Kinetics of intracellular, extracellular and production of pro-inflammatory cytokines in lipopolysaccharide-stimulated human peripheral blood mononuclear cells

Kumolosasi Msi Endang, Emil Salim, Ibrahim Jantan, Waqas Ahmad

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8 Citations (Scopus)

Abstract

Purpose: To determine the detailed time course of intracellular, extracellular and production of proinflammatory 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 37°C, 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.

Original languageEnglish
Pages (from-to)536-543
Number of pages8
JournalTropical Journal of Pharmaceutical Research
Volume13
Issue number4
DOIs
Publication statusPublished - 2014

Fingerprint

Lipopolysaccharides
Blood Cells
Interleukin-1
Interleukin-6
Cytokines
Interleukin-8
Anti-Inflammatory Agents
Enzyme-Linked Immunosorbent Assay

Keywords

  • Interleukin-1α
  • Interleukin-1β
  • Interleukin-6
  • Interleukin-8
  • Pro-inflammatory cytokine
  • Tumor necrosis factor-α

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Pharmacology (medical)

Cite this

@article{b3848179a1e54ee1a7230a6121fd1f12,
title = "Kinetics of intracellular, extracellular and production of pro-inflammatory cytokines in lipopolysaccharide-stimulated human peripheral blood mononuclear cells",
abstract = "Purpose: To determine the detailed time course of intracellular, extracellular and production of proinflammatory 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 37°C, 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 = "Interleukin-1α, Interleukin-1β, Interleukin-6, Interleukin-8, Pro-inflammatory cytokine, Tumor necrosis factor-α",
author = "Endang, {Kumolosasi Msi} and Emil Salim and Ibrahim Jantan and Waqas Ahmad",
year = "2014",
doi = "10.4314/tjpr.v13i4.8",
language = "English",
volume = "13",
pages = "536--543",
journal = "Tropical Journal of Pharmaceutical Research",
issn = "1596-5996",
publisher = "Pharmacotherapy Group",
number = "4",

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TY - JOUR

T1 - Kinetics of intracellular, extracellular and production of pro-inflammatory cytokines in lipopolysaccharide-stimulated human peripheral blood mononuclear cells

AU - Endang, Kumolosasi Msi

AU - Salim, Emil

AU - Jantan, Ibrahim

AU - Ahmad, Waqas

PY - 2014

Y1 - 2014

N2 - Purpose: To determine the detailed time course of intracellular, extracellular and production of proinflammatory 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 37°C, 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.

AB - Purpose: To determine the detailed time course of intracellular, extracellular and production of proinflammatory 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 37°C, 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.

KW - Interleukin-1α

KW - Interleukin-1β

KW - Interleukin-6

KW - Interleukin-8

KW - Pro-inflammatory cytokine

KW - Tumor necrosis factor-α

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U2 - 10.4314/tjpr.v13i4.8

DO - 10.4314/tjpr.v13i4.8

M3 - Article

AN - SCOPUS:84899689361

VL - 13

SP - 536

EP - 543

JO - Tropical Journal of Pharmaceutical Research

JF - Tropical Journal of Pharmaceutical Research

SN - 1596-5996

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