In vitro production of IL 1β, IL 1α, TNF and IL 2 in healthy subjects: distribution, effect of cyclooxygenase inhibition and evidence of independent gene regulation

Abstract

Numerous studies have reported altered in vitro cytokine production in various diseases. In the present study we used specific immunoassays to quantitate production of interleukin 1 beta (IL 1 beta), IL 1 alpha, tumor necrosis factor (TNF) and IL 2 from human peripheral blood mononuclear cells (PBMC). The distribution of cell-associated and secreted cytokines was studied in PBMC of 21 individuals; in response to lipopolysaccharide (LPS) the proportion of cell-associated IL 1 beta ranged from 13% to 56%, for IL 1 alpha 29% to 98%, and for TNF 2% to 17%. In a larger cohort of 32 subjects, the total amount of immunoreactive cytokines produced in response to LPS or phytohemagglutinin was normally distributed within the study group. Mean production of IL 1 alpha in response to LPS was 10.1 ng/ml and exceeded production of IL 1 beta (5.6 ng/ml) and TNF (2.2 ng/ml). The distribution pattern was characterized by high intersubject variability extending over two orders of magnitude and the presence of high and low "producers". Production of IL 1 alpha and IL 1 beta correlated (R = 0.69). In contrast, production of IL 1 beta did not correlate with production of TNF or IL 2. Indomethacin present during stimulation of PBMC increased the amount of IL 1 beta produced and showed a high correlation (R = 0.83) compared to cultures without indomethacin. Thus, low production of IL 1 beta in certain subjects appears not to be due to inhibitable levels of cyclooxygenase products. In a retrospective study, PBMC from 12 subjects who had taken oral cyclooxygenase inhibitors during the preceding 7 days produced 43% more IL 1 beta than subjects who did not take these drugs (p less than 0.05). These studies demonstrate that the amount of cytokine synthesized by PBMC (a) is regulated independently for IL 1, TNF and IL 2; (b) correlates for IL 1 beta and IL 1 alpha; (c) is intrinsic for low and high "producers", and (d) production of IL 1 beta increases with the use of oral cyclooxygenase inhibitors.

Identification of High and Low Responders to Lipopolysaccharide in Normal Subjects: An Unbiased Approach to Identify Modulators of Innate Immunity

“…For example, LPS-induced TNF-␣ production measured in the two samples showed a correlation (r) of 0.98 (Table I). These data are consistent with previous studies showing reproducible interindividual differences in LPS-induced IL-1␤ (16,18,19), TNF-␣ (15,16,20), and IL-10 production (15). These findings establish the temporal stability of the LPS-induced cytokine expression phenotype in whole blood.…”

Protein metabolism in rheumatoid arthritis and aging. Effects of muscle strength training and tumor necrosis factor α

“…Total body potassium was measured in a whole-body counter, as described previously (3). Cytokine production by PBMC was measured by specific, non-cross-reacting radioimmunoassay as described previously (19,20). The interassay variation of samples was <lo% and the intraassay variation was 5 5 % .…”

Risks of interleukin‐1 genetic polymorphisms and Helicobacter pylori infection in the development of gastric cancer

“…A polymorphic allele with a T instead of a C at position ) 511 of the regulatory region of the IL-1B gene is associated with increased IL-1b production. 16,22 IL-1RN encodes the IL-1 receptor antagonist (IL-1ra), which is an anti-inflammatory cytokine and competes with IL-1b by binding to IL-1 receptors. The IL-1RN gene has a penta-allelic 86-bp tandem repeat in the second intron, resulting in a short allele (IL-1RN*2, two repeats) or a long allele (IL-1RN*L, three to six repeats).…”