the geographic region (Hokkaido) showed odds ratios (ORs) of 4.08 and 2.57, whereas the HLA-DRB1∗04:05 allele showed ORs of 0.36 and 0.30 for positive CSF findings and OCB positivity, respectively. Conclusions: The results indicate that latitude and HLA-DRB1 alleles independently affect the emergence of CSF IgG abnormalities in Japanese patients with MS..
42. Lili Wang, Takuya Matsushita, Lohith Madireddy, Parvin Mousavi, Sergio E. Baranzini, PINBPA
Cytoscape app for network analysis of GWAS data, Bioinformatics, 10.1093/bioinformatics/btu644, 31, 2, 262-264, 2015.01, Protein interaction network-based pathway analysis (PINBPA) for genome-wide association studies (GWAS) has been developed as a Cytoscape app, to enable analysis of GWAS data in a network fashion. Users can easily import GWAS summary-level data, draw Manhattan plots, define blocks, prioritize genes with random walk with restart, detect enriched subnetworks and test the significance of subnetworks via a user-friendly interface..
43. Satoshi Yoshimura, Noriko Isobe, Takuya Matsushita, Katsuhisa Masaki, Shinya Sato, Yuji Kawano, Hirofumi Ochi, Jun-Ichi Kira, Genetic and infectious profiles influence cerebrospinal fluid IgG abnormality in japanese multiple sclerosis patients, PLoS One, 10.1371/journal.pone.0095367.t001, 9, 4, 2014.04, Background: Abnormal intrathecal synthesis of IgG, reflected by cerebrospinal fluid (CSF) oligoclonal IgG bands (OBs) and increased IgG index, is much less frequently observed in Japanese multiple sclerosis (MS) cohorts compared with Western cohorts. We aimed to clarify whether genetic and common infectious backgrounds influence CSF IgG abnormality in Japanese MS patients. Methodology: We analyzed HLA-DRB1 alleles, and IgG antibodies against Chlamydia pneumoniae, Helicobacter pylori, Epstein-Barr virus nuclear antigen (EBNA), and varicella zoster virus (VZV) in 94 patients with MS and 367 unrelated healthy controls (HCs). We defined CSF IgG abnormality as the presence of CSF OBs and/or increased IgG index (.0.658). Principal Findings: CSF IgG abnormality was found in 59 of 94 (62.8%) MS patients. CSF IgG abnormality-positive patients had a significantly higher frequency of brain MRI lesions meeting the Barkhof criteria compared with abnormality-negative patients. Compared with HCs, CSF IgG abnormality-positive MS patients showed a significantly higher frequency of DRB1&1501, whereas CSF IgG abnormality-negative patients had a significantly higher frequency of DRB1&0405. CSF IgG abnormality-positive MS patients had a significantly higher frequency of anti-C. pneumoniae IgG antibodies compared with CSF IgG abnormality-negative MS patients, although there was no difference in the frequency of anti-C. pneumoniae IgG antibodies between HCs and total MS patients. Compared with HCs, anti-H. pylori IgG antibodies were detected significantly less frequently in the total MS patients, especially in CSF IgG abnormality-negative MS patients. The frequencies of antibodies against EBNA and VZV did not differ significantly among the groups. Conclusions: CSF IgG abnormality is associated with Western MS-like brain MRI features. DRB1&1501 and C. pneumoniae infection confer CSF IgG abnormality, while DRB1&0405 and H. pylori infection are positively and negatively associated with CSF IgG abnormality-negative MS, respectively, suggesting that genetic and environmental factors differentially contribute to MS susceptibility according to the CSF IgG abnormality status..
44. Gulibahaer Ainiding, Yuji Kawano, Shinya Sato, Noriko Isobe, Takuya Matsushita, Satoshi Yoshimura, Tomomi Yonekawa, Ryo Yamasaki, Hiroyuki Murai, Jun-Ichi Kira, Interleukin 2 receptor α chain gene polymorphisms and risks of multiple sclerosis and neuromyelitis optica in southern Japanese, Journal of the Neurological Sciences, 10.1016/j.jns.2013.11.037, 337, 1-2, 147-150, 2014.02, Background Interleukin 2 receptor α subunit (IL2RA) is a genetic risk for multiple sclerosis (MS) in Caucasians. However, the association between MS and IL2RA in Japanese idiopathic demyelinating diseases of the central nervous system has not been examined. Objective To determine whether IL2RA gene polymorphisms confer risks of developing MS or neuromyelitis optica (NMO) in a Japanese population. Methods DNA samples were obtained from 115 MS patients, 75 NMO/NMO spectrum disorder (NMOSD) patients, and 238 healthy controls. The single nucleotide polymorphisms (SNPs) rs2104286, rs12722489, and rs7090512 were genotyped by real-time PCR using TaqMan SNP genotyping assays. Results No significant associations of the three IL2RA SNPs with the development of the diseases were observed. In MS patients only, the annualized relapse rates were significantly higher for the rs2104286-TT genotype than for the non-TT (CT+CC) genotype and for the rs12722489-CC genotype than for the non-CC genotype in females (p = 0.0138 for both), but not in males. Conclusions Although the possibility that IL2RA is a risk factor for MS development was not confirmed in this Japanese population, IL2RA gene polymorphisms were able to modify the disease activity in female MS patients, but had no influence on either susceptibility or disease phenotype in NMO/NMOSD patients..
45. Tomomi Yonekawa, Hiroyuki Murai, Satoshi Utsuki, Takuya Matsushita, Katsuhisa Masaki, Noriko Isobe, Ryo Yamasaki, Mari Yoshida, Susumu Kusunoki, Kiyomi Sakata, Kiyotaka Fujii, Jun-Ichi Kira, A nationwide survey of hypertrophic pachymeningitis in Japan, Journal of Neurology, Neurosurgery and Psychiatry, 10.1136/jnnp-2013-306410, 85, 7, 732-739, 2014.01, Objectives: To clarify the prevalence, frequent causes and distinct features of hypertrophic pachymeningitis (HP) according to background conditions in a nationwide survey in Japan. Methods: The study began with a preliminary survey to determine the approximate number of HP patients diagnosed from 1 January 2005 to 31 December 2009, and was followed by a questionnaire survey for clinical and laboratory findings. HP was defined as a condition with thickening of the cranial or spinal dura mater with inflammation, evidenced by MRI or histology. Results: Crude HP prevalence was 0.949/100 000 population. The mean age at onset was 58.3±15.8 years. Among 159 cases for whom detailed data were collated, antineutrophil cytoplasmic antibody (ANCA)-related HP was found in 54 cases (34.0%) and IgG4/multifocal fibrosclerosis (MFS)-related HP in 14 cases (8.8%). Seventy cases (44.0%) were classified as 'idiopathic' and 21 (13.2%) as 'others'. ANCA-related HP cases showed a female preponderance, a higher age of onset, and higher frequencies of otological symptoms and elevated systemic inflammatory biomarkers, but lower frequencies of diplopia compared with idiopathic HP. IgG4/MFS-related HP cases showed a marked male predominance; all had cranial HP while none had isolated spinal HP or decreased sensation. Conclusions: HP is not extremely rare. ANCA-related HP is the most frequent form, followed by IgG4/MFS-related HP. Both forms have unique features, which may help to differentiate background causes..
46. Yi Wen Cui, Yuji Kawano, Ryo Yamasaki, Nan Shi, Katsuhisa Masaki, Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Takahisa Tateishi, Shintaro Hayashi, Jun-Ichi Kira, Decreased CCR2 and CD62L expressions on peripheral blood classical monocytes in amyotrophic lateral sclerosis, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12088, 5, 1, 92-96, 2014.01, Objective Recent evidence has suggested the importance of an aberrantly activated monocyte system in amyotrophic lateral sclerosis (ALS) pathogenesis. However, the roles of each monocyte subset, namely CD14+CD16-classical monocytes, CD14dimCD16+ non-classical monocytes and CD14+CD16+ intermediate monocytes, in ALS remain unknown. We aimed to clarify the alterations in the monocyte subset proportions and the surface marker expressions on each monocyte subset in ALS. Methods Blood samples were collected from 19 ALS patients and 28 healthy controls (HC). The surface expressions of CCR2, CX3CR1, CD64 and CD62L were measured in the three monocyte subsets (classical, non-classical and intermediate) by flow cytometry. Results The percentages of CCR2 and CD62L on CD14+CD16-classical monocytes were significantly lower in ALS patients than in HC (P = 0.0012 and P = 0.0296, respectively). No differences were found in CX3CR1 and CD64 on each monocyte subset. The percentage of intermediate monocytes showed a significant negative correlation with the revised ALS functional rating scale score (r =-0.631, P = 0.0038). Conclusions Reductions in chemotaxis-and adhesion-related molecules on classical inflammatory monocytes are present in ALS, further suggesting the involvement of an aberrant innate immune system in ALS pathogenesis..
47. Hikaru Doi, Zi Ye Song, Satoshi Yoshimura, Takahisa Tateishi, Tomomi Yonekawa, Ryo Yamasaki, Hiroyuki Murai, Takuya Matsushita, Jun-Ichi Kira, Distinct cytokine and T helper cell profiles between patients with multiple sclerosis who had or had not received interferon-beta, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12138, 5, 3, 321-327, 2014.01, Objectives Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system, and is generally considered to be mediated by T helper (Th) 1/Th17 cells. Interferon-β (IFNβ) is widely used as a disease-modifying MS drug, but its effects on Th17 cells are still disputed. Furthermore, the effects of IFNβ on Th9 cells have not been elucidated. The present study aimed to clarify the effects of IFNβ on cytokines/growth factors in cerebrospinal fluid (CSF) and cytokine-producing Th cells in peripheral blood.
Methods First, the frequency of IFNγ, interleukin (IL)-17A, IL-9, and IL-4-producing Th cells in peripheral blood lymphocytes was analyzed by flow cytometry in 34 MS patients and 15 healthy volunteers enrolled in the cytokine-producing Th cell study. Second, levels of 27 cytokines/growth factors in the CSF were measured using a multiple fluorescence bead-based immunoassay in 34 MS patients enrolled in the cytokines/growth factors study.
Results We found a significantly higher frequency of IL-4-IL-9+CD4+ T cells and lower frequency of IFNγ+IL-17A-CD4+ cells in peripheral blood lymphocytes in the 10 MS patients who had received IFNβ than in the 24 MS patients who had not received IFNβ or the 15 healthy controls (P < 0.05). The seven MS patients who received IFNβ showed significantly lower IL-17A levels in CSF than did the 27 MS patients who had not received IFNβ (P < 0.05).
Conclusions The present results suggest the suppression of IL-17A production in the central nervous system and augmentation of Th9 cells in the peripheral blood by IFNβ in MS patients..
48. Hajime Arahata, Yasumasa Ohyagi, Kyoko M. Iinuma, Masahito Tanaka, Takahisa Tateishi, Ryo Yamasaki, Takuya Matsushita, Jun-Ichi Kira, Early inhibition of tumor necrosis factor-α and interleukin-6 in muscle tissue as a therapy for dystrophinopathy in mdx mice, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12111, 5, 3, 371-377, 2014.01, Objective Pro-inflammatory cytokines can exacerbate muscle fiber damage in dystrophinopathy. The aim of the present study was to identify cytokine/chemokine alterations in the muscle tissues of mdx mice, a model of dystrophinopathy, and the beneficial effects of anti-proinflammatory cytokine therapy. Methods A total of 23 cytokines and chemokines were quantitatively measured in muscle tissues from mdx mice by fluorescent bead-based immunoassay. The mdx mice were treated with anti-tumor necrosis factor-α (TNF-α) and anti-interleukin-6 (IL-6) drugs, and their physical condition was evaluated by Rotarod and muscle damage by histopathological analysis. Results Levels of TNF-α and IL-6 were elevated at 14 days (P14), before a transient increase of macrophage and neutrophil-activating pro-inflammatory cytokines/chemokines, such as C-C motif ligand 2 (CCL2), CCL4 and KC (mouse C-X-C motif ligand 8 homolog), at P20. Administration of an anti-TNF-α drug (etanercept) and an anti-IL-6 receptor antibody (MR16-1) from P7 improved physical performance, and reduced both the area of basophilic fibers that indicated degenerating/regenerating fibers and CD68-positive macrophage infiltration. Initiating therapy at P7 inhibited the elevation of CCL2, CCL4 and KC more effectively than at P13. Conclusions The early administration of anti-TNF-α and anti-IL-6 drugs attenuated muscle fiber degeneration in a mouse model of dystrophinopathy..
49. Satoshi Yoshimura, Tomomi Yonekawa, Noriko Isobe, Takuya Matsushita, Jun-Ichi Kira, Bacterial infectious burden and prevalence of idiopathic central nervous system demyelinating diseases in Japanese, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12047, 4, 3, 351-352, 2013.12.
50. Yuji Kawano, Takuya Matsushita, Yi Wen Cui, Noriko Isobe, Satoshi Yoshimura, Tomomi Yonekawa, Katsuhisa Masaki, Ryo Yamasaki, Hiroyuki Murai, Jun-Ichi Kira, Protein kinase C-η polymorphism rs2230500 does not confer disease susceptibility to multiple sclerosis or neuromyelitis optica, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12056, 4, 3, 283-287, 2013.12, Objectives To determine whether the non-synonymous 1425G/A polymorphism (rs2230500), an Asian-specific single nucleotide polymorphism that increases the kinase activity and affects the function of immune cells, of the protein kinase C-η gene (PRKCH) confers the risk of developing idiopathic demyelinating diseases of the central nervous system in a Japanese population. Methods Blood samples were collected from 96 multiple sclerosis (MS) patients, 52 neuromyelitis optica (NMO)/NMO spectrum disorder (NMOSD) patients and 151 healthy controls. The polymorphism rs2230500 was genotyped by sequencing. Results No significant association was observed between the PRKCH rs2230500 polymorphism and the risk of either MS or NMO/NMOSD. Clinical characteristics were also unaffected by the rs2230500 status. Conclusions Although the possibility that PRKCH has some effect on MS and NMO/NMOSD risk cannot be completely excluded because of the small study sample size, the polymorphism rs2230500 did not appear to confer disease susceptibility to MS or NMO/NMOSD in this Japanese population..
