Institute for Clinical Pharmacology, Medical Faculty, University of Rostock, Schillingallee 70, 18057, Rostock, Germany.
BMC Med Res Methodol. 2012 Aug 1;12:112. doi: 10.1186/1471-2288-12-112.
The variation of immune cell activities over time is an immanent property of the human immune system, as can be measured by the stimulated secretion of cytokines in cell cultures. However, inter-individual variability is considerably higher. Especially the latter is the major reason why it has not been possible to establish international standard values for cytokines as was possible for other parameters, such as leukocyte sub-population numbers. In this trial, a highly standardized whole-blood culture model (TrueCulture®), developed to characterise drug effects on cells of the human immune system in clinical trials, was used to analyse cytokine patterns in the blood samples of 12 healthy subjects over a period of one month.
After an overnight fast, 12 healthy subjects donated blood three times a week on three consecutive days over a period of 4 weeks. TruCulture® blood collection and whole-blood culture systems were used to measure whole-blood leukocyte stimulation. The levels of IL-2, IL-5, IL-13, IL-6, IL-8, IL-10, IFNγ, and MCP-1 in the culture supernatants were quantified by sandwich ELISA.
The pattern of cytokine concentrations in the supernatants of the stimulated whole-blood cultures was highly individual, but considerably stable over the whole observation period of 4 weeks.
By using TruCulture® it seems feasible to determine subject-specific cytokine reference patterns, for example under healthy conditions, or before starting an experimental treatment, e.g. during a clinical trial, against which changes in the behaviour of the immune system can be detected more accurately in future.
免疫细胞活性随时间的变化是人体免疫系统的固有特性,可以通过细胞培养中细胞因子的刺激分泌来测量。然而,个体间的可变性要高得多。特别是后者是迄今为止无法像其他参数(如白细胞亚群数量)那样为细胞因子建立国际标准值的主要原因。在这项试验中,使用了一种高度标准化的全血培养模型(TrueCulture®),该模型旨在研究药物对临床试验中人类免疫系统细胞的作用,用于分析 12 名健康受试者在一个月内的血液样本中的细胞因子模式。
12 名健康受试者在一夜禁食后,每周三次在连续三天内捐献血液,持续 4 周。使用 TruCulture®血液采集和全血培养系统测量全血白细胞刺激。通过夹心 ELISA 定量测定培养上清液中 IL-2、IL-5、IL-13、IL-6、IL-8、IL-10、IFNγ 和 MCP-1 的水平。
刺激全血培养物上清液中细胞因子浓度的模式具有高度个体性,但在整个 4 周的观察期内相当稳定。
使用 TruCulture®似乎可以确定特定于个体的细胞因子参考模式,例如在健康条件下,或在开始实验性治疗之前,例如在临床试验期间,可以更准确地检测未来免疫系统行为的变化。