High Mobility Group Box 1 Levels Research Articles

Identification of Neutralizing Anti-HMGB1 IgM Autoantibody and Its B Cell Producers

Abstract Extracellular High Mobility Group Box 1 (HMGB1) is a prototypic damage-associated molecular pattern (DAMP). While a homeostatic level of extracellular HMGB1 may be beneficial for immune defense, tissue repair and tissue regeneration, excessive HMGB1 is linked to chronic inflammation and inflammatory diseases. This prompts an intriguing question: How does a healthy body control the level of extracellular HMGB1? Here we have identified, in the plasma of both healthy humans and healthy mice, an anti-HMGB1 IgM autoantibody that neutralizes extracellular HMGB1 via binding specifically to a 100% conserved epitope, namely HMW4 (HMGB1 98–112). In mice, this anti-HMGB1 IgM (i.e., anti-HMW4 IgM) is produced by peritoneal B-1 cells, and concomitant triggering of their B cell receptor and toll-like receptor 4 by extracellular HMGB1 stimulates the production of anti-HMW4 IgM. The ability of extracellular HMGB1 to induce its own neutralizing antibody suggests a feedback loop limiting the level of this DAMP in a healthy body. Currently, our studies are ongoing to identify and characterize the human counterpart B cells that produce the anti-HMGB1 IgM, by first amplifying HMW4-reactive B cells from human peripheral blood mononuclear cells (PBMCs) in NSG recipient mice, in the presence of B cell-stimulating adjuvants (such as B cell mitogens). These studies serve as the prelude to exploring human anti-HMGB1 IgM down the road for treating patients with inflammatory diseases and provide insight pertaining to the role and regulation of other DAMP molecules.

Curcumin inhibits acute pulmonary embolism and inflammation by inhibiting high mobility group box 1 (HMGB1)

This study assessed the role of Curcumin in acute pulmonary embolism (APE) and inflammation. Male rats were assigned into sham operation group, APE group, low-dose and high-dose of Curcumin group for this study. Levels of IL-1β, TNF-α, and Interleukin-6 (IL-6) inflammatory indicators were analyzed, including also, pathological changes of lung tissue. We also assessed High mobility group box 1 (HMGB1), receptor for advanced glycation end product (RAGE) and Nuclear factor kappa B (NF-κB) protein expressions. Results showed that, inflammation indicators were reduced after treatment at 6 h, 24 h, and 72 h, when compared with APE group, but were still higher than sham operation group (P <0.05). Serum inflammation index was higher at 2 h after modeling (P <0.05). Moreover, treatment groups showed continuously reduced HMGB1 protein expression, with lower level of HMGB1 in high dose group (P <0.05). RAGE expression continued to increase in APE and treatment groups (P <0.05). However, its level in treatment groups was lower than APE group (P <0 05). The NF-KB expression continued to increase in APE and treatment groups (P <0.05) with a lower level in treatment group (P <0.05). Curcumin effectively suppressed inflammatory response in acute pulmonary embolism, by reducing RAGE/NF-KB signal activity and inhibiting inflammatory response by inhibiting HMGB1 expression.

High mobility group box 1 levels as potential predictors of asthma severity.

High mobility group box 1 levels as potential predictors of asthma severity.

Inhibition of macrophages inflammasome activation via autophagic degradation of HMGB1 by EGCG ameliorates HBV-induced liver injury and fibrosis.

