Development of DNA vaccine for canine lymphoma

犬淋巴瘤DNA疫苗的开发

• 批准号: 20780226
• 负责人: SETOGUCHI Asuka
• 金额: $ 2.75万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20780226/
• 关键词: 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

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

淋巴瘤是狗和猫中最常见的造血系统肿瘤，不可能用人类医学中的常规治疗方法治愈。淋巴瘤的一线治疗是化疗，二十年来人们一直在研究化疗以改善治疗效果。然而，肿瘤细胞获得性多重耐药等问题仍然突出。开发针对狗和猫淋巴瘤的新治疗策略似乎相当重要。在这项研究中，我们开发了使用 DNA 疫苗治疗犬淋巴瘤的新型免疫介导疗法。首先，我们尝试构建包含免疫球蛋白（Ig）或T细胞受体（TCR）高度可变病变序列的疫苗质粒，这些序列有望作为淋巴瘤细胞的肿瘤抗原。 2008年从鹿儿岛大学兽医教学医院诊断为低度淋巴瘤的两只狗获得的两种犬淋巴瘤细胞系（CL-1：T细胞淋巴瘤细胞系，GL-1：B细胞白血病细胞系）和肿瘤组织- 2009年使用。将通过PCR获得的Ig或TCR的序列插入表达质粒中，并将构建的疫苗质粒转染至COS-7细胞。通过间接免疫荧光或Western blotting检测细胞表面Ig或TCR蛋白的表达。同时，我们尝试建立利用实时PCR从外周血样本中获取最小残留区（MRD）来评估DNA疫苗效果的方法。利用该方法可以对9例临床病例化疗期间的MRD水平进行评估。这表明MRD可以作为评估淋巴瘤犬肿瘤细胞负荷的客观标志物。化疗结束时的 MRD 可能是预测化疗后缓解持续时间的预后因素

项目成果

リンパ腫の治療:きほんのき

淋巴瘤治疗：Kihonki

• 作者: Konnai S;Mekata H;Mingala C.N.;Abes N.S.;Gutierrez C.A.;Herrera R.V.;Dargantes A.P.;Witola W.H.;Cruz L.C.;Inoue N.;Onuma M.;Ohashi K.;瀬戸口明日香
• 通讯作者: 瀬戸口明日香

uantitative assessment of minimal residual disease (MRD) in canine lymphoma by using real-time polymerase chain reaction

使用实时聚合酶链反应定量评估犬淋巴瘤的微小残留病（MRD）

• 发表时间: 2008
• 期刊: Vet.Immunol.Immunopathol 126
• 作者: Yamazaki;J.;Baba;K.;Goto-Koshino;Y.;Setoguchi-Mukai;A.;Fujino;Y.;Ohno;K.;Tsujimoto;H
• 通讯作者: H

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

阿托伐醌和阿奇霉素治疗临床病例中可能出现耐药性吉氏巴贝虫及基于CYTB基因的Jar an流行病学调查

• 发表时间: 2008
• 作者: Sakuma M;Fukuda K;Takayama K;Kobayashi Y;Setoguchi-Mukai A;Endo Y
• 通讯作者: Endo Y

Monitoring of Minimal Residual Disease (MRD) after Multidrug Chemotherapy and Its Correlation to Outcome in Dogs with Lymphoma: A Proof-of-Concept Pilot Study

• DOI: 10.1111/j.1939-1676.2010.0536.x
• 发表时间: 2010-07-01
• 期刊: JOURNAL OF VETERINARY INTERNAL MEDICINE
• 影响因子: 2.6
• 作者: Yamazaki, J.;Takahashi, M.;Tsujimoto, H.
• 通讯作者: Tsujimoto, H.

Antiviral activity of membrane fusion inhibitor that target gp40 of the feline immunodeficiency virus envelope protein

靶向猫免疫缺陷病毒包膜蛋白 gp40 的膜融合抑制剂的抗病毒活性

• 发表时间: 2009
• 期刊: Vet Microbiol. 136
• 作者: Misukoshi F;Baba K;Goto Y;Setoguchi A;Fujino Y;Ohno K;Oishi S;Kodera Y;Fujii N;Tsujimoto H
• 通讯作者: Tsujimoto H