Emmendörffer A, Hecht M, Lohmann-Matthes M L, Roesler J
Fraunhofer Institute ITA, Department of Immunobiology, Hannover, F.R.G.
J Immunol Methods. 1990 Aug 7;131(2):269-75. doi: 10.1016/0022-1759(90)90198-5.
Analysis of the functional activity of phagocytes is of great importance in the differential diagnosis of patients with recurrent bacterial infections. Here we describe a method to determine the production of reactive oxygen intermediates (ROI) by microcytofluorometry using dihydrorhodamine 123, a derivative of rhodamine 123. Using this method the ROI production of erythrocyte-depleted whole blood samples can be measured without further time-consuming purification steps. Possible harmful manipulation of the isolated cells can also be avoided and highly reproducible and significant results are obtained in the minimum of time. This assay provides a very sensitive alternative to the clinically used NBT test in the diagnosis of patients with chronic granulomatous disease (CGD). Moreover, the analysis of oxygen-dependent effector functions of murine effector cells and cell lines may be important in investigating resistance to certain microbes (e.g., Candida albicans, Staphylococcus aureus or different protozoa such as Toxoplasma gondii or Leishmania species).
对复发性细菌感染患者进行鉴别诊断时，分析吞噬细胞的功能活性至关重要。在此，我们描述一种使用罗丹明123的衍生物二氢罗丹明123，通过微量细胞荧光测定法来测定活性氧中间体（ROI）产生的方法。采用该方法，无需进一步耗时的纯化步骤，即可测定去除红细胞的全血样本中ROI的产生。还可避免对分离细胞可能的有害操作，并能在最短时间内获得高度可重复且显著的结果。在慢性肉芽肿病（CGD）患者的诊断中，该检测方法为临床使用的NBT试验提供了一种非常灵敏的替代方法。此外，分析小鼠效应细胞和细胞系的氧依赖性效应功能，对于研究对某些微生物（如白色念珠菌、金黄色葡萄球菌或不同的原生动物，如弓形虫或利什曼原虫属）的抗性可能具有重要意义。
