Department of Research and Development, Celsense Inc., Pittsburgh, Pennsylvania, USA.
Cytotherapy. 2010 Apr;12(2):238-50. doi: 10.3109/14653240903446902.
BACKGROUND AIMS: Dendritic cells (DC) are increasingly being used as cellular vaccines to treat cancer and infectious diseases. While there have been some promising results in early clinical trials using DC-based vaccines, the inability to visualize non-invasively the location, migration and fate of cells once adoptively transferred into patients is often cited as a limiting factor in the advancement of these therapies. A novel perflouropolyether (PFPE) tracer agent was used to label human DC ex vivo for the purpose of tracking the cells in vivo by (19)F magnetic resonance imaging (MRI). We provide an assessment of this technology and examine its impact on the health and function of the DC. METHODS: Monocyte-derived DC were labeled with PFPE and then assessed. Cell viability was determined by examining cell membrane integrity and mitochondrial lipid content. Immunostaining and flow cytometry were used to measure surface antigen expression of DC maturation markers. Functional tests included bioassays for interleukin (IL)-12p70 production, T-cell stimulatory function and chemotaxis. MRI efficacy was demonstrated by inoculation of PFPE-labeled human DC into NOD-SCID mice. RESULTS: DC were effectively labeled with PFPE without significant impact on cell viability, phenotype or function. The PFPE-labeled DC were clearly detected in vivo by (19)F MRI, with mature DC being shown to migrate selectively towards draining lymph node regions within 18 h. CONCLUSIONS: This study is the first application of PFPE cell labeling and MRI cell tracking using human immunotherapeutic cells. These techniques may have significant potential for tracking therapeutic cells in future clinical trials.
背景目的:树突状细胞(DC)越来越多地被用作细胞疫苗来治疗癌症和传染病。虽然基于 DC 的疫苗在早期临床试验中取得了一些有希望的结果,但由于无法对患者体内过继转移的细胞的位置、迁移和命运进行非侵入性可视化,这通常被认为是这些疗法发展的一个限制因素。一种新型全氟聚醚(PFPE)示踪剂被用于体外标记人 DC,以便通过(19)氟磁共振成像(MRI)对细胞进行体内追踪。我们对这项技术进行了评估,并研究了它对 DC 健康和功能的影响。
方法:单核细胞来源的 DC 用 PFPE 标记,然后进行评估。通过检查细胞膜完整性和线粒体脂质含量来确定细胞活力。免疫染色和流式细胞术用于测量 DC 成熟标志物的表面抗原表达。功能测试包括白细胞介素(IL)-12p70 产生、T 细胞刺激功能和趋化性的生物测定。通过将 PFPE 标记的人 DC 接种到 NOD-SCID 小鼠中来证明 MRI 的功效。
结果:DC 被 PFPE 有效标记,而对细胞活力、表型或功能没有显著影响。PFPE 标记的 DC 可通过(19)F MRI 清晰地在体内检测到,成熟的 DC 在 18 小时内被证明选择性地向引流淋巴结区域迁移。
结论:这是首次应用 PFPE 细胞标记和 MRI 细胞追踪技术用于人类免疫治疗细胞。这些技术在未来的临床试验中可能具有追踪治疗细胞的重要潜力。
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