Basic study of polynucleotide vaccine for tumor immunotherapy

肿瘤免疫治疗多核苷酸疫苗基础研究

基本信息

批准号：
08457316
负责人：
KITAYAMA Joji
金额：
$ 2.24万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

项目摘要

[PURPOSE] Intramuscular injection of naked DNA such as plasmid-type expression vector leads to the expression of coded protein in the muscle, and host immune response against the protein is elicited when it is non-self like viral protein. In present study we investigate whether this new method is applicable to cancer immunotherapy or not. [METHOD] At first several kinds of CAT expression vectors were intramusculary injected to perform CAT assay with muscular extract and CAG promoter and CE promoter were found to be most efficient in muscle. cDNA designated as H52, which codes one of melanoma antigens expressed on murine melanoma cell line B16. was inserted downstreams of CAG promoter (pCAGH52). Another cDNA, which codes mutant p53 protein expressed in murine fibrosarcoma cell line Meth-A, was inserted downstreams of CE promoter (pCEp53). C57BL or BALB/c mice were intramusculary injected thrice at one week interval with 100 microgram of pCAGH52 or pCEp53 respectively. One week after the last injection those mice were injected subcutaneously with 100000 cells of B16 or Meth-A respectively. [RESULT] There was no difference in tumor incidence between treated group and control group and survival was not prolonged by DNA injection. [CONCLUSION] Simple DNA injection is not sufficient to elicit host immune response against autologous tumor. Becausae the amount of protein produced after DNA injection is very small, some adjuvant would be necessary to be recognized by antigen presenting cells.

[目的]肌肉内注射赤裸的DNA，例如质粒型表达载体会导致肌肉中编码蛋白的表达，而当它像病毒蛋白（病毒蛋白）一样，会引起对蛋白质的宿主免疫反应。在目前的研究中，我们研究了这种新方法是否适用于癌症免疫疗法。 [方法]首先，注射了几种类型的CAT表达载体，以肌肉提取物和CAG启动子和CE启动子进行CAT测定最有效。指定为H52的cDNA，它代码在鼠类黑色素瘤细胞系B16上表达的一种黑色素瘤抗原之一。被插入CAG启动子的下游（PCAGH52）。 CE启动子下游插入了另一种用鼠纤维肉瘤细胞系Meth-A表示突变体p53蛋白的cDNA（PCEP53）。 C57BL或BALB/C小鼠分别用100微克的PCAGH52或PCEP53分别为肌内注射三次。最后一个注射后一周，这些小鼠分别向100000个B16或Meth-A的细胞皮下注射。 [结果]治疗组和对照组之间的肿瘤发生率没有差异，而DNA注射不会延长生存率。 [结论]简单的DNA注射不足以引起宿主对自体肿瘤的免疫反应。因为DNA注射后产生的蛋白质量很小，有些辅助是必须被抗原呈递细胞识别。