1995 Fiscal Year Annual Research Report

運動ニューロン疾患発症機序の分子生物学的研究

Research Project

Project/Area Number	07044232
Research Institution	Tokyo Medical and Dental University
Principal Investigator	飯田静夫東京医科歯科大学, 医学部, 教授 (00009987)
Co-Investigator(Kenkyū-buntansha)	有賀敏夫 Medical College of Virginia, Virginia Com, 助教授 YU Robert K. Medical College of Virginia, Virginia Com, 教授瀧孝雄東京医科歯科大学, 医学部, 助教授 (10046295)
Keywords	ギラン・バレー症候群 / 糖脂質 / 末梢神経 / 運動ニューロン / フィッシャー症候群 / 先行感染 / 自己免疫疾患 / 抗糖脂質抗体
Research Abstract	運動ニューロン疾患の発症機序を分子レベルで解明すると云う目的に対して、多くの免疫性神経疾患で抗糖脂質抗体が出現し、この抗体が神経組織に作用して疾患を惹き起こすことをある程度実証することが出来た。ギランバレー症候群やフィッシャー症候群のかなりな数の患者では先行感染したC.jejuniの菌体成分LPSが末梢神経の構成成分である糖脂質のガンクリオシドCM1やGD1aと分子相同性を持ち、このため感染菌に対して産生された抗体が自己抗体として神経を傷害することが明らかにされた。またM血症で慢性脱髄性ニューロパチーを伴う疾患では硫酸化グルクロン酸を含む糖脂質SGPGにたいする抗体が産生され、同様に神経系に対する自己抗体として発症する事が明らかになった。SGPGは末梢神経だけでなく脳の微小血管内皮細胞にも存在することが明らかにされた。これらの成果を挙げるために極微量の糖脂質を薄層クロマトプレートからPVDF膜へ転写するTLC/Blotting法を開発し、PVDF膜上で試料の抗体との反応や化学的構造解析、機器分析が出来るようになり、神経系で疾患に関与する糖脂質の同定が非常に進んだ。また微小血管内皮細胞とアストロサイト細胞をコラーゲンとフィブロネクチンをコートした膜の両面に培養し、培養系での血液/脳・末梢神経関門モデルを作製することに成功した。このモデルを用いて、SGPGに対する抗体を添加すると培養系の血液/脳・末梢神経関門の透過性は変化し、この系を用いて神経傷害の起こる機序の解明が可能となった。SGPGは血管内皮細胞ではIL-1βによって発現が増強され、接着分子としても働いていることが明らかにされた。

Research Products
(15 results)

All Other

All Publications (15 results)

[Publications] Ishikawa D,Kato T,Handa S,Taki T.: "New methods using polyvinylidene difluoride membranes to detect enzymes involved in glycosphingolipid metabolism." Analitical Biochemistry. 231. 13-19 (1995)
[Publications] Matsumoto A,Yoshino H,Yuki N,et al.: "Ganglioside characterization of a cell line displaying motor neuron-like phenotype:GM2 as possible major ganglioside in motor neurons." Journal of the Neurological Sciences. 131. 111-118 (1995)
[Publications] Tadano-Aritomo K,Kubo H,Ireland P,et al.: "Structural analysis of mono-and bis-sulfated glycosphingolipids by negative liqud secondary ion mass spectrometry with high-and low-energy collision-induced dissociation." Carbohydrate Research. 273. 41-52 (1995)
[Publications] Taki T,Ishikawa D,Handa S,Kasama T.: "Direct mass spectrometric analysis of glycosphingolipid transferred to a polyvinylidene difluoride membrane by thinlayer chromatography blotting." Anal Biochem. 225(1). 24-27 (1995)
[Publications] Watarai S,Kushi Y,Shigeto R,et al.: "Production of monoclonal antibodies directed to Hanganutziu-Deicher active gangliosides,N-glycolylneuraminic acid-containing gangliosides." Journal of Biochemistry. 1995. 1062-1069 (1995)
[Publications] Yuki N,Handa S,Tai T,et al.: "Ganglioside-like epitopes of lipopolysaccharides from Campylobacter jejuni(PEN 19)in three isolates from patients with Guillain-Barre syndrome." J Neurol Sci. 130(1). 112-116 (1995)
[Publications] Yuki N,Tagawa Y,Handa S.: "Autoantibodies to periferal nerve glycosphingolipids SPG;SLPG,and SGPG in Guillain-Barre syndrome and chronic inflamatory demyelinating polyneuropathy." J.Neuroimmunol. (in press).
[Publications] Ariga T,Baht S,Kanda Y,et al.: "Expression and localization on Lewisx and GD1a ganglioside in human glioma cells." Glycoconjugate J.(in press).
[Publications] Kanda T,Yamawaki M,Ariga T,YuRK.: "Interleukin 1 beta up-regulates the expression of sulfoglucuronosyl paragloboside,a ligand for L-selectin,inbrain microvascular endothelial cells." Proc Natl Acad Sci USA. 92(17). 7897-7901 (1995)
[Publications] Ariga T,Kasai N,Miyoshi I,et al.: "Accumulation of isogloboside and ganglio-N-tetraosyl ceramide having blood group B determinant in the hepatomas of female LEC rats." Biochim Biophys Acta. 1254(3). 257-266 (1995)
[Publications] Freischutz B,Saito M,Rahmann H,Yu RK.: "Characterization of sialyltransferase-IV activity and its involvement in the c-pathway of brain ganglioside metabolism." J Neurochem. 6470(11424). 385-393 (1995)
[Publications] Gu X,Preuss U,Gu T,Yu RK.: "Regulation of sialyltransferase activities by phosphorylation and dephosphorylation." J Neurochem. 6416(56094). 2295-2302 (1995)
[Publications] Kanda T,Ariga T,Yamawaki M,Pal S,Katoh SR,Yu RK.: "Effect of nerve growth factor and forskolin on glycosyltransferase activities and expression of a globoseries glycosphingolipid in PC12D pheochromocytoma cells." J Neurochem. 6400(21274). 810-817 (1995)
[Publications] Yamayaki M,Ariga T,Bigbee JW,et al.: "Generation and charaqcterization of anti-sulfoglucuronosyl paragloboside monoclonal Antibody EGR50 and its immunoreactivity with peripheral nerve." J.Neurosci Res. (in press).
[Publications] Yamawaki M,Vasquez A,Ben A,et al.: "Sensitization of Lewis rats with sulfoglucuronosyl paragloboside;Electrophysiological and immunological studies of an animal model of peripheral neuropathy." J.Neurosci Res. (in press).

