嗜酸性粒细胞：与使用抗CD16偶联微珠进行分离相比，一种新型分离试剂盒的功能差异。
Eosinophil granulocytes: functional differences of a new isolation kit compared to the isolation with anti-CD16-conjugated MicroBeads.
作者信息
Schefzyk Matthias, Bruder Manuela, Schmiedl Andreas, Stephan Michael, Kapp Alexander, Wedi Bettina, Raap Ulrike
出版信息
Exp Dermatol. 2009 Jul;18(7):653-5. doi: 10.1111/j.1600-0625.2008.00824.x. Epub 2009 Jan 21.
BACKGROUND/PURPOSE: Using a new eosinophil isolation kit, we were not able to confirm our previous findings of a delayed apoptosis of eosinophils in atopic dermatitis. Thus, we investigated whether this new isolation kit modulates the functional activity of eosinophils.
METHODS
Peripheral blood eosinophils were isolated with the new isolation kit as well as conventionally with anti-CD16-conjugated MicroBeads. We analysed viability, apoptosis, CD69 and CD95 expression, streptavidin binding and superoxide anion release.
RESULTS
Purity of eosinophils was higher using the new isolation kit (P < 0.05). However, these eosinophils had a decreased survival (P < 0.05-0.01), presented morphological features of apoptosis, showed an increased percentage of apoptotic nuclei (P < 0.01), an increased release of superoxide anions (P < 0.05), a higher expression of CD69 and CD95 (P < 0.05) and an increased binding to streptavidin compared to eosinophils isolated with anti-CD16 conjugated MicroBeads.
CONCLUSION
The new eosinophil isolation kit should not be used for the investigation of eosinophils as it potently affects their functional activity.
背景/目的：使用一种新型嗜酸性粒细胞分离试剂盒时，我们无法证实之前关于特应性皮炎中嗜酸性粒细胞凋亡延迟的研究结果。因此，我们研究了这种新型分离试剂盒是否会调节嗜酸性粒细胞的功能活性。
方法
使用新型分离试剂盒以及传统的抗CD16偶联微珠法分离外周血嗜酸性粒细胞。我们分析了细胞活力、凋亡、CD69和CD95表达、抗生物素蛋白链菌素结合以及超氧阴离子释放情况。
结果
使用新型分离试剂盒时嗜酸性粒细胞的纯度更高（P < 0.05）。然而，与使用抗CD16偶联微珠法分离的嗜酸性粒细胞相比，这些嗜酸性粒细胞的存活率降低（P < 0.05 - 0.01），呈现出凋亡的形态学特征，凋亡细胞核的百分比增加（P < 0.01），超氧阴离子释放增加（P < 0.05），CD69和CD95表达更高（P < 0.05），与抗生物素蛋白链菌素的结合增加。
结论
新型嗜酸性粒细胞分离试剂盒不应被用于嗜酸性粒细胞的研究，因为它会显著影响其功能活性。
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