51. Yuko Nagara, Takahisa Tateishi, Ryo Yamasaki, Shintaro Hayashi, Mami Kawamura, Hitoshi Kikuchi, Kyoko Motomura Iinuma, Masahito Tanaka, Toru Iwaki, Takuya Matsushita, Yasumasa Ohyagi, Jun-Ichi Kira, Impaired cytoplasmic-nuclear transport of hypoxia-inducible factor-1α in amyotrophic lateral sclerosis, Brain Pathology, 10.1111/bpa.12040, 23, 5, 534-546, 2013.09, We investigated the mechanisms underlying abnormal vascular endothelial growth factor (VEGF) production in amyotrophic lateral sclerosis (ALS). We immunohistochemically studied VEGF, its receptors VEGFR1 and 2, and hypoxia-inducible factor-1α (HIF-1α) in autopsied ALS spinal cords. We also chronologically assessed the expression of HIF-1α, karyopherin β1, karyopherin β-cargo protein complex inhibitors and nuclear pore complex proteins in G93A mutant superoxide dismutase 1 (mSOD1) transgenic mice at presymptomatic, symptomatic and end stages. In ALS patients, compared with controls, HIF-1α immunoreactivity in the cytoplasm of anterior horn cells (AHCs) was significantly increased, while immunoreactivities for VEGF and VEGFRs were significantly decreased. Similar changes in HIF-1α and VEGF levels were observed in mSOD1 transgenic mice. HIF-1α co-localized with karyopherin β1 in the cytoplasm of AHCs and karyopherin β1 co-localized with nucleoporin 62 (Nup62) on the nuclear envelope. From the presymptomatic stage of mSOD1 transgenic mice, karyopherin β1 immunoreactivity in AHC nuclei significantly decreased and morphological irregularities of the Nup62-immunostained nuclear envelope became more pronounced with disease progression. Thus, in AHCs from mSOD1 transgenic mice, transport of cytoplasmic HIF-1α to the nuclear envelope and into the nucleus is impaired from the presymptomatic stage, suggesting that impaired cytoplasmic-nuclear transport of HIF-1α through the nuclear pore might precede motor neuron degeneration..
52. Yi Wen Cui, Yuji Kawano, Nan Shi, Katsuhisa Masaki, Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Takahisa Tateishi, Ryo Yamasaki, Hiroyuki Murai, Jun-Ichi Kira, Alterations in chemokine receptor expressions on peripheral blood monocytes in multiple sclerosis and neuromyelitis optica, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12039, 4, 2, 201-205, 2013.08, Objectives Human peripheral blood monocytes comprise three different subtypes: CD14+CD16- (classical type), CD14lowCD16+ (non-classical type) and CD14+CD16+ (intermediate type). These subsets are known to have different functions, but little is known about their roles in multiple sclerosis (MS), especially for maintaining remission. We aimed to clarify the alterations in monocyte subsets in patients with MS and neuromyelitis optica (NMO)/NMO spectrum disorder (NMOSD) in the remission phase. Methods Blood samples were collected from 19 MS patients and 10 NMO/NMOSD patients in the remission phase, and 42 healthy controls (HC). The surface expressions of CCR2, CX3CR1, CD64 (FcγR1) and CD62L were analyzed in the monocyte subsets by flow cytometry. Results CCR2 expression was significantly decreased in classical monocytes from MS patients, regardless of interferon-β (IFN-β) treatment, but not in those from NMO/NMOSD patients. CX3CR1 expression was also decreased in all monocyte subsets from MS patients receiving IFN-β, whereas CX3CR1 expression in classical monocytes was only decreased in NMO/NMOSD patients receiving prednisolone. In NMO/NMOSD patients on prednisolone, the percentages of CD14+CD16+ intermediate monocytes, CD14lowCD16+ non-classical monocytes and CD64+CD14+CD16+ monocytes among total monocytes were significantly lower than in HC. CD62L expression on the monocyte subsets showed no significant differences among the patients and HC. Conclusions Our findings suggest that alterations in chemokine receptor expressions on peripheral blood monocytes can occur in MS and NMO/NMOSD during the remission phase. Down-modulation of CCR2 in MS, and CX3CR1 in MS and NMO/NMOSD could partly contribute to sustained remission by preventing monocyte infiltration into the central nervous system..
53. Nobutoshi Kawamura, Ryo Yamasaki, Tomomi Yonekawa, Takuya Matsushita, Susumu Kusunoki, Shigemi Nagayama, Yasuo Fukuda, Hidenori Ogata, Dai Matsuse, Hiroyuki Murai, Jun-Ichi Kira, Anti-neurofascin antibody in patients with combined central and peripheral demyelination, Neurology, 10.1212/WNL.0b013e3182a1aa9c, 81, 8, 714-722, 2013.08, Objectives: We aimed to identify the target antigens for combined central and peripheral demyelination (CCPD). Methods: We screened target antigens by immunohistochemistry and immunoblotting using peripheral nerve tissues to identify target antigens recognized by serum antibodies from selected CCPD and chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) cases. We then measured the level of antibody to the relevant antigen in 7 patients with CCPD, 16 patients with CIDP, 20 patients with multiple sclerosis, 20 patients with Guillain-Barré syndrome, 21 patients with other neuropathies, and 23 healthy controls (HC) by ELISA and cell-based assays using HEK293 cells. Results: At the initial screening, sera from 2 patients with CCPD showed cross-like binding to sciatic nerve sections at fixed intervals, with nearly perfect colocalization with neurofascin immunostaining at the node and paranode. ELISA with recombinant neurofascin revealed significantly higher mean optical density values in the CCPD group than in other disease groups and HC. Anti-neurofascin antibody positivity rates were 86% in patients with CCPD, 10% in patients with multiple sclerosis, 25% in patients with CIDP, 15% in patients with Guillain-Barré syndrome, and 0% in patients with other neuropathies and HC. The cell-based assay detected serum anti-neurofascin antibody in 5 of 7 patients with CCPD; all others were negative. CSF samples examined from 2 patients with CCPD were both positive. In anti-neurofascin antibody- positive CCPD patients, including those with a limited response to corticosteroids, IV immunoglobulin or plasma exchange alleviated the symptoms. Conclusion: Anti-neurofascin antibody is frequently present in patients with CCPD. Recognition of this antibody may be important, because patients with CCPD who are antibody positive respond well to IV immunoglobulin or plasma exchange..
54. Katsuhisa Masaki, Satoshi Suzuki, Takuya Matsushita, Takeshi Matsuoka, Shihoko Imamura, Ryo Yamasaki, Makiko Suzuki, Toshihiko Suenaga, Toru Iwaki, Jun-Ichi Kira, Connexin 43 Astrocytopathy Linked to Rapidly Progressive Multiple Sclerosis and Neuromyelitis Optica, PLoS One, 10.1371/journal.pone.0072919, 8, 8, 2013.08, Background:Multiple sclerosis (MS) and neuromyelitis optica (NMO) occasionally have an extremely aggressive and debilitating disease course; however, its molecular basis is unknown. This study aimed to determine a relationship between connexin (Cx) pathology and disease aggressiveness in Asian patients with MS and NMO.Methods/Principal Findings:Samples included 11 autopsied cases with NMO and NMO spectrum disorder (NMOSD), six with MS, and 20 with other neurological diseases (OND). Methods of analysis included immunohistochemical expression of astrocytic Cx43/Cx30, oligodendrocytic Cx47/Cx32 relative to AQP4 and other astrocytic and oligodendrocytic proteins, extent of demyelination, the vasculocentric deposition of complement and immunoglobulin, and lesion staging by CD68 staining for macrophages. Lesions were classified as actively demyelinating (n=59), chronic active (n=58) and chronic inactive (n=23). Sera from 120 subjects including 30 MS, 30 NMO, 40 OND and 20 healthy controls were examined for anti-Cx43 antibody by cell-based assay. Six NMO/NMOSD and three MS cases showed preferential loss of astrocytic Cx43 beyond the demyelinated areas in actively demyelinating and chronic active lesions, where heterotypic Cx43/Cx47 astrocyte oligodendrocyte gap junctions were extensively lost. Cx43 loss was significantly associated with a rapidly progressive disease course as six of nine cases with Cx43 loss, but none of eight cases without Cx43 loss regardless of disease phenotype, died within two years after disease onset (66.7% vs. 0%, P=0.0090). Overall, five of nine cases with Cx43 loss and none of eight cases without Cx43 loss had distal oligodendrogliopathy characterized by selective myelin associated glycoprotein loss (55.6% vs. 0.0%, P=0.0296). Loss of oligodendrocytic Cx32 and Cx47 expression was observed in most active and chronic lesions from all MS and NMO/NMOSD cases. Cx43-specific antibodies were absent in NMO/NMOSD and MS patients.Conclusions:These findings suggest that autoantibody-independent astrocytic Cx43 loss may relate to disease aggressiveness and distal oligodendrogliopathy in both MS and NMO..
55. Jian Huang, Noriko Isobe, Takuya Matsushita, Satoshi Yoshimura, Shinya Sato, Tomomi Yonekawa, Ryo Yamasaki, Hiroyuki Murai, Jun-Ichi Kira, Distinct genetic profiles between Japanese multiple sclerosis patients with and without Barkhof brain lesions, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12017, 4, 2, 173-180, 2013.08, Objectives The frequency of brain lesions that fulfil the Barkhof criteria (Barkhof brain lesions) is low in Asian patients with multiple sclerosis (MS). Several genes are associated with MS in the Japanese, but their influence on brain lesion development is unknown. Here, we clarified the genetic profiles of Barkhof brain lesion-positive and -negative MS in the Japanese. Methods We genotyped the HLA-DRB1 and -DPB1 alleles, the NOTCH4 missense mutation rs422951, and the IL-7RA single nucleotide polymorphism rs6897932 in 123 non-neuromyelitis optica/neuromyelitis optica spectrum disorder MS patients and 367 healthy controls by annealing of sequence-specific oligonucleotide probes to polymerase chain reaction-amplified genomic DNA (for the HLA alleles), DNA sequencing (for rs422951), and real-time polymerase chain reaction using TaqMan genotyping assays (for rs6897932). Results Compared with the healthy controls, the frequency of DRB1*0405 was significantly higher in the Barkhof brain lesion-negative group, that of DPB1*0301 was significantly higher in the Barkhof brain lesion-positive group, and those of DRB1*0901 and DPB1*0401 were significantly lower in the Barkhof brain lesion-positive group. The frequency of NOTCH4 rs422951 G allele carriers was significantly lower in both groups compared with the healthy controls, whereas the frequency of the IL-7RA rs6897932 CC genotype was significantly higher in the Barkhof brain lesion-positive group. Haplotype analyses identified one susceptibility and three resistance haplotypes for Barkhof brain lesion-positive MS, and two susceptibility and three resistance haplotypes for Barkhof brain lesion-negative MS. Conclusions The genetic profiles of Japanese MS patients are distinct according to the presence or absence of Barkhof brain lesions..
56. Yasukiyo Araki, Masako Kinoshita, Rie Motoyama, Takuya Matsushita, Masanori Nakagawa, Jun-Ichi Kira, Masami Tanaka, Month of birth in multiple sclerosis with and without longitudinally extensive spinal cord lesions
A study of a Japanese national survey, Journal of the Neurological Sciences, 10.1016/j.jns.2013.04.006, 330, 1-2, 67-70, 2013.07, Objectives Month of birth has been associated with the environmental factors for multiple sclerosis (MS). This study aimed to investigate whether individuals with MS had significantly different frequencies of birth in a particular month of the year, and whether month-of-birth patterns were influenced by the longitudinally extensive spinal cord lesions (LESCL) status relative to the general population in the same Japanese birth cohort. Methods In this study, performed as a part of the fourth Japanese nationwide survey of MS, patients were divided into two groups according to the presence (n = 307) or absence (n = 906) of LESCLs that were diagnosed on the basis of magnetic resonance imaging findings. The number of births in every month was counted for both groups. Control data were obtained from birth records of the Japanese general population of the median years of birth of each group. Differences in the month-of-birth distributions between the patients and the general population were assessed using the chi-square test. Results In MS patients without LESCLs, significantly more patients were born in January and June, whereas significantly fewer patients were born in May, compared to the general population. The seasonal patterns of birth were not in association with those of ambient ultraviolet (UV) radiation. No significant differences were found for the month-of-birth distributions between MS patients with LESCLs and the general population. Conclusions A different distribution of month of birth in Japanese MS patients without LESCLs from the general population, but not in those with LESCLs, suggests some role for environmental factors in the pathogenesis of the former group. Environmental factors other than UV radiation should be further elucidated in Japanese and other Asian MS cohorts..
57. Kazutaka Sonoda, Ryo Yamasaki, Takuya Matsushita, Takeo Yoshimura, Hiroyuki Murai, Jun-Ichi Kira, Case of chronic lymphocytic inflammation with pontine perivascular enhancement responsive to steroids showing features common to multiple sclerosis, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12028, 4, 1, 104-106, 2013.06.
