Evaluation of Cell Proliferation and Apoptosis in Immunotoxicity Testing.
Evaluation of Cell Proliferation and Apoptosis in Immunotoxicity Testing.
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DOI：
10.1007/978-1-4939-8549-4_14
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发表时间：
2018
影响因子：
--
通讯作者：
M. Nagarkatti;S. Rieder;P. Nagarkatti
中科院分区：
文献类型：
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作者：
M. Nagarkatti;S. Rieder;P. Nagarkatti
免疫毒性测试在确定各种化学物质对免疫系统的毒性影响方面很重要。免疫系统是许多毒物的直接目标，其不良影响包括严重的健康并发症，如易受感染、癌症、过敏反应和自身免疫性疾病。研究不同化学物质的有害影响的一种方法是研究免疫细胞的凋亡和/或增殖。细胞凋亡被定义为细胞程序性死亡，一般而言，这一过程有助于发展和维持耐受和动态平衡。然而，在受到毒物侮辱的情况下，免疫细胞的凋亡增强可能会导致免疫抑制，从而导致癌症的发展和无法抵抗感染。细胞凋亡的特征是细胞皱缩、核固缩、细胞膜和线粒体改变、DNA断裂以及caspase对蛋白质的降解。各种方法被用来研究细胞凋亡，包括用流式细胞术和原位标记直接测量凋亡细胞，以及指示凋亡分子的RNA、DNA和蛋白质分析。除了细胞凋亡外，细胞增殖的量化还可以提供有关毒物对各种免疫细胞群影响的重要补充信息。在某些情况下，毒物可以作为有丝分裂原，推动免疫细胞进入细胞周期的不同阶段。活跃的细胞周期有四个阶段：G1、S、G2和M，细胞分裂发生在M期。细胞增殖可通过多种方法进行定量，包括Ki-67或CFSE染色、BrdU标记、四甲基偶氮唑盐(四甲基偶氮唑盐)比色法和/或三磷酸腺苷定量。
Immunotoxicity testing is important in determining the toxic effects of various chemicals on the immune system. The immune system is a direct target of numerous toxicants, and the adverse effects include serious health complications such as susceptibility to infections, cancer, allergic reactions, and autoimmune diseases. One way to investigate the harmful effects of different chemicals is to study apoptosis and/or proliferation in immune cells. Apoptosis is defined as programmed cell death, and in general, this process helps in development and maintenance of tolerance and homeostasis. However, in the case of an insult by a toxicant, enhanced apoptosis of immune cells may cause immunosuppression resulting in the development of cancer and the inability to fight infections. Apoptosis is characterized by cell shrinkage, nuclear condensation, changes in cell membrane and mitochondria, DNA fragmentation, and protein degradation by caspases. Various methods are employed to investigate apoptosis, including direct measurement of apoptotic cells with flow cytometry and in situ labeling, as well as RNA, DNA, and protein assays that are indicative of apoptotic molecules. In addition to apoptosis, quantification of cell proliferation can provide important additional information about the effect of a toxicant upon various immune cell populations. In some cases, a toxicant may act as a mitogen pushing the immune cell into the different stages of the cell cycle. There are four stages of the active cell cycle: G1, S, G2, and M, with cell division occurring in M stage. Proliferation can be quantified by numerous methods, including staining with ki-67 or CFSE, BrdU labeling, MTT assay, and/or ATP quantification.