Liver fibrosis is a reversible wound-healing response that can lead to end-stage liver diseases without effective treatment, in which HBV infection is a major cause. However, the underlying mechanisms for the development of HBV-induced fibrosis remains elusive, and efficacious therapies for this disease are still lacking. In present investigation, we investigated the effect and mechanism of green tea polyphenol epigallocatechin-3-gallate (EGCG) on HBV-induced liver injury and fibrosis. The effect of EGCG on liver fibrosis was examined in a recombinant cccDNA (rcccDNA) chronic HBV mouse model by immunohistochemical staining, Sirius red and Masson's trichrome staining. The functional relevance between high mobility group box 1 (HMGB1) and inflammasome activation and the role of EGCG in it were analyzed by Western blotting. The effect of EGCG on autophagic flux was determined by Western blotting and flow cytometric analysis. EGCG treatment efficiently was found to alleviate HBV-induced liver injury and fibrosis in a recombinant cccDNA (rcccDNA) chronic HBV mouse model, a proven suitable research platform for HBV-induced fibrosis. Mechanistically, EGCG was revealed to repress the activation of macrophage NLRP3 inflammasome, a critical trigger of HBV-induced liver fibrosis. Further study revealed that EGCG suppressed macrophage inflammasome through downregulating the level of extracellular HMGB1. Furthermore, our data demonstrated that EGCG treatment downregulated the levels of extracellular HMGB1 through activating autophagic degradation of cytoplasmic HMGB1 in hepatocytes. Accordingly, autophagy blockade was revealed to significantly reverse EGCG-mediated inhibition on extracellular HMGB1-activated macrophage inflammasome and thus suppress the therapeutic effect of EGCG on HBV-induced liver injury and fibrosis. EGCG ameliorates HBV-induced liver injury and fibrosis via autophagic degradation of cytoplasmic HMGB1 and the subsequent suppression of macrophage inflammasome activation. These data provided a new pathogenic mechanism for HBV-induced liver fibrosis involving the extracellular HMGB1-mediated macrophage inflammasome activation, and also suggested EGCG administration as a promising therapeutic strategy for this disease.

Canagliflozin attenuates thioacetamide-induced liver injury through modulation of HMGB1/RAGE/TLR4 signaling pathways

Thioacetamide (TAA), a classic liver toxic compound, is used to establish experimental models of liver injury via induction of inflammation and oxidative stress. The current study was employed to explore the effects of canagliflozin (CANA), a sodium glucose cotransporter 2 (SGLT-2) inhibitor and antidiabetic agent, on TAA-induced acute liver injury. MethodsA rat model of acute hepatic injury was established using single intraperitoneal injection of TAA (500 mg/kg) and rats received CANA (10 and 30 mg/kg, orally) once daily for 10 days prior to TAA challenge. Liver function, oxidative stress, and inflammatory parameters were measured in serum and hepatic tissues of rats. ResultsElevated levels of liver enzymes, hepatic malondialdehyde (MDA), and serum lactate dehydrogenase (LDH) were significantly attenuated by CANA. CANA also increased hepatic superoxide dismutase (SOD) and glutathione (GSH). Hepatic levels of high-mobility group box 1 (HMGB1), toll like receptor4 (TLR4), receptor for advanced glycation end products (RAGE), and pro-inflammatory cytokines (IL-6, and IL-1β) were normalized with CANA. Additionally, Hepatic expression of p-JNK/p-p38 MAPK was significantly attenuated by CANA compared to TAA-treated rats. CANA also decreased hepatic immunoexpression of NF-κB and TNF-α and attenuated hepatic histopathological alterations via reduction of inflammation and necrosis scores and collagen deposition. Moreover, mRNA expression levels of TNF-α and IL-6 were reduced upon CANA treatment. ConclusionCANA attenuates TAA-prompted acute liver damage, via suppressing HMGB1/RAGE/TLR4 signaling, regulation of oxidative stress and inflammation pathways.

Abstract 2633: Zanidatamab zovodotin (ZW49) induces hallmarks of immunogenic cell death and is active in patient-derived xenograft models of gastric cancer