58. Yuji Kanamori, Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Hiroshi Shigeto, Nobutoshi Kawamura, Ryo Yamasaki, Hiroyuki Murai, Shozo Tobimatsu, Jun-Ichi Kira, Multimodality evoked potentials for discrimination of atopic myelitis and multiple sclerosis, Clinical and Experimental Neuroimmunology, 10.1111/cen3.12018, 4, 1, 29-35, 2013.06, Objectives: To clarify the differences in multimodality evoked potential findings between patients with atopic myelitis (AM) and those with multiple sclerosis (MS). Methods: A retrospective chart review of 70 consecutive AM patients and 93 MS patients was carried out. All patients were negative for serum antiaquaporin- 4 antibody. Visual- (VEP), somatosensory- (SEP) and motor-evoked potentials (MEP) recorded at first examination, and magnetic resonance imaging (MRI) findings from the first examination were compared between AM and MS patients. Results: Compared with MS patients, AM patients showed male preponderance, lower the Expanded Disability Status Scale scores and less frequent spinal cord MRI lesions. Visual impairment and muscle weakness were also less severe in AM patients. Frequencies of abnormal VEP and prolonged central conduction time on lower limb MEP were significantly lower in AM patients than in MS patients (AM vs MS: 9.5% vs 55.6%, and 28.2% vs 54.4%, respectively), whereas frequencies of peripheral nerve involvement in upper and lower limb MEP and upper limb SEP were significantly higher in AM than in MS patients (AM vs MS: 12.8% vs 2.9%, 17.9% vs 2.9% and 33.3% vs 4.4%, respectively). When patients whose EP were examined within 5 years of disease onset were compared, lower frequencies of abnormal VEP and higher peripheral nerve involvement detected by MEP and SEP were observed in AM patients. Conclusions: AM patients have distinct physiological features compared with MS patients, even at the first examination of evoked potentials, which might suggest distinct immunological mechanisms between the two conditions. Multimodality evoked potentials might contribute to the early discrimination of these two disorders..
59. Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Katsuhisa Masaki, Satoshi Yoshimura, Jakub Fichna, Shu Chen, Jadwiga Furmaniak, Bernard Rees Smith, Jun-Ichi Kira, Clinical relevance of serum aquaporin-4 antibody levels in neuromyelitis optica, Neurochemical Research, 10.1007/s11064-013-1009-0, 38, 5, 997-1001, 2013.05, Neuromyelitis optica (NMO) is an inflammatory disease that selectively affects the optic nerves and spinal cord. The discovery of NMO-IgG targeting aquaporin-4 (AQP4) in NMO patients suggested that NMO is a distinct entity, with a fundamentally different etiology from that of multiple sclerosis (MS). Although NMO usually leads to grave disability because of the more severe tissue destruction compared with classical MS, there have been several reports describing a benign form of NMO over a long disease term. NMO-IgG/AQP4 antibodies show high specificity but medium sensitivity for NMO, while the clinical relevance of AQP4 antibody titers remains to be determined. We aimed to clarify the clinical relevance of AQP4 antibody levels determined by a bridging enzyme-linked immunosorbent assay in 38 patients with NMO or NMO spectrum disorder. The AQP4 antibody levels were higher in patients with optic neuritis (ON) than in those without ON (p = 0.0164). Among the 12 patients examined in a longitudinal study, four showed an increase in the ELISA values during some relapses, and eight showed no clear correlation between the ELISA values and relapse. Of the four patients who demonstrated a steady rise in the antibody levels over time, two patients had no concomitant relapses, despite elevation of the AQP4 antibody levels. We conclude that high AQP4 antibody levels are associated with the occurrence of ON, but that the antibody levels themselves are not closely correlated with the onset of relapse..
60. Takuya Matsushita, Takahisa Tateishi, Noriko Isobe, Tomomi Yonekawa, Ryo Yamasaki, Dai Matsuse, Hiroyuki Murai, Jun-Ichi Kira, Characteristic Cerebrospinal Fluid Cytokine/Chemokine Profiles in Neuromyelitis Optica, Relapsing Remitting or Primary Progressive Multiple Sclerosis, PLoS One, 10.1371/journal.pone.0061835, 8, 4, 2013.04, Background: Differences in cytokine/chemokine profiles among patients with neuromyelitis optica (NMO), relapsing remitting multiple sclerosis (RRMS), and primary progressive MS (PPMS), and the relationships of these profiles with clinical and neuroimaging features are unclear. A greater understanding of these profiles may help in differential diagnosis. Methods/Principal Findings: We measured 27 cytokines/chemokines and growth factors in CSF collected from 20 patients with NMO, 26 with RRMS, nine with PPMS, and 18 with other non-inflammatory neurological diseases (OND) by multiplexed fluorescent bead-based immunoassay. Interleukin (IL)-17A, IL-6, CXCL8 and CXCL10 levels were significantly higher in NMO patients than in OND and RRMS patients at relapse, while granulocyte-colony stimulating factor (G-CSF) and CCL4 levels were significantly higher in NMO patients than in OND patients. In NMO patients, IL-6 and CXCL8 levels were positively correlated with disability and CSF protein concentration while IL-6, CXCL8, G-CSF, granulocyte-macrophage colonystimulating factor (GM-CSF) and IFN-c were positively correlated with CSF neutrophil counts at the time of sample collection. In RRMS patients, IL-6 levels were significantly higher than in OND patients at the relapse phase while CSF cell counts were negatively correlated with the levels of CCL2. Correlation coefficients of cytokines/chemokines in the relapse phase were significantly different in three combinations, IL-6 and GM-CSF, G-CSF and GM-CSF, and GM-CSF and IFN-c, between RRMS and NMO/NMOSD patients. In PPMS patients, CCL4 and CXCL10 levels were significantly higher than in OND patients. Conclusions: Our findings suggest distinct cytokine/chemokine alterations in CSF exist among NMO, RRMS and PPMS. In NMO, over-expression of a cluster of Th17- and Th1-related proinflammatory cytokines/chemokines is characteristic, while in PPMS, increased CCL4 and CXCL10 levels may reflect on-going low grade T cell and macrophage/microglia inflammation in the central nervous system. In RRMS, only a mild elevation of proinflammatory cytokines/chemokines was detectable at relapse..
61. Jian Huang, Satoshi Yoshimura, Noriko Isobe, Takuya Matsushita, Tomomi Yonekawa, Shinya Sato, Ryo Yamasaki, Jun-Ichi Kira, A NOTCH4 missense mutation confers resistance to multiple sclerosis in Japanese, Multiple Sclerosis, 10.1177/1352458513482512, 19, 13, 1696-1703, 2013.01, Background: The G allele of NOTCH4 rs422951 is protective against demyelinating disease in Japanese. Objectives: The purpose of this study was to assess the relation of the G allele to neuromyelitis optica (NMO)/NMO spectrum disorder (NMOSD) and multiple sclerosis (MS) and the interaction between the G allele and HLA-DRB1 alleles, and to clarify any association of the G allele with clinical features. Methods: DNA sequencing was used to genotype 106 NMO/NMOSD patients, 118 MS patients and 152 healthy controls (HCs) for rs422951. Results: G allele frequency in MS patients, but not that in NMO/NMOSD patients, was lower than that in HCs (8.9% vs 21.7%, p<0.0001, odds ratio (OR)=0.35). HLA-DRB1*0405 was positively associated with MS (OR=2.22, pcorr=0.0380) while DRB1*0901 was negatively associated (OR=0.32, pcorr =0.0114). Logistic regression analyses revealed that, after adjusting for gender and either HLA-DRB1*0405 or DRB1*0901, rs422951 was associated with MS in the dominant model (OR=0.37, 95% confidence interval (CI)= 0.20-0.66, p=0.0012). Haplotype analyses identified two susceptible and three resistant haplotypes formed from rs422951 and either HLA-DRB1*0405 or DRB1*0901. There were no statistically significant differences in clinical features between G allele carriers and non-G allele carriers. Conclusion: This NOTCH4 missense mutation decreased the risk for developing MS in Japanese, but did not affect clinical features of those who had already developed the disease..
62. Satoshi Yoshimura, Noriko Isobe, Takuya Matsushita, Tomomi Yonekawa, Katsuhisa Masaki, Shinya Sato, Yuji Kawano, Jun-Ichi Kira, Katsuichi Miyamoto, Susumu Kusunoki, Yuji Nakatsuji, Hideki Mochizuki, Kazuhide Ochi, Masayasu Matsumoto, Takeshi Kanda, Hirofumi Ochi, Tetsuro Miki, Kazumasa Okada, Sadatoshi Tsuji, Distinct genetic and infectious profiles in Japanese neuromyelitis optica patients according to anti-aquaporin 4 antibody status, Journal of Neurology, Neurosurgery and Psychiatry, 10.1136/jnnp-2012-302925, 84, 1, 29-34, 2013.01, Objective: To clarify whether genetic and common infectious backgrounds are distinct, according to anti-aquaporin 4 (AQP4) antibody status in Japanese patients with neuromyelitis optica (NMO). Methods: We analysed human leucocyte antigen (HLA)-DRB1 and HLA-DPB1 alleles, and IgG antibodies against Helicobacter pylori, Chlamydia pneumoniae, varicella zoster virus and Epstein-Barr virus nuclear antigen (EBNA) in 116 patients with NMO, including 39 patients with neuromyelitis optica spectrum disorder (NMOSD), 145 multiple sclerosis (MS) patients and 367 unrelated healthy controls. 77 NMO/NMOSD patients were seropositive for AQP4 antibody while 39 were seronegative. Results: Compared with healthy controls, NMO/NMOSD patients showed a significantly lower frequency of DRB1&z.ast;0901 and significantly higher frequencies of DRB1&z.ast;1602 and DPB1&z.ast;0501 , which conferred susceptibility to anti-AQP4 antibody positive NMO/NMOSD, but not antibody negative NMO/NMOSD. DRB1&z.ast;0901 was a common protective allele, irrespective of the presence or absence of anti-AQP4 antibody. Anti-H pylori and anti- C pneumoniae antibodies were more commonly detected in anti-AQP4 antibody positive NMO/NMOSD patients than healthy controls. Antibody negative NMO/NMOSD patients did not differ from healthy controls regarding the presence of these antibodies. The presence or absence of antibodies against varicella zoster virus and EBNA did not vary among the groups. The frequencies of antibodies against these four pathogens were not significantly different between MS patients and healthy controls. Conclusions: Our results suggest that HLA-DRB1&z.ast;1602 and DPB1&z.ast;0501 alleles and H pylori and Chlamydia pneumonia infection are risk factors only for anti-AQP4 antibody positive NMO/NMOSD but not for anti-AQP4 antibody negative NMO/NMOSD..
63. Satoshi Yoshimura, Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Katsuhisa Masaki, Shinya Sato, Yuji Kawano, Ken Yamamoto, Jun-Ichi Kira, Genetic and Infectious Profiles of Japanese Multiple Sclerosis Patients, PLoS One, 10.1371/journal.pone.0048592, 7, 11, 2012.11, Background: 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. Methodology/Principal Findings: We analyzed HLA-DRB1 and -DPB1 alleles, and IgG antibodies specific for Helicobacter pylori, Chlamydia pneumoniae, varicella zoster virus, and Epstein-Barr virus nuclear antigen (EBNA) in 145 MS patients and 367 healthy controls (HCs). Frequencies of DRB1*0405 and DPB1*0301 were significantly higher, and DRB1*0901 and DPB1*0401 significantly lower, in MS patients as compared with HCs. MS patients with DRB1*0405 had a significantly earlier age of onset and lower Progression Index than patients without this allele. The proportion and absolute number of patients with DRB1*0405 successively increased with advancing year of birth. In MS patients without DRB1*0405, the frequency of the DRB1*1501 allele was significantly higher, while the DRB1*0901 allele was significantly lower, compared with HCs. Furthermore, DRB1*0405-negative MS patients were significantly more likely to be positive for EBNA antibodies compared with HCs. Conclusions: Our study suggests that MS patients harboring DRB1*0405, a genetic risk factor for MS in the Japanese population, have a younger age at onset and a relatively benign disease course, while DRB1*0405-negative MS patients have features similar to Western-type MS in terms of association with Epstein-Barr virus infection and DRB1*1501. The recent increase of MS in young Japanese people may be caused, in part, by an increase in DRB1*0405-positive MS patients..
64. Shinya Sato, Noriko Isobe, Satoshi Yoshimura, Yuji Kanamori, Katsuhisa Masaki, Takuya Matsushita, Jun-Ichi Kira, HLA-DPB1*0201 is associated with susceptibility to atopic myelitis in Japanese, Journal of Neuroimmunology, 10.1016/j.jneuroim.2012.07.007, 251, 1-2, 110-113, 2012.10, To determine the relationship between susceptibility to atopic myelitis (AM) and polymorphisms of the human leukocyte antigen (HLA)- DPB1 and - DRB1 alleles, we compared each phenotype frequency between 55 AM patients and 367 unrelated healthy controls in Japan. The HLA- DPB1* 0201 allele was significantly more frequent in AM patients than in healthy controls (54.5% vs. 31.9%, corrected P value = 0.0150, odds ratio = 2.564, 95% confidence interval = 1.444-4.554). Our result suggests that the immunogenetic background of AM differs from that of other CNS autoimmune diseases, such as multiple sclerosis and neuromyelitis optica, which show distinct HLA class II associations..
65. Katsuhisa Masaki, Satoshi Suzuki, Takuya Matsushita, Tomomi Yonekawa, Takeshi Matsuoka, Noriko Isobe, Kyoko Motomura, Xiao Mu Wu, Takeshi Tabira, Toru Iwaki, Jun-Ichi Kira, Extensive loss of connexins in Baló's disease
Evidence for an auto-antibody-independent astrocytopathy via impaired astrocyte-oligodendrocyte/ myelin interaction, Acta Neuropathologica, 10.1007/s00401-012-0972-x, 123, 6, 887-900, 2012.06, Extensive aquaporin-4 (AQP4) loss without perivascular deposition of either activated complement or immunoglobulins is a characteristic of Baló's disease. Our aim in this study was to investigate the relationship between astrocytopathy and demyelination in Baló's disease, focusing on connexins (Cx), which form gap junctions among glial cells and myelin. Autopsied specimens from four cases that provided seven actively demyelinating concentric lesions infiltrated with numerous CD68 + macrophages were immunohistochemically examined for the astrocyte markers glial fibrillary acidic protein (GFAP), AQP4, Cx43, Cx30 and megalencephalic leukoencephalopathy with subcortical cyst 1 (MLC1). Specimens were also stained for oligodendrocyte/ myelin markers, namely Cx32, Cx47, myelin-associated glycoprotein (MAG), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein (OSP) and Nogo-A. Serum samples from six patients that had undergone magnetic resonance imaging, confirming a diagnosis of Baló's disease, were assayed for the presence of anti-Cx43, -Cx32 and -AQP4 antibodies. Despite the presence of numerous GFAP- and MLC1-positive astrocytes, there was a marked decrease in the levels of Cx43, Cx32 and Cx47. At the leading edges, Cx43 and AQP4 were mostly absent despite positive GFAP, MLC1, Cx32, Cx47, MOG, MAG, and OSP immunoreactivity. Of the six Baló's disease patients, none were positive for anti-Cxs or -AQP4 antibodies. Baló's disease is characterized by extensive loss of Cxs and AQP4, and a lack of auto-antibodies to Cxs and AQP4. Loss of Cx43 and AQP4 in the presence of other oligodendrocyte/myelin proteins at the leading edges suggests the possibility that auto-antibody-independent astrocytopathy may contribute to disease pathology via the disruption of astrocyte-oligodendrocyte/myelin interactions..
