Development of DNA vaccine against intracellular protozoa bored on ubiquitin-proteasome system

基于泛素-蛋白酶体系统的细胞内原虫DNA疫苗的研制

• 批准号: 15390136
• 负责人: HIMENO Kunisuke
• 金额: $ 9.73万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390136/
• 关键词: intracellular protozoa; DNA vaccine; gene gun; ubiquitin; ubiquitin-proteasome; CD8^+ T cell; proteasome KO mice; proteasome inhibitor; 細胞内寄生原虫; ユビキチンプロテアソーム系; トキソプラズマ; SAG-1; ユビキチン融合遺伝子; PA28ノックアウトマウス

The intracellular protozoan Toxoplasma gondii (T.gondii) is a major opportunistic pathogen in the immunocompromized patient population worldwide. Among the vaccine candidates, the SAG1 protein is the best characterized. This 30-kDa protein is the major surface antigen of T.gondii tachyzoites, is highly conserved in T.gondii strains, inducing high antibody levels in humans, and is recognized by all the serum samples from infected subjects. CD8^+ T cells play an essential role in the protection against a highly virulent RH strain of T.gondii. Some CD8^+ CTL epitopes included in the SAG1 had been defined. However, most vaccine trials including DNA vaccine have resulted in failure agaist the high virulent RH strain except against the cultured and attenuated strain. In the present study, we constructed the vaccine vector encoding ubiquitinated SAG1 which is expected to be processed by "ubiquitin-proteasome pathway" and then be led to MHC class I molecules, resulting in the activation of SAG1-specific CD8^+ T cells.Mice immunized with the chimeric DNA developed potent protective immunity against Toxoplasma mediated by SAG1-specific CD8^+ T cells, while mice immunized with SAG1 gene alone did not. The enhanced immunity was dependent on the ubiquitination of SAG1 that enabled it to be efficiently processed by the proteasome pathway resulting in efficient induction of specific CD8^+T cells. The immunity was dependent on immunoproteasome LMP7 but independent on PA28a/b, a proteasome activator.

细胞内原生动物弓形虫弓形虫（T.Gondii）是全球免疫功能低下的患者人群中的主要机会病原体。在候选疫苗中，SAG1蛋白的特征是最好的。该30 kDa蛋白是t.gondii tachyzoites的主要表面抗原，在T.gondii菌株中高度保守，诱导了人类的高抗体水平，并且被感染受试者的所有血清样品识别。 CD8^+ T细胞在防止T.gondii的高毒性RH菌株的保护中起着至关重要的作用。已经定义了SAG1中包含的一些CD8^+ CTL表位。然而，包括DNA疫苗在内的大多数疫苗试验导致失败的毒剂高毒性RH菌株，除了针对培养和减弱的菌株。在本研究中，我们构建了编码泛素化SAG1的疫苗载体，该疫苗有望通过“泛素 - 蛋白酶体途径”处理，然后将其引入MHC I类分子，从而激活SAG1特异性CD8^+ T细胞。用嵌合DNA免疫的小鼠对通过SAG1特异性CD8^+ T细胞介导的毒质量产生了有效的保护性免疫，而单独用SAG1基因免疫的小鼠则没有。增强的免疫力取决于SAG1的泛素化，这使其能够通过蛋白酶体途径有效处理，从而有效诱导特定的CD8^+T细胞。免疫力取决于免疫蛋白酶体LMP7，但独立于PA28A/B（一种蛋白酶体激活剂）。

项目成果

Malaria vaccines that inhibit parasite spread by mosquito vectors.

