Inflammation impacts androgen receptor signaling in basal prostate stem cells through interleukin 1 receptor antagonist.

Chronic prostate inflammation in patients with benign prostate hyperplasia (BPH) correlates with the severity of symptoms. How inflammation contributes to prostate enlargement and/or BPH symptoms and the underlying mechanisms remain unclear. In this study, we utilize a unique transgenic mouse model that mimics chronic non-bacterial prostatitis in men and investigate the impact of inflammation on androgen receptor (AR) in basal prostate stem cells (bPSC) and their differentiation in vivo. We find that inflammation significantly enhances AR levels and activity in bPSC. More importantly, we identify interleukin 1 receptor antagonist (IL-1RA) as a crucial regulator of AR in bPSC during inflammation. IL-1RA is one of the top molecules upregulated by inflammation, and inhibiting IL-1RA reverses the enhanced AR activity in organoids derived from inflamed bPSC. Additionally, IL-1RA appears to activate AR by counteracting IL-1α's inhibitory effect. Furthermore, using a lineage tracing model, we observe that inflammation induces bPSC proliferation and differentiation into luminal cells even under castrate conditions, indicating that AR activation driven by inflammation is sufficient to promote bPSC proliferation and differentiation. Taken together, our study uncovers mechanisms through which inflammation modulates AR signaling in bPSC and induces bPSC luminal differentiation that may contribute to prostate hyperplasia.

Spatially controllable fluid hydrogel with in-situ electrospinning PCL/chitosan fiber for treating irregular wounds.

Skin injuries sustained during exercise are often irregular in shape and frequently accompanied by infections. Bacteria residing in the crevices of these wounds can lead to persistent infections. Routine wound monitoring, which requires removing the wound dressing to assess recovery, is inconvenient and increases the risk of infection. To address this, we prepared a polyvinyl alcohol/polyhydroxylated fullerenes ((PVA/PHF) hydrogel with good fluidity and photothermal antibacterial properties, which can penetrate into the crevices of irregular wounds. After the hydrogel was applied to the wound, the hydrogel was locally defined by the polycaprolactone/Chitosan (PCL/CS) membrane of in-situ electrospinning, which effectively killed bacteria, and the healing efficiency was increased by 240 % in the wound healing experiment. The PVA/PHF hydrogel exhibits excellent electrical conductivity, making it suitable for real-time monitoring of human body motion as a strain sensor. This capability provides doctors with an accurate basis for wound assessment. At the same time, the hydrogel can achieve self-healing within 1.5 s and withstand up to 2200 % tensile strain, which can be used for large-scale motion monitoring of the human body. This flowable hydrogel, capable of penetrating wound crevices, offers a dual function of both treatment and monitoring.

DDX5 deficiency drives non-canonical NF-κB activation and NRF2 expression, influencing sorafenib response and hepatocellular carcinoma progression.

In advanced hepatocellular carcinoma (HCC), RNA helicase DDX5 regulates the Wnt/ß-catenin-ferroptosis axis, influencing the efficacy of the multi-tyrosine kinase inhibitor (mTKI) sorafenib. DDX5 inhibits Wnt/ß-catenin signaling, preventing sorafenib-induced ferroptosis escape. Sorafenib/mTKIs reduce DDX5 expression, correlating with poor patient survival post-sorafenib treatment. Notably, DDX5-knockout in HCC cells activates Wnt/ß-catenin signaling persistently. Herein, we investigate the mechanistic impact of Wnt/ß-catenin activation resulting from DDX5 downregulation in the progression and treatment of HCC. RNAseq analyses identified shared genes repressed by DDX5 and upregulated by sorafenib, including Wnt signaling genes, NF-κB-inducing kinase (NIK) essential for non-canonical NF-κB (p52/RelB) activation, and cytoprotective transcription factor NRF2. We demonstrate, Wnt/ß-catenin activation induced NIK transcription, leading to non-canonical NF-κB activation, which subsequently mediated NRF2 transcription. Additionally, DDX5 deficiency extended NRF2 protein half-life by inactivating KEAP1 through p62/SQSTM1 stabilization. In a preclinical HCC mouse model, NRF2 knockdown or DDX5 overexpression restricted tumor growth upon sorafenib treatment, via induction of ferroptosis. Importantly, DDX5-knockout HCC cells exhibited elevated expression of Wnt signaling genes, NIK, p52/RelB, and NRF2-regulated genes, regardless of sorafenib treatment. Transcriptomic analyses of HCCs from TCGA and the Stelic Animal Model (STAM) of non-alcoholic steatohepatitis revealed elevated expression of these interconnected pathways in the context of DDX5 downregulation. In conclusion, DDX5 deficiency triggers Wnt/ß-catenin signaling, promoting p52/RelB and NRF2 activation, thereby enabling ferroptosis evasion upon sorafenib treatment. Similarly, independent of sorafenib, DDX5 deficiency in liver tumors enhances activation and gene expression of these interconnected pathways, underscoring the clinical relevance of DDX5 deficiency in HCC progression and therapeutic response.