66. Gulibahaer Ainiding, kenichiro yamashita, Takako Torii, Konosuke Furuta, Noriko Isobe, Takuya Matsushita, Katsuhisa Masaki, Shoji Matsumoto, Jun-Ichi Kira, Clinical disability progression and platelet GP IIb/IIIa values in patients with atopic myelitis, Journal of Neuroimmunology, 10.1016/j.jneuroim.2012.03.009, 246, 1-2, 108-112, 2012.05, We aimed to clarify the disability progression and platelet aggregative function in atopic myelitis (AM). Seventeen AM patients and 35 healthy controls were subjected to clinico-allergological evaluations and glycoprotein IIb/IIIa (GP IIb/IIIa) measurements using a VerifyNow assay system. In AM patients, the disease duration had significant positive correlations with the Kurtzke Expanded Disability Status Scale scores and Sensory Functional Scale scores. The GP IIb/IIIa values were significantly higher in AM patients than in controls as well as in females compared with males. AM is essentially a progressive disease affecting the sensory system, and involves an increased platelet aggregative function..
67. Noriko Isobe, Yuji Kanamori, Tomomi Yonekawa, Takuya Matsushita, Hiroshi Shigeto, Nobutoshi Kawamura, Jun-Ichi Kira, First diagnostic criteria for atopic myelitis with special reference to discrimination from myelitis-onset multiple sclerosis, Journal of the Neurological Sciences, 10.1016/j.jns.2012.02.007, 316, 1-2, 30-35, 2012.05, Objective: To establish the first evidence-based diagnostic criteria for atopic myelitis (AM) enabling it to be discriminated from myelitis-onset multiple sclerosis (MS), which is a difficult differential diagnosis. Methods: Sixty-nine consecutive AM patients examined from 1996 to 2010 at Kyushu University hospital, who fulfilled the empirical definition of AM (2003), and 51 myelitis-onset MS patients in whom allergen-specific IgE was measured, were enrolled. The first available brain MRI findings were compared between the two. Then, we compared the clinical and laboratory features between the 16 AM cases who did not meet the Barkhof brain MRI criteria for MS after more than 5 years follow-up and 51 myelitis-onset MS cases. Based on the discriminative findings, we established diagnostic criteria for AM and calculated the sensitivity and specificity. Results: AM patients had a significantly lower frequency of Barkhof brain lesions on baseline MRI than myelitis-onset MS patients. AM patients had a significantly higher frequency of present and/or past history of atopic disease and hyperIgEemia, and higher cerebrospinal fluid levels of interleukin 9 and CCL11/eotaxin, but a lower frequency of oligoclonal IgG bands than myelitis-onset MS patients. Our proposed diagnostic criteria for AM demonstrated 93.3% sensitivity and 93.3% specificity for AM against myelitis-onset MS, with 82.4% positive predictive value and 97.7% negative predictive value. Conclusion: Our first evidence-based criteria for AM show high sensitivity and specificity, and would be useful clinically..
68. Mami Fukunaga Kawamura, Ryo Yamasaki, Nobutoshi Kawamura, Takahisa Tateishi, Yuko Nagara, Takuya Matsushita, Yasumasa Ohyagi, Jun-Ichi Kira, Impaired recruitment of neuroprotective microglia and T cells during acute neuronal injury coincides with increased neuronal vulnerability in an amyotrophic lateral sclerosis model, Experimental Neurology, 10.1016/j.expneurol.2012.01.015, 234, 2, 437-445, 2012.04, Non-cell-autonomous motor neuronal death is suggested in a mutant Cu/Zn superoxide dismutase 1 (mSOD1)-mediated amyotrophic lateral sclerosis (ALS) model, in which microglia and T cells play significant roles in disease progression. However, it remains unknown whether these cells are toxic or protective. The present study aimed to clarify the developmental age-related alterations of neuronal, glial and T cell responses to acute neuron injury in non-transgenic (N-Tg) mice, and the in vivo effects of mSOD1 on these changes by studying N-Tg and mSOD1-Tg mice subjected to unilateral hypoglossal nerve axotomy at young (8. weeks) and adult (17. weeks) ages. Adult N-Tg mice showed increased neuronal viability on day 21 after axotomy and trends toward increased numbers of recruited microglia on day 3 and T cells on day 7, in the hypoglossal nucleus, compared with young N-Tg mice. Quantitative comparisons between mSOD1-Tg and N-Tg mice at the same ages, on day 3 after axotomy, showed that microglial recruitment was significantly lower in mSOD1-Tg mice than in 17-week-old N-Tg mice (the disease progression stage), but the same difference was not seen in 8-week-old mice (the presymptomatic stage), despite good preservation of hypoglossal neurons. Infiltration of CD3-positive T cells, mostly CD4-positive, on day 7 and the viability rate of hypoglossal neurons on the operated side compared with the contralateral side on day 21 were significantly decreased in mSOD1-Tg mice compared with N-Tg mice aged 17. weeks, but the same difference was not seen in mice aged 8. weeks. On day 3 after axotomy, expression levels of IGF-1 mRNA in the operated hypoglossal nucleus were significantly lower in mSOD1-Tg mice than N-Tg mice at 17. weeks of age. The observation that depressed microglial and T cell responses and expression of neurotrophic factors coincided with reduced neuronal viability in adult mSOD1-Tg mice suggests that diminished neuroprotective functions of mSOD1 microglia and T cells may contribute to exaggerated neuronal death..
69. Riwanti Estiasari, Takuya Matsushita, Katsuhisa Masaki, Takuya Akiyama, Tomomi Yonekawa, Noriko Isobe, Jun-Ichi Kira, Comparison of clinical, immunological and neuroimaging features between anti-aquaporin-4 antibody-positive and antibody-negative Sjögren's syndrome patients with central nervous system manifestations, Multiple Sclerosis, 10.1177/1352458511431727, 18, 6, 807-816, 2012.01, Background and objective: The objective of this study is to clarify clinical, immunological, and neuroimaging features in anti-aquaporin-4 (AQP4) antibody-positive and antibody-negative Sjögren's syndrome (SS) patients with central nervous system (CNS) involvement.Methods: Medical records and MRI scans were retrospectively analyzed in 22 consecutive SS patients with CNS manifestations.Results: Seven (31.8%) patients were positive for anti-AQP4 antibodies. The frequency of visual impairment was higher in anti-AQP4 antibody-positive patients than in antibody-negative patients (71.4% vs. 0.0%, p = 0.0008). Brain MRI showed that discrete lesions were more commonly found in the cerebrum, brainstem, and optic nerve in anti-AQP4 antibody-positive patients than in antibody-negative patients (p = 0.002, p = 0.006, and p = 0.004, respectively), while spinal cord MRI showed that posterior column lesions in the cervical spinal cord were more frequent in anti-AQP4 antibody-positive patients than in antibody-negative patients (71.4% vs. 14.3%, p = 0.01). SS-A antibody titers were higher in anti-AQP4 antibody-positive patients than in antibody-negative patients (p = 0.012) and were also higher in patients with longitudinally extensive spinal cord lesions (LESCLs) than in those without LESCLs (p = 0.019).Conclusions: In SS, the presence of anti-AQP4 antibodies is associated with involvement of the optic nerve, cerebrum and brainstem, and with cervical posterior column lesions in the spinal cord..
70. Noriko Isobe, Tomomi Yonekawa, Takuya Matsushita, Yuji Kawano, Katsuhisa Masaki, Satoshi Yoshimura, Jakub Fichna, Shu Chen, Jadwiga Furmaniak, Bernard Rees Smith, Jun-Ichi Kira, Quantitative assays for anti-aquaporin-4 antibody with subclass analysis in neuromyelitis optica, Multiple Sclerosis, 10.1177/1352458512443917, 18, 11, 1541-1551, 2012.01, Background: To clarify the clinical relevance of anti-aquaporin-4 (anti-AQP4) antibody titers and immunoglobulin (IgG) subclass. Methods: Using a bridging enzyme-linked immunosorbent assay (ELISA), a flow cytometric assay (FCMA) and an immunofluorescence assay (IFA) for anti-AQP4 antibodies, sera from 142 patients with multiple sclerosis (MS) as defined by the McDonald criteria (2005), 29 with neuromyelitis optica (NMO) who fulfilled the 1999 criteria, 19 with recurrent and/or longitudinally extensive myelitis (RM/LM), 86 with other non-inflammatory neurological diseases (OND) and 28 healthy controls (HC) were studied. Results: Anti-AQP4 antibody positivity rates by IFA, FCMA, and ELISA were 41.4%, 51.7% and 48.3%, respectively, in NMO (1999) patients, and 0% in the OND and HC groups. Twenty-six MS patients (18.3%) were positive for the antibody; 17 met the 2006 NMO criteria, including positivity for anti-AQP4 antibody, and five had longitudinally extensive myelitis (LM). Among the cases with anti-AQP4 antibody detected by FCMA, IgG1, 2, 3, and 4 anti-AQP4 antibodies were found in 97.8%, 37.0%, 6.5% and 6.5% respectively. There was no association of either antibody positivity or level of anti-AQP4 antibody IgG subclasses with clinical parameters after adjustment of p values for multiple comparisons. Conclusions: FCMA and bridging ELISA are useful for detecting and quantifying anti-AQP4 antibodies..
71. Nobutoshi Kawamura, Hua Piao, Motozumi Minohara, Takuya Matsushita, Susumu Kusunoki, Hiroshi Matsumoto, Kazuhiro Ikenaka, Yoshimitsu Mizunoe, Jun-Ichi Kira, Campylobacter jejuni DNA-binding protein from starved cells in Guillain-Barré syndrome patients, Journal of Neuroimmunology, 10.1016/j.jneuroim.2011.09.004, 240-241, 74-78, 2011.12, Campylobacter jejuni enteritis is frequently associated with an axonal form of Guillain-Barré syndrome (GBS) and C. jejuni DNA-binding protein from starved cells (C-Dps) induces paranodal myelin detachment and axonal degeneration through binding with sulfatide in vivo. Here we investigated the invasion of C-Dps into hosts with C. jejuni-related GBS. Our analyses of patient sera found that both C-Dps and anti-C-Dps antibodies were most commonly detected in sera from C. jejuni-related GBS patients (5/27, 14.8% and 15/24, 62.5%; respectively). These findings suggest that C-Dps invades the host and may potentially contribute to the peripheral nerve damage in C. jejuni-related GBS..
72. J. P. McElroy, Noriko Isobe, P. A. Gourraud, S. J. Caillier, Takuya Matsushita, T. Kohriyama, K. Miyamoto, Y. Nakatsuji, T. Miki, S. L. Hauser, J. R. Oksenberg, Jun-Ichi Kira, SNP-based analysis of the HLA locus in Japanese multiple sclerosis patients, Genes and Immunity, 10.1038/gene.2011.25, 12, 7, 523-530, 2011.10, Although several major histocompatibility complex (MHC)-wide single-nucleotide polymorphism (SNP) studies have been performed in populations of European descent, none have been performed in Asian populations. The objective of this study was to identify human leukocyte antigen (HLA) loci associated with multiple sclerosis (MS) in a Japanese population genotyped for 3534 MHC region SNPs. Using a logistic regression model, two SNPs (MHC Class III SNP rs422951 in the NOTCH4 gene and MHC Class II SNP rs3997849, susceptible alleles A and G, respectively) were independently associated with MS susceptibility (204 patients; 280 controls), two (MHC Class II SNP rs660895 and MHC Class I SNP rs2269704 in the NRM gene, susceptible alleles G and G, respectively) with aquaporin-4 (AQP4) MS susceptibility (149 patients; 280 controls) and a single SNP (MHC Class II SNP rs1694112, susceptible allele G) was significant when contrasting AQP4 against AQP4 patients. Haplotype analysis revealed a large susceptible association, likely DRB104 or a locus included in the DRB104 haplotype, with AQP4 MS, which excluded DRB115:01. This study is the largest study of the HLA's contribution to MS in Japanese individuals..
73. Arnold Angelo M Pineda, Motozumi Minohara, Nobutoshi Kawamura, Takuya Matsushita, Ryo Yamasaki, Xiaojia Sun, Hua Piao, Hiroaki Shimokawa, Jun-Ichi Kira, Preventive and therapeutic effects of the selective Rho-kinase inhibitor fasudil on experimental autoimmune neuritis, Journal of the Neurological Sciences, 10.1016/j.jns.2011.03.031, 306, 1-2, 115-120, 2011.07, We studied the effects of fasudil, a selective Rho-kinase inhibitor, on experimental autoimmune neuritis (EAN). Continuous parenteral administration of fasudil prevented the development of EAN induced by P0 peptide 180-199 in Lewis rats while it also reduced EAN severity when administered after disease onset. Immunohistochemical examination disclosed a marked decrease in the amount of inflammatory cell infiltration and attenuation of demyelination and axonal degeneration. Specific proliferation of lymphocytes from fasudil-treated rats in response to P0 peptide was significantly reduced as compared with those from phosphate-buffered saline (PBS)-treated rats. Fasudil treatment was associated with a significant reduction in secretion of IFN-γ; by contrast, secretion of IL-4 was almost the same in the fasudil- and PBS-treated groups. As a result, the IFN-γ/IL-4 ratio in the supernatant was significantly deceased in fasudil-treated rats compared with PBS-treated ones. Therefore, our results indicate a beneficial effect of selective blockade of Rho-kinase in animals with autoimmune inflammation of the peripheral nerves, and may provide a rationale for the selective blockade of Rho-kinase as a new therapy for Guillain-Barré syndrome..
