Dipeptidyl peptidase 4 interacts with porcine coronavirus PHEV spikes and mediates host range expansion.

Porcine hemagglutinating encephalomyelitis virus (PHEV), a neurotropic betacoronavirus, is prevalent in natural reservoir pigs and infects mice. This raises concerns about host jumping or spillover, but little is known about the cause of occurrence. Here, we revealed that dipeptidyl peptidase 4 (DPP4) is a candidate binding target of PHEV spikes and works as a broad barrier to overcome. Investigations of the host breadth of PHEV confirmed that cells derived from pigs and mice are permissive to virus propagation. Both porcine DPP4 and murine DPP4 have high affinity for the viral spike receptor-binding domain (RBD), independent of their catalytic activity. Loss of DPP4 expression results in limited PHEV infection. Structurally, PHEV spike protein binds to the outer surface of blades IV and V of the DPP4 ß-propeller domain, and the DPP4 residues N229 and N321 (relative to human DPP4 numbering) participate in RBD binding via its linked carbohydrate entities. Removal of these N-glycosylations profoundly enhanced the RBD-DPP4 interaction and viral invasion, suggesting they act as shielding in PHEV infection. Furthermore, we found that glycosylation, rather than structural differences or surface charges, is more responsible for DPP4 recognition and species barrier formation. Overall, our findings shed light on virus-receptor interactions and highlight that PHEV tolerance to DPP4 orthologs is a putative determinant of its cross-species transmission or host range expansion.IMPORTANCEPHEV is a neurotropic betacoronavirus that is circulating worldwide and has raised veterinary and economic concerns. In addition to being a reservoir species of pigs, PHEV can also infect wild-type mice, suggesting a "host jump" event. Understanding cross-species transmission is crucial for disease prevention and control but remains to be addressed. Herein, we show that the multifunctional receptor DPP4 plays a pivotal role in the host tropism of PHEV and identifies the conserved glycosylation sites in DPP4 responsible for this restriction. These findings highlight that the ability of PHEV to utilize DPP4 orthologs potentially affects its natural host expansion.

Development and evaluation of inactivated vaccines incorporating a novel Senecavirus A strain-based Immunogen and various adjuvants in mice.

Porcine idiopathic vesicular disease (PIVD), one of several clinically indistinguishable vesicular diseases of pigs, is caused by the emerging pathogen Senecavirus A (SVA). Despite the widespread prevalence of porcine SVA infection, no effective commercial vaccines for PIVD prevention and control are available, due to high costs associated with vaccine testing in pigs, considerable SVA diversity, and SVA rapid evolution. In this study, SVA CH/JL/2022 (OP562896), a novel mutant SVA strain derived from an isolate obtained from a pig farm in Jilin Province, China, was inactivated then combined with four adjuvants, MONTANIDETM GEL02 PR (GEL 02), MONTANIDETM ISA 201 VG (ISA 201), MONTANIDETM IMG 1313 VG N (IMS1313), or Rehydragel LV (LV). The resulting inactivated SVA CH/JL/2022 vaccines were assessed for efficacy in mice and found to induce robust in vivo lymphocyte proliferation responses and strong IgG1, IgG2a, and neutralizing antibody responses with IgG2a/IgG1 ratios of <1. Furthermore, all vaccinated groups exhibited significantly higher levels of serum cytokines IL-2, IL-4, IL-6, and IFN as compared to unvaccinated mice. These results indicate that all vaccines elicited both Th1 and Th2 responses, with Th2 responses predominating. Moreover, vaccinated mice exhibited enhanced resistance to SVA infection, as evidenced by reduced viral RNA levels and SVA infection-induced histopathological changes. Collectively, our results demonstrate that the SVA-GEL vaccine induced more robust immunological responses in mice than did the other three vaccines, thus highlighting the potential of SVA-GEL to serve an effective tool for preventing and controlling SVA infection.

Online photography intervention reduces internet addiction during COVID-19: A randomised controlled trial.

The COVID-19 pandemic has escalated concerns regarding internet addiction. This study investigated the impact of an online photograph intervention on internet addiction and identified its effectiveness mechanisms. Individuals diagnosed with internet addiction (internet addiction test score > 36, Mage = 22.85, 78.6% women) were randomly allocated to either the intervention (n = 64) or control (n = 81) groups. For 10 days, the intervention group was tasked with photographing "things that make you feel a sense of control in life" and writing 100-word descriptions explaining the reason for the sense of control elicited by the image. Meanwhile, the control group participated only in the assessments. Assessments were conducted before the intervention, immediately afterward and 7 days later. A significant interaction between group and time was observed, with the intervention group showing a notable reduction in internet addiction and an increase in sense of control. The sense of control at post-test significantly mediated the intervention's effect on reducing internet addiction at follow-up, but only among individuals with high perceived social support. The findings indicate that online photography interventions can effectively diminish internet addiction, with a sense of control acting as a potent mechanism for intervention, particularly in individuals with substantial perceived social support.

[Applications of ion chromatography for the analysis of Chinese herbal medicine components].

