Development of DNA vaccine for canine lymphoma

• Project/Area Number: 20780226
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Clinical veterinary science
• Research Institution: Kagoshima University
• Principal Investigator: SETOGUCHI Asuka Kagoshima University, 農学部, 講師 (00396813)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2008: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 腫瘍 / 免疫 / ワクチン / 犬 / リンパ腫 / DNAワクチン / 遺伝子再構成 / 免疫グロブリン / T細胞受容体 / プラスミドベクター

Research Abstract

Lymphoma is a most common hematopoietic tumor in dogs and ca ts and it is impossible to cure with the conventional treatment as in human medicine. The first line therapy for lymphoma is chemotherapy that has been investigated to i mprove the treatment result over two decades. However, there have been some outstand ing problem such as acquired multi-drug resistance of tumor cells. The development of new treatment strategy for lymphoma in dogs and cats seems to be quite important. In this study, we developed the novel immune-mediated treatment for canine lymphoma using DNA vaccine. At first, we tried to construct the vaccine plasmid including the sequence of highly variable lesion of immunoglobline (Ig) or T cell receptor (TCR) which are encouraging candidate as tumor antigen for lymphoma cell. Two canine lymph oma cell line (CL-1 : T cell lymphoma cell line, GL-1: B cell leukemia cell line) and tumor tissues obtained from two dogs diagnosed as low-grade lymphoma at Kagoshima U niversity Veterinary Teaching Hospital for 2008-2009 were used. The obtained sequenc e of Ig or TCR by PCR were inserted to expression plasmid and this constructed vacci ne plasmid was transfected to COS-7 cells. Expression of Ig or TCR protein on cell s urface was examined by indirect immunofluorecence or Western blotting. At the same time, we tried to establish the method to evaluate the effect of DNA vaccine using m inimal residual region (MRD) from peripheral blood samples using real-time PCR. Using this method, the MRD levels in clinical cases could evaluate during chemotherapy i n 9 clinical cases. And it is indicated that MRD could be an objective marker to eva luate tumor cell burden in dogs with lymphoma. MRD at the end of chemotherapy could be a prognostic factor to predict remission duration after chemotherapy

Research Products

• [Journal Article] Monitoring of minimal residual disease (MRD) after multidrug chemotherapy and its correlation to outcome in dogs with lymphoma: a proof-of-concept pilot study (2010)
  • Author(s): Yamazaki, J., Takahashi, M., Setoguchi, A., Fujino, Y., Ohno, K., Tsujimoto, H
  • Journal Title: J.Vet.Intern.Med 24 Pages: 897-903
• [Journal Article] Disseminated histiocytic sarcoma with excessive hemophagocytosis in a cat (2009)
  • Author(s): Ide, K., Setoguchi-Mukai, A., Nakagawa, T., Uetsuka, K., Nakayama, H., Fujino, Y., Ohno, K., Tsujimoto, H
  • Journal Title: J.Vet.Med.Sci 71 Pages: 817-820
• [Journal Article] Antiviral activity of membrane fusion inhibitor that target gp40 of the feline immunodeficiency virus envelope protein (2009)
  • Author(s): Misukoshi F, Baba K, Goto Y, Setoguchi A, Fujino Y, Ohno K, Oishi S, Kodera Y, Fujii N, Tsujimoto H
  • Journal Title: Vet Microbiol. 136 Pages: 155-159
  • Peer Reviewed
• [Journal Article] uantitative assessment of minimal residual disease (MRD) in canine lymphoma by using real-time polymerase chain reaction (2008)
  • Author(s): Yamazaki, J., Baba, K., Goto-Koshino, Y., Setoguchi-Mukai, A., Fujino, Y., Ohno, K., Tsujimoto, H
  • Journal Title: Vet.Immunol.Immunopathol 126 Pages: 321-331
• [Journal Article] Effect of tyrosine kinase inhibition by imatinib mesyla to on mast cell tumors in dogs (2008)
  • Author(s): Isotani M, Ishida N, Tominaga M, Tamura K, Yagihara H, Ochi S, Kato R, Kobayashi T, Fujita M, Fujino Y, Seto guchi A, Ono K, Washizu T, Bonkobara M
  • Journal Title: J Vet Intern Med. 22 Pages: 985-988
  • Peer Reviewed
• [Presentation] Possible emergence of drug-resistant Babesia gibsoni in clinical cases treated with Atovaquone and Azithromyci n, and epidemiological survey based on CYTB gene in Jar an (2008)
  • Author(s): Sakuma M, Fukuda K, Takayama K, Kobayashi Y, Setoguchi-Mukai A, Endo Y
  • Organizer: American College of Veterinary Internal Medicine Forum 2008
  • Place of Presentation: San Antonio, TX, USA
• [Presentation] ミニチュアダックスフントの消化器型リンパ腫
  • Author(s): 瀬戸口明日香
  • Organizer: 平成19年度日本獣医師会学会年次大会シンポジウム
  • Place of Presentation: 高松
• [Presentation] リンパ腫の治療:きほんのき
  • Author(s): 瀬戸口明日香
  • Organizer: 第5回日本獣医内科学アカデミー総会総会教育講演
  • Place of Presentation: 東京