Isolation and characterization of tumor-specific antigen toward cancer gene therapy

用于癌症基因治疗的肿瘤特异性抗原的分离和表征

基本信息

批准号：
09044306
负责人：
KANEDA Yasufumi
金额：
$ 2.3万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

We isolated melanoma specific antigen cDNA from melanoma patient cDNA library by PCR.MAGE-1 and 3 were introduced into mouse muscle by HVJ-liposomes and could induce antibody rtesponse tu tumor antigen. Then, we tested preventive effects of cancer gene vaccination in tumorbearing mice. Op 100 is immunogenic and shown to induce both antibody and cytotoxic T cell (CTL) responses in humans. The gplOO DNA vaccine was used in a mouse melanoma model to assess immunity against the B16 melanoma of C57BL/6 mice. Intrasmuscular injection of the DNA-HVJ-AVE liposomes induced both anti-gp100 anyibody and CTL responses. Gp 100 DNA-HVJ-AVE liposome immunization delayed tumor development in mice challenged with B16 melanoma cells. Mice limmunized with gplOO DNA-HVJ-AVE liposomes survived longer compared with control mice immunized with HVJ-AVE liposome alone. Next, an RNA melanoma vaccine was investigated to induce protective immunity in a mouse-melanoma model when administered by various routes. The human melanoma-associated antigen gplOO mRNA was in vitro synthesized by pSFV3 expression vector and encapsulated in HVJ- liposomes. Immunization by either intramuscular or intradermal injection of the mRNA-HVJ-liposomes was ineffective for the induction of protective immunity against B16 melanoma challenge. However, immunization by direct injection of the gplOO mRNA HVJ-liposomes into mouse spleen induced both anti-gp 100 Ab and CTL responses against melanoma. Immunizations with gplOO mRNA into the spleen delayed tumor growth and significantly prolonged survival compared to control treated mice. These pre-clinical studies demonstrate that an RNA tumor antigen vaccine strategy has potential application for human cancer treatment and prevention.

我们通过pCR.mage-1和3从黑色素瘤患者cDNA文库中分离出黑色素瘤特异性抗原cDNA，通过HVJ-脂质体引入小鼠肌肉中，并可能诱导抗体Rtespse rtesponse tu肿瘤抗原。然后，我们测试了癌基因疫苗接种在肿瘤小鼠中的预防作用。 OP 100具有免疫原性，并证明可以诱导人类中的抗体和细胞毒性T细胞（CTL）反应。 Gploo DNA疫苗用于小鼠黑色素瘤模型中，以评估针对C57BL/6小鼠B16黑色素瘤的免疫力。 DNA-HVJ-ave脂质体的内刺肌注入诱导抗GP100任何分体和CTL反应。 GP 100 DNA-HVJ-ave脂质体免疫延迟了受B16黑色素瘤细胞挑战的小鼠的肿瘤发育。与单独用HVJ-ave脂质体免疫的对照小鼠相比，用Gploo DNA-HVJ-ave脂质体燃烧的小鼠存活更长。接下来，研究了一种RNA黑色素瘤疫苗，以诱导小鼠黑色素模型中的保护性免疫。人类黑色素瘤相关的抗原Gploo mRNA由PSFV3表达载体体外合成，并封装在HVJ-脂质体中。通过肌肉内或皮内注射mRNA-HVJ-脂质体的免疫免疫，无法诱导针对B16黑色素瘤挑战的保护性免疫。然而，通过直接注射Gploo mRNA HVJ-脂质体在小鼠脾脏中进行免疫，诱导了抗GP 100 AB和CTL反应对黑色素瘤。与对照治疗的小鼠相比，用Gploo mRNA的免疫接种脾脏延迟肿瘤生长，生存率显着延长。这些临床前研究表明，RNA肿瘤抗原疫苗策略具有潜在的人类癌症治疗和预防应用。