Analysis of Antitumor Immunity Using Dendritic Cells Treated with Various Protein-transduction Domain (PTD)-containing Protein Antigen

使用经各种含蛋白转导域 (PTD) 的蛋白抗原处理的树突状细胞进行抗肿瘤免疫分析

• 批准号: 15390339
• 负责人: SHIBAGAKI Naotaka
• 金额: $ 9.6万
• 依托单位: University of Yamanashi
• 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 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390339/
• 关键词: Vaccine; Dendritic Cells; Tumor Immunity; Protein Transduction; TAT; Polyarginine

Protein transduction domains (PTDs) have been used increasingly to deliver reagents, such as proteins, oligonucleotides to a variety of cell types in vitro and in vivo. We have previously shown that HIV TAT-PTD-containing whole protein antigens (Ags)-transduced dendritic cells (DC) stimulated Ag-specific CD8+ and CD4+ T cells by processing and presenting Ag-epitopes onto MHC class I and class II. Although the CTL activity generated was sufficient to prevent engraftment of mice with Ag-expressing tumors, treatment of tumor-bearing mice with TAT-PTD Ag-transduced DC resulted in tumor regression in some, but not all animals. Recently, several other PTDs were reported their higher transduction efficiencies than TAT-PTD. To evaluate the PTD for induction of more efficient immune responses in tumor vaccination study, we engineered several recombinant fusion OVA proteins which contain PTDs including polyarginine (R9), known as the most efficacious PTD. Our results demonstrated that R9-PTD showed higher transduction efficiency to DC among PTDs studied, and that efficacy was closely correlated with the potency of Ag-specific CD4+, CD8+ T cell activations in vitro and in vivo. Twice vaccination with R9-PTD-OVA-transduced DC in (OVA-expressing) tumor-bearing mice induced higher antitumor immunity by strong boosting effects, and elicited complete rejection of tumor mass when co-injection with LPS or OK432. This approach should be clinically applicable, and offers theoretical and practical advantages to those that are in current use, such as peptide therapy.

蛋白质转导结构域（PTD）已越来越多地用于在体外和体内将试剂如蛋白质、寡核苷酸递送至多种细胞类型。我们以前已经表明，HIV TAT-PTD-含有全蛋白抗原（Ag）转导的树突状细胞（DC）刺激Ag特异性的CD 8+和CD 4 + T细胞的加工和提出的Ag-表位的MHC I类和II类。尽管产生的CTL活性足以防止具有Ag表达肿瘤的小鼠的移植，但用TAT-PTD Ag转导的DC治疗荷瘤小鼠导致一些但不是所有动物的肿瘤消退。最近，几种其他PTD被报道其比TAT-PTD更高的转导效率。为了评价PTD在肿瘤疫苗接种研究中诱导更有效的免疫应答，我们工程化了几种重组融合OVA蛋白，其含有PTD，包括聚精氨酸（R9），被称为最有效的PTD。我们的研究结果表明，R9-PTD在所研究的PTD中显示出更高的DC转导效率，并且该功效与体外和体内Ag特异性CD 4+、CD 8 + T细胞活化的效力密切相关。在（表达OVA的）荷瘤小鼠中两次接种R9-PTD-OVA转导的DC，通过强烈的加强作用诱导了更高的抗肿瘤免疫力，并且当与LPS或OK 432共注射时，引起了对肿瘤块的完全排斥。这种方法应该是临床上适用的，并提供理论和实践的优势，目前使用的那些，如肽治疗。