74. L. Fang, Noriko Isobe, S. Yoshimura, T. Yonekawa, Takuya Matsushita, K. Masaki, H. Doi, K. Ochi, K. Miyamoto, Y. Kawano, Jun-Ichi Kira, Interleukin-7 receptor alpha gene polymorphism influences multiple sclerosis risk in Asians, Neurology, 10.1212/WNL.0b013e31821f466c, 76, 24, 2125-2127, 2011.06.
75. Tomomi Yonekawa, Takuya Matsushita, Motozumi Minohara, Noriko Isobe, Katsuhisa Masaki, Satoshi Yoshimura, Yasuharu Nishimura, Jun-Ichi Kira, T cell reactivities to myelin protein-derived peptides in neuromyelitis optica patients with anti-aquaporin-4 antibody, Neurology Asia, 16, 2, 139-142, 2011.06, We previously reported the establishment of major myelin protein-derived T cell lines from 11 patients with multiple sclerosis. In the present study, we determined anti-aquaporin-4 (AQP4) antibody status in these patients and classified them into five patients with anti-AQP4 antibody who met the criteria for neuromyelitis optica (NMO) or NMO spectrum disorders, and six patients without anti-AQP4 antibody who fulfilled the revised McDonald criteria for multiple sclerosis. T cell lines reactive to myelin oligodendrocyte glycoprotein, proteolipid protein and myelin basic protein were detected in 5/5, 3/5 and 3/5 of the anti-AQP4 antibody-positive patients, respectively, and in 5/6, 4/6 and 4/6 of the anti-AQP4 antibody-negative ones, respectively. T cell lines from most of these patients showed inter- or intra-molecular epitope spreading, irrespective of anti-AQP4 antibody status. These findings suggest that T cells are stimulated in vivo against major myelin proteins in anti-AQP4 antibody-positive patients with NMO/NMO spectrum disorders..
76. Hikaru Doi, Takuya Matsushita, Noriko Isobe, Takahisa Tateishi, Jun-Ichi Kira, Analysis of cerebrospinal fluid cytokines and growth factors in multiple sclerosis patients with and without chronic headaches, Neurology Asia, 16, 1, 65-70, 2011.03, Background: We previously reported that, in Japanese patients with multiple sclerosis (MS), the frequency of chronic headaches was signifi cantly higher after administration of interferon beta (IFNβ). However, the mechanisms underlying IFNβ-related chronic headaches were unknown. Objective: To clarify the mechanisms underlying IFNβ-induced chronic headaches in MS patients by analyzing cytokine levels in cerebrospinal fluid (CSF). Methods: We measured the levels of 27 CSF cytokines and growth factors using a fluorescent bead-based immunoassay, during a headache-free period, in 34 MS patients enrolled in our previous survey on chronic headaches. Results: There were no signifi cant differences in CSF cytokine levels between the 21 MS patients with chronic headaches and the 13 without chronic headaches. Among the 14 patients receiving IFNβ therapy, the 5 patients with chronic headaches showed signifi cantly lower levels of interleukin (IL) 15, IL17 and chemokine (C-C motif) ligand 2 (CCL2) (also known as monocyte chemoattractant protein 1; MCP1) compared with the 9 patients without chronic headaches (P < 0.05). Conclusions: The present survey revealed that in MS, chronic headache sufferers on IFNβ therapy had decreased levels of IL15, IL17 and CCL2 in CSF. This suggests that chronic headaches may tend to develop in good responders to IFNβ..
77. Hua Piao, Motozumi Minohara, Nobutoshi Kawamura, Wei Li, Takuya Matsushita, Ryo Yamasaki, Yoshimitsu Mizunoe, Jun-Ichi Kira, Tissue binding patterns and in vitro effects of campylobacter jejuni DNA-binding protein from starved cells, Neurochemical Research, 10.1007/s11064-010-0263-7, 36, 1, 58-66, 2011.01, Campylobacter jejuni (C. jejuni) is frequently associated with axonal Guillain-Barré syndrome (GBS). We reported that C. jejuni DNA-binding protein from starved cells (C-Dps) binds to and damages myelinated nerves in vivo. We studied the binding patterns of C-Dps to nervous tissues and its in vitro effects on neural cells. Immunohistochemically, C-Dps labeled the nodes of Ranvier, the outermost parts of internodal myelin and the basement membrane in the peripheral nerves, and neurons and myelin in the central nervous tissues. Its binding was blocked by sulfatide. C-Dps bound to the cell surfaces of nerve growth factor (NGF)-treated PC12 cells leading to dose-dependent LDH release, which was inhibited by either heat-denaturation of C-Dps or coincubation with an anti-C-Dps mAb. However, its binding to the surfaces of cultured NSC34 cells, S16 cells, or dorsal root ganglion cells, did not induce cytotoxicity. These findings suggest a possible involvement of C-Dps in C. jejuni-related GBS..
78. Koji Shinoda, Takuya Matsushita, Konosuke Furuta, Noriko Isobe, Tomomi Yonekawa, Yasumasa Ohyagi, Jun-Ichi Kira, Wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) syndrome in a patient with neuromyelitis optica spectrum disorder and anti-aquaporin-4 antibody, Multiple Sclerosis, 10.1177/1352458510391690, 17, 7, 885-887, 2011.01, This report describes, for the first time, an occurrence of wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) in a 19-year-old female with neuromyelitis optica (NMO) spectrum disorder, who had anti-aquaporin-4 (AQP4) antibody. A high signal intensity lesion on T2-weighted MRI was detected in the midbrain tegmentum adjacent to the aqueduct, and presumably involved the medial longitudinal fasciculus bilaterally at the caudal levels. Plasma exchange resolved both WEBINO syndrome and the midbrain lesion. Although WEBINO syndrome is occasionally reported in multiple sclerosis patients, diagnosis of NMO should not be excluded in patients with WEBINO syndrome, because AQP4 is expressed abundantly around the periaqueductal region..
79. Takuya Matsushita, Katsuhisa Masaki, Satoru Suzuki, Takeshi Matsuoka, Tomomi Yonekawa, Xiao Mu Wu, Takeshi Tabira, Toru Iwaki, Jun-Ichi Kira, Astrocytopathy in neuromyelitis optica, multiple sclerosis and Baló's disease, Clinical Neurology, 10.5692/clinicalneurol.51.898, 51, 11, 898-900, 2011, Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) while neuromyelitis optica (NMO) is an inflammatory disease of the CNS that selectively affects the optic nerves and spinal cord. An anti-body for aquaporin-4 (AQP4), which is a water channel located in astrocyte foot process, is specifically positive for NMO and antibody and complement dependent astrocytic damage is thought to be a main cause of NMO. Baló's disease is characterized by alternating rings of demyelination and preserved myelin. We pathologically compared the astrocytic changes among autopsied cases with these CNS demyelinating diseases. NMO, MS and Baló's disease shared with reduced AQP4 immunoreactivity independent of antibodies and complements. The pathological finding was accompanied with a reduced immunoreactivity of connexin 43 and perivascular lymphocytic cuffing predominantly composed by T cells. The loss of astrocytic proteins such as AQP4 and connexin 43 preceded the loss of myelin proteins in some lesions. These features suggest astrocyte damages resulting in the loss of connexin 43 cause demyelination through the impairment of interaction between astrocytes and oligodendrocytes and the pathomechanism involves a T cell reaction..
80. Ryo Yamasaki, Masahito Tanaka, Mami Fukunaga, Takahisa Tateishi, Hitoshi Kikuchi, Kyoko Motomura, Takuya Matsushita, Yasumasa Ohyagi, Jun-Ichi Kira, Restoration of microglial function by granulocyte-colony stimulating factor in ALS model mice, Journal of Neuroimmunology, 10.1016/j.jneuroim.2010.07.002, 229, 1-2, 51-62, 2010.12, We studied the effects of G-CSF on microglial reactions in mutant SOD1 (mSOD1)-Tg (G93A) ALS model mice. Following hypoglossal axotomy, the numbers of neurons and microglia expressing GDNF were significantly lower in mSOD1-Tg mice than in non-transgenic (NTG) littermates. This decrease in the number of neurons after axotomy and a decrease in the number of large myelinated axons in mSOD1-Tg mice over the disease course were improved by G-CSF, which also increased microglial recruitment. Impaired migration of cultured mSOD1-Tg microglia to MCP-1 was recovered following G-CSF treatment. Restoration of microglial responses by G-CSF may contribute to its neuroprotective effects..
81. Satoshi Yoshimura, Hirofumi Ochi, Noriko Isobe, Takuya Matsushita, Kyoko Motomura, Takeshi Matsuoka, Motozumi Minohara, Jun-Ichi Kira, Altered production of brain-derived neurotrophic factor by peripheral blood immune cells in multiple sclerosis, Multiple Sclerosis, 10.1177/1352458510375706, 16, 10, 1178-1188, 2010.10, Background: Within multiple sclerosis lesions, brain-derived neurotrophic factor is detected in neurons and immunocytes. Objective: To clarify brain-derived neurotrophic factor production by peripheral blood immunocytes and its relationship with clinical parameters in multiple sclerosis. Methods: Serum brain-derived neurotrophic factor levels were measured by conventional enzyme-linked immunosorbent assay while brain-derived neurotrophic factor production by immunocytes was determined by an in situ enzyme-linked immunosorbent assay in 74 multiple sclerosis patients, 32 healthy controls, and 86 patients with other neurological diseases. The tyrosine kinase receptor TrkB expression level in peripheral blood mononuclear cells was measured by real-time polymerase chain reaction. Results: Multiple sclerosis patients showed significantly higher serum brain-derived neurotrophic factor levels than healthy controls and patients with other neurological diseases. Multiple sclerosis patients with high brain-derived neurotrophic factor levels were younger, and showed fewer relapse numbers than those with low brain-derived neurotrophic factor levels. Brain-derived neurotrophic factor production by T cells increased with age in healthy controls, but not in multiple sclerosis patients. Interferon beta induced a significant increase in serum brain-derived neurotrophic factor levels. Brain-derived neurotrophic factor production from T cells and TrkB expression levels in peripheral blood mononuclear cells were significantly enhanced in interferon beta-treated multiple sclerosis patients compared with untreated ones. Conclusions: A high brain-derived neurotrophic factor level is related to early mild disease in young multiple sclerosis patients. Interferon beta potentiates brain-derived neurotrophic factor production and brain-derived neurotrophic factor receptor expression in peripheral blood mononuclear cells, which may act beneficially..
82. Takuya Matsushita, Noriko Isobe, M. Kawajiri, M. Mogi, K. Tsukuda, M. Horiuchi, Y. Ohyagi, Jun-Ichi Kira, CSF angiotensin II and angiotensin-converting enzyme levels in anti-aquaporin-4 autoimmunity, Journal of the Neurological Sciences, 10.1016/j.jns.2010.05.014, 295, 1-2, 41-45, 2010.08, Background: Anti-aquaporin-4 (AQP4) antibody targets perivascular astrocyte foot processes, which contain abundant angiotensinogen, a precursor of angiotensin II, angiotensin-converting enzyme (ACE) and ACE2. Objective: To disclose any abnormality in the intrathecal angiotensin II metabolic pathway in Japanese patients with neuromyelitis optica (NMO) or NMO spectrum disorders (NMOs) and positive for anti-AQP4 antibody. Methods: We measured CSF angiotensin II, ACE and ACE2 levels in 15 anti-AQP4 antibody-positive patients with NMO or NMOs, 21 anti-AQP4 antibody-negative multiple sclerosis (MS) patients, 32 patients with other neurological diseases (OND) and 24 non-neurologic controls, using established ELISAs. Results: CSF angiotensin II levels were lower in patients with NMO/NMOs (2.01 ± 1.82 pg/ml) and those with MS (3.15 ± 1.67 pg/ml) than in the OND (5.41 ± 2.34 pg/ml) and control groups (6.71 ± 2.65 pg/ml) (Pcorr < 0.005). The difference in CSF angiotensin II levels between NMO/NMOs and MS patients was nearly significant (Puncorr = 0.052). In NMO/NMOs and MS patients, angiotensin II levels were negatively correlated with CSF/serum albumin ratio (P < 0.05). ACE levels in CSF were lower in patients with NMO/NMOs (34.3 ± 5.61 ng/ml) than in MS patients (42.5 ± 8.19 ng/ml, Pcorr = 0.035) and controls (44.7 ± 4.02 ng/ml, Pcorr < 0.0003) while ACE2 levels were lower in NMO/NMOs (1.13 ± 0.49 ng/ml) and MS (1.75 ± 0.86 ng/ml) patients than in controls (2.76 ± 0.23 ng/ml, Pcorr < 0.001 for both). Conclusion: CSF angiotensin II, ACE, and ACE2 levels are decreased in NMO/NMOs patients with anti-AQP4 antibody, reflecting severe destruction of perivascular astrocytes..
