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Quantitative intracellular cytokine measurement: age-related changes in proinflammatory cytokine production.

Abstract：

:The proinflammatory cytokines play a central role in mediating cellular and physiological responses, and levels may reflect immune system effectiveness. In this study, the effect of ageing on the inflammatory response was examined using a novel method to detect production of the proinflammatory cytokines, i.e. tumour necrosis factor-alpha (TNF-alpha), IL-6 and IL-1beta. Peripheral blood mononuclear cells (PBMC) obtained from healthy donors of different ages were incubated for 0, 24, 48 and 72 h with or without phorbol 12-myristate 13-acetate (PMA) stimulation. At each time point these cells were permeabilized and incubated with secondary conjugated FITC MoAbs specific for each cytokine. A flow cytometric system was developed to quantify specific intracellular fluorescence in T cells (CD3+) and monocytes (CD14+). TNF-alpha, IL-6 and IL-1beta production in cell culture supernatants was also measured using ELISAs. In older subjects, flow cytometry detected significant increases in intracellular T cell TNF-alpha and IL-6 (P < 0.05). IL-1beta was not detected in any of the T cell samples. Likewise, the monocytes of older subjects demonstrated increased intracellular levels of all three cytokines, but these increases were not significant (P > 0.05). These changes in intracellular proinflammatory cytokine levels may explain some of the exaggerated inflammatory responses seen in elderly patients.

journal_name

Clin Exp Immunol

journal_title

Clinical and experimental immunology

authors

O'Mahony L,Holland J,Jackson J,Feighery C,Hennessy TP,Mealy K

doi

10.1046/j.1365-2249.1998.00641.x

subject

Has Abstract

pub_date

1998-08-01 00:00:00

pages

213-9

issue

2

eissn

0009-9104

issn

1365-2249

journal_volume

113

pub_type

杂志文章

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