Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Kinetics of Intracellular, Extracellular and Production of Pro-Inflammatory Cytokines in Lipopolysaccharide-Stimulated Human Peripheral Blood Mononuclear Cells

Endang Kumolosasi , Emil Salim, Ibrahim Jantan, Waqas Ahmad

Drug and Herbal Research Center, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia;

For correspondence:- Endang Kumolosasi Email: ekumolosasi@yahoo.co.id Tel:+60149208598

Received: 17 April 2013 Accepted: 19 February 2014 Published: 23 April 2014

Citation: Kumolosasi E, Salim E, Jantan I, Ahmad W. Kinetics of Intracellular, Extracellular and Production of Pro-Inflammatory Cytokines in Lipopolysaccharide-Stimulated Human Peripheral Blood Mononuclear Cells. Trop J Pharm Res 2014; 13(4):536-543 doi: 10.4314/tjpr.v13i4.8

© 2014 The authors.
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Abstract

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 37ºC, 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-^5;, Interleukin-1^5;, Interleukin-1^6;, Interleukin-6, Interleukin-8