Academic MS Center Limburg, Zuyderland Medical Center, Sittard, the Netherlands; School for Mental Health and Neuroscience, Maastricht University Medical Center, Maastricht, the Netherlands.
Academic MS Center Limburg, Zuyderland Medical Center, Sittard, the Netherlands; Department of Neurology, Canisius Wilhelmina Hospital, Nijmegen, the Netherlands.
Mol Immunol. 2019 Jan;105:198-204. doi: 10.1016/j.molimm.2018.12.001. Epub 2018 Dec 11.
Different laboratory approaches have been exploited to analyze an effect of vitamin D supplements on T cell cytokine profiles in multiple sclerosis, with poorly reproducible results. We assessed the correlation between intra-cellular flowcytometry analysis of CD4 T cell-enriched CD3CD8 lymphocytes after PMA/ionomycin stimulation directly ex-vivo or after 72 h pre-stimulation with anti-CD3, and cytokine levels excreted in culture supernatants. Pre-stimulation with anti-CD3 resulted in higher proportions of cells positive for IFN-γ, IL-17 A, IL-4, IL-10 and GM-CSF (all P < 0.001), but not TNF-α. Positive correlation between approaches was highly variable, but most eminent for IFN- γ and IL-4 (R = 0.608-0.612 and R = 0.677-0.777, resp., all P < 0.001). No effect of 16-weeks vitamin D supplements on any outcome was found except for a decreased TNF-α concentration in culture supernatants. Choice of immune-assay is, apparently, a relevant confounder for the reproducibility of individual studies.
不同的实验室方法被用于分析维生素 D 补充剂对多发性硬化症 T 细胞细胞因子谱的影响，但结果重复性较差。我们评估了 PMA/离子霉素刺激后直接离体或用抗 CD3 预刺激 72 小时后 CD4 T 细胞富集的 CD3CD8 淋巴细胞内流式细胞术分析与培养上清液中细胞因子水平之间的相关性。用抗 CD3 预刺激导致 IFN-γ、IL-17A、IL-4、IL-10 和 GM-CSF 的阳性细胞比例更高（均 P<0.001），但 TNF-α 则不然。方法之间的正相关性差异很大，但 IFN-γ 和 IL-4 最为明显（R=0.608-0.612 和 R=0.677-0.777，均 P<0.001）。除了培养上清液中 TNF-α 浓度降低外，16 周的维生素 D 补充剂对任何结果都没有影响。显然，免疫测定的选择是个体研究重复性的一个重要混杂因素。
