IMMUNOTHERAPY WITH ELECTROFUSED DENDRITIC TUMOR HYBRIDS

电融合树突状肿瘤杂交体的免疫治疗

• 批准号: 6514261
• 负责人: SUYU SHU
• 金额: $ 33.3万
• 依托单位: CLEVELAND CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6514261
• 关键词: antigen presentation; beta galactosidase; biomedical equipment development; cell fusion; colony stimulating factor; dendritic cells; disease /disorder model; electric field; hybrid cells; interleukin 4; ionophores; laboratory mouse; model design /development; neoplasm /cancer immunotherapy; neoplasm /cancer vaccine; neoplastic cell; nonhuman therapy evaluation; vaccine development

DESCRIPTION: (Applicant's Abstract) The efficacy of cancer immunotherapy depends highly on the potency and duration of the induced immune response. To date, cancer vaccinations have utilized intact tumor cells, naive proteins, CTL-defined peptides, nucleic acids, cell membranes and recombinant viruses as well as genetically modified tumors. Although some approaches have been to increase the antigen presenting capacity of tumor cells, in most cases, the dominant role of host antigen-presenting cells (APCs) has been demonstrated. Among various APCs, dendritic cells (DCs) seem to have the essential properties required for eliciting T-cell responses: migration and homing, antigen processing and presentation and co-stimulation. Recently, DCs have been used to present tumor antigens. This is generally achieved by pulsing DCs with peptides, tumor lysates or RNA derived from neoplastic cells. It has also been demonstrated that immunization with DC-tumor fused chimeric cells results in the regression of established metastases. Fused cells should have the ability to elicit both MHC class I and II-restricted responses by processing and presenting known and undefined tumor antigens. In most reported cases, however, fusion has been accomplished with the use of PEG, resulting in low efficiency and high toxicity. With this approach, it may be the technical rather than conceptual aspects that limit its application. The applicant has recently demonstrated that fusion of DCs and tumor cells by applying electric field pulses is at least 10-fold more efficient than that by PEG. Interestingly, electrofusion of immature DCs with tumor cells resulted in hybrid cells having characteristics of mature DCs with high expression of MHC class II, B7.1 and B7.2 molecules. To analyze biological and immunological functions of these artificially generated chimeric cells, the current application will utilize the well-characterized model tumor-associated antigen, B-galactosidase, to address fundamental issues. Subsequent experiments will use several weakly and poorly immunogenic tumors. The goal of the current application is to develop in murine models, the principles and methodologies for utilizing DCs fused with entire tumor cells for active and adoptive immunotherapy of cancer. The specific aims are: 1) To optimize electrofusion techniques; 2) To characterize antigen processing and presentation, MHC restriction and trafficking patterns of fused cells; 3) To analyze characteristics of immune responses elicited by DC-tumor fused cells; 4) To explore the therapeutic potential of the chimeric cells; and 5) To develop methods for primary in vitro immunization for generating tumor-specific T cells.

描述：（申请人的摘要）癌症免疫疗法的功效 在很大程度上取决于诱导的免疫反应的效力和持续时间。到 日期，癌症疫苗已经使用了完整的肿瘤细胞，天真的蛋白质， CTL定义的肽，核酸，细胞膜和重组病毒作为 以及转基因肿瘤。虽然有些方法已经 在大多数情况下，增加肿瘤细胞的抗原表现能力 已经证明了宿主抗原抗原细胞（APC）的主要作用。 在各种APC中，树突状细胞（DC）似乎具有必不可少的特性 引起T细胞响应所需的需要：迁移和归巢，抗原 处理，表现和共同刺激。最近，DC已习惯 目前的肿瘤抗原。这通常是通过与DC一起使用的 从肿瘤细胞衍生的肽，肿瘤裂解物或RNA。也一直 证明通过DC肿瘤融合的嵌合细胞免疫导致 已建立转移的回归。融合细胞应具有能力 通过处理和 呈现已知和未定义的肿瘤抗原。但是，在大多数报道的情况下， 融合已经通过使用PEG完成，导致低效率 和高毒性。使用这种方法，可能是技术而不是 限制其应用的概念方面。申请人最近有 证明DC和肿瘤细胞通过施加电场的融合 脉冲的效率至少比PEG高10倍。有趣的是， 未成熟DC与肿瘤细胞的电渗，导致杂化细胞具有 MHC II，B7.1和 B7.2分子。分析这些生物学和免疫学功能 人为生成的嵌合细胞，当前应用将利用 特征良好的模型肿瘤相关抗原B-半乳糖苷酶，以解决 基本问题。随后的实验将使用几个弱且较差 免疫原性肿瘤。当前应用的目的是在鼠中发展 模型，用于利用与整个DC的原理和方法论 肿瘤细胞用于癌症的主动和继发性免疫疗法。具体目标 为：1）优化电渗渗技术； 2）表征抗原 融合的处理和表现，MHC限制和贩运模式 细胞； 3）分析DC-tumor引起的免疫反应的特征 融合细胞； 4）探索嵌合细胞的治疗潜力；和 5）开发用于产生初级体外免疫的方法 肿瘤特异性T细胞。