診断の困難なイヌリンパ腫における新規臨床マーカーの開発

• Project/Area Number: 10J40178
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Clinical veterinary science
• Research Institution: The University of Tokyo
• Principal Investigator: 越野 裕子 (後藤 裕子) 東京大学, 大学院農学生命科学研究科, 特別研究員(RPD)
• Project Period (FY): 2010 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2013: ¥500,000 (Direct Cost: ¥500,000); Fiscal Year 2012: ¥1,200,000 (Direct Cost: ¥1,200,000); Fiscal Year 2011: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2010: ¥1,000,000 (Direct Cost: ¥1,000,000)
• Keywords: 犬 / リンパ腫 / イヌ / 臨床マーカー / 増殖細胞マーカー

Research Abstract

1. 蛍光プライマーとキャピラリー電気泳動を用いた免疫レセプター遺伝子再構成の検出
免疫レセプター遺伝子再構成の検出においては、従来ポリアクリルアミドゲルによる電気泳動が用いられてきた。この方法は比較的安価で特別な機器を必要としない一方、分解能に限界があり、非常に接近した複数のバンドを識別できない問題点がある。とくに腸のサンプルを用いた場合には不明瞭な結果が得られることがしばしばあり、判断が難しい。本研究では消化管型リンパ腫の確実な診断を行うため、蛍光プライマーとキャピラリー電気泳動を用いた検出方法を立ち上げた。この方法により、ポリアクリルアミドゲル電気泳動では不明瞭であったPCR結果を確実に判断できる。また、増幅産物の正確な長さを知ることができるため、PBMC中腫瘍細胞のモニタリングにも応用可能である。本研究の成果は現在、投稿準備中であり、また、臨床検査としての応用に向けて準備中である。
2. リンパ腫組織のKi-67染色
増殖細胞マーカーの一種であるKi-67は様々な腫瘍において高い発現が認められることが知られているが、近年、イヌの多中心リンパ腫においても高い発現が報告された。この報告においてはhigh-gradeを主体とした多中心型リンパ腫の針吸引生検サンプルが対象となっていたが、本研究では対象を消化管型リンパ腫の内視鏡サンプルおよびlow-gradeリンパ腫細胞とし、診断の困難なこれら疾患の新規治療マーカーとしての可能性を模索することとする。現在、症例サンプルの収集および腸内視鏡サンプルの染色条件検討をおこなっており、良好な染色像が得られている。

Strategy for Future Research Activity

(抄録なし)