1995 Fiscal Year Annual Research Report

運動ニューロン疾患発症機序の分子生物学的研究

Principal Investigator

飯田 静夫 東京医科歯科大学, 医学部, 教授 (00009987)

Research Products

[Publications] Ishikawa D,Kato T,Handa S,Taki T.: "New methods using polyvinylidene difluoride membranes to detect enzymes involved in glycosphingolipid metabolism." Analitical Biochemistry. 231. 13-19 (1995)

[Publications] Matsumoto A,Yoshino H,Yuki N,et al.: "Ganglioside characterization of a cell line displaying motor neuron-like phenotype:GM2 as possible major ganglioside in motor neurons." Journal of the Neurological Sciences. 131. 111-118 (1995)

[Publications] Tadano-Aritomo K,Kubo H,Ireland P,et al.: "Structural analysis of mono-and bis-sulfated glycosphingolipids by negative liqud secondary ion mass spectrometry with high-and low-energy collision-induced dissociation." Carbohydrate Research. 273. 41-52 (1995)

[Publications] Taki T,Ishikawa D,Handa S,Kasama T.: "Direct mass spectrometric analysis of glycosphingolipid transferred to a polyvinylidene difluoride membrane by thinlayer chromatography blotting." Anal Biochem. 225(1). 24-27 (1995)

[Publications] Watarai S,Kushi Y,Shigeto R,et al.: "Production of monoclonal antibodies directed to Hanganutziu-Deicher active gangliosides,N-glycolylneuraminic acid-containing gangliosides." Journal of Biochemistry. 1995. 1062-1069 (1995)

[Publications] Yuki N,Handa S,Tai T,et al.: "Ganglioside-like epitopes of lipopolysaccharides from Campylobacter jejuni(PEN 19)in three isolates from patients with Guillain-Barre syndrome." J Neurol Sci. 130(1). 112-116 (1995)

[Publications] Yuki N,Tagawa Y,Handa S.: "Autoantibodies to periferal nerve glycosphingolipids SPG;SLPG,and SGPG in Guillain-Barre syndrome and chronic inflamatory demyelinating polyneuropathy." J.Neuroimmunol. (in press).

[Publications] Ariga T,Baht S,Kanda Y,et al.: "Expression and localization on Lewisx and GD1a ganglioside in human glioma cells." Glycoconjugate J.(in press).

[Publications] Kanda T,Yamawaki M,Ariga T,YuRK.: "Interleukin 1 beta up-regulates the expression of sulfoglucuronosyl paragloboside,a ligand for L-selectin,inbrain microvascular endothelial cells." Proc Natl Acad Sci USA. 92(17). 7897-7901 (1995)

[Publications] Ariga T,Kasai N,Miyoshi I,et al.: "Accumulation of isogloboside and ganglio-N-tetraosyl ceramide having blood group B determinant in the hepatomas of female LEC rats." Biochim Biophys Acta. 1254(3). 257-266 (1995)

[Publications] Freischutz B,Saito M,Rahmann H,Yu RK.: "Characterization of sialyltransferase-IV activity and its involvement in the c-pathway of brain ganglioside metabolism." J Neurochem. 6470(11424). 385-393 (1995)

[Publications] Gu X,Preuss U,Gu T,Yu RK.: "Regulation of sialyltransferase activities by phosphorylation and dephosphorylation." J Neurochem. 6416(56094). 2295-2302 (1995)

[Publications] Kanda T,Ariga T,Yamawaki M,Pal S,Katoh SR,Yu RK.: "Effect of nerve growth factor and forskolin on glycosyltransferase activities and expression of a globoseries glycosphingolipid in PC12D pheochromocytoma cells." J Neurochem. 6400(21274). 810-817 (1995)

[Publications] Yamayaki M,Ariga T,Bigbee JW,et al.: "Generation and charaqcterization of anti-sulfoglucuronosyl paragloboside monoclonal Antibody EGR50 and its immunoreactivity with peripheral nerve." J.Neurosci Res. (in press).

[Publications] Yamawaki M,Vasquez A,Ben A,et al.: "Sensitization of Lewis rats with sulfoglucuronosyl paragloboside;Electrophysiological and immunological studies of an animal model of peripheral neuropathy." J.Neurosci Res. (in press).

飯田静夫東京医科歯科大学, 医学部, 教授 (00009987)