ヒト腸管切除検体を用いた炎症性腸疾患の免疫学的発症機序の解明と新規治療法の開発

• Project/Area Number: 12J05920
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Gastroenterology
• Research Institution: Keio University
• Principal Investigator: 三上 洋平 慶應義塾大学, 医学部, 特別研究員(PD)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,630,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2012: ¥1,100,000 (Direct Cost: ¥1,100,000)
• Keywords: 炎症性疾患 / サイトカイン / マクロファージ / 自然リンパ球 / 遺伝子発現 / 炎症性腸疾患 / Innate lympohoid cell / Lymphoid tissue inducer cells / Th17細胞 / Th1細胞 / Gut-associated lymphoid tissue

Outline of Annual Research Achievements

前年度までにマウスを用いて行った基盤的研究から、炎症状態、定常状態ともヘルパーT細胞(Th1, Th17細胞)および、Innate lymphoid cell (ILC)は独立した制御機構が存在することが示唆された。IBD患者腸管切除検体において、ILCの細胞数の増減は認められない物の、粘膜修復能に関与するIL-22産生能を炎症部と非炎症部において比較したところ、ILCのIL-22産生能は炎症部と比して非炎症部において有意に高値を示した。また、IL-22産生性のILCはCD127(+)CD56(-)分画(CD127SP)およびCD127(+)CD56(+)分画(DP)において認められた。次にCD127SPおよびDP分画を単離し定量PCRを実施したところ、Flowcytometryの結果と一致してCD127SPおよびDP分画はRORγt陽性であり、これらはそれぞれマウスにおけるNCR- ILC3, NCR+ILC3に一致する分画と考えられる。これらILC3分画からのIL-22産生機構を検討するために、マウスの腸管を用いて、ILC3分画における網羅的遺伝子発現解析(RNA-seq)および、クロマチン構造(ATAC-seq)を検討した。まず初めに、ILC3の各分画を効率よく分離するための細胞調整方法および分離方法を検討した。CCR6+NCR-ILC3はLTiに、CCR6-NCR-ILC3はT-betを発現しNCR+ILC3に近い性質を持っていることを確認しNCR+ILC3とLTiの遺伝子発現をRNA-seqを用いて比較検討した。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Risk and Management of Intra-Abdominal Abscess in Crohn's Disease Treated with Infliximab (2014)
  • Author(s): Yoneno K, Hisamatsu T, Matsuoka K, Okamoto S, Takayama T, Ichikawa R, Sujino T, Miyoshi J, Takabayashi K, Mikami Y, Mizuno S, Wada Y, Yajima T, Naganuma M, Inoue N, Iwao Y, Ogata H, Hasegawa H, Kitagawa Y, Hibi T, Kanai T
  • Journal Title: Digestion Volume: 89 Issue: 3 Pages: 201-208
  • DOI: 10.1159/000360618
  • Peer Reviewed / Open Access
• [Journal Article] Cross-talk between RORγt+ innate lymphoid cells and intestinal macrophages induces mucosal IL-22 production in Crohn's disease (2014)
  • Author(s): Mizuno S, Mikami Y, Kamada N, Handa T, Hayashi A, Sato T, Matsuoka K, Matano M, Ohta Y, Sugita A, Koganei K, Sahara R, Takazoe M, Hisamatsu T, Kanai T
  • Journal Title: Inflamm Bowel Dis. Volume: 20 Pages: 1426-1434
  • DOI: 10.1007/82_2014_371
  • ISBN: 9783319073941, 9783319073958
  • Peer Reviewed
• [Journal Article] Classical Th1 cells obtain colitogenicity by co-existence of RORγt-expressing T cells in experimental colitis (2014)
  • Author(s): Saigusa K, Hisamatsu T, Handa T, Sujino T, Mikami Y, Hayashi A, Mizuno S, Takeshita K, Sato T, Matsuoka K, Kanai T
  • Journal Title: Inflamm Bowel Dis Volume: 20 Issue: 10 Pages: 1820-1827
  • DOI: 10.1097/mib.0000000000000149
  • Peer Reviewed
• [Journal Article] Macrophages and dendritic cells emerge in the liver during intestinal inflammation and predjspose the liver to inflammation. (2014)
  • Author(s): Mikami Y, Mizuno S, Nakamoto N, Hayashi A, Sujino T, Sato T, Kamada N, Matsuoka K, Hisamatsu T, Ebinuma H, Hibi T, Yoshimura A, Kanai T.
  • Journal Title: PLoS One Volume: 9 Issue: 1 Pages: 1-1
  • DOI: 10.1371/journal.pone.0084619
  • Peer Reviewed
• [Journal Article] Dendritic cells administered intrarectally penetrate the intestinal barrier to break intestinal tolerance via Th2-medeiated colitis in mice. (2013)
  • Author(s): Handa T, Kanai T, Sato T, Mikami Y, Sujino T, Hayashi. A, Mizuno S, Matsumoto A, Hisamatsu T, Hibi T.