Research Products

• [Journal Article] Differential expression of CD45 isoforms in canine leukocytes (2014)
  • Author(s): Goto-Koshino Y, Tomiyasu H, Suzuki H, Tamamoto T, Mizutani N, FujinoY, Ohno K, Tsujimoto H.
  • Journal Title: Vet Immunol Immunopathol Volume: 160 Issue: 1-2 Pages: 118-122
  • DOI: 10.1016/j.vetimm.2014.03.011
  • Peer Reviewed
• [Journal Article] Plasma Transferrin Concentration as a Nutritional Marker in Malnourished Dogs with Nutritional Treatment (2014)
  • Author(s): Nakajima M, Ohno K, Goto-Koshino Y, Fujino Y, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 4 Pages: 539-543
  • DOI: 10.1292/jvms.13-0583
  • NAID: 130003382416
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] Phase I Dose-Escalation Study of Nimustine in Tumor-Bearing Dogs (2014)
  • Author(s): Takahashi M, Goto-Koshino Y, Fukushima K, Kanemoto H, Nakashima K, Fujino Y, Ohno K, Endo Y, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 6 Pages: 895-899
  • DOI: 10.1292/jvms.13-0345
  • NAID: 130003391282
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Prognostic Analyses on Anatomical and Morphological Classification of Feline Lymphoma (2014)
  • Author(s): Sato H, Fujino Y, Chino J, Takahashi M, Fukushima K, Goto-Koshino Y, Uchida K, Ohno K, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 6 Pages: 807-811
  • DOI: 10.1292/jvms.13-0260
  • NAID: 130003391281
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Prognostic significance of the expression levels of the p16, p15, and p14genes in dogs with high-grade lymphoma (2014)
  • Author(s): Fujiwara-Igarashi A, Goto-Koshino Y, Sato M, Maeda S, Igarashi H, Takahashi M, Fujino Y, Ohno K, Tsujimoto H.
  • Journal Title: Vet J Volume: 199(2) Issue: 2 Pages: 236-244
  • DOI: 10.1016/j.tvjl.2013.11.004
  • Peer Reviewed
• [Journal Article] Epigenetic regulation of the ABCBl gene in drug-sensitive and drug-resistant lymphoid tumour cell lines obtained from canine patients (2014)
  • Author(s): Tomiyasu H, Goto-Koshino Y, Fujino Y, Ohno K, Tsujimoto H.
  • Journal Title: Vet J Volume: 199(1) Issue: 1 Pages: 103-109
  • DOI: 10.1016/j.tvjl.2013.10.022
  • Peer Reviewed
• [Journal Article] The Regulation of the Expression of <i>ABCG2</i> Gene through Mitogen-Activated Protein Kinase Pathways in Canine Lymphoid Tumor Cell Lines (2014)
  • Author(s): Tomiyasu H, Gato-Koshino Y, Fujino Y, Ohno K, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 2 Pages: 237-242
  • DOI: 10.1292/jvms.13-0337
  • NAID: 130003382366
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Regulations of ABCBl and ABCG2 expression through MAPK pathways in acute lymphoblastic leukemia cell lines (2013)
  • Author(s): Tomiyasu H, Watanabe M, Sugita K, Goto-Koshino Y, Fujino Y, Ohno K, Sugano S, Tsujimoto H.
  • Journal Title: Anticancer Res Volume: 33(12) Pages: 5317-5323
  • Peer Reviewed
• [Journal Article] Feline serum amyloid A protein as an endogenous Toll-like receptor 4 agonist (2013)
  • Author(s): Tamamoto T, Ohno K, Goto-Koshino Y, Tsujimoto H.
  • Journal Title: Vet Immunol Immunopathol Volume: 155(3) Issue: 3 Pages: 190-196
  • DOI: 10.1016/j.vetimm.2013.06.010
  • Peer Reviewed
• [Journal Article] Simultaneous Inactivation of the <i>p16, p15</i> and <i>p14</i> Genes Encoding Cyclin-Dependent Kinase Inhibitors in Canine T-Lymphoid Tumor Cells (2013)
  • Author(s): Fujiwara-Igarashi A, Goto-Koshino Y, Mochizuki H, Maeda S, Fujino Y, Ohno K, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 75 Issue: 6 Pages: 733-742
  • DOI: 10.1292/jvms.12-0351
  • NAID: 130003362118
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Analyses on Activation of NF-&kappa;B and Effect of Bortezomib in Canine Neoplastic Lymphoid Cell Lines (2013)
  • Author(s): Kojima K, Fujino Y, Goto-Koshino Y, Ohno K, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 75 Issue: 6 Pages: 727-731
