IMMUNITY TO MELANOMA WITH IL-2-SECRETING ALLOGENEIC CELL

分泌 IL-2 的同种异体细胞对黑色素瘤具有免疫力

• 批准号: 3200161
• 负责人: EDWARD P. COHEN
• 金额: $ 12.97万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-03 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3200161
• 关键词: T lymphocyte; cell population study; cellular oncology; fibroblasts; homologous transplantation; interleukin 2; laboratory mouse; melanoma; monoclonal antibody; neoplasm /cancer immunotherapy; transfection; transplant rejection; tumor antigens

In spite of the fact that they express immunogenic determinants, B16 melanoma cells grow progressively in immunocompetent C57BL/6 mice (H-2b). The mechanism of "escape" is not established. Since cytotoxic T lymphocytes are known to recognize an extraordinarily wide array of small peptides (in the context of histocompatibility class I), including neoantigens associated with malignant cells, it may be that the growth of antigeneic tumors results from a failure of immune recognition, not from the absence of tumor associated determinants. An understanding of the cellular "defect" that enables tumor cells to grow in immunocompetent recipients could have important implications for the specific immunotherapy of cancer. As an experimental approach, we used transfection to construct IL-2-secreting, allogeneic mouse fibroblasts that express melanoma associated antigens. We then tested the cells' immunogenic properties in terms of their ability to elicit an anti melanoma immune response in mice syngeneic with the tumor. The construct was prepared by transfecting genomic DNA from B16 cells (H-2b) into LM cells (H-2k) which are allogeneic with C57BL/6 mice. Colonies of transfected cells expressing melanoma- associated determinants were isolated, and then infected with an expression competent plasmid carrying the gene for IL-2. In our experiments, C57BL/6 mice rejecting the IL-2-secreting, melanoma antigen-positive allogeneic mouse cells developed cellular immunity to the melanoma. This immunity exceeded that following immunization with non-IL-2-secreting constructs, or with B16 cells. The objectives of the proposal are to determine the specific cell types activated for rejection of B16 melanoma following immunization of C57BL/6 mice with the IL-2-secreting cell constructs, and compare them with cell types activated following immunization of mice with constructs that lack one or more of the immunogenic properties. In addition, we plan to determine if exogenous IL-2 can substitute for the direct cell-to-cell (paracrine) transfer of IL-2 in the induction of an anti melanoma immune response. Finally, we plan to determine if immunizations of tumor-bearing mice with the IL-2-secreting constructs leads to increased survival.

尽管它们表达了免疫原性的决定因素，但B16 黑色素瘤细胞在免疫能力的C57BL/6小鼠（H-2B）中逐渐生长。 “逃脱”的机制尚未确定。 由于细胞毒性t 众所周知，淋巴细胞识别出非常宽的小阵列 肽（在组织相容性I类的背景下），包括 与恶性细胞相关的新抗原，可能是 抗替代性肿瘤是由免疫识别失败引起的，而不是来自 缺乏相关的决定因素。 对 细胞“缺陷”，使肿瘤细胞在免疫能力中生长 接受者可能对特定免疫疗法具有重要意义 癌症。 作为一种实验方法，我们使用转染来构建 表达黑色素瘤的IL-2分泌，同种异体小鼠成纤维细胞 相关抗原。 然后，我们在细胞中测试了细胞的免疫原性 他们引起小鼠抗黑色素瘤免疫反应的能力的术语 与肿瘤合成素。 该结构是通过转染制备的 来自B16细胞（H-2B）到LM细胞（H-2K）的基因组DNA，这些DNA是同种异体的 与C57BL/6鼠。 表达黑色素瘤的转染细胞的菌落 分离相关的决定因素，然后用表达感染 携带IL-2基因的合格质粒。 在我们的实验中，C57BL/6 拒绝分泌IL-2分泌的小鼠，黑色素瘤抗原阳性同种异体 小鼠细胞对黑色素瘤产生了细胞免疫。 这种免疫力 超过了通过非IL-2分泌结构进行免疫后，或 与B16细胞。 该提案的目标是确定 激活拒绝B16黑色素瘤的特定细胞类型之后 用IL-2分泌细胞构建体对C57BL/6小鼠的免疫接种，并 将它们与小鼠免疫后激活的细胞类型进行比较 缺乏一种或多种免疫原性的构造。 在 此外，我们计划确定外源IL-2是否可以代替 IL-2的直接细胞向细胞转移（旁）诱导 抗黑色素瘤免疫反应。 最后，我们计划确定是否 用IL-2分泌构建体对肿瘤小鼠的免疫接种 导致生存增加。