Selection and Characterization of a DNA Aptamer Recognizing High Mobility Group Box 1 Protein (HMGB1) and Enhancing Its Pro-inflammatory Activity
| Author | Hanchao Li | en |
| Author | Wengang Sun | en |
| Author | Yanhua Huang | en |
| Author | Qian Li | en |
| Author | Hong Tian | en |
| Author | Zhiming Hao | en |
| Author | Yongwei Huo | en |
| Orcid | Yongwei Huo [0000-0003-2408-4264] | en |
| Issued Date | 2024-12-31 | en |
| Abstract | Background: High mobility group box 1 (HMGB1) plays an essential role in various pathological conditions, including inflammation, fibrosis, autoimmune diseases, and carcinogenesis. The quantification of HMGB1 in body fluids holds promise for clinical applications. Objectives: This study aimed to isolate high-affinity single-stranded DNA (ssDNA) aptamers that target HMGB1. Methods: In this study, ssDNA aptamers were selected using Systematic Evolution of Ligands by Exponential Enrichment (SELEX). The affinity and specificity of the aptamers were evaluated through South-Western blot analysis, enzyme-linked aptamer sorbent assay (ELASA), and aptamer-based histochemistry staining. The impact of the aptamers on the biological activity of HMGB1 was tested in the human acute monocytic leukemia cell line, THP-1. Results: An aptamer (H-ap25, dissociation constant = 8.20 ± 0.53 nmol/L) with high affinity for the HMGB1 B box was generated. Further experiments verified that H-ap25 can be used to detect HMGB1 in South-Western blot analysis, ELASA, and aptamer-based histochemistry staining. Moreover, H-ap25 significantly augmented HMGB1-induced expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, Toll-like receptor 9 (TLR9), and activation of NF-κB in THP-1 cells. Conclusions: Our results demonstrated that H-ap25 can be used both as an enhancer of HMGB1 and as a probe in research. | en |
| DOI | https://doi.org/10.5812/ijpr-147246 | en |
| Keyword | Toll-Like Receptor 9 (TLR9) | en |
| Keyword | High Mobility Group Box 1 (HMGB1) | en |
| Keyword | Aptamer | en |
| Keyword | Systematic Evolution of Ligands by Exponential Enrichment (SELEX) | en |
| Keyword | Inflammation | en |
| Publisher | Brieflands | en |
| Title | Selection and Characterization of a DNA Aptamer Recognizing High Mobility Group Box 1 Protein (HMGB1) and Enhancing Its Pro-inflammatory Activity | en |
| Type | Research Article | en |