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首页> 美国卫生研究院文献>PLoS Clinical Trials >An Optimized Method for Manufacturing a Clinical Scale Dendritic Cell-Based Vaccine for the Treatment of Glioblastoma
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An Optimized Method for Manufacturing a Clinical Scale Dendritic Cell-Based Vaccine for the Treatment of Glioblastoma

机译：一种制备临床规模的以树突细胞为基础的疫苗治疗胶质母细胞瘤的优化方法

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Immune-based treatments represent a promising new class of therapy designed to boost the immune system to specifically eradicate malignant cells. Immunotherapy may generate specific anti-tumor immune responses, and dendritic cells (DC), professional antigen-presenting cells, are widely used in experimental cancer immunotherapy. Several reports describe methods for the generation of mature, antigen-pulsed DC for clinical use. Improved quality and standardization are desirable to obtain GMP-compliant protocols. In this study we describe the generation of DC from 31 Glioblastoma (GB) patients starting from their monocytes isolated by immunomagnetic CD14 selection using the CliniMACS® device. Upon differentiation of CD14+ with IL-4 and GM-CSF, DC were induced to maturation with TNF-α, PGE2, IL-1β, and IL-6. Whole tumor lysate was obtained, for the first time, in a closed system using the semi-automated dissociator GentleMACS®. The yield of proteins improved by 130% compared to the manual dissociation method. Interestingly the Mean Fluorescence Intensity for CD83 increased significantly in DC pulsed with “new method” lysate compared to DC pulsed with “classical method” lysate. Our results indicate that immunomagnetic isolation of CD14+ monocytes using the CliniMACS® device and their pulsing with whole tumor lysate proteins is a suitable method for clinical-scale generation of high quality, functional DC under GMP-grade conditions.
机译：基于免疫的疗法代表了一种有前途的新型疗法，旨在增强免疫系统以特异性根除恶性细胞。免疫疗法可能会产生特异性的抗肿瘤免疫反应，树突状细胞（DC）是专业的抗原呈递细胞，广泛用于实验性癌症免疫疗法中。一些报告描述了用于临床使用的成熟的，抗原脉冲的DC的产生方法。需要改进质量和标准化以获得符合GMP的协议。在这项研究中，我们描述了通过使用CliniMACS®设备通过免疫磁CD14选择分离出的单核细胞，从31名胶质母细胞瘤（GB）患者中产生DC。用IL-4和GM-CSF分化CD14 +后，DC被TNF-α，PGE2，IL-1β和IL-6诱导成熟。使用半自动解离器，在封闭系统中首次获得了完整的肿瘤裂解物。与手动解离方法相比，蛋白质的产量提高了130％。有趣的是，与“传统方法”裂解物脉冲产生的直流脉冲相比，“新方法”裂解物脉冲产生的直流脉冲中CD83的平均荧光强度显着增加。我们的结果表明，使用CliniMACS®装置对CD14 + 单核细胞进行免疫磁分离，并对其进行全肿瘤溶解产物脉冲处理，是在GMP级条件下临床规模生成高质量，功能性DC的合适方法。 。

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