Tox induces T cell IL-10 production in a BATF-dependent manner.

Tox is a member of the high mobility group (HMG)-Box transcription factors and plays important roles in thymic T cell development. Outside of the thymus, however, Tox is also highly expressed by CD8 and CD4 T cells in various states of activation and in settings of cancer and autoimmune disease. In CD4 T cells, Tox has been primarily studied in T follicular helper (TFH) cells where it, along with Tox2, promotes TFH differentiation by regulating key TFH-associated genes and suppressing CD4 cytotoxic T cell differentiation. However, the role of Tox in other T helper (Th) cell subtypes is less clear. Here, we show that Tox is expressed in several physiologically-activated Th subtypes and its ectopic expression enhances the in vitro differentiation of Th2 and T regulatory (Treg) cells. Tox overexpression in unpolarized Th cells also induced the expression of several genes involved in cell activation (Pdcd1), cellular trafficking (Ccl3, Ccl4, Xcl1) and suppressing inflammation (Il10) across multiple Th subtypes. We found that Tox binds the regulatory regions of these genes along with the transcription factors BATF, IRF4, and JunB and that Tox-induced expression of IL-10, but not PD-1, is BATF-dependent. Based on these data, we propose a model where Tox regulates Th cell chemotactic genes involved in facilitating dendritic cell-T cell interactions and aids in the resolution or prevention of inflammation through the production of IL-10.

RNA helicase DDX5 modulates sorafenib sensitivity in hepatocellular carcinoma via the Wnt/ß-catenin-ferroptosis axis.

Reduced expression of the RNA helicase DDX5 associated with increased hepatocellular carcinoma (HCC) tumor grade and poor patient survival following treatment with sorafenib. While immunotherapy is the first-line treatment for HCC, sorafenib and other multi-tyrosine kinase inhibitors (mTKIs) are widely used when immunotherapy is contra-indicated or fails. Herein, we elucidate the role of DDX5 in sensitizing HCC to sorafenib, offering new therapeutic strategies. Treatment of various human HCC cell lines with sorafenib/mTKIs downregulated DDX5 in vitro and in preclinical HCC models. Conversely, DDX5 overexpression reduced the viability of sorafenib-treated cells via ferroptosis, suggesting a role for DDX5 in sorafenib sensitivity. RNAseq of wild-type vs. DDX5-knockdown cells treated with or without sorafenib identified a set of common genes repressed by DDX5 and upregulated by sorafenib. This set significantly overlaps with Wnt signaling genes, including Disheveled-1 (DVL1), an indispensable Wnt activator and prognostic indicator of poor survival for sorafenib-treated patients. DDX5-knockout (DDX5KO) HCC cells exhibited DVL1 induction, Wnt/ß-catenin pathway activation, and ferroptosis upon inhibition of canonical Wnt signaling. Consistently, xenograft HCC tumors exhibited reduced growth by inhibition of Wnt/ß-catenin signaling via induction of ferroptosis. Significantly, overexpression of DDX5 in HCC xenografts repressed DVL1 expression and increased ferroptosis, resulting in reduced tumor growth by sorafenib. We conclude that DDX5 downregulation by sorafenib mediates adaptive resistance by activating Wnt/ß-catenin signaling, leading to ferroptosis escape. Conversely, overexpression of DDX5 in vivo enhances the anti-tumor efficacy of sorafenib by suppressing Wnt/ß-catenin activation and induction of ferroptosis. Thus, DDX5 overexpression in combination with mTKIs is a promising therapeutic strategy for HCC.

