Damsgaard Camilla T, Lauritzen Lotte, Calder Philip C, Kjaer Tanja M R, Frøkiaer Hanne
Department of Human Nutrition, Faculty of Life Sciences, University of Copenhagen, 1958 Frederiksberg C, Denmark.
J Immunol Methods. 2009 Jan 30;340(2):95-101. doi: 10.1016/j.jim.2008.10.005. Epub 2008 Nov 8.
Whole-blood and peripheral blood mononuclear cell (PBMC) cultures are used as non-validated surrogate measures of monocytic cytokine production. The aim of this investigation was to compare ex vivo cytokine production from human whole-blood and PBMC with that from isolated monocytes. We also assessed the intra- and inter-individual variation in cytokine production. In 64 healthy men (age 19-40 years) IL-6, TNF and IL-10 were measured by enzyme-linked immunosorbent assay in supernatants from whole-blood, PBMC and monocytes cultured 24 h with lipopolysaccharide (LPS) or UV-killed L. acidophilus. Cytokines produced from whole-blood was found to be more strongly correlated with monocytic cytokines than cytokines from PBMC, particularly after LPS-stimulation: r=0.57, P<0.001 versus r=0.33, P=0.01 for IL-6 and r=0.43, P<0.001 versus r=0.30, P=0.02 for TNF-alpha. Adjustment for a preceding 8-week dietary fatty acid-intervention did not change any of the associations. Based on measurements at three time-points 8 weeks apart the intra-individual variation was > or = 50% smaller than the inter-individual variation (P<0.05) in most whole-blood cytokine responses and LPS-stimulated IL-6 from PBMC. We conclude that whole-blood cultures are well-suited low-cost proxy-measures of monocytic cytokine production. Moreover, large inter-individual variation in cytokine production was demonstrated whereas the individual responses in whole-blood were reproducible even over long time-periods.
全血和外周血单核细胞(PBMC)培养被用作单核细胞细胞因子产生的未经验证的替代指标。本研究的目的是比较人全血和PBMC与分离的单核细胞的体外细胞因子产生情况。我们还评估了细胞因子产生的个体内和个体间差异。在64名健康男性(年龄19 - 40岁)中,通过酶联免疫吸附测定法测量了全血、PBMC和单核细胞与脂多糖(LPS)或紫外线灭活的嗜酸乳杆菌培养24小时后的上清液中的IL - 6、TNF和IL - 10。发现全血产生的细胞因子与单核细胞细胞因子的相关性比PBMC产生的细胞因子更强,特别是在LPS刺激后:IL - 6的r = 0.57,P < 0.001,而PBMC的r = 0.33,P = 0.01;TNF - α的r = 0.43,P < 0.001,而PBMC的r = 0.30,P = 0.02。对之前8周饮食脂肪酸干预进行调整后,任何关联均未改变。基于相隔8周的三个时间点的测量,在大多数全血细胞因子反应和PBMC中LPS刺激的IL - 6中,个体内差异比个体间差异小≥50%(P < 0.05)。我们得出结论,全血培养是适合单核细胞细胞因子产生的低成本替代指标。此外,证明了细胞因子产生存在较大的个体间差异,而全血中的个体反应即使在很长时间内也是可重复的。