Effects of mimicked acetylated HMGB1 on macrophages and dendritic cells

Chen,Xiaohong; Xu,Yong; Xiong,Ping; Tan,Zheng; Gong,Feili; Hou,Xiaohua; Zheng,Fang

doi:10.3892/mmr.2018.9584

Effects of mimicked acetylated HMGB1 on macrophages and dendritic cells

Authors:
- Xiaohong Chen
- Yong Xu
- Ping Xiong
- Zheng Tan
- Feili Gong
- Xiaohua Hou
- Fang Zheng
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Affiliations: Division of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China, Department of Immunology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China
Published online on: October 24, 2018 https://doi.org/10.3892/mmr.2018.9584
Pages: 5527-5535
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Extracellular high mobility group box 1 (HMGB1) serves a critical role in inflammatory diseases. HMGB1 is released into the extracellular environment mainly by passive release from necrotic cells or active secretion from monocytes/macrophages following stimulation. However, the translocation of actively secreted HMGB1 from the nucleus to the cytoplasm with post‑translational modifications such as acetylation is required; HMGB1 is then released into the extracellular space. Whether acetylation influences the extracellular function of HMGB1 remains unknown. In the present study, an optimized method of gene mutation by using well‑designed primers in particular, which were employed to identify the mutant gene. The substitution of six lysine residues for glutamines was conducted to mimic acetylated HMGB1 (HMGB1‑M) and observe the effects of HMGB1‑M on macrophages and dendritic cells (DCs). Tumor necrosis factor (TNF)‑α production in RAW 264.7 cells was assessed by ELISA. The phagocytic potential of RAW 264.7 cells, DC maturation and CXCR4 expression were analyzed by flow cytometry. The results of the present study revealed that HMGB1‑M increased cytoplasmic translocation. Compared with HMGB1, HMGB1‑M increased TNF‑α production within RAW 264.7 cells and decreased the mean fluorescence intensity (MFI) of integrin α X, and the percentage and MFI of major histocompatibility complex‑II on DCs. HMGB1‑M exhibited no significant effects on phagocytosis of macrophages and expression frequency of cluster of differentiation 80 and chemokine receptor type 4 on DCs. These results suggested that HMGB1‑M may partly promote inflammation and decrease DC maturation. Thus, the findings of the present study may provide insight into the complex role of HMGB1 in inflammatory diseases.

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