selection and characterization of a dna aptamer recognizing high mobility group box 1 protein (hmgb1) and enhancing its pro-inflammatory activity

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.