Distinct use of super-enhancer elements controls cell type-specific CD25 transcription and function.

The IL-2 receptor α chain (IL-2Rα/CD25) is constitutively expressed on double-negative (DN2/DN3 thymocytes and regulatory T cells (Tregs) but induced by IL-2 on T and natural killer (NK) cells, with Il2ra expression regulated by a STAT5-dependent super-enhancer. We investigated CD25 regulation and function using a series of mice with deletions spanning STAT5-binding elements. Deleting the upstream super-enhancer region mainly affected constitutive CD25 expression on DN2/DN3 thymocytes and Tregs, with these mice developing autoimmune alopecia, whereas deleting an intronic region decreased IL-2-induced CD25 on peripheral T and NK cells. Thus, distinct super-enhancer elements preferentially control constitutive versus inducible expression in a cell type-specific manner. The mediator-1 coactivator colocalized with specific STAT5-binding sites. Moreover, both upstream and intronic regions had extensive chromatin interactions, and deletion of either region altered the super-enhancer structure in mature T cells. These results demonstrate differential functions for distinct super-enhancer elements, thereby indicating previously unknown ways to manipulate CD25 expression in a cell type-specific fashion.

MXene@Hydrogel composite nanofibers with the photo-stimulus response and optical monitoring functions for on-demand drug release.

The photo-stimulus response has the advantage of non-invasiveness, which could be used to control the "on" and "off" of drug release achieving on-demand release. Herein, we design a heating electrospray during electrospinning to prepare photo-stimulus response composite nanofibers consisting of MXene@Hydrogel. This heating electrospray enables to spray MXene@Hydrogel during the electrospinning process, and the hydrogel is uniformly distributed which cannot be achieved by the traditional soaking method. In addition, this heating electrospray can also overcome the difficulty that hydrogels are hard to be uniformly distributed in the inner fiber membrane.The "on" and "off" state of drug release could be controlled by light. Not only near infrared (NIR) light but also sunlight could trigger the drug release, which could benefit outdoor use when cannot find NIR light. Evidence by hydrogen bond has been formed between MXene and Hydrogel, the mechanical property of MXene@Hydrogel composite nanofibers is significantly enhanced, which is conducive to the application of human joints and other parts that need to move. These nanofibers also possess fluorescence property, which is further used to real-time monitor the in-vivo drug release. No matter the fast or slow release, this nanofiber can achieve sensitive detection, which is superior to the current absorbance spectrum method.

Stretchable, flexible and breathable polylactic acid/polyvinyl pyrrolidone bandage based on Kirigami for wounds monitoring and treatment.

Chronic wounds are slow to recover. During treatment, the dressing needs to be removed to check the recovery status, a process that often results in wound tears. Traditional dressings lack stretching and flexing properties and are not suitable using on wounds in joints, which require movement from time to time. In this study, we present a stretchable, flexible and breathable bandage consisting of three layers, including Mxene coating on the top, the polylactic acid/polyvinyl pyrrolidone (PLA/PVP) layer designed as Kirigami in the middle, and the f-sensor at the bottom. By the way, the f-sensor is in contact with the wound sensing real-time microenvironmental changes due to infection. When the infection intensifies, the Mxene coating at the top is utilized to enable anti-infection treatment. And Kirigami structure of PLA/PVP ensures that this bandage has stretchability, bendability, and breathability. The stretch of the smart bandage increases to 831 % compared to the original structure, and the modulus reduces to 0.04 %, which adapts extremely well to the movement of the joints and relieves the pressure on the wound. This monitoring-treatment closed-loop working mode, eliminating the need to remove dressings and avoid tissue tearing, shows a promising capability in the field of surgical wound care.

