Corsini Emanuela, House Robert V
Methods Mol Biol. 2010;598:283-302. doi: 10.1007/978-1-60761-401-2_20.
One of the most potentially useful tools in immunotoxicology is the assessment of cytokines, the proteins/peptides that are responsible for regulating a variety of processes including immunity, inflammation, apoptosis, and hematopoiesis. Cytokine production measurements offer an outstanding promise and may eventually substitute for other more laborious procedures. The particular profile of cytokine production may provide an important information regarding the nature of many immunotoxic responses.Recent expansion in the knowledge of cytokine biology and the realization that cytokines play a role in human diseases have created a need for the precise assessment and accurate interpretation of their presence and activity in the body fluids, tissues and cells. Proper evaluation of cytokines requires attention to several technical details. Multi-cytokine analysis still needs to be standardized in terms of optimum source for analysis, protocols and quality control issues, such as the use of reference standards and the expression of results.Important practical details and considerations will be discussed in this chapter, including the source of the sample to be tested (circulating fluids, or ex vivo/in vitro isolated cells), the potential effects of collection, processing, and storage of the results of the assays, as well as potential variables associated with the source material (matrix effects, relevance, inhibitory substances), and factors influencing the choice of assay used (bioassay, immunoassay, molecular biology technique, flow cytometry).
免疫毒理学中最具潜在实用价值的工具之一是细胞因子评估,细胞因子是负责调节包括免疫、炎症、细胞凋亡和造血等多种过程的蛋白质/肽。细胞因子产生量的测量前景广阔,最终可能会替代其他更繁琐的程序。细胞因子产生的特定模式可能提供有关许多免疫毒性反应性质的重要信息。
近年来,细胞因子生物学知识不断扩展,人们也认识到细胞因子在人类疾病中发挥作用,这就需要对其在体液、组织和细胞中的存在及活性进行精确评估和准确解读。对细胞因子进行正确评估需要关注几个技术细节。在分析的最佳来源、方案以及质量控制问题(如参考标准的使用和结果的表达)方面,多细胞因子分析仍需标准化。
本章将讨论重要的实际细节和注意事项,包括待检测样品的来源(循环液或离体/体外分离细胞)、检测结果的收集、处理和储存的潜在影响,以及与来源材料相关的潜在变量(基质效应、相关性、抑制物质),还有影响所采用检测方法选择的因素(生物测定、免疫测定、分子生物学技术、流式细胞术)。