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: 6103748
• 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/6103748
• 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.

培养用于过继治疗的抗肿瘤CD 8 T细胞的培养技术是 可用于小鼠肿瘤模型和人黑素瘤患者;相反， 可靠的培养技术来培养抗肿瘤CD 4 T细胞， 治疗特征尚不可用。老鼠的数据表明 Th 1型抗肿瘤CD 4 + T细胞（白细胞介素-2和干扰素- γ分泌）可能增强目前可能的治疗效果 与抗肿瘤CD 8 + T细胞;因此，一个主要的努力， 可重复的CD 4+培养技术正在进行中。 在小鼠实验中，我们已经证明，树突状细胞（DC）脉冲与肿瘤 裂解物是培养物中肿瘤特异性CD 4 + T细胞的有效活化剂。 已经建立了CD 4 + T细胞系，其特异性识别 肿瘤，具有“Th 1”特征，给予时具有治疗作用 to tumor肿瘤-荷瘤mice小鼠.提高了培养的可重复性 通过从培养物中去除外来物种蛋白并通过改善DC 通过共刺激分子的差异上调的性能 B7-1和B7-2。我们正在优化CD 4 + T细胞的过继治疗， 在有或没有CD 8 + T细胞共施用的情况下使肿瘤排斥最大化。 建立的小鼠抗肿瘤CD 4 + T细胞也提供了T细胞来源， 受体的分子研究;西村利用抗肉瘤 CD 4 + T细胞作为T细胞杂交瘤融合伴侣以辅助肿瘤抗原 隔离在平行的工作中，我们试图培养抗原特异性的 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），其允许从人类富集DC 血液;这些DC可用于处理和呈现回忆 抗原和-肿瘤抗原与自体CD 4 + T细胞的结合。CD 4应答 回忆抗原在正常供体和黑色素瘤中很容易被证实， 患者，但在结肠癌患者中受损。此外， 很难证明抗肿瘤反应。我们已经确定 这些困难的几个原因：（1）培养的人DC需要 培养物中的细胞因子活化以增加57和MHC II类表达（2） 结肠癌患者似乎有一个额外的损害。 黑色素瘤患者缺乏CD 4+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Th 1- CD 4 T细胞在癌症患者中的作用