Immunotherapy with Dendritic-Allogeneic Tumor Cells

树突状同种异体肿瘤细胞的免疫治疗

• 批准号: 6928007
• 负责人: SUYU SHU
• 金额: $ 31.37万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6928007
• 关键词: T lymphocyte; antigen presenting cell; beta galactosidase; breast neoplasms; dendritic cells; enzyme linked immunosorbent assay; flow cytometry; histocompatibility antigens; immune response; laboratory mouse; melanoma; neoplasm /cancer immunotherapy; neoplasm /cancer vaccine; passive immunization; vaccine development; vaccine evaluation

DESCRIPTION (provided by applicant): Dendritic cells (DCs) induce primary T cell responses and are being extensively evaluated as vehicles for antigen (Ag) delivery in cancer immunotherapy. One strategy employs the use of hybrid cells generated by fusing DCs with tumor cells. Technically, fusion using chemical fusogens such as polyethylene glycol (PEG) or viruses has been plagued by low efficiency, toxicity and poor reproducibility. We recently developed an electrofusion protocol with which heterokaryons are generated with high fusion rates. In pre-clinical studies, fusion cells stimulated IFN? secretion from both CD4 and CD8 immune T cells, and for immunotherapy, a single vaccination resulted in tumor regression. Based on these observations, we have recently designed a clinical trial for the treatment of metastatic melanoma by vaccination with fusion of autologous DCs and allogeneic melanoma cell lines (PMCV or CanvaxinTU). In this protocol, 3 selected allogeneic tumor cells provide a source of tumor-associated Ags. However, little laboratory analysis has focused on defining the immunogenicity and potential therapeutic effects of this approach. Because of the immune response to histo-incompatible allo-Ags on allogeneic tumor cells, immune responses to tumor Ags may be subjected to down- or up-regulation. The mechanisms of allogeneic tumor vaccine need to be analyzed. In this proposal, two animal tumor systems will be used for the development of allogeneic vaccine. The first system will utilize beta-galactosidase (beta-gal) as a surrogate tumor rejection Ag that has been introduced into the B16/F10 (H-2b) melanoma and 4T1 (H-2d) breast carcinoma. The second system consists of three murine melanomas [B16 (H-2b), K1735 (H-2k) and cloudman S91 (H-2d)] of different haplotypes but sharing a number of natural tumor-associated Ags. These allogeneic tumors will be electrofused with syngeneic DCs. Their molecular characteristics, immunogenicities, and therapeutic effects will be analyzed against syngeneic tumors. The specific aims are: 1) To determine the immunogenicity and therapeutic efficacy of syngeneic DC-allogeneic tumor (allo-DC-tumor) hybrids generated by electrofusion; 2) To characterize and analyze immune responses to tumor-associated Ags and to allogeneic histocompatibility Ags; 3) Active immunotherapy against advanced tumors; and 4) To generate tumor-immune T cells from mice immunized with allo-DC-tumor hybrid cells for adoptive immunotherapy.

描述（由申请人提供）：树突状细胞 (DC) 诱导初级 T 细胞反应，并作为癌症免疫治疗中抗原 (Ag) 递送的载体被广泛评估。一种策略是利用 DC 与肿瘤细胞融合产生的杂交细胞。从技术上讲，使用聚乙二醇（PEG）等化学融合剂或病毒进行融合一直受到效率低、毒性大和重现性差的困扰。我们最近开发了一种电融合方案，通过该方案以高融合率生成异核体。在临床前研究中，融合细胞刺激IFN？ CD4 和 CD8 免疫 T 细胞分泌，对于免疫治疗，单次疫苗接种即可导致肿瘤消退。基于这些观察，我们最近设计了一项通过自体 DC 和同种异体黑色素瘤细胞系（PMCV 或 CanvaxinTU）融合疫苗接种来治疗转移性黑色素瘤的临床试验。在此协议中，3 个选定的同种异体肿瘤细胞提供了肿瘤相关 Ag 的来源。然而，很少有实验室分析集中于确定这种方法的免疫原性和潜在治疗效果。由于同种异体肿瘤细胞上对组织不相容的同种异体抗原的免疫反应，对肿瘤抗原的免疫反应可能会下调或上调。同种异体肿瘤疫苗的机制需要分析。在该提案中，两种动物肿瘤系统将用于同种异体疫苗的开发。第一个系统将利用 β-半乳糖苷酶 (β-gal) 作为替代肿瘤排斥 Ag，该抗原已被引入 B16/F10 (H-2b) 黑色素瘤和 4T1 (H-2d) 乳腺癌中。第二个系统由三种小鼠黑色素瘤 [B16 (H-2b)、K1735 (H-2k) 和 cloudman S91 (H-2d)] 组成，它们具有不同的单倍型，但共享许多天然肿瘤相关 Ag。这些同种异体肿瘤将与同种树突状细胞进行电融合。将针对同基因肿瘤分析它们的分子特征、免疫原性和治疗效果。具体目标是：1）确定电融合产生的同基因DC-同种异体肿瘤（allo-DC-tumor）杂交体的免疫原性和治疗效果； 2) 表征和分析对肿瘤相关抗原和同种异体组织相容性抗原的免疫反应； 3）针对晚期肿瘤的主动免疫治疗； 4) 从用同种异体 DC 肿瘤杂交细胞免疫的小鼠中产生肿瘤免疫 T 细胞，用于过继免疫治疗。