Deep learning models for hepatitis E incidence prediction leveraging meteorological factors.

BACKGROUND: Infectious diseases are a major threat to public health, causing serious medical consumption and casualties. Accurate prediction of infectious diseases incidence is of great significance for public health organizations to prevent the spread of diseases. However, only using historical incidence data for prediction can not get good results. This study analyzes the influence of meteorological factors on the incidence of hepatitis E, which are used to improve the accuracy of incidence prediction. METHODS: We extracted the monthly meteorological data, incidence and cases number of hepatitis E from January 2005 to December 2017 in Shandong province, China. We employ GRA method to analyze the correlation between the incidence and meteorological factors. With these meteorological factors, we achieve a variety of methods for incidence of hepatitis E by LSTM and attention-based LSTM. We selected data from July 2015 to December 2017 to validate the models, and the rest was taken as training set. Three metrics were applied to compare the performance of models, including root mean square error(RMSE), mean absolute percentage error(MAPE) and mean absolute error(MAE). RESULTS: Duration of sunshine and rainfall-related factors(total rainfall, maximum daily rainfall) are more relevant to the incidence of hepatitis E than other factors. Without meteorological factors, we obtained 20.74%, 19.50% for incidence in term of MAPE, by LSTM and A-LSTM, respectively. With meteorological factors, we obtained 14.74%, 12.91%, 13.21%, 16.83% for incidence, in term of MAPE, by LSTM-All, MA-LSTM-All, TA-LSTM-All, BiA-LSTM-All, respectively. The prediction accuracy increased by 7.83%. Without meteorological factors, we achieved 20.41%, 19.39% for cases in term of MAPE, by LSTM and A-LSTM, respectively. With meteorological factors, we achieved 14.20%, 12.49%, 12.72%, 15.73% for cases, in term of MAPE, by LSTM-All, MA-LSTM-All, TA-LSTM-All, BiA-LSTM-All, respectively. The prediction accuracy increased by 7.92%. More detailed results are shown in results section of this paper. CONCLUSIONS: The experiments show that attention-based LSTM is superior to other comparative models. Multivariate attention and temporal attention can greatly improve the prediction performance of the models. Among them, when all meteorological factors are used, multivariate attention performance is better. This study can provide reference for the prediction of other infectious diseases.

Impaired long-term anti-HBs responses in choronic hepatitis C patients: Results from a five-year follow-up study with healthy control.

Although hepatitis B virus (HBV) vaccination is recommended for hepatitis C virus (HCV)-infected individuals to avoid HBV superinfection, the persistence of their humoral and cell-mediated immunity responses to HBV vaccination is still under investigation. Patients with chronic hepatitis C (CHC) and matched healthy controls, who completed three doses of hepatitis B vaccine (HepB) in 2014, were followed up five years later. One booster dose of HepB was given to those with antibody against hepatitis B surface antigen (anti-HBs) lower than 10mIU/mL. Anti-HBs was tested at follow-up and on the 14th day after the booster dose, as well as HBsAg specific spot-forming cells of interferon γ and interleukin (IL) 2, 4, 5, and 6. At five years, only 56.58% of the CHC patients had sero-protective titers (≥10mIU/mL) of anti-HBs, compared to 70.83% in the controls (P < .05). Similarly, the geometric mean concentration (GMC) of anti-HBs in CHC patients was significantly lower than that in controls (16.95 vs 37.34 mIU/mL, P < .05). After the booster, both GMC and the rate of anamnestic response increased to a very high level in the two groups and the difference between them disappeared (P > .05). Multivariable analysis showed that HCV infection was an independent predictor factor to anti-HBs level at follow-up. HBsAg specific IL-6 was stronger in the CHC patients compared to the controls (P < .05). The data indicate that the durability of protective anti-HBs is poorer in CHC patients compared to healthy individuals, and impaired long-term anti-HBs responses might be associated with the increased HBsAg specific IL-6 responses.

