Clinical Manifestations and Genetic Analysis of 17 Patients with Autosomal Dominant Hyper-IgE Syndrome in Mainland China: New Reports and a Literature Review.

PURPOSE: Autosomal dominant hyper-IgE syndrome (AD-HIES) is a rare complicated primary immunodeficiency disease (PID). Signal transducer and activator of transcription 3 (STAT3) gene mutation is found to cause AD-HIES. The distribution of AD-HIES patients with STAT3 deficiency in the Chinese population is not clear. Herein, we retrospectively report 17 AD-HIES patients with STAT3 deficiency and demonstrate their clinical, immunological, and genetic features. METHODS: Patients' clinical data were collected from their medical records. Routine laboratory testing results included lymphocyte subset analysis and immunoglobulin quantification. STAT3 mutations were investigated by sequencing of genomic DNA. RESULTS: Among 575 patients with PID, 28 (4.87%) were clinically diagnosed as HIES. Among them, 17 (2.96%) were confirmed as STAT3 mutant AD-HIES. The ratio of male to female patients was 8:9. All of the 17 patients had NIH scores over 40 points. The mean ages at onset and diagnosis were 1.05 and 10.35 years, respectively. Three patients (17.65%, 3/17) died with a mean age of 13.33 years. Eczema, recurrent skin infection, and respiratory tract infection were the most common clinical symptoms and are present in all of the 17 patients in this study. Six patients (37.5%, 6/16) suffered complication from BCG vaccination. Noninfection symptoms are characteristic facial features in 17 patients (100%, 17/17), retention of primary teeth in 10 patients (90.91%, 10/11), and abnormal bone fractures in 7 patients (41.18%, 7/17). Eleven types of STAT3 mutations were identified in 17 patients, including 1 novel mutation. CONCLUSIONS: We here retrospectively report the largest Chinese cohort of AD-HIES patients with STAT3 mutation. Unique features, when compared to existing literature reports, include (1) later age of diagnosis, (2) significantly higher rate of BCG complications, and (3) lower rate of candidiasis and chronic otitis media.

The bioeffects of degradable products derived from a biodegradable Mg-based alloy in macrophages via heterophagy.

Biodegradable magnesium alloys are promising candidates for use in biomedical applications. However, degradable particles (DPs) derived from Mg-based alloys have been observed in tissue in proximity to sites of implantation, which might result in unexpected effects. Although previous in vitro studies have found that macrophages can take up DPs, little is known about the potential phagocytic pathway and the mechanism that processes DPs in cells. Additionally, it is necessary to estimate the potential bioeffects of DPs on macrophages. Thus, in this study, DPs were generated from a Mg-2.1Nd-0.2Zn-0.5Zr alloy (JDBM) by an electrochemical method, and then macrophages were incubated with the DPs to reveal the potential impact. The results showed that the cell viability of macrophages decreased in a concentration-dependent manner in the presence of DPs due to effects of an apoptotic pathway. However, the DPs were phagocytosed into the cytoplasm of macrophages and further degraded in phagolysosomes, which comprised lysosomes and phagosomes, by heterophagy instead of autophagy. Furthermore, several pro-inflammatory cytokines in macrophages were upregulated by DPs through the induction of reactive oxygen species (ROS) production. To the best of our knowledge, this is the first study to show that DPs derived from a Mg-based alloy are consistently degraded in phagolysosomes after phagocytosis by macrophages via heterophagy, which results in an inflammatory response owing to ROS overproduction. Thus, our research has increased the knowledge of the metabolism of biodegradable Mg metal, which will contribute to an understanding of the health effects of biodegradable magnesium metal implants used for tissue repair. STATEMENT OF SIGNIFICANCE: Biomedical degradable Mg-based alloys have great promise in applied medicine. Although previous studies have found that macrophages can uptake degradable particles (DPs) in vitro and observed in the sites of implantation in vivoin vivo, few studies have been carried out on the potential bioeffects relationship between DPs and macrophages. In this study, we analyzed the bioeffects of DPs derived from a Mg-based alloy on the macrophages. We illustrated that the DPs were size-dependently engulfed by macrophages via heterophagy and further degraded in the phagolysosome rather than autophagosome. Furthermore, DPs were able to induce a slight inflammatory response in macrophages by inducing ROS production. Thus, our research enhances the knowledge of the interaction between DPs of Mg-based alloy and cells, and offers a new perspective regarding the use of biodegradable alloys.

Degradation Products of Polydopamine Restrained Inflammatory Response of LPS-Stimulated Macrophages Through Mediation TLR-4-MYD88 Dependent Signaling Pathways by Antioxidant.