Abstract Zanidatamab zovodotin (ZW49) is an antibody drug conjugate (ADC) comprised of a bispecific anti-HER2 IgG1 antibody (ZW25, zanidatamab) conjugated to a microtubule inhibitor auristatin payload (ZD02044) via a protease cleavable linker. Clinical studies of ZW49 in HER2-expressing advanced solid tumors are currently underway (NCT03821233). We have previously presented data illustrating the rapid internalization of ZW49 and the anti-tumor activity of ZW49 in multiple patient-derived xenograft models originating from breast carcinomas. Here we present ZW49’s ability to induce in vitro hallmarks of immunogenic cell death (ICD) including increased calreticulin exposure, increased high mobility group box-1 (HMGB1) exposure, and increased extracellular ATP secretion in a HER2 dependent manner. Additionally, we present the results from a study evaluating the anti-tumor activity of ZW49 in a panel of patient-derived xenografts of gastric cancer. HER2-positive SK-BR-3 and HER2-negative MDA-MB-468 tumor cell lines were treated with ZW49 and cell surface calreticulin was assessed by flow cytometry. ZW49 treatment of SKBR-3 cells induced a significantly higher percentage of cells staining positive for cell surface calreticulin compared to untreated cells. In contrast, ZW49 treatment of MDA-MB-468 cells resulted in no significant difference in the percentage of cells staining positive for calreticulin compared to untreated cells. ZW49 treated HER2-positive cancer cell lines SK-BR-3 and SK-OV-3 induced higher levels of extracellular ATP compared to untreated cells. In the HER2-negative cell line MDA-MB-468, treatment with ZW49 did not alter levels of extracellular ATP compared to untreated cells. ZW49 induced higher levels of HMGB1 in the HER2-positive cancer cell lines SKBR3 and N87 compared to untreated cells, whereas in the HER2-negative cell line MDA-MB-468, ZW49 induced similar levels of HMGB1 to untreated cells. Anti-tumor activity was observed in 5/7 (71%) PDX models of gastric cancer after a single i.v. dose of 6 mg/kg, including in models with moderate and weak HER2 expression. The strong anti-tumor activity of ZW-49 in vivo, together with its ability to induce ICD and potential adaptive immune responses, support ZW49 as a promising ADC for the treatment of HER2-expressing cancers warranting further investigation, including potential combination with checkpoint inhibitors. Citation Format: Stuart D. Barnscher, Andrea Hernández Rojas, Kevin J. Hamblett, Nichole Escalante. Zanidatamab zovodotin (ZW49) induces hallmarks of immunogenic cell death and is active in patient-derived xenograft models of gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2633.

Abstract 1366: HMGB1 localization and its effect on the immune response in the lung tumor microenvironment

Abstract Lung adenocarcinoma is the leading cause of cancer mortality. High Mobility Group Box 1 (HMGB1) is a master regulator of innate immunity that is elevated in lung cancer tissues. Furthermore, HMGB1 has been previously demonstrated to suppress functional anti-tumor immune responses in this context. However, beyond demonstrating that HMGB1 is elevated and modulates inflammation, studies to date have not comprehensively investigated the influence of HMGB1 on the immune response to lung cancer. HMGB1 contributes to PD-L1 expression in melanoma and promotes secretion of inhibitory cytokines TGF-α and IL-10, which have roles in the differentiation of regulatory T cells. Therefore, HMGB1 sits atop an immunomodulatory signaling cascade in the tumor microenvironment. While treatment of lung cancers with immune checkpoint inhibitors (ICI) (anti-PD-1/PD-L1) has had success, there are limitations. One major limitation is the failure of patients with advanced or refractory disease to generate a strong anti-tumor immune response. Recently, we have identified HMGB1 as a protein that is functionally dependent on monounsaturated fatty acid (MUFA) availability. Utilizing a combination of molecular assays and 3-dimensional cultures, we now show that HMGB1 is sequestered intracellularly, rather than being secreted extracellularly in a MUFA-dependent fashion. We hypothesized that MUFAs promote an anti-tumor immune response via modulating HMGB1 release from cancer cells. Thus, by restricting extracellular HMGB1 levels, HMGB1-responsive proinflammatory/tumor signaling pathways (e.g., NF-kB) are suppressed. Taken together, these observations suggest a novel approach to decreasing maladaptive immune and inflammatory signaling from innate immune cells. Exploiting the MUFA/HMGB1 signaling axis could be a novel approach to improve the efficacy of therapies in populations that are not currently benefiting from current strategies for eliminating tumors. Citation Format: Kisa Jafri, Nicole M. Andre, Glenn E. Simmons. HMGB1 localization and its effect on the immune response in the lung tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1366.