抑制蚊媒传播寄生虫的疟疾疫苗。

• 发表时间: 2003
• 期刊: Recent Research Developments in Immunology 5
• 作者: Toyooka;K.;et al.;Kaneko O;Yano K;Kaneko 0;Yano K;Arakawa T;Sawasaki T;Arakawa T;Sawasaki T;Arakawa T;Kaneko O;Rungruang T;Sattabongkot J;Li F;Li F;Ling IT;Tsuboi T;Li F;Kongkasuriyachai D;Win TT;Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Hisaeda H. et al.;Zhang M. et al.;Zhang M. et al.;Hisaeda H. et al.;Okamoto M. et al.;Obata C. et al.;Onishi K. et al.;Yamanaka A. et al.;Zhang M. et al.;Li Y.et al.;Hisaeda H. et al.;Okamoto M. et al.;Obata C. et al.;Onishi K. et al.;Yamanaka A. et al.;Zhang M. et al.;Li Y.et al.;Hisaeda H. et al.;Onishi K. et al.;Yamanaka A. et al.;Hamano S. et al.;Seki YI. et al.;Katunuma N. et al.;Hisaeda H. et al.;Sakai T. et al.;Sakai T. et al.;Hamano S. et al.;Seki YI. et al.;Katunuma N. et al.;Hisaeda H. et al.
• 通讯作者: Hisaeda H. et al.

The ubiquitin-proteasome system plays an essential role in presenting an 8-mer CTL epitope expressed in APC to corresponding CD8^+T cells.

泛素-蛋白酶体系统在将APC中表达的8聚体CTL表位呈递给相应的CD8+T细胞方面发挥重要作用。

• 发表时间: 2006
• 期刊: Int.Immunol. (in press)
• 作者: Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Duan X.et al.
• 通讯作者: Duan X.et al.

Cathepsin L is crucial for Th1-type immune response during Leishmania major infection.

组织蛋白酶 L 对于利什曼原虫重大感染期间的 Th1 型免疫反应至关重要。

• 发表时间: 2004
• 期刊: Microbes and Infection Apr 6(5)
• 作者: Toyooka;K.;et al.;Kaneko O;Yano K;Kaneko 0;Yano K;Arakawa T;Sawasaki T;Arakawa T;Sawasaki T;Arakawa T;Kaneko O;Rungruang T;Sattabongkot J;Li F;Li F;Ling IT;Tsuboi T;Li F;Kongkasuriyachai D;Win TT;Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Hisaeda H. et al.;Zhang M. et al.;Zhang M. et al.;Hisaeda H. et al.;Okamoto M. et al.;Obata C. et al.;Onishi K. et al.;Yamanaka A. et al.;Zhang M. et al.;Li Y.et al.;Hisaeda H. et al.;Okamoto M. et al.;Obata C. et al.;Onishi K. et al.
• 通讯作者: Onishi K. et al.

Gene gun-based co-immunijzation of merozoite surface protein-1 cDNA with IL-12 expression plasmid confers protection against lethal Plasmodium yoelii in A/J mice.

基于基因枪的裂殖子表面蛋白 1 cDNA 与 IL-12 表达质粒的联合免疫可在 A/J 小鼠中提供针对致命约氏疟原虫的保护。

• 发表时间: 2003
• 期刊: Vaccine 21
• 作者: Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Duan X.et al.;Ishii K. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Zhang M. et al.;Hisaeda H. et al.;Okamoto M. et al.;Obata C. et al.;Onishi K. et al.;Yamanaka A. et al.;Zhang M. et al.;Li Y.et al.;Hisaeda H. et al.;Okamoto M. et al.;Obata C. et al.;Onishi K. et al.;Yamanaka A. et al.;Zhang M. et al.;Li Y.et al.;Hisaeda H. et al.;Obata C. et al.;Onishi K. et al.;Yamanaka A. et al.;Hamano S. et al.;Seki YI. et al.;Katunuma N. et al.;Hisaeda H. et al.;Sakai T. et al.
• 通讯作者: Sakai T. et al.

M.Zhang, K.Himeno et al.: "Melanoma immunotherapy by a DNA vaccine based on the ubiquitin-proteasome pathway."Gene Therapy. (in press). (2004)

M.Zhang、K.Himeno 等人：“通过基于泛素-蛋白酶体途径的 DNA 疫苗进行黑色素瘤免疫治疗。”基因治疗。