  • DOI: 10.1292/jvms.12-0168
  • NAID: 130003362111
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Cytomorphological and Immunological Classification of Feline Lymphomas: Clinicopathological Features of 76 Cases (2013)
  • Author(s): Chino J, Fujino Y, Kobayashi T, Kariya K, Goto-Koshino Y, Ohno K, Nakayama H, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 75 Issue: 6 Pages: 701-707
  • DOI: 10.1292/jvms.12-0246
  • NAID: 130003362114
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] The prognostic significance of minimal residual disease in the early phases of chemotherapy in dogs with high-grade B-cell lymphoma. (2013)
  • Author(s): Sato M, Yamzaki J, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H.
  • Journal Title: Vet J Volume: 195 Pages: 319-324
  • Peer Reviewed
• [Journal Article] Robo4 plays a role in bone marrow homing and mobilization, but is not essential in the long-term repopulating capacity of hematopoietic stem cells. (2012)
  • Author(s): Goto-Koshino Y, Fukuchi Y, Shibata F, Abe D, Kuroda K, Okamoto S, Kitamura T, Nakajima 〓
  • Journal Title: Plos One Volume: 7
  • Peer Reviewed
• [Journal Article] Regulation of the expression of ABCB1 and LRP genes by MAPK/ERK pathway and its role in the generation of side population cells in canine lymphoma cell lines. (2012)
  • Author(s): Tomiyasu H, Watanabe M, Goto-Koshino Y, Fujino Y, Ohno K, Sugano S, Tsujimoto H.
  • Journal Title: Leuk Lymphoma Volume: (Epub ahead of print) Issue: 6 Pages: 1309-1315
  • DOI: 10.3109/10428194.2012.751529
  • Peer Reviewed
• [Journal Article] Serum amyloid A uptake by feline peripheral macrophages. (2012)
  • Author(s): Tamamoto T, Ohno K, Goto-Koshino Y, Fujino Y, Tsujimoto H.
  • Journal Title: Vet Immunol Immunopathol Volume: 150 Issue: 1-2 Pages: 47-52
  • DOI: 10.1016/j.vetimm.2012.08.005
  • Peer Reviewed
• [Journal Article] Increased expression of fractalkine and its receptor CX3CR1 in canine inflammatory bowel disease and their possible role in recruitment of intraepithelial lymphocytes (2012)
  • Author(s): Shingo Maeda
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 148 Issue: 3-4 Pages: 226-235
  • DOI: 10.1016/j.vetimm.2012.04.021
  • Peer Reviewed
• [Journal Article] Comparison of the antitumor effects of an MDM2 inhibitor, nutlin-3, in feline lymphoma cell lines with or without p53 mutation. (2012)
  • Author(s): Mochizuki H, Goto-Koshino Y, Sato M, Fujino Y, Ohno K,
  • Journal Title: Vet Immunol Immunopathol Volume: 147 Issue: 3-4 Pages: 187-194
  • DOI: 10.1016/j.vetimm.2012.04.017
  • Peer Reviewed
• [Journal Article] Mucosal imbalance of interleukin-1 beta and interleukin-1 receptor antagonist in canine inflammatory bowel disease (2012)
  • Author(s): Maeda, S., Ohno, K., Nakamura, K., Uchida, K., Nakasima, K., Fukushima, K., Tsukamoto, A., Goto-Koshino, Y., Fujino, Y., Tsujimoto, H
  • Journal Title: The Veterinary Journal Volume: (In press) Issue: 1 Pages: 66-70
  • DOI: 10.1016/j.tvjl.2012.02.026
  • Peer Reviewed
• [Journal Article] X-Chromosome Inactivation Pattern Analysis for the Assessment of Cell Clonality in Cats. (2012)
  • Author(s): Mochizuki H, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H.
  • Journal Title: Vet Pathol Volume: 49 Issue: 6 Pages: 963-970
  • DOI: 10.1177/0300985812439216
  • Peer Reviewed
• [Journal Article] GeneScan analysis to detect clonality of T-cell receptor γ gene rearrangement in feline lymphoid neoplasms. (2012)
  • Author(s): Mochizuki H, Nakamura K, Sato H, Goto-Koshino Y, Sato M, Takahashi M, Fukushima K, Nakashima K, Fuj ino Y, Ohno K, Uchida K, Nakayama H, Tsujimoto
  • Journal Title: Vet Immunol Imunopathol Volume: 145 Issue: 1-2 Pages: 402-409
  • DOI: 10.1016/j.vetimm.2011.12.015
  • Peer Reviewed
