GENERATION OF THERAPEUTIC MURINE AND HUMAN ANTI-TUMOR CD4+ T CELLS

治疗性小鼠和人类抗肿瘤 CD4 T 细胞的产生

• 批准号: 3731876
• 负责人: P A COHEN
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3731876
• 关键词: T cell receptor; biotechnology; cytokine; growth inhibitors; helper T lymphocyte; human subject; human tissue; laboratory mouse; melanoma; neoplasm /cancer immunotherapy; neoplasm /cancer remission /regression; neoplastic cell; tissue /cell culture; tumor antigens

Culture techniques to grow anti-tumor CD8 T cells for adoptive therapy are available for mouse tumor models and human melanoma patients; in contrast, reliable culture techniques to grow anti-tumor CD4 T cells with therapeutic characteristics are not yet available. Mouse data indicate that anti-tumor CD4+ T cells of "Th1-type" (interleukin-2 and interferon- gamma secreting) may potentiate the therapeutic effects currently possible with anti-tumor CD8+ T cells; hence, a major effort to develop reproducible CD4+ culture techniques is underway. In mouse work we have shown that dendritic cells (DC) pulsed with tumor lysates are potent activators of tumor-specific CD4+ T cells in culture. CD4+ T cell lines have been established which specifically recognize tumor, have "Th1" characteristics, and have therapeutic effects when given to tumor-bearing mice. Improved culture reproducibility is being achieved by removing foreign species proteins from culture and by improving DC performance through differential upregulation of costimulatory molecules B7-1 and B7-2. We are optimizing adoptive therapy with CD4+ T cells to maximize tumor rejection with or without coadministration of CD8+ T cells. Established mouse anti-tumor CD4+ T cells also provides a source of T cell receptor for molecule studies; Dr. M. Nishimura has utilized anti-sarcoma CD4+ T cells as T cell hybridoma fusion partners to aid in tumor antigen isolation. In parallel work we have attempted to grow antigen-specific human CD4 T cells from melanoma and colon cancer patients as well as from normal donors. We have perfected elutration techniques in collaboration with Charles Carter (DTM) which permit the enrichment of DC from human blood; these DC can be utilized to process and present both recall antigens and-tumor antigen to autologous CD4+ T cells. CD4 responses to recall antigens are readily demonstrable in normal donors and melanoma patients, but impaired in colon cancer patients. In addition, it has been difficult to demonstrate an anti-tumor response. We have identified several reasons for these difficulties: (1) cultured human DC require cytokine activation in culture to increase 57 and MHC Class II express (2) colon cancer patients appear to have an additional impairment at the. level of CD4+ function lacking in melanoma patients. We are.addressing and correcting these dysfunctional components to generate tumor-specific Th1- type CD4 T cells in cancer patients.

用于培养抗肿瘤 CD8 T 细胞用于过继治疗的培养技术是 可用于小鼠肿瘤模型和人类黑色素瘤患者；相比之下， 可靠的培养技术来培养抗肿瘤 CD4 T 细胞 治疗特征尚不清楚。鼠标数据表明 “Th1 型”抗肿瘤 CD4+ T 细胞（白细胞介素 2 和干扰素 γ分泌）可能会增强目前可能的治疗效果 具有抗肿瘤 CD8+ T 细胞；因此，大力发展 可重复的 CD4+ 培养技术正在进行中。 在小鼠实验中，我们发现树突状细胞 (DC) 会受到肿瘤的脉冲 裂解物是培养物中肿瘤特异性 CD4+ T 细胞的有效激活剂。 已经建立了特异性识别 CD4+ T 细胞系 肿瘤，具有“Th1”特征，给予后具有治疗作用 对荷瘤小鼠。培养的再现性正在提高 通过从培养物中去除外来物种蛋白质并改善 DC 通过共刺激分子的差异上调来提高性能 B7-1和B7-2。我们正在优化 CD4+ T 细胞的过继疗法 无论是否同时使用 CD8+ T 细胞，都能最大限度地抑制肿瘤。 建立的小鼠抗肿瘤CD4+ T细胞也提供了T细胞来源 分子研究的受体； M. Nishimura 博士使用抗肉瘤药物 CD4+ T 细胞作为 T 细胞杂交瘤融合伴侣，有助于肿瘤抗原 隔离。在并行工作中，我们尝试培养抗原特异性 来自黑色素瘤和结肠癌患者以及来自 正常捐赠者。我们通过合作完善了洗脱技术 与 Charles Carter (DTM) 合作，可从人类中富集 DC 血;这些 DC 可用于处理和呈现召回 自体 CD4+ T 细胞的抗原和肿瘤抗原。 CD4 反应 正常供体和黑色素瘤中很容易证明记忆抗原 患者，但结肠癌患者受损。此外，还曾 很难证明抗肿瘤反应。我们已经确定 造成这些困难的几个原因：（1）培养的人类 DC 需要 培养物中的细胞因子激活可增加 57 和 MHC II 类表达 (2) 结肠癌患者似乎还有额外的损伤。 黑色素瘤患者缺乏 CD4+ 功能水平。我们正在解决和 纠正这些功能失调的成分以产生肿瘤特异性 Th1- 癌症患者体内的 CD4 型 T 细胞。