Vroblova V, Smolej L, Krejsek J
Faculty of Medicine and University Hospital, Charles University, Hradec Kralove, Czech Republic.
Hematology. 2012 Sep;17(5):268-74. doi: 10.1179/1607845412Y.0000000015.
Zeta-associated protein of 70 kDa (ZAP-70) is a tyrosine kinase that plays a role in signal transduction from the T-cell receptor. ZAP-70 is expressed in normal T-cells and NK-cells. Increased expression of ZAP-70 has been identified in chronic lymphocytic leukemia (CLL). CLL patients with increased ZAP-70 expression have significantly worse prognosis in terms of both progression-free survival and overall survival. There are several methods to quantify ZAP-70: polymerase chain reaction (PCR), immunoblotting, immunohistochemistry, and flow cytometry. Use of flow cytometry for ZAP-70 detection seems to be advantageous as this technique enables us to assess the presence of ZAP-70 separately on CLL clone, T-cells, and NK-cells. On the other hand, detection of ZAP-70 by flow cytometry is substantially influenced by many variables. The principal drawback of flow cytometry is the absence of consensus regarding selection of optimal anti-ZAP-70 antibody, fluorochrome conjugate, the most reliable staining technique, and optimal positivity threshold. This article summarizes pitfalls of flow cytometric analysis of ZAP-70 in CLL.
70 kDa的ζ相关蛋白（ZAP-70）是一种酪氨酸激酶，在T细胞受体的信号转导中发挥作用。ZAP-70在正常T细胞和自然杀伤细胞（NK细胞）中表达。在慢性淋巴细胞白血病（CLL）中已发现ZAP-70表达增加。ZAP-70表达增加的CLL患者在无进展生存期和总生存期方面的预后明显更差。有几种方法可用于定量ZAP-70：聚合酶链反应（PCR）、免疫印迹、免疫组织化学和流式细胞术。使用流式细胞术检测ZAP-70似乎具有优势，因为该技术使我们能够分别评估CLL克隆、T细胞和NK细胞上ZAP-70的存在情况。另一方面，流式细胞术检测ZAP-70受到许多变量的显著影响。流式细胞术的主要缺点是在选择最佳抗ZAP-70抗体、荧光染料偶联物、最可靠的染色技术和最佳阳性阈值方面缺乏共识。本文总结了CLL中ZAP-70流式细胞术分析的陷阱。
