可透析白细胞提取物对细菌成分激活的白细胞和内皮细胞中肿瘤坏死因子α、白细胞介素-6和白细胞介素-8的产生具有不同的调节作用。

Dialyzable leukocyte extract differentially regulates the production of TNFalpha, IL-6, and IL-8 in bacterial component-activated leukocytes and endothelial cells.

作者信息

Ojeda Miriam Ojeda, van't Veer Cornelis, Fernández Ortega Celia B, Araña Rosainz Manuel de J, Buurman Wim A

机构信息

Division of Physical Chemistry, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana, 10600, Cuba.

出版信息

Inflamm Res. 2005 Feb;54(2):74-81. doi: 10.1007/s00011-004-1326-5.

DOI:10.1007/s00011-004-1326-5

PMID:15750714

Abstract

OBJECTIVE

To investigate i) whether the Dialyzable Leukocyte Extract (DLE) modulates the production of proinflammatory cytokines in leukocytes activated by the bacterial cell wall components lipopolysaccharide (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN); ii) the effect of DLE on LPS-stimulated endothelial cells; and iii) whether the regulatory effect of DLE on inflammatory mediators is related to the modulation of Toll-like receptors (TLRs), NF-kappaB and cAMP signaling pathways.

METHODS

Leukocytes were stimulated with LPS, LTA, and PGN in the presence of DLE. Endothelial cells were stimulated with LPS and treated with DLE. The levels of Tumor Necrosis Factor-alpha(TNFalpha), Interleukin-6 (IL-6), and IL-8 in culture supernatants were evaluated by ELISA. The expression of Toll-like receptor 2 (TLR2) and 4 (TLR4), NF-kappaB activity and cAMP levels were evaluated by flow cytometry, EMSA, and EIA, respectively.

RESULTS

The addition of DLE to leukocytes stimulated with cell wall constituents suppressed the production of TNFalpha. However, DLE induced IL-8 release in monocytes and enhanced IL-6 and IL-8 production by activated monocytes and endothelial cells. Also, DLE induced TLR2 and TLR4 expression, and increased cAMP levels, whereas NF-kappaB activity was inhibited.

CONCLUSIONS

The present data indicate the differential regulation by DLE of the production of TNFalpha, IL-6, and IL-8 cytokines, associated with effects on TLR2 and TLR4 expression and NF-kappaB and cAMP activities. We suggest a putative mechanism for the biological effects of DLE in activated leukocytes and endothelial cells.

摘要

目的

研究 i）可透析白细胞提取物（DLE）是否能调节由细菌细胞壁成分脂多糖（LPS）、脂磷壁酸（LTA）和肽聚糖（PGN）激活的白细胞中促炎细胞因子的产生；ii）DLE 对 LPS 刺激的内皮细胞的影响；iii）DLE 对炎症介质的调节作用是否与 Toll 样受体（TLR）、核因子κB（NF-κB）和环磷酸腺苷（cAMP）信号通路的调节有关。

方法

在 DLE 存在的情况下，用 LPS、LTA 和 PGN 刺激白细胞。用 LPS 刺激内皮细胞并用 DLE 处理。通过酶联免疫吸附测定（ELISA）评估培养上清液中肿瘤坏死因子-α（TNFα）、白细胞介素-6（IL-6）和 IL-8 的水平。分别通过流式细胞术、电泳迁移率变动分析（EMSA）和酶免疫分析（EIA）评估 Toll 样受体 2（TLR2）和 4（TLR4）的表达、NF-κB 活性和 cAMP 水平。