Release of High-Mobility Group Box-1 after a Raynaud's Attack Leads to Fibroblast Activation and Interferon-γ Induced Protein-10 Production: Role in Systemic Sclerosis Pathogenesis.

Raynaud's Phenomenon (RP) leading to repetitive ischemia and reperfusion (IR) stress, is the first recognizable sign of systemic sclerosis (SSc) leading to increased oxidative stress. High-mobility group box-1 (HMGB1) is a nuclear factor released by apoptotic and necrotic cells after oxidative stress. Since HMGB1 can signal through the receptor for advanced glycation end products (RAGE), we investigated whether an RP attack promotes the release of HMGB1, leading to fibroblast activation and the upregulation of interferon (IFN)-inducible genes. A cold challenge was performed to simulate an RP attack in patients with SSc, primary RP (PRP), and healthy controls. We measured levels of HMGB1 and IFN gamma-induced Protein 10 (IP-10) at different time points in the serum. Digital perfusion was assessed by photoplethysmography. In vitro, HMGB1 or transforming growth factor (TGF-β1) (as control) was used to stimulate healthy human dermal fibroblasts. Inflammatory, profibrotic, and IFN-inducible genes, were measured by RT-qPCR. In an independent cohort, sera were obtained from 20 patients with SSc and 20 age- and sex-matched healthy controls to determine HMGB1 and IP-10 levels. We found that HMGB1 levels increased significantly 30 min after the cold challenge in SSc compared to healthy controls. In vitro stimulation with HMGB1 resulted in increased mRNA expression of IP-10, and interleukin-6 (IL-6) while TGF-β1 stimulation promoted IL-6 and Connective Tissue Growth Factor (CTGF). In serum, both HMGB1 and IP-10 levels were significantly higher in patients with SSc compared to healthy controls. We show that cold challenge leads to the release of HMGB1 in SSc patients. HMGB1 induces IP-10 expression in dermal fibroblasts partly through the soluble RAGE (sRAGE) axis suggesting a link between RP attacks, the release of HMGB1 and IFN-induced proteins as a putative early pathogenetic mechanism in SSc.

Meta-analysis of HMGB1 levels in the cerebrospinal fluid and serum of patients with epilepsy.

Epilepsy pathogenesis and progression are strongly influenced by inflammation. High-mobility group box-1 (HMGB1) is a key proinflammatory factor. The purpose of this study was to quantify and assess the relationship between HMGB1 level and epilepsy. We searched Embase, Web of Science, PubMed, and the Cochrane Library for studies examining the relationship between HMGB1 and epilepsy. Two independent researchers extracted data and assessed quality using the Cochrane Collaboration tool. Data extracted were analyzed using Stata 15 and Review Manager 5.3. The study protocol was registered prospectively at INPLASY, ID: INPLASY2021120029. A total of 12 studies were eligible for inclusion. After exclusion of one study with reduced robustness, 11 studies were included, with a total of 443 patients and 333 matched controls. Two of the articles included cerebrospinal fluid and serum HMGB1 data, which were distinguished by "a" and "b," respectively. The meta-analysis indicated that in comparison with the control group, the HMGB1 level was higher in epilepsy patients (SMD = 0.56, 95% CI = 0.27-0.85, P = 0.0002). Subgroup analysis of specimen types indicated that both serum HMGB1 and cerebrospinal fluid HMGB1 were higher in epilepsy patients than in the control group, with the increase in cerebrospinal fluid HMGB1 being more obvious. Subgroup analysis of disease types demonstrated that the serum HMGB1 level of epileptic seizure patients (including febrile and nonfebrile seizures) was significantly higher than that of matched controls. However, serum HMGB1 levels did not differ significantly between mild epilepsy patients and severe epilepsy patients. Patient age subgroup analysis showed higher HMGB1 in adolescents with epilepsy. Begg's test did not indicate publication bias. This is the first meta-analysis to summarize the association between HMGB1 level and epilepsy. The results of this meta-analysis indicate that epilepsy patients have elevated HMGB1. Large-scale studies with a high level of evidence are needed to reveal the exact relationship between HMGB1 level and epilepsy.

