| Project/Area Number |
19K07975
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Review Section |
Basic Section 52020:Neurology-related
|
|---|
| Research Institution | Yamaguchi University |
|---|
Principal Investigator |
Yukio Takeshita 山口大学, 大学院医学系研究科, 助教 (70749829)
|
|---|
| Project Period (FY) |
2019-04-01 – 2022-03-31
|
| Project Status |
Completed (Fiscal Year 2021)
|
| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
|
|---|
| Keywords | 血液脳関門 / 多発性硬化症 / 炎症細胞浸潤 / Blood-Brain Barrier / 炎症細胞浸潤機構 / 自己免疫性中枢神経疾患 / ヒト由来BBB構成細胞株 |
|---|
| Outline of Research at the Start |
ヒト由来Blood-Brain Barrier構成細胞株による生理的流速負荷型In Vitro系Blood-Brain Barrierモデルを用いて、難治性自己免疫性中枢神経疾患の患者における疾患特異的な炎症細胞浸潤の分子機構を解明する。自己免疫性中枢神経疾患における液性因子を介した炎症細胞浸潤機序を明らかにすることができれば、各自己免疫性中枢神経疾患の病態に則した新規治療法の開発に寄与することが出来る。
|
| Outline of Final Research Achievements |
Sera from the patient of RRMS which can downregulate BBB function of HBMECs was selected. The membrane of HBMECs were biotinylated and its plasma membrane fraction was isolated by ultracentrifugation. The extracted membrane-protein were immunoprecipitated with IgG from the RRMS or normal subjects . After separating eluted proteins by SDS-PAGE, specific bands in the RRMS were determined by mass spectrometry.
We identified the 21 membrane-proteins as targeted auto-antigens in the RRMS. One of them was known to be relevant to BBB function. In 10 RRMS patients, three patients had the auto-antibody against this molecule.
We succeeded in construction of the method that allows us to efficiently investigate targeted antigens from plasma membrane fraction of HBMEC, using the biotinylation of HBMECs and the isolation of membrane fraction by ultracentrifugation. This identified antigen could be an immunological target molecule, contributing to the BBB breakdown in RRMS.
|
| Academic Significance and Societal Importance of the Research Achievements |
今回、微小脳血管内皮細胞株の膜蛋白を抽出し、RRMS患者中の自己抗体で標識される膜蛋白分子を同定する方法を確立した。本手法により他の神経炎症性疾患における血管内皮細胞の標的膜蛋白分子の同定が可能となりうる。また、同定した分子群においてBBB調整機能を持つ標的分子Xは、複数のRRMS患者で自己抗体が検出され、RRMSの発症マーカーになるだけでなく、自己抗体が標的となる膜蛋白Xに作用することでBBBの炎症細胞浸潤を誘発し、RRMSの発症に関与している可能性がある。
|