Ion chromatography (IC) is a novel high performance liquid chromatographic technique that is suitable for the separation and analysis of ionic substances in different matrix samples. Since 1975, it has been widely used in many fields, such as the environment, energy, food, and medicine. IC compensates for the separation limitations of traditional gas chromatography and high performance liquid chromatography and can realize the qualitative analysis and quantitative detection of strongly polar components. This chromatographic technique features not only simple operations but also rapid analysis. The sensors used in IC are characterized by high sensitivity and selectivity, and the technique can simultaneously separate and determine multiple components. Several advances in IC instrumentation and chromatographic theories have been developed in recent years. IC can analyze various types of samples, including ions, sugars, amino acids, and organic acids (bases). Chinese herbal medicines are typically characterized by highly complex chemical compositions and may contain carbohydrates, proteins, alkaloids, and other active components. They also contain toxic residues such as sulfur dioxide, which may be produced during the processing of medicinal materials. Therefore, the analysis and elucidation of the precise chemical constituents of Chinese herbal medicines present key problems that must be resolved in modern Chinese herbal medicine research. In this context, IC has become an important method for analyzing and identifying the complex components of Chinese herbal medicines because this method is suitable for detecting a single active ingredients among complex components. This paper introduces the different types and principles of IC as well as research progress in this technique. As the applications of IC-based methods in pharmaceutical science, cell biology, and microbiology increase, further development is necessary to expand the applications of this technique. The development of innovative techniques has enabled IC technologies to achieve higher analytical sensitivity, better selectivity, and wider application. The components of Chinese herbal medicines can be divided into endogenous and exogenous components according to their source: endogenous components include glycosides, amino acids, and organic acids, while exogenous components include toxic residues such as sulfur dioxide. Next, the applications of IC to the complex components of Chinese herbal medicines in recent decades are summarized. The most commonly used IC technologies and methods include ion exchange chromatography and conductivity detection. The advantages of IC for the analysis of alkaloids have been demonstrated. This method exhibits better characteristics than traditional analytical methods. However, the applications of IC for the speciation analysis of inorganic anions are limited. Moreover, few reports on the direct application of the technique for the determination of the main active substances in Chinese herbal medicines, including flavonoids, phenylpropanoids, and steroids, have been reported. Finally, this paper reviews new IC technologies and their application progress in Chinese herbal medicine, focusing on their prospects for the effective separation and analysis of complex components. In particular, we discuss the available sample (on-line) pretreatment technologies and explore possible technologies for the selective and efficient enrichment and separation of different components. Next, we assess innovative research on solid-phase materials that can improve the separation effect and analytical sensitivity of IC. We also describe the features of multidimensional chromatography, which combines the advantages of various chromatographic techniques. This review provides a theoretical reference for the further development of IC technology for the analysis of the complex chemical components of Chinese herbal medicines.

Benign mixed mammary tumor with sebaceous differentiation in a dog.

We describe here a case of canine mammary benign mixed tumor with sebaceous metaplasia in the right fifth mammary gland of an eight-year-old, intact female Poodle dog. Grossly, the mass was firm with off-white, poorly lobulated cut surfaces. Histologically, the luminal epithelial cells and myoepithelial cells proliferated with cartilage formation and focal squamous metaplasia. Moreover, a large number of nests of various sizes, which were filled with foamy cells in the center and associated with basaloid reserve-like cells in the periphery, showed sebaceous gland-like structures. Immunohistochemically, myoepithelial cells and reserve-like cells in the metaplastic sebaceous gland-like structures were CK14, α-smooth muscle actin (α-SMA) and p63 positive, suggesting a possibility that these two components may have a common cell of origin.

Genistein is effective in inhibiting Orf virus infection in vitro by targeting viral RNA polymerase subunit RPO30 protein.

Orf virus (ORFV), a typical member of the genus Parapoxvirus, Poxvirus family, causes a contagious pustular dermatitis in sheep, goats, and humans. Poxviruses encode a multisubunit DNA-dependent RNA polymerase (vRNAP) that carries out viral gene expression in the host cytoplasm, which is a viral factor essential to poxvirus replication. Due to its vital role in viral life, vRNAP has emerged as one of the potential drug targets. In the present study, we investigated the antiviral effect of genistein against ORFV infection. We provided evidence that genistein exerted antiviral effect through blocking viral genome DNA transcription/replication and viral protein synthesis and reducing viral progeny, which were dosedependently decreased in genistein-treated cells. Furthermore, we identified that genistein interacted with the vRNAP RPO30 protein by CETSA, molecular modeling and Fluorescence quenching, a novel antiviral target for ORFV. By blocking vRNAP RPO30 protein using antibody against RPO30, we confirmed that the inhibitory effect exerted by genistein against ORFV infection is mediated through the interaction with RPO30. In conclusion, we demonstrate that genistein effectively inhibits ORFV transcription in host cells by targeting vRNAP RPO30, which might be a promising drug candidate against poxvirus infection.

CD97 negatively regulates the innate immune response against RNA viruses by promoting RNF125-mediated RIG-I degradation.