Establishment of a visual gene knockout system based on CRISPR/Cas9 for the rare actinomycete Nonomuraea gerenzanensis.

OBJECTIVES: To develop a modified CRISPR/Cas9 system with the ß-glucuronidase (GusA) reporter and a dual sgRNA cassette for Nonomuraea gerenzanensis (N. gerenzanensis). RESULTS: With the aid of a visual GusA reporter, the complicated and tedious process of cloning and gene identification could be abandoned entirely in the genetic editing of N. gerenzanensis. Moreover, introducing a dual sgRNA cassette into the CRISPR/Cas9 system significantly improved gene deletion efficiency compared to the single sgRNA element. Furthermore, the length of the homologous flanking sequences set to the lowest value of 500 bp in this system could still reach the relatively higher conjugation transfer frequency. CONCLUSIONS: The enhanced CRISPR/Cas9 system could efficiently perform genetic manipulation on the rare actinomycete N. gerenzanensis.

A comprehensive single cell data analysis of lymphoblastoid cells reveals the role of super-enhancers in maintaining EBV latency.

We probed the lifecycle of Epstein-Barr virus (EBV) on a cell-by-cell basis using single cell RNA sequencing (scRNA-seq) data from nine publicly available lymphoblastoid cell lines (LCLs). While the majority of LCLs comprised cells containing EBV in the latent phase, two other clusters of cells were clearly evident and were distinguished by distinct expression of host and viral genes. Notably, both were high expressors of EBV LMP1/BNLF2 and BZLF1 compared to another cluster that expressed neither gene. The two novel clusters differed from each other in their expression of EBV lytic genes, including glycoprotein gene GP350. The first cluster, comprising GP350- LMP1hi cells, expressed high levels of HIF1A and was transcriptionally regulated by HIF1-α. Treatment of LCLs with Pevonedistat, a drug that enhances HIF1-α signaling, markedly induced this cluster. The second cluster, containing GP350+ LMP1hi cells, expressed EBV lytic genes. Host genes that are controlled by super-enhancers (SEs), such as transcription factors MYC and IRF4, had the lowest expression in this cluster. Functionally, the expression of genes regulated by MYC and IRF4 in GP350+ LMP1hi cells were lower compared to other cells. Indeed, induction of EBV lytic reactivation in EBV+ AKATA reduced the expression of these SE-regulated genes. Furthermore, CRISPR-mediated perturbation of the MYC or IRF4 SEs in LCLs induced the lytic EBV gene expression, suggesting that host SEs and/or SE target genes are required for maintenance of EBV latency. Collectively, our study revealed EBV-associated heterogeneity among LCLs that may have functional consequence on host and viral biology.

IL-1ß promotes IL-9-producing Th cell differentiation in IL-2-limiting conditions through the inhibition of BCL6.

IL-9-producing CD4+ T helper cells, termed Th9 cells, differentiate from naïve precursor cells in response to a combination of cytokine and cell surface receptor signals that are elevated in inflamed tissues. After differentiation, Th9 cells accumulate in these tissues where they exacerbate allergic and intestinal disease or enhance anti-parasite and anti-tumor immunity. Previous work indicates that the differentiation of Th9 cells requires the inflammatory cytokines IL-4 and TGF-ß and is also dependent of the T cell growth factor IL-2. While the roles of IL-4 and TGF-ß-mediated signaling are relatively well understood, how IL-2 signaling contributes to Th9 cell differentiation outside of directly inducing the Il9 locus remains less clear. We show here that murine Th9 cells that differentiate in IL-2-limiting conditions exhibit reduced IL-9 production, diminished NF-kB activation and a reduced NF-kB-associated transcriptional signature, suggesting that IL-2 signaling is required for optimal NF-kB activation in Th9 cells. Interestingly, both IL-9 production and the NF-kB transcriptional signature could be rescued by addition of the NF-kB-activating cytokine IL-1ß to IL-2-limiting cultures. IL-1ß was unique among NF-kB-activating factors in its ability to rescue Th9 differentiation as IL-2 deprived Th9 cells selectively induced IL-1R expression and IL-1ß/IL-1R1 signaling enhanced the sensitivity of Th9 cells to limiting amounts of IL-2 by suppressing expression of the Th9 inhibitory factor BCL6. These data shed new light on the intertwined nature of IL-2 and NF-kB signaling pathways in differentiating Th cells and elucidate the potential mechanisms that promote Th9 inflammatory function in IL-2-limiting conditions.