Age- and Sex- Dependent Changes in Serum Levels of TAS, TOS, TLR2, TLR4, HSP60, HSP90, and HMGB1

Objective: Cellular and physiological functions may be affected in an age- and sex-specific manner. The aim of this study is to investigate sex- and age-specific differences in the serum levels of Total Antioxidant Status (TAS), Total Oxidant Status (TOS), Oxidative Stress Index (OSI), Toll-like Receptor 2 (TLR2), Toll-Like Receptor 4 (TLR4), Heat Shock Protein 60 (HSP60), Heat Shock Protein 90 (HSP90), and High Mobility Group Box 1 (HMGB1) as well as to examine the correlation between them. 
 Methods: Four groups of mice, each including seven animals, were used in the present study: young males and females (6 months old); old males and females (24 months old). Blood samples were taken from the heart and serum was used to assay the levels of TLR2, TLR4, HSP60, HSP90, HMGB1, TAS and TOS. 
 Results: HGMB1, TOS and OSI were higher in old females than in young females (p

The Fifth Bioelectronic Medicine Summit Hosted by the Feinstein Institutes for Medical Research (Manhasset, NY) and Columbia Engineering (NY, NY) at The Garden City Hotel, Garden City, NY 11530 on October 11-12th, 2022- Bioelectronic Medicine: Today’s Tools, Tomorrow’s Therapies

The Fifth Bioelectronic Medicine Summit Hosted by the Feinstein Institutes for Medical Research (Manhasset, NY) and Columbia Engineering (NY, NY) at The Garden City Hotel, Garden City, NY 11530 on October 11-12th, 2022- Bioelectronic Medicine: Today’s Tools, Tomorrow’s Therapies

Association between high-mobility group box 1 levels and febrile seizures in children: a systematic review and meta-analysis

The relationship between High-mobility group box 1 (HMGB1) and febrile seizures (FS) in children remains unclear. This study aimed to apply meta-analysis to reveal the correlation between HMGB1 levels and FS in children. Databases including PubMed, EMBASE, Web of science, Cochrane library, CNKI, SinoMed and WanFangData were searched for relevant studies. Pooled standard mean deviation and 95% confidence interval were calculated as effect size since the random-effects model was used when I2 > 50%. Meanwhile, between-study heterogeneity was determined by performing subgroup and sensitivity analyses. A total of 9 studies were finally included. Meta-analysis showed that the children with FS had significantly higher HMGB1 levels compared with healthy children and children with fever but no seizures (P<0.05). Additionally, subgroup analysis showed that the HMGB1 level in children with complex FS was higher than those with simple FS (P<0.05), and children with duration >15 min were higher than those with duration ≤15min (P<0.05). There were no statistical differences between children with or without a family history of FS (P>0.05). Finally, children with FS who converted to epilepsy exhibited higher HMGB1 levels than those who did not convert to epilepsy (P<0.05). The level of HMGB1 may be implicated in the prolongation, recurrence and development of FS in children. Thus, it was necessary to evaluate the precise concentrations of HMGB1 in FS patients and to further determine the various activities of HMGB1 during FS by well-designed, large-scale, and case-controlled trials.

Elevated HMGB1 and sRAGE levels in cerebrospinal fluid of aneurysmal subarachnoid hemorrhage patients

