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用超顺磁性氧化铁颗粒标记的抗原特异性人细胞毒性T淋巴细胞的可视化。

Visualization of antigen-specific human cytotoxic T lymphocytes labeled with superparamagnetic iron-oxide particles.

作者信息

Beer Ambros J, Holzapfel Konstantin, Neudorfer Juliana, Piontek Guido, Settles Marcus, Krönig Holger, Peschel Christian, Schlegel Jürgen, Rummeny Ernst J, Bernhard Helga

机构信息

Department of Nuclear Medicine, Klinikum rechts der Isar, Technical University of Munich (TUM), Munich, Germany.

出版信息

Eur Radiol. 2008 Jun;18(6):1087-95. doi: 10.1007/s00330-008-0874-4. Epub 2008 Feb 20.

Abstract

New technologies are needed to characterize the migration and survival of antigen-specific T cells in vivo. In this study, we developed a novel technique for the labeling of human cytotoxic T lymphocytes with superparamagnetic iron-oxide particles and the subsequent depiction with a conventional 1.5-T magnetic resonance scanner. Antigen-specific CD8(+) T lymphocytes were labeled with ferucarbotran by lipofection. The uptake of ferucarbotran was confirmed by immunofluorescence microscopy using a dextran-specific antibody, and the intracellular enrichment of iron was measured by atomic absorption spectrometry. The imaging of T cells was performed by magnetic resonance on day 0, 2, 7 and 14 after the labeling procedure. On day 0 and 2 post labeling, a pronounced shortening of T2*-relaxation times was observed, which diminished after 7 days and was not detectable anymore after 14 days, probably due to the retained mitotic activity of the labeled T cells. Of importance, the antigen-specific cytolytic activity of the T cells was preserved following ferucarbotran labeling. Efficient ferucarbotran labeling of functionally active T lymphocytes and their detection by magnetic resonance imaging allows the in vivo monitoring of T cells and, subsequently, will impact the further development of T cell-based therapies.

摘要

需要新技术来表征抗原特异性T细胞在体内的迁移和存活情况。在本研究中,我们开发了一种新技术,用超顺磁性氧化铁颗粒标记人细胞毒性T淋巴细胞,随后用传统的1.5-T磁共振扫描仪进行描绘。通过脂质转染用 ferucarbotran 标记抗原特异性CD8(+) T淋巴细胞。使用葡聚糖特异性抗体通过免疫荧光显微镜确认 ferucarbotran 的摄取,并通过原子吸收光谱法测量细胞内铁的富集。在标记程序后的第0、2、7和14天通过磁共振对T细胞进行成像。在标记后的第0天和第2天,观察到T2*弛豫时间明显缩短,7天后缩短程度减小,14天后不再可检测到,这可能是由于标记的T细胞保留了有丝分裂活性。重要的是,ferucarbotran 标记后T细胞的抗原特异性溶细胞活性得以保留。对功能活跃的T淋巴细胞进行有效的 ferucarbotran 标记并通过磁共振成像对其进行检测,能够在体内监测T细胞,随后将影响基于T细胞的疗法的进一步发展。

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