Purpose: To determine the detailed
time course of intracellular, extracellular and
production of pro-inflammatory cytokines (TNF-α, IL-1β,
IL-1α, IL-6 and IL-8) in normal human peripheral blood
mononuclear cells (PBMC).
Methods: PBMC were isolated from
human whole blood, stimulated by 0.1 μg/ml
lipopolysaccharide (LPS) and incubated at 37șC, 5% CO2.
Samples were harvested at different time intervals (4,
8, 12, 16, 20 and 24 h) after stimulation. ELISA was
employed for the measurement of the extracellular and
intracellular cytokine levels of the samples.
Results: The release of TNF-α, IL-6
and IL-8 on LPS-stimulated PBMC were significantly
higher with concentrations in the range of 3161 ± 162.5
to 4027 ± 361.5 pg/ml (p < 0.001), 3921.5 ± 879.3 to
11628.3 ± 2647.3 pg/ml (p ≤ 0.030), and 4122.0 ± 382.9
to 5898.6±115.8 pg/ml (p ≤ 0.049), respectively compared
to intracellular levels that were very low (TNF-α, 23.5
± 5.0 to 69.5 ± 13.8 pg/ml; IL-6, 22.5 ± 16.5 to 96.5 ±
9.6 pg/ml; and IL-8, 501.1 ± 221.0 to 1452.5 ± 415.7
pg/ml) and remained unchanged during 24 h. In contrast,
both IL-1α and IL-1β were secreted gradually. Secretion
and production of IL-6 was significantly higher at 8 h
(9394.4 ± 846.3 pg/ml; p = 0.002) and at 20 h (11628.3 ±
2647.3 pg/ml; p = 0.014) than other cytokines.
Conclusion: The differences in the
characteristic kinetics of cytokines may be caused by
different mechanisms of secretion and function. IL-1,
TNF-α and IL-8 play a role as proinflammatory cytokines,
whereas IL-6 consecutively plays a dual role as
pro-inflammatory cytokine and anti-inflammatory.
Keywords: Pro-inflammatory cytokine,
Tumor necrosis factor-α, Interleukin-1α, Interleukin-1β,
Interleukin-6, Interleukin-8.