  • Journal Title: Immunology Letters Volume: 150 Issue: 1-2 Pages: 123-9
  • DOI: 10.1016/j.imlet.2013.01.010
  • Peer Reviewed
• [Journal Article] CCR2 knockout exacerbates cerulein-induced chronic pancreatitis with hyperglycemia via decreased GLP-1 receptor expression and insulin secretion. (2013)
  • Author(s): Nakamura Y, Kanai T, Saeki K, Takabe M, Irie J, Miyoshi J, Mikami Y, Teratani T, Suzuki T, Miyata N, Hisamatsu T, Nakamoto N, Yamagishi Y, Higuchi H, Ebinuma H, Hozawa S, Saito H, Itoh H, Hibi T.
  • Journal Title: Am J Physiol Gastrointest Liver Physiol Volume: 304 Issue: 8 Pages: G700-G707
  • DOI: 10.1152/ajpgi.00318.2012
  • Peer Reviewed
• [Journal Article] C-C motif chemokine receptor 9 positive macrophages activate hepatic stellate cells and promote liver fibrosis in mice. (2013)
  • Author(s): Chu PS, Nakamoto N, Ebinuma H, Usui S, Saeki K, Matsumoto A, Mikami Y, Sugiyama K, Tomita K, Kanai T, Saito H, Hibi T.
  • Journal Title: Hepatology Volume: 58 Issue: 1 Pages: 337-350
  • DOI: 10.1002/hep.26351
  • Peer Reviewed
• [Journal Article] IL-22-Producing RORyt-Dependent Innate Lymphoid Cells Play a Novel Protective Role in Murine Acute Hepatitis. (2013)
  • Author(s): Matsumoto A, Kanai T, Mikami Y, Chu PS, Nakamoto N, Ebinuma H, Saito H, Sato T, Yagita H, Hibi T.
  • Journal Title: PLoS One Volume: 8 Issue: 4 Pages: e62853-e62853
  • DOI: 10.1371/journal.pone.0062853
  • Peer Reviewed
• [Journal Article] A Single Strain of Clostridium butyricum Induces Intestinal I L-10-Producing Macrophages to Suppress Acute Exlperimental Colitis in Mice. (2013)
  • Author(s): Hayashi A, Sato T, Kamada N, Mikami Y, Matsuoka K, Hisamatsu T, Hlbi T, Roers A, Yagita H, Ohteki T, Yoshimura A, Kanai T.
  • Journal Title: Cell Host Microbe Volume: 13 Issue: 6 Pages: 711-22
  • DOI: 10.1016/j.chom.2013.05.013
  • Peer Reviewed
• [Journal Article] ROR_Yt-dependent IL-17A-producing cells in the pathogenesis of intestinal inflammation. (2012)
  • Author(s): Kanai T, Mikami Y, Sujino T, Hisamatsu T, Hibi T.
  • Journal Title: Mucosal Immunology Volume: 5 Issue: 3 Pages: 240-247
  • DOI: 10.1038/mi.2012.6
  • Peer Reviewed
• [Journal Article] CCR9(+) plasmacytoid dendritic cells in the small intestine suppress development of intestinal inflammation in mice. (2012)
  • Author(s): Mizuno S, Kanai T, Mikami Y, Sujino T, Ono Y, Hayashi A, Handa T, Matsumoto A, Nakamoto N, Matsuoka K, Hisamatsu T, Takaishi H, Hibi T.
  • Journal Title: Immunology letters Volume: 146 Issue: 1-2 Pages: 64-69
  • DOI: 10.1016/j.imlet.2012.05.001
  • Peer Reviewed
• [Journal Article] Long-term prognosis of patients with ulcerative colitis treated with cytapheresis therapy. (2012)
  • Author(s): Takayama T, Kanai T, Matsuoka K, Okamoto S, Sujino T, Mikami Y, Hisamatsu T, Yajima T, Iwao Y, Ogata H, Hibi T.
  • Journal Title: Journal of Crohns and Colitis Volume: 7 Issue: 2 Pages: e49-e54
  • DOI: 10.1016/j.crohns.2012.05.005
  • Peer Reviewed
• [Journal Article] CCR9(+) macrophages are required for eradication of peritoneal bacterial infections and prevention of polymicrobial sepsis. (2012)
  • Author(s): Mizukami T, Kanai T, Mikami Y, Hayashi A, Doi T, Handa T, Matsumoto A, Jun L, Matsuoka K, Sato T, Hisamatsu T, Hibi T.
  • Journal Title: Immunology Letters Volume: 147 Issue: 1-2 Pages: 75-79
  • DOI: 10.1016/j.imlet.2012.06.006
  • Peer Reviewed
• [Journal Article] T-helper 17 and Interleukin-17-Producing Lymphoid Tissue Inducer-Like T Cells Make Different Contributions to Colitis in Mice. (2012)
  • Author(s): Ono Y, Kanai T, Sujino T, Nemoto Y, Kanai Y, Mikami Y, Hayashi A, Matsumoto A, Takaishi H, Ogata H, Matsuoka K, Hisamatsu T, Watanabe M, Hibi T.
