PolyI:C Upregulated CCR5 and Promoted THP-1-Derived Macrophage Chemotaxis via TLR3/JMJD1A Signalling

(Pages: 325-333)
Xiaoxiao Yu, Ph.D, 1Huayang Wang, M.D, 2Hongjia Shao, M.M, 1Cuijuan Zhang, Ph.D, 3Xiuli Ju, Ph.D, 1Jie Yang, Ph.D., 1,*
Department of Paediatrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, Shandong, China
Department of Pathology, Qilu Hospital of Shandong University, Jinan, Shandong, China
Department of Paediatrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, Shandong, China
Department of Pathology, Qilu Hospital of Shandong University, Jinan, Shandong, China
*Corresponding Address: Department of Paediatrics Qilu Hospital of Shandong University Jinan Shandong China Email:yangj6466@163.com
The Cell Journal (Yakhteh) is an open access journal which means the articles are freely available online for any individual author to download and use the providing address. The journal is licensed under a Creative Commons Attribution-Non Commercial 3.0 Unported License which allows the author(s) to hold the copyright without restrictions that is permitting unrestricted use, distribution, and reproduction in any medium provided the original work is properly cited. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Yu X, Wang H, Shao H, Zhang C, Ju X, Yang J. PolyI:C upregulated CCR5 and promoted THP-1-Derived macrophage chemotaxis via TLR3/ JMJD1A signalling. Cell J. 2020; 22(3): 325-333. doi: 10.22074/cellj.2020.6713.

Abstract

Objective

This study aimed to evaluate the specific roles of polyinosinic:polycytidylic acid (polyI:C) in macrophage chemotaxis and reveal the potential regulatory mechanisms related to chemokine receptor 5 (CCR5).

Materials and Methods

In this experimental study, THP-1-derived macrophages (THP1-Mφs) induced from THP- 1 monocytes were treated with 25 μg/mL polyI:C. Toll-like receptor 3 (TLR3), Jumonji domain-containing protein (JMJD)1A, and JMJD1C small interfering RNA (siRNAs) were transfected into THP1-Mφs. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was used to detect the expression levels of TLR3, CCR5, 23 Jumonji C domain-containing histone demethylase family members, JMJD1A, and JMJD1C in THP1-Mφs with different siRNAs transfections. Western blot was performed to detect JMJD1A, JMJD1C, H3K9me2, and H3K9me3 expressions. A transwell migration assay was conducted to detect THP1-Mφ chemotaxis toward chemokine ligand 3 (CCL3). A chromatin immunoprecipitation (ChIP) assay was performed to detect H3K9me2-CCR5 complexes in THP1- Mφs.

Results

PolyI:C significantly upregulated CCR5 in THP1-Mφs and promoted chemotaxis toward CCL3 (P<0.05); these effects were significantly inhibited by TLR3 siRNA (P<0.01). JMJD1A and JMJD1C expression was significantly upregulated in polyI:C-stimulated THP1-Mφs, while only JMJD1A siRNA decreased CCR5 expression (P<0.05). JMJD1A siRNA significantly increased H3K9me2 expression in THP1-Mφs but not in polyI:C-stimulated THP1-Mφs. The ChIP result revealed that polyI:C significantly downregulated H3K9me2 in the promoter region of CCR5 in THP1- Mφs.

Conclusion

PolyI:C can enhance THP1-Mφ chemotaxis toward CCL3 regulated by TLR3/JMJD1A signalling and activate CCR5 expression by reducing H3K9me2 in the promoter region of CCR5.