Abstract:
BACKGROUND:Two assays--isolated peripheral blood mononuclear cells (PBMC) and a whole blood assay (WBA)--are commonly used to study TNF-alpha production by an individual in order to distinguish between high and low cytokine producers. We assessed the reliability and reproducibility of these assays. METHODS:The PBMC assays (n=5) were performed weekly over a period of 6 weeks and the WBAs (n=4) weekly over a 4-week period. Polymethylmethacrylate particles (approx. 6 x 10(2) particles/cell) and optimal concentrations of endotoxin (6.25 and 12.5 ng/ml) were used as the stimulatory agents in PBMC and WBAs, respectively. TNF-alpha production was measured by ELISA. RESULTS:There was a high degree of both intra- and inter-individual variability of TNF-alpha secretion, with unpredictable changes in the amount of the cytokine produced by cells from the same donor. This variability could not be eliminated by correcting for cell numbers. CONCLUSION:The PBMC and WBA models of TNF-alpha production by human peripheral blood cells cannot be used for the evaluation of inter-individual variability in cytokine secretion due to the high intra-individual variability observed. In the case of PBMC this is partly due to differences in the confluency of the cells between individuals.
journal_name
Cytokinejournal_title
Cytokineauthors
Bakiyeva LT,Brooks RA,Rushton Ndoi
10.1016/j.cyto.2004.11.011keywords:
subject
Has Abstractpub_date
2005-04-07 00:00:00pages
35-40issue
1eissn
1043-4666issn
1096-0023pii
S1043-4666(05)00019-0journal_volume
30pub_type
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