  • Journal Title: Gastroenterology Volume: 143 Issue: 5 Pages: 1288-1297
  • DOI: 10.1053/j.gastro.2012.07.108
  • Peer Reviewed
• [Journal Article] IgA plasma cells express the negative regulatory co-stimulatory molecule programmed cell death 1 ligand and have a potential tolerogenic role in the intestine. (2012)
  • Author(s): Doi T, Kanai T, Mikami Y, Sujino T, Jun L, Ono Y, Hayashi A, Hibi T.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 425 Issue: 4 Pages: 918-923
  • DOI: 10.1016/j.bbrc.2012.08.010
  • Peer Reviewed
• [Journal Article] Dysregulated balance of retinoid-related orphan receptor _Yt-dependent innate lymphoid cells is involved in the pathogenesis of chronic DSS-induced colitis. (2012)
  • Author(s): Kimura K, Kanai T, Hayashi A, Mikami Y, Sujino T, Mizuno S, Handa T, Matsuoka K, Hisamatsu T, Sato T, Hibi T.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 427 Issue: 4 Pages: 694-700
  • DOI: 10.1016/j.bbrc.2012.09.091
  • Peer Reviewed
• [Journal Article] MyD88-dependent interleukin-10 production from regulatory CD11b(+)Gr-1(high) cells suppresses development of acute cerulein pancreatitis in mice. (2012)
  • Author(s): Koike Y, Kanai T, Saeki K, Nakamura Y, Nakano M, Mikami Y, Yamagishi Y, Nakamoto N, Ebinuma H, Hibi T.
  • Journal Title: Immunology Letters Volume: 148 Issue: 2 Pages: 172-177
  • DOI: 10.1016/j.imlet.2012.08.008
  • Peer Reviewed
• [Journal Article] Immune aspects of the pathogenesis of inflammatory bowel disease. (2012)
  • Author(s): Hisamatsu T, Kanai T, Mikami Y, Yoneno K, Matsuoka K, Hibi T.
  • Journal Title: Pharmacology & Therapeutics Volume: 137 Issue: 3 Pages: 283-297
  • DOI: 10.1016/j.pharmthera.2012.10.008
  • Peer Reviewed
• [Journal Article] Anticoagulation therapy dramatically improved severe sigmoiditis with findings resembling inflammatory bowel disease, which was caused by mesenteric venous thrombosis. (2012)
  • Author(s): Mikami Y, Kanai T, Iwasaki E, Naganuma M, Yamagishi Y, Shimoda M, Matsuoka K, Hisamatsu T, Iwao Y, Ogata H, Nakatsuka S, Mukai M and Hibi T.
  • Journal Title: Clinical Journal of Gastroenterology Volume: 5 Pages: 377-382
  • Peer Reviewed
• [Presentation] Distinct roles of miR221/222 in balancing T helper cell homeostasis and function. (2014)
  • Author(s): Yohei Mikami, Hayato Takahashi, Yuka Kanno, Amanda Poholek, Kan Jiang and John J. O’Shea
  • Organizer: NIAMS IRP Retreat
  • Place of Presentation: Washington D.C. USA
  • Year and Date: 2014-05-29 – 2014-05-30
• [Presentation] Intestinal Inflammation Instructs Increasing Liver Macrophages and Dendritic Cells to Dispose Liver to the Inflammation (2013)
  • Author(s): Yohei Mikami
  • Organizer: JSICR-MMCB2013 Joint Symposium in Tokyo
  • Place of Presentation: 東京
  • Year and Date: 2013-05-21
• [Presentation] Germ free systemを用いた腸内フローラに起因する肝免疫異常の検討 (2013)
  • Author(s): 三上洋平
  • Organizer: 第9回日本消化管学会総会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2013-01-25
• [Presentation] Colitogenic ROR gamma t dependent Th1 cells may help the generation of colitogenic ROR gamma t independent Th1 cells (2012)
  • Author(s): Yohei Mikami
  • Organizer: Asian IBD Symposium -Seoul 2012
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2012-11-03
• [Presentation] Yohei Mikami (2012)
  • Author(s): Yohei Mikami
  • Organizer: Intestinal Inflammation Instructs Increasing Liver Macrophages and Dendritic Cells to Trigger Inflammation in the Liver
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2012-11-02
• [Presentation] New Immunosuppressive System by Myeloid-Derived Suppressor Cells in the Lamina Propria of Ulcerative Colitis Patients (2012)
  • Author(s): Yohei Mikami
  • Organizer: Digestive disease week 2012
  • Place of Presentation: San Diego (USA)
  • Year and Date: 2012-05-21