83. Takahisa Tateishi, Ryo Yamasaki, Masahito Tanaka, Takuya Matsushita, Hitoshi Kikuchi, Noriko Isobe, Yasumasa Ohyagi, Jun-Ichi Kira, CSF chemokine alterations related to the clinical course of amyotrophic lateral sclerosis, Journal of Neuroimmunology, 10.1016/j.jneuroim.2010.03.004, 222, 1-2, 76-81, 2010.05, We measured the levels of 27 cytokines/chemokines and growth factors in cerebrospinal fluid (CSF) from 42 patients with sporadic amyotrophic lateral sclerosis (ALS), 12 patients with lower motor neuron disease (LMND), and 34 control patients with non-inflammatory neurological diseases (OND), using a multiplexed fluorescent bead-based immunoassay. Among cytokines/chemokines elevated in ALS, CCL2 and CXCL8 levels were negatively correlated with the revised ALS functional rating scale (ALSFRS-R) score, while CCL4 showed a positive correlation with ALSFRS-R score. CCL4 and CXCL10 showed negative correlations with disease progression rate. These chemokine alterations are assumed to somehow correlate with the clinical course of ALS..
84. Koichi Hagiwara, Tsuyoshi Okamoto, Hiroshi Shigeto, Katsuya Ogata, Yuko Somehara, Takuya Matsushita, Jun-Ichi Kira, Shozo Tobimatsu, Oscillatory gamma synchronization binds the primary and secondary somatosensory areas in humans, NeuroImage, 10.1016/j.neuroimage.2010.02.001, 51, 1, 412-420, 2010.05, Induced gamma activity has a key role in the temporal binding of distributed cortico-cortical processing. To elucidate the neural synchronization in the early-stage somatosensory processing, we studied the functional connectivity between the primary and secondary somatosensory cortices (SI and SII) in healthy subjects using magnetoencephalography (MEG) with excellent spatiotemporal resolution. First, somatosensory-evoked magnetic fields were recorded to determine the locations of each cortical activity. Then we analyzed the phase-locking values (PLVs) of the induced gamma activity to assess neural synchrony within the somatosensory cortical network. We also assessed PLVs in patients with multiple sclerosis (MS) to validate our PLV analysis in evaluating the inter-areal functional connectivity, which can often be impaired in MS. The PLVs of the induced gamma activity were calculated for each pair of unaveraged MEG signals that represented the activities of the contralateral SI and bilateral SII areas. Analysis of PLVs between the SI and SII areas showed significantly increased PLVs for gamma-band activities, starting at an early post-stimulus stage in normal controls, whereas this increase in PLVs was apparently diminished in MS. The PLV analysis provided evidence for early-latency, gamma-band neuronal synchronization between the SI and SII areas in normal controls. Our study first demonstrates the gamma-band synchrony in the early-stage human somatosensory processing..
85. Takuya Matsushita, Noriko Isobe, Hua Piao, Takeshi Matsuoka, Takaaki Ishizu, Hikaru Doi, Katsuhisa Masaki, Takashi Yoshiura, Ryo Yamasaki, Yasumasa Ohyagi, Jun-Ichi Kira, Reappraisal of brain MRI features in patients with multiple sclerosis and neuromyelitis optica according to anti-aquaporin-4 antibody status, Journal of the Neurological Sciences, 10.1016/j.jns.2010.01.009, 291, 1-2, 37-43, 2010.04, Brain lesions are not uncommon in neuromyelitis optica (NMO) patients with anti-aquaporin-4 (AQP4) antibody; however, the appearance of these lesions is said to be different from that of those in Western patients with multiple sclerosis (MS). To clarify the similarities and dissimilarities of brain lesions in anti-AQP4 antibody-positive and -negative MS and NMO patients, we examined the presence of anti-AQP4 antibody in the sera of 148 consecutive patients fulfilling Poser's criteria for clinically definite MS, of whom 38 also met the revised NMO criteria, using an immunofluorescence method, and analyzed brain lesions by magnetic resonance imaging (MRI). Brain lesions fulfilling the Barkhof criteria were significantly more common in 121 patients without anti-AQP4 antibody than in 27 patients with anti-AQP4 antibody (57.0% vs. 33.3%, P = 0.033), while the frequency of those that met the Paty criteria was not different between the two groups (74.4% vs. 73.5%). Ovoid lesions were detected more commonly in patients without anti-AQP4 antibody than in those with the antibody (72.3% vs. 48.2%, P = 0.022). The anti-AQP4 antibody-positive patients had significantly more atypical brain lesions, such as extensive brain lesions, than the anti-AQP4 antibody-negative ones (18.5% vs. 1.7%, P = 0.0023). Thus, although MS-like brain lesions are more common in anti-AQP4 antibody-negative patients than anti-AQP4 antibody-positive patients, approximately 30 to 50% of patients with anti-AQP4 antibody harbour brain MRI lesions indistinguishable from those present in typical MS patients, such as periventricular ovoid lesions, suggesting the existence of considerable overlap in brain MRI features between anti-AQP4 antibody-positive and -negative Asian patients. In the present study, NMO patients with brain lesions showed a significantly higher annualized relapse rate (Pcorr = 0.017) and higher frequency of anti-AQP4 antibody (Pcorr < 0.0001) than typical NMO patients without brain lesions, suggesting that development of brain lesions in NMO may reflect high disease activity and thus be a warning sign..
86. Jun-Ichi Kira, Motozumi Minohara, Wei Li, Takuya Matsushita, Authors? reply to ?Helicobacter pylori with or without its neutrophil-activating protein may be the common denominator associated with multiple sclerosis and neuromyelitis optica?, Multiple Sclerosis, 10.1177/1352458510361994, 16, 3, 378-379, 2010.03.
87. Noriko Isobe, Takuya Matsushita, Ryo Yamasaki, S. V. Ramagopalan, Y. Kawano, Y. Nishimura, G. C. Ebers, Jun-Ichi Kira, Influence of HLA-DRB1 alleles on the susceptibility and resistance to multiple sclerosis in Japanese patients with respect to anti-aquaporin 4 antibody status, Multiple Sclerosis, 10.1177/1352458509355067, 16, 2, 147-155, 2010.02, Background: Epistatic interactions between human leukocyte antigen (HLA)-DRB1 alleles alter multiple sclerosis (MS) risk in Caucasians. Such interactions have never been studied in Asian MS patients. Objective: To investigate the influence of HLA-DRB1 alleles, including epistatic interactions at this locus, in Japanese MS patients with and without the anti-aquaporin 4 (AQP4) antibody. Methods: The HLA-DRB1 locus was genotyped in 108 MS patients and 127 healthy controls. MS patients were further classified into two groups according to anti-AQP4 antibody status (27 positive and 81 negative). Results: HLA-DRB1*09 (adjusted odds ratio (OR) = 0.243, 95% confidence interval (CI) 0.099-0.533) and HLA-DRB1*01 (adjusted OR = 0.327, 95% CI 0.103-0.873) decreased the incidence of anti-AQP4 antibody-negative MS. By contrast, HLA-DRB1*12 increased the risk of anti-AQP4 antibody-positive MS (adjusted OR = 3.691, 95% CI 1.233-10.565). Individuals with HLA-DRB1*09/15 decreased the risk of anti-AQP4 antibody-negative MS (adjusted OR = 0.164, 95% CI 0.026-0.593), while those with HLA-DRB1*12/15 increased the risk of anti-AQP4 antibody-positive MS (adjusted OR = 10.870, 95% CI 2.004-81.752). Conclusions: The ability of HLA-DRB1*09 to reduce the risk of anti-AQP4 antibody-negative MS may arise from an interaction with HLA-DRB1*15. By contrast, HLA-DRB1*12 increases susceptibility to anti-AQP4 antibody-positive MS, possibly via an interaction with HLA-DRB1*15..
88. Kazumasa Okada, Takuya Matsushita, Jun-Ichi Kira, Sadatoshi Tsuji, B-cell activating factor of the tnf family is upregulated in neuromyelitis optica, Neurology, 10.1212/WNL.0b013e3181c919ee, 74, 2, 177-178, 2010.01.
89. Hua Piao, Motozumi Minohara, Nobutoshi Kawamura, Wei Li, Yoshimitsu Mizunoe, Fujio Umehara, Yoshinobu Goto, Susumu Kusunoki, Takuya Matsushita, Kazuhiro Ikenaka, Takashi Maejima, Jun ichi Nabekura, Ryo Yamasaki, Jun-Ichi Kira, Induction of paranodal myelin detachment and sodium channel loss in vivo by Campylobacter jejuni DNA-binding protein from starved cells (C-Dps) in myelinated nerve fibers, Journal of the Neurological Sciences, 10.1016/j.jns.2009.10.007, 288, 1-2, 54-62, 2010.01, In an axonal variant of Guillain-Barré syndrome (GBS) associated with Campylobacter jejuni (C. jejuni) enteritis, the mechanism underlying axonal damage is obscure. We purified and characterized a DNA-binding protein from starved cells derived from C. jejuni (C-Dps). This C-Dps protein has significant homology with Helicobacter pylori neutrophil-activating protein (HP-NAP), which is chemotactic for human neutrophils through binding to sulfatide. Because sulfatide is essential for paranodal junction formation and for the maintenance of ion channels on myelinated axons, we examined the in vivo effects of C-Dps. First, we found that C-Dps specifically binds to sulfatide by ELISA and immunostaining of thin-layer chromatograms loaded with various glycolipids. Double immunostaining of peripheral nerves exposed to C-Dps with anti-sulfatide antibody and anti-C-Dps antibody revealed co-localization of them. When C-Dps was injected into rat sciatic nerves, it densely bound to the outermost parts of the myelin sheath and nodes of Ranvier. Injection of C-Dps rapidly induced paranodal myelin detachment and axonal degeneration; this was not seen following injection of PBS or heat-denatured C-Dps. Electron microscopically, C-Dps-injected nerves showed vesiculation of the myelin sheath at the nodes of Ranvier. Nerve conduction studies disclosed a significant reduction in compound muscle action potential amplitudes in C-Dps-injected nerves compared with pre-injection values, but not in PBS-, heat-denatured C-Dps-, or BSA-injected nerves. However, C-Dps did not directly affect Na+ currents in dissociated hippocampal neurons. Finally, when C-Dps was intrathecally infused into rats, it was deposited in a scattered pattern in the cauda equina, especially in the outer part of the myelin sheath and the nodal region. In C-Dps-infused rats, but not in BSA-infused ones, a decrease in the number of sodium channels, vesiculation of the myelin sheath, axonal degeneration and infiltration of Iba-1-positive macrophages were observed. Thus, we consider that C-Dps damages myelinated nerve fibers, possibly through interference with paranodal sulfatide function, and may contribute to the axonal pathology seen in C. jejuni-related GBS..
90. W. Qiu, J. S. Wu, M. N. Zhang, Takuya Matsushita, Jun-Ichi Kira, W. M. Carroll, F. L. Mastaglia, A. G. Kermode, Longitudinally extensive myelopathy in Caucasians
A West Australian study of 26 cases from the Perth Demyelinating Diseases Database, Journal of Neurology, Neurosurgery and Psychiatry, 10.1136/jnnp.2009.172973, 81, 2, 209-212, 2010.01, Objectives: To characterise West Australian cases of longitudinally extensive myelopathy (LEM). Methods: Twenty six patients with LEM were identified from a cohort of 983 patients with demyelinating disease. Clinical and MRI data and AQP4-IgG results were reviewed. Results: LEM cases were classified as conventional MS (CMS) 13, neuromyelitis optica (NMO) 7, and isolated LEM 6. LEM was the initial presentation in 13/26 cases. In CMS cases lesions were mainly in the lower cervical cord (C4-C7) whereas in NMO and isolated LEM they were more often thoracic and were longer. The severity of disability was highly variable but was greater in the NMO than the CMS group. Only one of 20 patients tested was seropositive for AQP4-IgG. Conclusion LEM occurred as part of CMS or NMO or in isolation. Patients with LEM had highly heterogeneous clinical characteristics and a low rate of AQP4-IgG seropositivity..
91. Wei Li, Motozumi Minohara, Hua Piao, Takuya Matsushita, Katsuhisa Masaki, Takeshi Matsuoka, Noriko Isobe, Jen Jen Su, Yasumasa Ohyagi, Jun-Ichi Kira, Association of anti-Helicobacter pylori neutrophil-activating protein antibody response with anti-aquaporin-4 autoimmunity in Japanese patients with multiple sclerosis and neuromyelitis optica, Multiple Sclerosis, 10.1177/1352458509348961, 15, 12, 1411-1421, 2009.12, There are two distinct subtypes of multiple sclerosis (MS) in Asians: opticospinal (OSMS) and conventional (CMS). OSMS has similar features to neuromyelitis optica (NMO) and half of OSMS patients have the NMO-Immunoglobulin G (IgG)/ anti-aquaporin-4 (AQP4) antibody. We reported that Helicobacter pylori (H. pylori) infection was significantly less common in CMS patients than controls. To reveal the immune responses to the H. pylori neutrophil-activating protein (HP-NAP) in Japanese MS patients, according to anti-AQP4 antibody status, sera from 162 MS patients, 37 patients with other inflammatory neurological diseases (OIND), and 85 healthy subjects were assayed for anti-H. pylori antibodies, anti-HP-NAP antibodies, and myeloperoxidase (MPO) by enzyme immunoassays. H. pylori seropositivity rates were significantly higher in anti-AQP4 antibody-positive MS/NMO (AQP4 + /MS) patients (19/27, 70.4%) than anti-AQP4 antibody-negative CMS (AQP4 - /CMS) patients (22/83, 26.5%). Among H. pylori-infected individuals, the anti-HP-NAP antibody was significantly more common in AQP4 + /MS and AQP4 - /OSMS patients than healthy subjects (36.8%, 34.6% versus 2.8%). Among the AQP4 + /MS patients, a significant positive correlation between anti-HP-NAP antibody levels and the final Kurtzkes Expanded Disability Status Scale scores was found, and MPO levels were higher in anti-HP-NAP antibody-positive patients than anti-HP-NAP antibody-negative ones. Therefore, HP-NAP may be associated with the pathology of anti-AQP4 antibody-related neural damage in MS/NMO patients..