BackgroundNeuroinflammation after aneurysmal subarachnoid hemorrhage (aSAH) leads to poor outcome of patients. High mobility group box 1 (HMGB1) contributes to inflammation through binding to receptors for advanced glycation end-products (RAGE) in various diseases. We aimed to determine the production of these two factors after aSAH and their relationship with clinical features. MethodsHMGB1 and soluble RAGE (sRAGE) levels in cerebrospinal fluid (CSF) of aSAH patients and controls were measured, and their temporal courses were observed. The correlation between early concentrations (days 1-3) and clinical symptoms assessed by disease severity scores, neuroinflammation estimated by CSF IL-6 levels, as well as prognosis evidenced by delayed cerebral ischemia (DCI) and 6-month adverse outcome was investigated. Finally, combined analysis of early levels for predicting prognosis was confirmed. ResultsCSF HMGB1 and sRAGE levels were higher in aSAH patients than in controls (P < 0.05), and the levels decreased from higher early to lower over time. Their early concentrations were positively associated with disease severity scores, IL-6 levels, DCI and 6-month poor outcome (P < 0.05). HMGB1 ≥ 6045.5 pg/ml (OR = 14.291, P = 0.046) and sRAGE ≥ 572.0 pg/ml (OR = 13.988, P = 0.043) emerged as independent predictors for DCI, while HMGB1 ≥ 5163.2 pg/ml (OR = 7.483, P = 0.043) and sRAGE ≥ 537.3 pg/ml (OR = 12.653, P = 0.042) were predictors for 6-month poor outcome. Combined analysis of them improved predictive values of adverse prognosis. ConclusionCSF HMGB1 and sRAGE levels of aSAH patients were increased early and then varied dynamically, which might act as potential biomarkers for poor outcome, especially when co-analyzed.

O6-1 - Inappropriate inflammation in ovarian endometrial cysts correlates with excessive T-cell activation and increased level of high-mobility group box-1

O6-1 - Inappropriate inflammation in ovarian endometrial cysts correlates with excessive T-cell activation and increased level of high-mobility group box-1

The effects of etomidate on expression of high mobility group box 1 via the nuclear factor kappa B pathway in rat model of sepsis

ABSTRACT Etomidate is an anesthetic agent used in hemodynamically unstable patients, but its use has been controversial in septic patients. The response of high-mobility group box 1 (HMGB1), a late-phase lethal cytokine in sepsis, to etomidate has not been reported. This study investigated the effects of etomidate on the expression and release of HMGB1 and the underlying mechanism using a cecal ligation and puncture (CLP) model. Thirty-six male Sprague–Dawley rats were divided into sham, CLP, and Etomi groups. Sepsis was induced in the CLP and Etomi groups, and intravenous etomidate (4 mg/kg) was infused for 40 min immediately after operation in the Etomi group. Serum creatinine, alanine aminotransferase (ALT), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and HMGB1 levels were measured 6 and 24 hours after surgery. Activation of nuclear factor (NF)-ĸB and HMGB1 mRNA expression in the liver, lung, kidney, and ileum tissues were measured, and immunohistochemical staining of HMGB1 was implemented. Increases of the TNF-α level 6 h after CLP and ALT and IL-6 levels 24 h after CLP were significantly inhibited by etomidate treatment. Etomidate treatment also significantly attenuated the increase in serum HMGB1 level at 6 and 24 h after CLP and suppressed the NF-ĸB and HMGB1 mRNA in multiple organs 24 h after CLP. Immunohistochemical staining also revealed that etomidate treatment inhibited HMGB1 expression. Etomidate inhibited the systemic release of HMGB1 and its expression in various organs. The mechanism may be associated with the inhibitory effects of etomidate on pro-inflammatory cytokine release and NF-ĸB activity.

Serum HMGB1 levels and its clinical significance in elderly patients with intertrochanteric fractures after intramedullary fixation surgery.