Polydopamine (PDA) has a promising application as coating of biomaterials due to its favorable degradability and bioadaptability. However, its bioactivity, such as anti-inflammatory capacity, was still little known. Herein, we investigated whether degradable products of PDA could affect inflammatory response in lipopolysaccharide (LPS)-stimulated human THP-1-derived macrophages. The supernatants containing degradation products of PDA, annotated as PDA extracts, were collected after PDA being immersed in cell culture medium for 3 days. Wherein, the composition of the degradation products was analyzed by HPLC assay. Collected PDA extracts were diluted into 100%, 50%, and 25% of original concentration, respectively, to evaluate their anti-inflammatory ability on LPS-induced macrophages from the expression levels of pro-inflammatory cytokines to associated molecular mechanism. Our results showed that the PDA extracts were mainly composed of dopamine, quinine, and PDA segments. Furthermore, macrophages showed no cytotoxicity after PDA extract treatment with or without LPS, while the release levels of TNF-α and IL-6 by LPS-induced macrophages were decreased in dose-dependent by PDA extract treatment. Additionally, TLR-4 and MYD88 expression in protein and RNA level were downregulated by PDA extracts in LPS-induced macrophages. Similarly, PDA extracts effectively inhibited LPS-induced NF-κB trans-locating into nuclear by inactivation of the phosphorylation of IKK-α/ß and IKß-α. Of note, the production of LPS-induced ROS was reduced by PDA extracts in macrophages, while HO-1 expression, a critical protein of antioxidant signaling pathway, was increased. Based on these results, we proposed a potential mechanism by which degradation products of PDA suppressed inflammation of macrophages via downregulation TLR-4-MYD88-NFκB pathway and simultaneous activation HO-1 pathway, which might be a possible therapeutic target.

A Biodegradable Mg-Based Alloy Inhibited the Inflammatory Response of THP-1 Cell-Derived Macrophages Through the TRPM7-PI3K-AKT1 Signaling Axis.

Mg-based alloys might be ideal biomaterials in clinical applications owing to favorable mechanical properties, biodegradability, biocompatibility, and especially their anti-inflammatory properties. However, the precise signaling mechanism underlying the inhibition of inflammation by Mg-based alloys has not been elucidated. Here, we investigated the effects of a Mg-2.1Nd-0.2Zn-0.5Zr alloy (denoted as JDBM) on lipopolysaccharide (LPS)-induced macrophages. THP-1 cell-derived macrophages were cultured on JDBM, Ti-6Al-4V alloy (Ti), 15% extract of JDBM, and 7.5 mM of MgCl2 for 1 h before the addition of LPS for an indicated time; the experiments included negative and positive controls. Our results showed JDBM, extract, and MgCl2 could decrease LPS-induced tumor necrosis factor (TNF) and interleukin (IL)-6 expression. However, there were no morphologic changes in macrophages on Ti or JDBM. Mechanically, extract and MgCl2 downregulated the expression of toll-like receptor (TLR)-4 and MYD88 compared with the positive control and inhibited LPS-induced nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways by inactivation of the phosphorylation of IKK-α/ß, IKß-α, P65, P38, and JNK. Additionally, the LPS-induced reactive oxygen species (ROS) expression was also decreased by extract and MgCl2. Interestingly, the expression of LPS-induced TNF and IL-6 could be recovered by knocking down TRPM7 of macrophages, in the presence of extract or MgCl2. Mechanically, the activities of AKT and AKT1 were increased by extract or MgCl2 with LPS and were blocked by a PI3K inhibitor, whereas siRNA TRPM7 inhibited only AKT1. Together, our results demonstrated the degradation products of Mg-based alloy, especially magnesium, and resolved inflammation by activation of the TRPM7-PI3K-AKT1 signaling pathway, which may be a potential advantage or target to promote biodegradable Mg-based alloy applications.

In vitro study of the inflammatory cells response to biodegradable Mg-based alloy extract.

Biodegradable Mg-based alloys have shown great potential as bone fixation devices or vascular stents. As implant biomaterials, the foreign body reaction (FBR) is an important issue to be studied, where the inflammatory cells play a key role. Here, we used two inflammatory cell lines i.e. THP-1 cells and THP-1 macrophages, to evaluate the effect of Mg-Nd-Zn-Zr alloy (denoted as JDBM) extracts on cell viability, death modes, cell cycle, phagocytosis, differentiation, migration and inflammatory response. The results showed that high-concentration extract induced necrosis and complete damage of cell function. For middle-concentration extract, cell apoptosis and partially impaired cell function were observed. TNF-α expression of macrophages was up-regulated by co-culture with extract in 20% concentration, but was down-regulated in the same concentration in the presence of LPS stimulation. Interestingly, the production of TNF-α decreased when macrophages were cultured in middle and high concentration extracts independent of LPS. Cell viability was also negatively affected by magnesium ions in JDBM extracts, which was a potential factor affecting cell function. Our results provide new information about the impact of Mg alloy extracts on phenotype of immune cells and the potential mechanism, which should be taken into account prior to clinical applications.