A community-wide epidemic of hepatitis A virus genotype IA associated with consumption of shellfish in Yantai, eastern China, January to March 2020.

During the first quarter of 2020, a considerable increase in reports of symptomatic hepatitis A cases was noted in Yantai, a coastal city in eastern China. This study aimed to characterize the epidemic and identify the probable source. Serum samples from cases with onsets from 1 January to 31 March 2020 and suspected bivalve mollusk samples from the local seafood market were screened for hepatitis A virus (HAV) RNA by PCR amplification and sequencing of the VP1/2A region. We also analyzed the characteristics and risk exposures of these cases. In total, 110 confirmed cases were notified during the epidemic. Among the 103 cases investigated, the median age was 41 years (range: 25-70 years), and 74 (71.8%) were male. Eighty-eight cases (85.4%) reported having eaten shellfish and 72 (69.9%) specifically oysters. HAV RNA was detected and sequenced successfully in 80.2% (69/86) of the cases, as well as in one oyster out of 20 shellfish samples. Phylogenetic analysis revealed that all isolates belonged to a single genotype IA but presented the co-circulation of five distinct genomic sub-lineages. The oyster-derived HAV strain shared over 98.2% nucleotide identity with all clinical strains obtained during the epidemic, particularly 100% homology with the strains of seven cases. These data indicated that contaminated oyster consumption was probably a common source of this epidemic, although multiple HAV strains were involved. We recommend strengthening shellfish surveillance, changing dietary habits in seafood consumption, and encouraging vaccination for target adults in coastal areas with a high prevalence of hepatitis A.

Cool and hot chitosan/platelet-derived growth factor nanofibers for outdoors burns.

Burns and scalds are thermal injuries caused by a large amount of heat accumulation in local tissues. The first cooling emergency is a key step. However, it is hard that in outdoors to find clean water to cool the scald tissue sites. Moreover, most dressings are concentrated on the treatment process today, neglecting the emergency treatment of temperature reduction. In this study, we imported refrigeration in the electrospinning process while using dirty water, rainwater and even urine of outdoors, so that the cooled sterile fibers were directly deposited on the burn and scald wounds, and the cooling emergency was achieved through the dual cooling mechanism. Since this fiber which is made up of cheap fish gelatin contains CuS adopting the green method, it can generate heat and effectively kill bacteria under the irradiation of an illumination lamp at the front end of a spinning device. As a result of the direct deposition, there is an excellent fit between the fibrous membrane and the skin, which reduces the air gap to achieve a better and quick cooling and heating effects. On the same Chitosan/Platelet-derived Growth Factor fiber membrane, this method of cooling first and heating second can shorten the recovery time from 30 days to 21 days. Thus, this treatment strategy has a great potential application prospect in the field of outdoor burn treatment.

Production enhancement of the glycopeptide antibiotic A40926 by an engineered Nonomuraea gerenzanensis strain.

OBJECTIVE: To improve the production of A40926, a combined strategy of constructing the engineered strain and optimizing the medium was implemented. RESULTS: The engineered strain lcu1 with the genetic features of dbv23 deletion and dbv3-dbv20 coexpression increased by 30.6% in the production of A40926, compared to the original strain. In addition, a combined medium called M9 was designed to be further optimized by the central composite design method. The optimized M9 medium was verified to significantly improve the A40926 yield from 257 to 332 mg l-1. CONCLUSIONS: The engineered strain lcu1 could significantly promote A40926 production in the optimized M9 medium, which indicated that the polygenic genetic manipulation and the media optimization played an equally important role in increasing the A40926 yield.

