Université Reims Champagne-Ardenne, EA Unité Interactions Animal-Environnement, Moulin de Housse, BP 1039, 51687 Reims, France.
Environ Sci Pollut Res Int. 2013 May;20(5):2692-704. doi: 10.1007/s11356-012-1410-2. Epub 2013 Jan 4.
The neutral red retention assay has been proposed to determine the lysosomal membrane stability in immune cells. Nevertheless, this assay implies many examinations under a microscope at short time intervals and therefore the analysis of few samples. The present study proposes two more rapid, efficient, and sensitive sample analyses using flow cytometry method. Lysosomal presence and lysosomal membrane integrity (LMI) were evaluated on the three-spined stickleback, Gasterosteus aculeatus (L.), a well-described model fish species for aquatic ecotoxicology studies. After development of the two biomarkers, they were validated by ex vivo contamination with endosulfan and copper and by in situ sampling. These immunomarkers were clearly modulated by pollutants and their variations seemed to be correlated with leucocyte mortality. Thus, from a practical point of view, lysosomal presence and LMI may provide novel and efficient means of evaluating immune capacities and indicating the toxic effects of environmental pollution.
中性红保留试验被提议用于测定免疫细胞中的溶酶体膜稳定性。然而，该试验需要在短时间间隔内进行多次显微镜检查，因此只能分析少量样本。本研究提出了两种使用流式细胞术更快速、高效和敏感的样本分析方法。溶酶体的存在和溶酶体膜完整性（LMI）在三刺棘鱼（Gasterosteus aculeatus（L.））中进行了评估，三刺棘鱼是水生生态毒理学研究中一种描述良好的鱼类模型物种。在开发这两种生物标志物后，通过用硫丹和铜进行离体污染和原位采样对其进行了验证。这些免疫标志物被污染物明显调节，其变化似乎与白细胞死亡率相关。因此，从实际的角度来看，溶酶体的存在和 LMI 可能提供评估免疫能力和指示环境污染的毒性影响的新的有效手段。
