کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902399 | 1545803 | 2005 | 14 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A novel four-colour flow cytometric assay to determine natural killer cell or T-cell-mediated cellular cytotoxicity against leukaemic cells in peripheral or bone marrow specimens containing greater than 20% of normal cells
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کلمات کلیدی
PC5AMLGvHDmAbFITCFSCSSCDLIECDJMMLANKIL-2natural killer - (سلول های) کشنده طبیعیE:T - E: TMonoclonal antibody - آنتی بادی مونوکلونالInterleukin-2 - اینترلوکین-۲Donor lymphocyte infusions - تزریق لنفوسیت دونرcomplete media - رسانه کاملT cells - سلول های تیNatural killer cells - سلولهای کشنده طبیعیCytotoxicity - سمیت سلولیphycoerythrine - فایکوئیدرینFlow cytometry - فلوسیتومتریAcute lymphoblastic leukaemia - لوسمی لنفوبلاستی حادAcute myeloid leukaemia - لوسمی میلوئید حادConjugate - مزدوجeffector:target - نمایشگر: هدفALL - همهMonoclonal antibodies - پادتنهای تَکتیرهside scatter - پراکندگی سمتPropidium iodide - پروتئین یدید
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
To be able to determine the cytotoxic activity of NK cells or T cells against leukaemic cells in patient samples containing >20% of normal peripheral blood cells, we have developed a four-colour flow cytometric cytotoxicity assay. The assay is based on differential immunostaining of both leukaemic cells and effector cells in combination with propidium iodide (PI). The cytometer is set for measuring the fluorescence of the monoclonal antibody (mAb) bound fluorochromes, with moderate overcompensation of the third and fourth fluorescence signals. PI-positive events were excluded from analysis by their characteristic uncompensated signal on these two detectors. Thus, all four fluorescence ranges can be used for detection of mAb-derived signals and this allows discrimination between various populations contained in effector and target cell samples. The cytotoxic activity in our assay is calculated by the absolute loss of vital leukaemic cells. For this purpose, fluorescent beads are included as an internal standard. When calculating the effector concentrations after coculture, characteristic changes can be seen which yield additional information about the presence of cytotoxic activity and the active effector cell subset. With this assay, we present a versatile tool that combines minimum cell manipulation before coculture with maximum information from a sample. The assay is suitable for the analysis of complex samples with regard to different cell subsets, their decrease or increase, and conjugate formation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 296, Issues 1â2, January 2005, Pages 63-76
Journal: Journal of Immunological Methods - Volume 296, Issues 1â2, January 2005, Pages 63-76
نویسندگان
Stefanie-Yvonne Zimmermann, Ruth Esser, Eckhard Rohrbach, Thomas Klingebiel, Ulrike Koehl,