Intramedullary fixation is a valuable alternative for the treatment of intertrochanteric fractures. However, further development of new biomarkers to predict the prognosis of the patient is still needed for timely and effective treatment and intervention. The present study aimed to explore the serum high-mobility group box 1 (HMGB1) levels in the prognosis of intertrochanteric fracture patients and its correlation with clinical results. The present prospective cohort study recruited 115 intertrochanteric fracture patients who were admitted from January 2015 to December 2019. All patients were evaluated preoperatively and treated (proximal femoral nail antirotation or intramedullary proximal femoral nail) by the same team. The serum HMGB1, interleukin-6, interleukin-1β, tumor necrosis factor α, and C-reactive protein levels were measured by enzyme-linked immunosorbent assay. Demographic and clinical data of all patients were collected. Harris score was used to assess the prognosis of intertrochanteric fracture patients after 6 months of treatment. Statistical analysis was conducted using SPSS software with P < .05 as statistically different. The time of the operation and the amount of bleeding in intramedullary proximal femoral nail were remarkably elevated compared with the proximal femoral nail antirotation group (P < .05). The age, proportion of complications and visual analogue score VAS after 72 hours of surgery in the Harris score < 80 group were remarkably increased compared with Harris score ≥ 80 group (P < .05). In addition, we found that the serum HMGB1 levels in Harris score < 80 group were markedly elevated than the patients in Harris score ≥ 80 group at all time points (P < .05). The results showed that the serum HMGB1 levels at postoperative 48 hours had the highest predictive value for predicting poor prognosis in intertrochanteric fracture patients. It was found that HMGB1, age and VAS after 72 hours of surgery were the risk factors for poor prognosis of intertrochanteric fracture patients. This study showed that the serum HMGB1 levels was significantly decreased in intertrochanteric fracture patients with bad prognoses. This study may provide a new approach to screening intertrochanteric fracture patients with worse prognoses in advance.

Correlation of HMGB1, TLR2 and TLR4 with left ventricular diastolic dysfunction in sepsis patients.

Left ventricular diastolic dysfunction (LVDD) is a common consequence of sepsis due to dysregulated inflammatory responses. Here we aim to investigate high mobility group box 1 (HMGB1), toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) as serum biomarkers to assess LVDD risk of patients with sepsis. We recruited 120 patients with sepsis, among which 52 had ultrasonically confirmed LVDD and 68 were without LVDD. Blood samples were collected, and enzyme-linked immunosorbent assay (ELISA) was used to analyse levels of HMGB1, TLR2 and TLR4 in serum. Multivariate analysis was performed to assess the odds ratio of the serum biomarkers. Spearman's correlation analysis was conducted to evaluate the correlation between the serum biomarkers to B-type natriuretic peptide (BNP) and cardiac troponin I (cTnl) levels and the ratios of early diastolic mitral inflow velocity to early diastolic mitral annulus velocity (E/e' ratios) in ultrasound. Receiver operating curve was used to measure the sensitivity and specificity of HMGB1, TLR2 and TLR4 individually and in combination as diagnostic markers. Elevated HMGB1, TLR2 and TLR4 had significant values in predicting LVDD suggested by high odds ratio (all P < .05). A significant correlation was found between these values and cTnl, the current gold standard for LVDD analysis. HMGB1, TLR2 and TLR4 also showed a high diagnostic sensitivity and specificity in ROC analysis. HMGB1, TLR2 and TLR4 are potentially valuable in predicting LVDD risk among patients with sepsis, providing additional tools with the capability of potentially assisting the clinical management of patients with sepsis.

CXCL12/CXCR4 Axis is Involved in the Recruitment of NK Cells by HMGB1 Contributing to Persistent Airway Inflammation and AHR During the Late Stage of RSV Infection.

We previously showed that both high-mobility group box-1 (HMGB1) and natural killer (NK) cells contribute to respiratory syncytial virus (RSV)-induced persistent airway inflammation and airway hyperresponsiveness (AHR). Meanwhile, Chemokine (C-X-C motif) ligand 12 (CXCL12) and its specific receptor (chemokine receptor 4, CXCR4) play important roles in recruitment of immune cells. CXCL12 has been reported to form a complex with HMGB1 that binds to CXCR4 and increases inflammatory cell migration. The relationship between HMGB1, NK cells and chemokines in RSV-infected model remains unclear. An anti-HMGB1 neutralizing antibody and inhibitor of CXCR4 (AMD3100) was administered to observe changes of NK cells and airway disorders in nude mice and BALB/c mice. Results showed that the mRNA expression and protein levels of HMGB1 were elevated in late stage of RSV infection and persistent airway inflammation and AHR were diminished after administration of anti-HMGB1 antibodies, with an associated significant decrease in CXCR4+ NK cells. In addition, CXCL12 and CXCR4 were reduced after HMGB1 blockade. Treatment with AMD3100 significantly suppressed the recruitment of NK cells and alleviated the airway disorders. Thus, CXCL12/CXCR4 axis is involved in the recruitment of NK cells by HMGB1, contributing to persistent airway inflammation and AHR during the late stage of RSV infection.