92. Yasuyuki Kihara, Takuya Matsushita, Yoshihiro Kita, Satoshi Uematsu, Shizuo Akira, Jun-Ichi Kira, Satoshi Ishii, Takao Shimizu, Targeted lipidomics reveals mPGES-1-PGE2 as a therapeutic target for multiple sclerosis, Proceedings of the National Academy of Sciences of the United States of America, 10.1073/pnas.0906891106, 106, 51, 21807-21812, 2009.12, The arachidonic acid (AA) cascade produces eicosanoids, such as prostaglandins (PGs), that regulate physiological and pathological functions. Although various nonsteroidal anti-inflammatory drugs have been developed, blocking upstream components (cyclooxygenase-1 and -2) of the AA cascade leads to severe side effects, including gastrointestinal ulcers and cardiovascular events, respectively, due to the complexity of the AA cascade. Here, using anAAcascade-targeted lipidomics approach, we report that microsomal PGE synthase 1 (mPGES-1) plays a key role in experimental autoimmune encephalomyelitis (EAE). Eicosanoids (mainly PGD2) are produced constitutively in the spinal cord of naive mice. However, in EAE lesions, the PGE2 pathway is favored and the PGD2, PGI2, and 5-lipoxygenase pathways are attenuated. Furthermore, mPGES-1-/- mice showed less severe symptoms of EAE and lower production of IL-17 and IFN-γ than mPGES-1 +/+ mice. Expression of PGE2 receptors (EP1, EP2, and EP4) was elevated in EAE lesions and correlated with clinical symptoms. Immunohistochemistry on central nervous systems of EAE mice and multiple sclerosis (MS) patients revealed overt expression of mPGES-1 protein in microglia/macrophages. Thus, the mPGES-1-PGE2-EPs axis of the AA cascade may exacerbate EAE pathology. Our findings have important implications for the design of therapies for MS..
93. Hikaru Doi, Takuya Matsushita, Noriko Isobe, Takaaki Ishizu, Yasumasa Ohyagi, Jun-Ichi Kira, Frequency of chronic headaches in japanese patients with multiple sclerosis
With special reference to opticospinal and common forms of multiple sclerosis, Headache, 10.1111/j.1526-4610.2009.01427.x, 49, 10, 1513-1520, 2009.11, Background. - Headache is common in Western patients with multiple sclerosis (MS), but its frequency has not been reported in Asian patients. In Asians, the opticospinal form of MS, showing similar characteristics to relapsing neuromyelitis optica in Westerners, is regarded as a different subtype from conventional MS. Objectives. - The aim of this study was to clarify the frequency of primary and chronic secondary headaches in Japanese patients with MS and the factors associated with the emergence of such headaches. Methods. - We investigated 127 consecutive patients with clinically definite MS. Frequencies of primary and chronic secondary headaches were compared according to clinical subtype, administration of interferon beta, and anti-aquaporin-4 antibody status. Results. - The frequency of patients with primary and chronic secondary headaches at the time of interview was 64/127 (50.4%); the frequency of migraine was 26/127 (20.4%) and that of tension-type headache was 38/127 (29.9%). The frequencies of patients with primary and chronic secondary headaches and migraine without aura after the onset of MS were higher in patients undergoing interferon beta therapy than in those not on the therapy (42.4% vs 23.4%, P <.05 and 15.1% vs 4.3%, P =.05, respectively). There were no significant differences in the frequency of primary and chronic secondary headaches based on clinical subtype of MS. However, among patients not receiving interferon beta, the occurrence of migraine with aura after the onset of MS was significantly higher in patients with anti-aquaporin-4 antibody than in patients without the antibody (13.3% vs 0.0%, P <.05). Conclusions. - In Japanese patients with MS, the frequency of primary and chronic secondary headaches, especially migraine, was higher than in the general Japanese population. Administration of interferon beta was related to a higher frequency of primary and chronic secondary headaches, especially migraine without aura, irrespective of clinical subtype of MS..
94. Takuya Matsushita, Noriko Isobe, T. Matsuoka, T. Ishizu, Y. Kawano, T. Yoshiura, Y. Ohyagi, Jun-Ichi Kira, Extensive vasogenic edema of anti-aquaporin-4 antibody-related brain lesions, Multiple Sclerosis, 10.1177/1352458509106613, 15, 9, 1113-1117, 2009.08, Objective: Using neuroimaging, we analyzed the nature of extensive brain lesions in five anti-aquaporin-4 (AQP4) antibody-positive patients with neuromyelitis optica spectrum disorders. Results: Extensive brain lesions involved white matter in three, and basal ganglia and corpus callosum in one each. Four patients showed high diffusivity on apparent diffusion coefficient maps and three demonstrated increased choline/creatine ratios and decreased N-acetyl-aspartate/creatine ratios on 1H-magnetic resonance spectroscopy. These findings suggested that the lesions were vasogenic edema associated with inflammation. Unusual brain symptoms associated with such lesions included recurrent limbic encephalitis, parkinsonism, and coma. Conclusion: Anti-AQP4 antibody is considered to be associated with the neuroimaging appearances of vasogenic edema..
95. Takuya Matsushita, Noriko Isobe, T. Matsuoka, N. Shi, Y. Kawano, X. M. Wu, T. Yoshiura, Y. Nakao, T. Ishizu, Jun-Ichi Kira, Aquaporin-4 autoimmune syndrome and anti-aquaporin-4 antibody-negative opticospinal multiple sclerosis in Japanese, Multiple Sclerosis, 10.1177/1352458509104595, 15, 7, 834-847, 2009.07, Background: Antibodies to aquaporin-4 (AQP4) are found in a fraction of Japanese opticospinal multiple sclerosis (OSMS) patients. However, it remains unknown whether anti-AQP4 antibody-positive and negative OSMS patients possess an identical disease. Objective: The objective of the current study was to clarify immunological differences between the two groups of patients. Methods: We studied the serum antibody titers against AQP4 in 191 patients with idiopathic central nervous system demyelinating diseases and clarified their relationships with immunological parameters. Results: Anti-AQP4 antibody positivity rate was higher in patients with OSMS (21/58, 36.2%), idiopathic recurrent myelitis (4/17, 23.5%), and recurrent optic neuritis (7/26, 26.9%), than in conventional MS (CMS) patients (6/90, 6.7%) and patients with other diseases (0/87). Anti-AQP4 antibody titer was significantly higher in patients with SS-A/B antibodies than in those without them. Anti-AQP4 antibody-negative OSMS patients showed significantly higher CD4+ IFN-γ+IL-4γT cell percentages and intracellular IFN-γ/IL-4 ratios than anti-AQP4 antibody-positive patients, anti-AQP4 antibody-negative CMS patients, and healthy controls, and CD4+IFN-γ+IL-4-T cell percentages were negatively correlated with anti-AQP4 antibody titers. Conclusion: Anti-AQP4 antibody-positive patients are immunologically distinct from anti-AQP4 antibody-negative OSMS patients owing to a Th2 shift in the former group in comparison to a Th1 shift in the latter..
96. Akihiro Watanabe, Takuya Matsushita, Hikaru Doi, Takashi Matsuoka, Hiroshi Shigeto, Noriko Isobe, Yuji Kawano, Shozo Tobimatsu, Jun-Ichi Kira, Multimodality-evoked potential study of anti-aquaporin-4 antibody-positive and -negative multiple sclerosis patients, Journal of the Neurological Sciences, 10.1016/j.jns.2009.02.371, 281, 1-2, 34-40, 2009.06, Neuromyelitis optica (NMO) is claimed to be a distinct disease entity from multiple sclerosis (MS) because of its strong association with NMO-IgG/anti-AQP4 antibody; however, the in vivo role of the antibody remains unknown. Therefore, we aimed to clarify whether the presence of anti-AQP4 antibody is associated with any abnormalities in multimodality-evoked potentials in 111 patients with relapsing-remitting or relapsing-progressive MS, including the opticospinal form of MS, 18 of whom were seropositive for anti-AQP4 antibody. More patients with anti-AQP4 antibody showed a lack of the P100 component on visual-evoked potentials (VEPs) than those without the antibody (11/17, 64.7% vs. 20/84, 23.8%, p = 0.003), whereas the frequency of delayed P100 latency was significantly higher in the latter group than in the former (1/17, 5.9% vs. 28/84, 33.3%, p = 0.021). The frequencies of non-responses and delayed central sensory conduction times in median and posterior tibial nerve somatosensory-evoked potentials (SEPs) were not significantly different between anti-AQP4 antibody-positive and -negative patients. In terms of upper and lower limb motor-evoked potentials (MEPs), the frequencies of non-responses and delayed central motor conduction times did not differ significantly based on the presence or absence of anti-AQP4 antibody. The frequency of optic nerve lesions on MRI was significantly higher in anti-AQP4 antibody-positive patients than in anti-AQP4 antibody-negative patients (p = 0.0137). Multiple logistic analyses revealed that anti-AQP4 antibody positivity (OR = 8.406, p = 0.02) and unevoked VEP responses (OR = 35.432, p < 0.001) were significantly related to development of severe visual impairment. Such an association of anti-AQP4 antibody with disability was not found for either severe motor or sensory impairment. These findings suggest a distinctive nature of optic nerve lesions between anti-AQP4 antibody-positive and -negative patients; lesions are supposed to be more necrotic in the former group and more demyelinating in the latter..
97. Takuya Matsushita, Takeshi Matsuoka, Takaaki Ishizu, Hitoshi Kikuchi, Manabu Osoegawa, Yuji Kawano, Futoshi Mihara, Yasumasa Ohyagi, Jun-ichi Kira, Anterior periventricular linear lesions in optic-spinal multiple sclerosis: a combined neuroimaging and neuropathological study., Multiple Sclerosis, 10.1177/1352458507084085, 14, 3, 343-353, 2008.04, There are two distinct subtypes of multiple sclerosis (MS) in Asians, optic-spinal (OSMS) and conventional (CMS). In OSMS, severe spinal cord lesions are characteristic while brain lesions are scant. We sought to clarify atypical brain lesions in OSMS by neuroimaging and pathological studies. For brain MRI, 124 consecutive Japanese patients with clinically definite MS based on Poser criteria were enrolled, 57 with OSMS and 67 with CMS. Ten autopsied cases (seven OSMS and three CMS) were studied pathologically. Although the frequency of brain lesions fulfilling Barkhof criteria was significantly higher in CMS than in OSMS, periventricular linear lesions along with the anterior portion of the corpus callosum and the lateral ventricles were significantly more common in OSMS than in CMS. Pathologically, periventricular lesions in CMS extended deeply into the white matter, while those in OSMS were confined to periventricular areas. T cell infiltration in lesions was prominent in CMS but not in OSMS. Although severe axonal loss and cavity formation were commonly seen in periventricular and spinal cord lesions in OSMS, lymphocytic infiltrates and vessel wall thickening were observed only in the latter. Thus, we suggested that anterior periventricular linear lesions without ovoid ones are characteristic of OSMS..
98. Masahito Tanaka, Takuya Matsushita, Takahisa Tateishi, Hirofumi Ochi, Yuji Kawano, F-J Mei, Motozumi Minohara, Hiroyuki Murai, Jun-ichi Kira, Distinct CSF cytokine/chemokine profiles in atopic myelitis and other causes of myelitis., Neurology, 10.1212/01.wnl.0000326589.57128.c3, 71, 13, 974-981, 2008.09, BACKGROUND:We reported the emergence of a distinct myelitis in patients with atopic diathesis (atopic myelitis [AM]) by a nationwide survey throughout Japan. Similar cases have recently been reported in Caucasians. Pathologic studies of biopsied spinal cord specimens revealed chronic active inflammation with eosinophilic infiltration.OBJECTIVE:To clarify the cytokine/chemokine alterations in CSF from patients with AM in comparison to other causes of myelitis.METHODS:We measured 27 cytokines, chemokines, and growth factors simultaneously in CSF from 22 patients with AM, 20 with opticospinal multiple sclerosis (OSMS), 11 with HTLV-1-associated myelopathy (HAM), 9 with Sjögren syndrome-related myelitis (SM), and 20 with other noninflammatory neurologic diseases (OND), using a fluorescent bead-based immunoassay. RESULTS:In patients with AM, CCL11 and interleukin (IL)-9 were significantly increased as compared with patients with OND and other myelitis while in patients with OSMS interferon-gamma and granulocyte-colony stimulating factor levels were significantly higher than in patients with OND and other causes of myelitis. Significant increase of IL-17 in comparison to patients with OND was found only in patients with OSMS, irrespective of presence or absence of anti-aquaporin-4 (AQP4) antibody. In patients with HAM, CXCL10 and CCL5 were higher than in patients with OND and other myelitis. In patients with SM, CCL3 and CCL4 were higher than in patients with OND. In patients with AM, CCL11, IL-9, and IL-1 receptor antagonist (IL-1ra) showed positive correlations with the final Kurtzke Expanded Disability Status Scale scores while IL-1ra and IL-12(p70) had positive correlations with disease duration. CONCLUSION:Intrathecal upregulation of CCL11 and Th2 cytokines is characteristic of atopic myelitis, which is distinct from interleukin-17/interferon-gamma-related autoimmune condition of opticospinal multiple sclerosis..