A nanofiber hydrogel derived entirely from ocean biomass for wound healing.

Crustaceans and fish scales in the marine food industry are basically thrown away as waste. This not only wastes resources but also causes environmental pollution. While reducing pollution and waste, biological activity and storage of materials are urgent issues to be solved. In this study, by first preparing dry fibers and then making hydrogels, we prepared a fish scale/sodium alginate/chitosan nanofiber hydrogel (FS-P) by cross-linking the nanofibers in situ. From fish and other organisms, fish gelatin (FG), collagen and CaCO3 were extracted. Fish scale (FS)/sodium alginate/chitosan nanofibers were cross-linked with copper sulfide nanoparticles prepared by a one-step green method to obtain FS-P nanofiber hydrogels under mild conditions without catalyst and additional procedures. These fiber hydrogels not only have good tissue adhesion and tensile properties, but also have the antibacterial effect of natural antibacterial and CuS photothermal synergism, which can achieve 51.32% and 49.96% of the antibacterial effect against Staphylococcus aureus and Escherichia coli respectively, avoiding the generation of superbacteria. The nanofiber hydrogels have 87.56% voidage and 52.68% degradability after 14 days. The combined strategy of using marine bio-based fibers to prepare gels promoted angiogenesis and tissue repair.

Autocrine vitamin D signaling switches off pro-inflammatory programs of TH1 cells.

The molecular mechanisms governing orderly shutdown and retraction of CD4+ type 1 helper T (TH1) cell responses remain poorly understood. Here we show that complement triggers contraction of TH1 responses by inducing intrinsic expression of the vitamin D (VitD) receptor and the VitD-activating enzyme CYP27B1, permitting T cells to both activate and respond to VitD. VitD then initiated the transition from pro-inflammatory interferon-γ+ TH1 cells to suppressive interleukin-10+ cells. This process was primed by dynamic changes in the epigenetic landscape of CD4+ T cells, generating super-enhancers and recruiting several transcription factors, notably c-JUN, STAT3 and BACH2, which together with VitD receptor shaped the transcriptional response to VitD. Accordingly, VitD did not induce interleukin-10 expression in cells with dysfunctional BACH2 or STAT3. Bronchoalveolar lavage fluid CD4+ T cells of patients with COVID-19 were TH1-skewed and showed de-repression of genes downregulated by VitD, from either lack of substrate (VitD deficiency) and/or abnormal regulation of this system.

Mitochondrial C5aR1 activity in macrophages controls IL-1ß production underlying sterile inflammation.

While serum-circulating complement destroys invading pathogens, intracellularly active complement, termed the "complosome," functions as a vital orchestrator of cell-metabolic events underlying T cell effector responses. Whether intracellular complement is also nonredundant for the activity of myeloid immune cells is currently unknown. Here, we show that monocytes and macrophages constitutively express complement component (C) 5 and generate autocrine C5a via formation of an intracellular C5 convertase. Cholesterol crystal sensing by macrophages induced C5aR1 signaling on mitochondrial membranes, which shifted ATP production via reverse electron chain flux toward reactive oxygen species generation and anaerobic glycolysis to favor IL-1ß production, both at the transcriptional level and processing of pro­IL-1ß. Consequently, atherosclerosis-prone mice lacking macrophage-specific C5ar1 had ameliorated cardiovascular disease on a high-cholesterol diet. Conversely, inflammatory gene signatures and IL-1ß produced by cells in unstable atherosclerotic plaques of patients were normalized by a specific cell-permeable C5aR1 antagonist. Deficiency of the macrophage cell-autonomous C5 system also protected mice from crystal nephropathy mediated by folic acid. These data demonstrate the unexpected intracellular formation of a C5 convertase and identify C5aR1 as a direct modulator of mitochondrial function and inflammatory output from myeloid cells. Together, these findings suggest that the complosome is a contributor to the biologic processes underlying sterile inflammation and indicate that targeting this system could be beneficial in macrophage-dependent diseases, such as atherosclerosis.