Prenatal Sevoflurane Exposure Impairs the Learning and Memory of Rat Offspring via HMGB1-Induced NLRP3/ASC Inflammasome Activation.

The neurotoxic effects of sevoflurane anesthesia on the immature nervous system have aroused public concern, but the specific effects and mechanism remain poorly understood. Pyroptosis caused by the activation of the NLRP3 inflammasome is pivotal for cell survival and acts as a key player in cognitive impairment. This study was carried out to determine the critical role of the NLRP3 inflammasome and high-mobility group box 1 (HMGB1) in sevoflurane-induced cognitive impairment. On gestational day 20 (G20), 3% sevoflurane was administered for 4 h to pregnant rats. The hippocampus and cerebral cortex of the offspring were harvested at postnatal day 1 (P1) for Western blotting and immunofluorescence staining. Pregnant rat sevoflurane exposure increased the protein levels of NLRP3, ASC, cleaved-caspase 1 (p20), mature-IL-1β (m-IL-1β), and HMGB1 in the cerebral cortex and hippocampus of offspring rats. More microglial cells of offspring were also observed after sevoflurane anesthesia. The Morris water maze (MWM) test was implemented to evaluate cognitive function from postnatal day 30 (P30) to postnatal 35 (P35) of offspring. The sevoflurane-treated offspring took longer than the control rats to find the MWM platform during the learning phase. Furthermore, they had a longer travel distance and less time in the target quadrant than the control rats in the probe trial. Maternal intraperitoneal injection of glycyrrhizin (an inhibitor of HMGB1) attenuated the sevoflurane-induced microglia and NLRP3/ASC inflammasome activation and cognitive impairment of offspring. Simultaneously, the sevoflurane-induced increase in Toll-like receptors (TLR4) and nuclear factor-κB (NF-κB) was significantly reduced by glycyrrhizin. We concluded that the HMGB1 inhibitor may repress the sevoflurane-induced activation of the NLRP3/ASC inflammasome and cognitive dysfunction and that TLR4/NF-κB signaling maybe the key pathway, at least in part.

Correlation of serum-soluble mesothelin-related protein, HMGB1 and CA125 in diffuse malignant peritoneal mesothelioma and their combined measurement for prognosis

Diagnosis of diffuse malignant peritoneal mesothelioma (DMPM) is challenging due to the lack of efficient biomarkers for early-stage DMPM. This study was designed to characterize three serum-soluble mesothelium-related proteins, including soluble mesothelin-related protein (SMRP), high mobility group box 1 (HMGB1), and cancer antigen 125 (CA125) in diagnosing DMPM. Serum samples of DMPM patients and healthy controls were collected and an enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of HMGB1, CA125, and SMRP. Correlations between these three serum proteins were examined and the diagnostic values of the biomarkers were assessed by receiver operating curve (ROC) analysis. The combined expression levels of the three markers were also analyzed in terms of predicting patient survival. Higher levels of CA125, SMRP, and HMGB1 was found to be a prominent characteristic of DMPM patients (with > two-fold higher for all levels in DMPM patients compared to control patients, all p < .001), particularly for those with higher-stage DMPM (stage III-IV) compared with lower-stage DMPM (stage I-II) (all p < .05). HMGB1, CA125, and SMRP were all significantly inter-correlated with each other (all p < .05), the combination of the three serum markers had high sensitivity and specificity for diagnosing DMPM. Combined values of the three markers demonstrated a high AUC of 0.85, sensitivity of 78.95%, specificity of 82.75% for identifying DMPM. The combined level of the three markers also demonstrated a significant positive correlation with poor survival of DMPM patients (p = .022). CA125, SMRP, and HMGB1 are potentially valuable diagnostic biomarkers to facilitate the diagnosis of DMPM.