title: Whole-blood culture is a valid low-cost method to measure monocytic cytokines - a comparison of cytokine production in cultures of human whole-blood, mononuclear cells and monocytes.
abstract: 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.
authors.affiliation: Department of Human Nutrition, Faculty of Life Sciences, University of Copenhagen, 1958 Frederiksberg C, Denmark. ctd@life.ku.dk
authors.forename: Camilla T
authors.initials: CT
authors.lastname: Damsgaard
authors.forename: Lotte
authors.initials: L
authors.lastname: Lauritzen
authors.forename: Philip C
authors.initials: PC
authors.lastname: Calder
authors.forename: Tanja M R
authors.initials: TM
authors.lastname: Kjaer
authors.forename: Hanne
authors.initials: H
authors.lastname: Frøkiaer
mesh_terms: D000328:Adult; D001773:Blood Cells / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000191:economics* / Q000379:methods*; D016207:Cytokines / Q000032:analysis* / Q000096:biosynthesis*; D006801:Humans; D016753:Interleukin-10 / Q000032:analysis / Q000096:biosynthesis; D015850:Interleukin-6 / Q000032:analysis / Q000096:biosynthesis; D008297:Male; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000096:biosynthesis
chemical_list: D016207:Cytokines; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10