• [Journal Article] Molecular cloning and expression analysis of the canine chemokine receptor CCR9 (2012)
  • Author(s): Maeda S., Ohno, K., Tsukamoto, A., Nakashima, K., Fukushima, K., Goto-Koshino, Y., Fujino, Y., Tsujimoto H.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 145 Issue: 1-2 Pages: 534-539
  • DOI: 10.1016/j.vetimm.2011.12.004
  • Peer Reviewed
• [Journal Article] Molecular cloning and characterization of canine fractalkine its receptor CX3CR1 (2012)
  • Author(s): Maeda S., Ohno, K., Nakashima, K., Fukushima, K., Tsukamoto, A., Suzuki, H., Fujiwara, A., Goto-Koshino, Y., Fujino, Y., Tsujimoto H.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 145 Issue: 1-2 Pages: 100-109
  • DOI: 10.1016/j.vetimm.2011.10.018
  • Peer Reviewed
• [Journal Article] Quantification of chemokine receptor gene expression in duodenel mucosa of dogs with inflammatory bowel disease (2011)
  • Author(s): Maeda, S., Ohno, K., Nakamura, K., Uchida, K., Nakasima, K., Fukushima, K., Tsukamoto, A., Goto-Koshino, Y., Fujino, Y., Tsujimoto, H.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 144 Issue: 3-4 Pages: 290-298
  • DOI: 10.1016/j.vetimm.2011.08.020
  • Peer Reviewed
• [Journal Article] A Rapid and Simple Method to Obtain Canine Peripheral Blood-Derived Macrophages (2011)
  • Author(s): Goto-Koshino Y, et al.
  • Journal Title: The Journal of Veterinary Medical Science Volume: In press(掲載確定)
  • Peer Reviewed
• [Journal Article] Anti-Tumor Effect of Adenoviral Vector-Mediate p53 Gene Transfer on the Growth of Canine Osteosarcoma Xenografts in Nude Mice. (2011)
  • Author(s): Kanaya N, et al.
  • Journal Title: The Journal of Veterinary Medical Science Volume: In press(掲載確定)
  • Peer Reviewed
• [Journal Article] Evaluation of cytored uctive efficacy of vincristine, cyclophosp hamide, and Doxorubicin in dogs with lymphoma by measuring the number of neoplastic lymphoid cells with real-time polymerase chain reaction. (2011)
  • Author(s): Sato M, et al.
  • Journal Title: Journal of Veterinary Internal Medicine Volume: 25 Pages: 285-291
  • Peer Reviewed
• [Journal Article] Increase in Minimal Residual Disease in Peripheral Blood before Clinical Relapse in Dogs with Lymphoma that Achieved Complete Remission after Chemotherapy. (2011)
  • Author(s): Sato M, et al.
  • Journal Title: Journal o Veterinary Internal Medicine Volume: 25 Pages: 292-296
  • Peer Reviewed
• [Journal Article] Evaluation of the prognostic significance of BCL6 gene expression in canine high-grade B-cell lymphoma. (2011)
  • Author(s): Sato M, et al.
  • Journal Title: The Veterinary Journal Volume: In press
  • Peer Reviewed
• [Journal Article] Establishment of novel feline leukemia virus (FeLV)-negative B-cell cell line from a cat with B-cell lymphoma. (2011)
  • Author(s): Mochizuki H, et al.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 140 Pages: 307-311
  • Peer Reviewed
• [Journal Article] Characterization of canine BCL6 cDNA and detection systems for its protein expression. (2011)
  • Author(s): Sato M, et al.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 140 Pages: 166-169
  • Peer Reviewed
• [Journal Article] Eosinophilia and eosinophilic infiltration into splenic B-cell high-grade lymphoma in a dog. (2010)
  • Author(s): Tomiyasu H, et al.
  • Journal Title: The Journal o Veterinary Medical Science Volume: 72 Pages: 1367-1370
  • Peer Reviewed
• [Journal Article] Quantitative analysis of mRNA for 10 different drug resistance factors in dogs with lymphoma (2010)
  • Author(s): Tomiyasu H, et al.
  • Journal Title: The Journal of Veterinary Medical Science Volume: 72 Pages: 1165-1172
  • Peer Reviewed
• [Journal Article] A case of canine lymphomatoid granulomatosis with cutaneous lesions. (2010)
  • Author(s): Shimazaki T, et al.
  • Journal Title: The Journal of Veterinary Medical Science Volume: 72 Pages: 1067-1069
  • Peer Reviewed