CTLA4-Ig 治疗后人类巨噬细胞内 NF-kB 减少和 IKBα 增加。

Intracellular NF-kB-decrease and IKBα increase in human macrophages following CTLA4-Ig treatment.

机构信息

Research Laboratory and Academic Unit of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Genova, Italy.

出版信息

Clin Exp Rheumatol. 2013 Nov-Dec;31(6):943-6. Epub 2013 Dec 2.

PMID:24295474

Abstract

OBJECTIVES

The transcription factor NF-kB is involved in the expression of several genes linked to the immune response, including those of pro-inflammatory cytokines. We investigated cytokine production and NF-kB expression following CTLA4-Ig (abatacept) treatment of cultured human macrophages.

METHODS

Human THP1 cells, differentiated in macrophages, were treated with CTLA4-Ig (100, 500 μg/ml; 3,12,24 hours). Quantitative RT-PCR analysis (qRT-PCR) of mRNA for NF-kB, IKBα and for IL-6, TNF-α, IL-1β, was performed after 3 and 12 hours from treatment. Western blot (WB) analysis for NF-kB and IKBα was performed after 24 hours from treatment.

RESULTS

NF-kB gene expression was significantly downregulated (p<0.05), at 3 and 12 hours from CTLA4-Ig treatment, vs. untreated cells (cnt). IKBα resulted significantly increased vs. cnt (p<0.05), at 12 hours from CTLA4-Ig [500 μg/ml] treatment. After 3 hours, CTLA4-Ig [100 μg/ml] induced a significant decrease of TNF-α and IL-6 (p<0.05), vs. cnt and CTLA4-Ig [500 μg/ml] further reduced TNF-α (p<0.001), vs. cnt. After 12 hours from CTLA4-Ig treatment, a significant downregulation for IL-6 and IL1β expression (p<0.001), vs. cnt, was still evident. Results were confirmed by WB.

CONCLUSIONS

NF-kB pathway seems to be implicated in the CTLA4-Ig modulation of macrophage cytokine expression. NF-kB expression resulted downregulated while its cytoplasmatic inhibitor IKBα was increased.

摘要

目的

转录因子 NF-kB 参与了几种与免疫反应相关基因的表达，包括促炎细胞因子的基因表达。我们研究了 CTLA4-Ig（abatacept）处理培养的人巨噬细胞后细胞因子的产生和 NF-kB 的表达。

方法

人 THP1 细胞分化为巨噬细胞后，用 CTLA4-Ig（100、500μg/ml；3、12、24 小时）处理。用定量 RT-PCR 分析（qRT-PCR）检测治疗后 3 和 12 小时 NF-kB、IKBα 和 IL-6、TNF-α、IL-1β 的 mRNA。用 Western blot（WB）分析治疗后 24 小时 NF-kB 和 IKBα。

结果

与未处理细胞（cnt）相比，CTLA4-Ig 处理后 3 和 12 小时 NF-kB 基因表达显著下调（p<0.05）。IKBα 与 cnt 相比显著增加（p<0.05），在 CTLA4-Ig [500μg/ml]处理后 12 小时。3 小时时，CTLA4-Ig [100μg/ml]诱导 TNF-α和 IL-6 显著降低（p<0.05），与 cnt 和 CTLA4-Ig [500μg/ml]相比，TNF-α进一步降低（p<0.001），与 cnt 相比。CTLA4-Ig 处理后 12 小时，IL-6 和 IL1β 的表达仍显著下调（p<0.001），与 cnt 相比。结果通过 WB 得到了证实。