کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8417880 | 1545704 | 2014 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
An impedance-based cytotoxicity assay for real-time and label-free assessment of T-cell-mediated killing of adherent cells
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کلمات کلیدی
Effector to target ratioPBMCCRAE:T ratioCPMOvCaxCELLigence - xCelligenceChromium release assay - آزمایش آزاد شدن کرومHuman leukocyte antigen - آنتی ژن لوسکسی انسانHLA - آنتیژن گلبول سفید انسانیImpedance - امپدانسReal time - به موقعOvarian carcinoma - سرطان تخمدانcell index - شاخص سلولcounts per minute - شمار در هر دقیقهmajor histocompatibility complex - مجموعه سازگاری بافتی اصلیMHC - مجموعه سازگاری بافتی اصلی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The in vitro assessment of T-cell-mediated cytotoxicity plays an important and increasingly relevant role both in preclinical target evaluation and during immunomonitoring to accompany clinical trials employing targeted immunotherapies. For a long time, the gold standard for this purpose has been the chromium release assay (CRA). This end point assay, however, shows several disadvantages including the inevitable use of radioactivity. Based on electrical impedance measurements (using the xCELLigence system), we have established a label-free assay, facilitating the real-time monitoring of T-cell-mediated cytotoxicity. The coculture of peptide-specific T-cell lines with peptide-loaded target cells reproducibly led to a decrease in impedance due to induced apoptosis and detachment of target cells. Comparing our results to the standard CRA assay, we could demonstrate that impedance-based measurements show comparable results after short incubation periods (6Â h) but outperform the CRA both in reproducibility and sensitivity after prolonged incubation (24Â h), enabling the detection of target cell lysis with an effector to target ratio as low as 0.05:1. The impedance-based assay represents a valuable and highly sensitive tool for label-free real-time high throughput analysis of T-cell-mediated cytotoxicity.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 405, March 2014, Pages 192-198
Journal: Journal of Immunological Methods - Volume 405, March 2014, Pages 192-198
نویسندگان
Janet Kerstin Peper, Heiko Schuster, Markus W. Löffler, Barbara Schmid-Horch, Hans-Georg Rammensee, Stefan StevanoviÄ,