调节 THP-1 单核细胞和巨噬细胞中 CXCL12 及其受体 CXCR4、7 的转录和翻译解偶联。

Transcriptional and translational-uncoupling in regulation of the CXCL12 and its receptors CXCR4, 7 in THP-1 monocytes and macrophages.

机构信息

Department of Nutrition and Food Science, University of Maryland, College Park, Maryland, 20742, USA.

USDA, Diet, Genomics and Immunology Laboratory, Beltsville Human Nutrition Research Center, ARS, Beltsville, Maryland, 20705, USA.

出版信息

Immun Inflamm Dis. 2018 Mar;6(1):106-116. doi: 10.1002/iid3.199. Epub 2017 Nov 3.

DOI:10.1002/iid3.199

PMID:29105376

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5818454/

Abstract

INTRODUCTION

The chemokine CXCL12 and its receptors CXCR4 and 7 play crucial roles in the immune system. In the present study, regulation of this pathway was further examined using the in-vitro model of undifferentiated human THP-1 monocytes (u-THP-1) and phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophages (d-THP-1), to assess the effects of differentiation and the TLR4 ligand lipopolysaccharide (LPS) on the pathway.

METHODS/RESULTS: Differentiation did not affect the CXCR4, 7 mRNA levels. Interestingly, the CXCL12 and CXCR7 proteins but not CXCR4 were found to be up-regulated during differentiation. LPS, through CD14-dependent pathway, induced CXCL12 and CXCR4, 7 mRNA levels to a greater magnitude in d- than u-THP-1. The induction effect on CXCL12 stimulated by LPS was confirmed using ELISA. Increased migration of u-THP-1 was observed using conditioned medium from LPS-treated d-THP-1. Additionally, d-THP-1, although expressed higher CXCR7 protein levels, failed to migrate toward CXCL12. In contrast, LPS did not affect CXCR4, 7 protein levels.

CONCLUSION

Hence, this study indicated that CXCL12, CXCR4, and CXCR7 were differentially expressed and regulated in u-THP-1 and d-THP-1 cells in response to external stimuli. Importantly, we reported here a novel observation that uncoupling exists between transcriptional and translational regulation of CXCR4, 7 expressions by differentiation and TLR stimuli.

摘要

简介

趋化因子 CXCL12 及其受体 CXCR4 和 7 在免疫系统中发挥着至关重要的作用。在本研究中，我们使用未分化的人 THP-1 单核细胞（u-THP-1）和佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）分化的 THP-1 巨噬细胞（d-THP-1）的体外模型进一步研究了该途径的调节，以评估分化和 TLR4 配体脂多糖（LPS）对该途径的影响。

方法/结果：分化不会影响 CXCR4、7 mRNA 水平。有趣的是，在分化过程中发现 CXCL12 和 CXCR7 蛋白而非 CXCR4 上调。LPS 通过 CD14 依赖性途径，在 d-THP-1 中诱导 CXCL12 和 CXCR4、7 mRNA 水平的增加幅度大于 u-THP-1。通过 ELISA 证实了 LPS 对 CXCL12 刺激的诱导作用。用 LPS 处理的 d-THP-1 的条件培养基观察到 u-THP-1 的迁移增加。此外，尽管 d-THP-1 表达更高水平的 CXCR7 蛋白，但未能向 CXCL12 迁移。相反，LPS 不影响 CXCR4、7 蛋白水平。