GENE-MODIFIED DENDRITIC CELLS FOR TUMOR IMMUNOTHERAPY

用于肿瘤免疫治疗的基因修饰树突状细胞

• 批准号: 2896111
• 负责人: Saswati Chatterjee
• 金额: $ 23.72万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2002-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2896111
• 关键词: MHC class I antigen; adeno associated virus group; antigen presenting cell; antiviral antibody; autologous transplantation; cervix neoplasms; cytotoxic T lymphocyte; dendritic cells; female; gene therapy; human papillomavirus; human tissue; laboratory mouse; neoplasm /cancer immunotherapy; tissue /cell culture; transfection /expression vector; tumor antigens

DESCRIPTION: (Applicant's Abstract) Infection with human papilloma viruses (HPV) has become a major public health concern because of their high prevalence, increasing incidence, and oncogenicity. HPV-mediated oncogenicity has been mapped to the E6 and E7 open reading frames and has been associated with more than 90% of cervical carcinomas. Although E6 and E7 expressed from HPV-transformed cells can serve as effective targets for cytotoxic responses in animal models, natural cell mediated immunity to HPV in humans is surprisingly poor. Recently, dendritic cells (DC) have been identified as potent, professional APC which stimulate T cells to recognize and respond to specific antigens. Peptide pulsed DC have been shown to generate potent antitumor cytotoxic responses in animal models and one human trial. Peptide epitopes are presented by DC in association with HLA molecules, with different HLA alleles associating with different peptides. However, the identification of peptides with immunogenic epitopes which can be presented in association with the vast array of HLA alleles in the population is an arduous task. Thus the introduction of genes encoding the immunogen of interest into DC and the subsequent selection of optimal epitopes by DC T cell interactions is attractive for the generation of immunity. However gene transfer into human DC by transfection or retroviral transduction has proven difficult due to their non-proliferative status. Adeno-associated virus (AAV) vectors have recently been identified as highly promising vehicles for gene therapy because of their wide host range, lack of cytopathogenicity, stable integration, and ability to transduce nonproliferating cellular targets. The applicant has demonstrated sustained and efficient transduction of primary human monocyte-macrophages, also APCs closely related to DCs in function and ontogeny. In this study she will test the induction of HLA-restricted, antigen-specific CTL following introduction of the HPV E7 transforming gene into DC by AAV vector transduction. In vivo transplantation of syngeneic transduced murine DC will be tested for protection from tumor challenge and effects on previously existing tumors as well as for the induction of memory responses. Lastly, DCs from patients with HPV16+ cervical cancers will be transduced with AAV-E7 and tested for their ability to generate CTL which lyse autologous tumor cells. These studies will 1) test the generation of tumor-specific immunity following AAV vector transduction of the gene encoding the immunogen into DCs, 2) potentially simplify and facilitate the development of molecular vaccines, 3) provide important preclinical information regarding the use of AAV vectors for tumor immunotherapy and may form the groundwork for the development of gene-based immune strategies against cancer.

描述：(申请人摘要)感染人乳头瘤病毒 (HPV)已成为一个主要的公共卫生问题，因为它们的高水平 患病率、发病率和致瘤性。人乳头瘤病毒感染 致癌性已被映射到E6和E7开放阅读框架，并已 与90%以上的宫颈癌有关。虽然E6和 人乳头瘤病毒转化细胞表达的E7可作为有效靶点 动物模型中的细胞毒反应、自然细胞介导的HPV免疫 在人类中是令人惊讶的贫穷的。最近，树突状细胞(DC)已经被 被确认为有效的、专业的APC，刺激T细胞识别 并对特定的抗原做出反应。多肽致敏的DC已被证明 在动物模型和一名人类中产生有效的抗肿瘤细胞毒反应 审判。与人类白细胞抗原相关的多肽表位由DC呈递 分子，具有不同的等位基因与不同的多肽相关联。 然而，具有免疫原性表位的多肽的鉴定可以 与大量的人类白细胞抗原等位基因一起出现 人口是一项艰巨的任务。因此，编码基因的引入 将感兴趣的免疫基因导入DC并随后选择最优的 DC T细胞相互作用表位的产生具有吸引力 豁免权。然而，通过转基因或逆转录病毒将基因转移到人树突状细胞中 由于其非增殖状态，转导已被证明是困难的。 腺相关病毒(AAV)载体最近被鉴定为高度 由于其广泛的宿主范围，有望成为基因治疗的载体 具有细胞致病性、稳定的整合和转导能力 非增殖性细胞靶点。申请人已经证明了他的持续支持 和高效转导原代人单核巨噬细胞，也就是APC 在功能和个体发育上与DC密切相关。在这项研究中，她将 检测人类白细胞抗原限制性CTL的诱导情况 人乳头瘤病毒E7基因AAV载体转化DC的研究 转导。同基因转导小鼠DC的体内移植 将测试针对肿瘤挑战的保护和对以前 对于现有的肿瘤以及对记忆反应的诱导。最后， HPV16阳性宫颈癌患者的DC将被转导 AAV-E7，并测试其产生CTL的能力，该CTL可溶解自体 肿瘤细胞。这些研究将1)测试肿瘤特异性基因的产生 AAV载体转导编码该基因的免疫应答 DC中的免疫基因，2)潜在地简化和促进开发 分子疫苗，3)提供重要的临床前信息 关于使用AAV载体进行肿瘤免疫治疗，并可能形成 为开发基于基因的免疫策略奠定基础 癌症。

项目成果

Immunogenicity of a p210(BCR-ABL) fusion domain candidate DNA vaccine targeted to dendritic cells by a recombinant adeno-associated virus vector in vitro.

• 发表时间: 2002-06
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Ji-Yao Sun;R. Krouse;S. Forman;D. Senitzer;I. Sniecinski;S. Chatterjee;K. Wong

Immunogenic issues concerning recombinant adeno-associated virus vectors for gene therapy.

用于基因治疗的重组腺相关病毒载体的免疫原性问题。

• DOI: 10.2174/1566523023347616
• 发表时间: 2002
• 期刊: Current gene therapy
• 影响因子: 3.6
• 作者: Sun,JY;Chatterjee,S;WongJr,KK
• 通讯作者: WongJr,KK