99. H. Doi, Takuya Matsushita, Noriko Isobe, T. Matsuoka, M. Minohara, H. Ochi, Jun-Ichi Kira, Hypercomplementemia at relapse in patients with anti-aquaporin-4 antibody, Multiple Sclerosis, 10.1177/1352458508099139, 15, 3, 304-310, 2009.03, Objective: Because Asian patients with opticospinal multiple sclerosis (OSMS) frequently have anti-aquaporin-4 (AQP4) antibody, complement-mediated disruption of astrocyte foot processes is proposed but not yet proven. We aimed to clarify whether complement consumption occurs at relapse in anti-AQP4 antibody-positive patients. Methods: We analyzed serum CH50, C3, C4, and C-reactive protein (CRP) levels and their relation to clinical phases in 118 MS patients with or without anti-AQP4 antibody. Serum CH50 levels were higher in 24 patients with anti-AQP4 antibody than in 39 OSMS and 54 conventional form of MS (CMS) patients without anti-AQP4 antibody at relapse Pcorr < 0.05) but not in remission. The frequency of hypercomplementemia at relapse was also higher in anti-AQP4 antibody-positive patients than in anti-AQP4 antibody-negative CMS patients (70.4% vs 29.0%, Pcorr < 0.05). C3 and C4 levels did not differ significantly among the three groups at relapse. In patients with anti-AQP4 antibody, the coexistence of hypercomplementemia and high CRP values was more common at relapse than in the remission phase (36.0% vs 10.5%, P < 0.05). In patients with extensive central nervous system lesions, hypercomplementemia was significantly more common in anti-AQP4 antibody-positive patients than anti-AQP4 antibody-negative ones (88.9% vs 16.7%, P < 0.01). We consider that hypercomplementemia in anti-AQP4 antibody-positive patients may reflect a systemic inflammatory reaction at relapse..
100. Takuya Matsushita, T. Matsuoka, Noriko Isobe, Y. Kawano, M. Minohara, N. Shi, Y. Nishimura, H. Ochi, Jun-Ichi Kira, Association of the HLA-DPB1*0501 allele with anti-aquaporin-4 antibody positivity in Japanese patients with idiopathic central nervous system demyelinating disorders, Tissue Antigens, 10.1111/j.1399-0039.2008.01172.x, 73, 2, 171-176, 2009.02, There are two subtypes of multiple sclerosis (MS) in Asians: the opticospinal (OSMS) form that shows a selective involvement of the optic nerve and the spinal cord and the conventional (CMS) form that has disseminated lesions in the central nervous system including the cerebrum, cerebellum and brainstem. Both show distinct human leukocyte antigen (HLA) class II associations. OSMS has similar features to the relapsing form of neuromyelitis optica (NMO) in Western populations. Recently, it was shown that antibodies to aquaporin-4 (AQP4) are specifically detected in NMO patients and in some Japanese patients with OSMS or recurrent optic neuritis or myelitis. To clarify the immunogenetic background of anti-AQP4 antibody production, we studied HLA-DRB1 and -DPB1 gene polymorphisms in anti-AQP4 antibody-positive and -negative patients with idiopathic demyelinating diseases, such as MS, recurrent optic neuritis and recurrent myelitis. The phenotypic frequency of the HLA-DPB1*0501 allele was significantly increased in anti-AQP4 antibody-positive patients (89.5%, odds ratio = 4.8; 95% confidence interval = 1.6-14.3, n = 38, Pcorr = 0.032) compared with controls (64.0%, n = 125) but not in either anti-AQP4 antibody-negative OSMS (75.0%, n = 32) or CMS (69.2%, n = 52) patients. There was no significant correlation between any HLA-DRB1 allele and the existence of anti-AQP4 antibody. These findings suggest that the emergence of anti-AQP4 antibody is reinforced by the presence of the HLA-DPB1*0501 allele in Japanese..
101. T. Matsuoka, Takuya Matsushita, M. Osoegawa, Y. Kawano, M. Minohara, F. Mihara, Y. Nishimura, Y. Ohyagi, Jun-Ichi Kira, Association of the HLA-DRB1 alleles with characteristic MRI features of Asian multiple sclerosis, Multiple Sclerosis, 10.1177/1352458508097818, 14, 9, 1181-1190, 2008.12, Background: In Asian patients with multiple sclerosis (MS), a paucity of brain lesions and longitudinally extensive spinal cord lesions (LESCLs) extending three or more vertebral segments are characteristic findings on magnetic resonance imaging (MRI). We aimed to disclose possible factors contributing to the development of such MRI features. Method: Genotyping of HLA-DRB1 and -DPB1 alleles was performed in 121 consecutive Japanese patients with clinically definite MS based on the Poser criteria and 125 healthy controls. Possible factors associated with MRI features were determined by multiple logistic analysis. Patients with MS were classified based on the presence or absence of brain lesions fulfilling the Barkhof criteria (Barkhof brain lesions) and LESCLs. Barkhof brain lesion-negative (-) patients had a markedly lower frequency of HLA-DRB1ß0901 than controls (Pcorr < 0.05), whereas the frequency of DRB1ß1501 was increased in the Barkhof brain lesion-positive (+) group, although this increase was not significant after correction. No Barkhof(-)LESCL(+) patients carried DRB1ß0901 (Pcorr < 0.05), despite this being the most common allele in Japanese. The Barkhof(-)LESCL(-) group showed a significant increase in the frequency of DRB1ß0405 compared with controls (Pcorr < 0.05). None of the DPB1 alleles were significantly different among the groups. Using multiple logistic analysis, the absence of oligoclonal bands was positively associated with an absence of Barkhof brain lesions, whereas a higher EDSS score was positively associated with the presence of LESCLs; however, the presence of anti-aquaporin-4 antibodies was not associated with either feature. Conclusion: The characteristic MRI features in Asians are partly related to distinct HLA-DRB1 gene alleles and an absence of oligoclonal bands..
102. Takeshi Matsuoka, Takuya Matsushita, Manabu Osoegawa, Hirofumi Ochi, Yuji Kawano, Futoshi Mihara, Yasumasa Ohyagi, Jun-ichi Kira, Heterogeneity and continuum of multiple sclerosis in Japanese according to magnetic resonance imaging findings., Journal of the Neurological Sciences, 10.1016/j.jns.2007.09.010, 266, 1-2, 115-125, 2008.03, There are two distinct subtypes of multiple sclerosis (MS) in Asians: optic-spinal (OSMS) and conventional (CMS). Longitudinally extensive spinal cord lesions (LESCLs) extending over three or more vertebral segments are characteristic of patients with OSMS, yet in Asians, one-fourth of CMS patients also have LESCLs. To clarify the distinction between LESCLs in OSMS and CMS, and to characterize the relationship between the presence of LESCLs and brain magnetic resonance imaging (MRI) findings, we studied 142 patients with clinically definite MS of relapsing-remitting onset and 12 patients with primary progressive MS (PPMS) by MRI of the whole spinal cord and brain. The former was diagnosed by Poser criteria, including 57 with OSMS, 67 with CMS and 18 with brainstem-spinal form of MS, while the latter by McDonald criteria. The presence of LESCLs throughout the entire clinical course was significantly more common in OSMS patients than in CMS patients, while brain lesions fulfilling the Barkhof criteria (Barkhof brain lesions) were significantly more common in CMS patients than OSMS patients. LESCLs in OSMS patients most frequently affected the upper to middle thoracic cord, with either holocord or central gray matter involvement. By contrast, 70% of LESCLs in CMS patients predominantly affected the peripheral white matter of the mid-cervical cord. LESCLs in patients with PPMS also showed preferential involvement of the peripheral white matter of the mid-cervical cord. One-third of OSMS patients had neither LESCLs nor Barkhof brain lesions more than 10 years after disease onset, and showed significantly milder disability than OSMS patients with LESCLs. These findings suggest that LESCLs are heterogeneous between OSMS and CMS patients, and that there are distinct subtypes of MS in Japanese, according to clinical and MRI findings..
103. Jun-Ichi Kira, Takuya Matsushita, Noriko Isobe, Takaaki Ishizu, Opticospinal multiple sclerosis in Japanese, Neurology Asia, 13, 2, 167-175, 2008, Antibodies to aquaporin-4 (AQP4) are found in a number of Japanese opticospinal multiple sclerosis (OSMS) patients. Whether anti-AQP4 antibody-positive and -negative OSMS patients are afflicted with an identical disease remains unknown. To clarify immunological differences between the two groups of patients, we studied serum antibody titres against AQP4 in 191 patients with idiopathic central nervous system demyelinating diseases and clarified any relationships with immunological parameters. The anti-AQP4 antibody positivity rate was higher in patients with OSMS (36.2%), idiopathic recurrent myelitis (23.5%), and recurrent optic neuritis (26.9%) than in conventional MS patients (8.0%), and those with other diseases (0%). Anti-AQP4 antibody titre was significantly higher in patients with SS-A/B antibodies than in those without. Anti-AQP4 antibody-negative OSMS patients showed significantly higher CD4+IFN-γ+IL-4-T cell percentages and intracellular IFN-γ/IL-4 ratios than anti-AQP4 antibody-positive patients, anti-AQP4 antibody-negative conventional MS patients, and healthy controls. As well, CD4+IFN-γ +IL-4-T cell percentages were negatively correlated with anti-AQP4 antibody titres. In CSF, OSMS patients had significantly higher levels of IFN-γ and granulocyte colony-stimulating factor levels than patients with non-inflammatory neurological diseases and other causes of myelitis. A significant increase of IL-17 compared with non-inflammatory neurological diseases patients was only found in OSMS patients, irrespective of the presence or absence of anti-AQP4 antibody. These findings suggest that high titres of anti-AQP4 antibodies are produced as a result of heightened humoral autoimmunity, and that they are likely to contribute to extensive lesion development through disturbed resolution of vasogenic oedema. Moreover, since intrathecal up-regulation of IL-17 and IFN-γ is characteristic of OSMS, Th17/ Th1 cells may be critical for the initiation of inflammation and the disruption of blood-brain barrier (BBB); rendering anti-AQP4 antibody get across the BBB..
104. Takeshi Matsuoka, Takuya Matsushita, Yuji Kawano, Manabu Osoegawa, Hirofumi Ochi, Takaaki Ishizu, Motozumi Minohara, Hitoshi Kikuchi, Futoshi Mihara, Yasumasa Ohyagi, Jun-ichi Kira, Heterogeneity of aquaporin-4 autoimmunity and spinal cord lesions in multiple sclerosis in Japanese., Brain, 10.1093/brain/awm027, 130, 5, 1206-1223, 2007.05, Opticospinal multiple sclerosis (OSMS) in Asians has similar features to the relapsing-remitting form of neuromyelitis optica (NMO) seen in Westerners. OSMS is suggested to be NMO based on the frequent detection of specific IgG targeting aquaporin-4 (AQP4), designated NMO-IgG. The present study sought to clarify the significance of anti-AQP4 autoimmunity in the whole spectrum of MS. Sera from 113 consecutive Japanese patients with clinically definite MS, based on the Poser criteria, were assayed for anti-AQP4 antibodies by immunofluorescence using GFP-AQP4 fusion protein-transfected HEK-293T cells. Sensitivity and specificity of the anti-AQP4 antibody assay, 83.3 and 100%, respectively, were calculated using serum samples with NMO-IgG status predetermined at the Mayo Clinic. The anti-AQP4 antibody positivity rate was significantly higher in OSMS patients (13/48, 27.1%) than those with CMS (3/54, 5.6%), other neurological diseases (0/52) or healthy controls (0/35). None of the 11 patients tested with a brainstem-spinal form of MS were positive. Among OSMS patients, the antibody positivity rate was highest in OSMS patients with longitudinally extensive spinal cord lesions (LESCLs) extending over three vertebral segments and brain lesions that fulfilled the Barkhof criteria (5/9, 55.6%). Multiple logistic analyses revealed that emergence of the anti-AQP4 antibody was positively associated only with a higher relapse rate, but not with optic-spinal presentation or LESCLs. Compared with anti-AQP4 antibody-negative CMS patients, anti-AQP4 antibody-positive MS patients showed significantly higher frequencies of severe optic neuritis, acute transverse myelitis and LESCLs while most conditions were also common to anti-AQP4 antibody-negative OSMS patients. The LESCLs in anti-AQP4 antibody-positive patients were located at the upper-to-middle thoracic cord, while those in anti-AQP4 antibody-negative OSMS patients appeared throughout the cervical-to-thoracic cord. On axial planes, the former most frequently showed central grey matter involvement, while holocord involvement was predominant in the latter. In contrast, LESCLs in anti-AQP4 antibody-negative CMS patients preferentially involved the mid-cervical cord presenting a peripheral white matter-predominant pattern, as seen in the short lesions. Anti-AQP4 antibody-positive MS patients fulfilling definite NMO criteria showed female preponderance, higher relapse rate, greater frequency of brain lesions and less frequent responses to interferon beta-1b than anti-AQP4 antibody-negative OSMS patients with LESCLs. These findings suggested that LESCLs are distinct in anti-AQP4 antibody positivity and clinical phenotypes. There were cases of anti-AQP4 antibody-positive MS/NMO distinct from CMS, and anti-AQP4 antibody-negative OSMS with LESCLs in Japanese. This indicated that the mechanisms producing LESCLs are also heterogeneous in cases with optic-spinal presentation, namely AQP4 autoimmunity-related and -unrelated..
105. Takuya Matsushita, Hiroyuki Murai, Masakazu Kawajiri, Hiroshi Muratani, Toru Iwaki, Takayuki Taniwaki, Jun-Ichi Kira, Character changes from idiopathic cranial pachymeningoencephalitis, Journal of the Neurological Sciences, 10.1016/j.jns.2006.01.008, 244, 1-2, 163-166, 2006.05, A 66-year-old man with idiopathic cranial pachymeningoencephalitis was described. He suffered from left orbital pain, and character changes. He became short tempered, and was very attached to trifles. Two years prior to these symptoms, he had developed transient left abducent nerve palsy. Head MRI showed a thickening and enhancement of the dura mater on gadolinium-enhanced T1-weighted images, and high signal intensity lesions at bilateral frontal lobes predominantly in the white matter on T2-weighted images. Biopsies revealed microglial proliferation in the cerebral parenchyma, and mild lymphocytic perivascular infiltration. No evidence of intracranial infection was detected. We therefore treated him with methylprednisolone pulse therapy followed by oral prednisolone. His character became gradually normalized, and bilateral frontal lobe lesions seen on MRI disappeared. This is the first case to describe recurrent pachymeningoencephalitis with character changes, and symptoms were probably due to frontal lobe dysfunction..