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IMMUNOTHERAPY WITH ELECTROFUSED DENDRITIC TUMOR HYBRIDS

电融合树突状肿瘤杂交体的免疫治疗

基本信息

批准号：
6763097
负责人：
SUYU SHU
金额：
$ 34.43万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-07-01 至 2006-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6763097
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

项目摘要

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这在很大程度上取决于诱导的免疫应答的效力和持续时间。到迄今为止，癌症疫苗已经利用了完整的肿瘤细胞，天然蛋白， CTL定义的肽、核酸、细胞膜和重组病毒，以及转基因肿瘤。虽然有些方法已经被用于增加肿瘤细胞的抗原呈递能力，在大多数情况下，已经证明了宿主抗原呈递细胞（APC）的主导作用。在各种APC中，树突状细胞（Dendritic cells，DC）似乎具有基本的特性，引发T细胞反应所需的：迁移和归巢，抗原加工、呈现和共刺激。最近，DC已被用于呈现肿瘤抗原。这通常是通过脉冲DC来实现的，肽、肿瘤裂解物或源自肿瘤细胞的RNA。还已经证明用DC-肿瘤融合嵌合细胞免疫导致已经确定的转移的消退。融合细胞应该有能力通过加工和诱导MHC I类和II类限制性反应，呈现已知和未定义的肿瘤抗原。然而，在大多数报告的案件中，融合已经用PEG完成，导致效率低且毒性高。通过这种方法，它可能是技术性的，而不是限制其应用的概念方面。申请人最近证明了通过施加电场使DC和肿瘤细胞融合，脉冲的效率至少是PEG的10倍。有趣的是，未成熟DC与肿瘤细胞的电融合产生具有以下特征的杂交细胞：具有高表达MHC II类、B7.1和 B7.2分子。分析其生物学和免疫学功能人工产生的嵌合细胞，本申请将利用良好表征的模型肿瘤相关抗原，B-半乳糖苷酶，以解决根本问题。后续的实验将使用几个弱和差免疫原性肿瘤本申请的目标是在鼠中开发利用发展中国家的原则和方法肿瘤细胞用于癌症的主动和过继免疫治疗。具体目标目的：1）优化电融合技术; 2）表征抗原融合蛋白的加工和呈递、MHC限制和运输模式分析DC-肿瘤诱导的免疫应答特点 4）探索嵌合细胞的治疗潜力;以及 5)建立一级体外免疫方法，肿瘤特异性T细胞。