The G protein-coupled receptor ADGRE5 (CD97) binds to various metabolites that play crucial regulatory roles in metabolism. However, its function in the antiviral innate immune response remains to be determined. In this study, we report that CD97 inhibits virus-induced type-I interferon (IFN-I) release and enhances RNA virus replication in cells and mice. CD97 was identified as a new negative regulator of the innate immune receptor RIG-I, and RIG-1 degradation led to the suppression of the IFN-I signaling pathway. Furthermore, overexpression of CD97 promoted the ubiquitination of RIG-I, resulting in its degradation, but did not impact its mRNA expression. Mechanistically, CD97 upregulates RNF125 expression to induce RNF125-mediated RIG-I degradation via K48-linked ubiquitination at Lys181 after RNA virus infection. Most importantly, CD97-deficient mice are more resistant than wild-type mice to RNA virus infection. We also found that sanguinarine-mediated inhibition of CD97 effectively blocks VSV and SARS-CoV-2 replication. These findings elucidate a previously unknown mechanism through which CD97 negatively regulates RIG-I in the antiviral innate immune response and provide a molecular basis for the development of new therapeutic strategies and the design of targeted antiviral agents.

The role of lysosomes as intermediates in betacoronavirus PHEV egress from nerve cells.

IMPORTANCE: Betacoronaviruses, including severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and mouse hepatitis virus (MHV), exploit the lysosomal exocytosis pathway for egress. However, whether all betacoronaviruses members use the same pathway to exit cells remains unknown. Here, we demonstrated that porcine hemagglutinating encephalomyelitis virus (PHEV) egress occurs by Arl8b-dependent lysosomal exocytosis, a cellular egress mechanism shared by SARS-CoV-2 and MHV. Notably, PHEV acidifies lysosomes and activates lysosomal degradative enzymes, while SARS-CoV-2 and MHV deacidify lysosomes and limit the activation of lysosomal degradative enzymes. In addition, PHEV release depends on V-ATPase-mediated lysosomal pH. Furthermore, this is the first study to evaluate ßCoV using lysosome for spreading through the body, and we have found that lysosome played a critical role in PHEV neural transmission and brain damage caused by virus infection in the central nervous system. Taken together, different betacoronaviruses could disrupt lysosomal function differently to exit cells.

A Dual Blind Watermarking Method for 3D Models Based on Normal Features.

Digital watermarking technology is an important means to effectively protect three-dimensional (3D) model data. Among them, "blind detection" and "robustness" are key and difficult points in the current research of digital watermarking technology based on 3D models. In order to realize the blind detection of a watermark and improve its robustness against various common attacks at the same time, this paper proposes a dual blind watermarking method based on the normal feature of the centroid of first-ring neighboring points. The local spherical coordinate system is constructed by calculating two different normal vectors, and the first pattern watermark and the second random binary sequence watermark are embedded, respectively. The experimental results show that this method can not only realize the blind detection of dual watermarks, but also have the ability to resist common attacks such as translation, rotation, scaling, cropping, simplification, smoothing, noise, and vertex reordering to a certain extent.

Experimental Investigation of Tool Wear and Machining Quality of BTA Deep-Hole Drilling in Low-Carbon Alloy Steel SA-5083.

Nuclear power tube plates are made from the high-strength, low-carbon alloy steel SA-5083, which has high values of toughness and plasticity, though it is forged with poor consistency and entails serious work hardening. It requires a large number of deep holes with a high machining accuracy and high surface quality to be processed. However, the quality of the processed holes is often not up to the standard of the Boring and Trepanning Association (BTA) for the deep-hole drilling of tube plates; this has led to deep-hole processing becoming a bottleneck in the manufacture of steam generators for the main equipment of nuclear power islands. The variation laws of the diameter, roundness, perpendicularity, roughness, microhardness, and residual stress in relation to the feed, speed, and drilling depth are explored in the macro- and micro-dimensions; also explored is the wear morphology of BTA drills. The internal influence mechanisms between them are revealed in order to provide a scientific basis for the control of surface quality and machining accuracy as well as the optimization of process parameters. Our research results indicate that the guide block wear is mainly concentrated at the top 1-2 mm and that the drilling depth and feed have a great influence on the machining diameter. The hole wall roughness is between 0.3 and 0.6 µm, the maximum microhardness is about 2.15 times the hardness of the matrix material, and the residual stress is compressive stress. With increases in the feed and drilling depth, the hole diameter and the roughness increase. With an increase in the speed, the roughness decreases and the compressive stress of the BTA deep-hole drilling wall increases.

Nafamostat mesilate prevented caerulein-induced pancreatic injury by targeting HDAC6-mediated NLRP3 inflammasome activation.

OBJECTIVE: Nafamostat mesilate (NM), a synthetic broad-spectrum serine protease inhibitor, has been commonly used for treating acute pancreatitis (AP) and other inflammatory-associated diseases in some East Asia countries. Although the potent inhibitory activity against inflammation-related proteases (such as thrombin, trypsin, kallikrein, plasmin, coagulation factors, and complement factors) is generally believed to be responsible for the anti-inflammatory effects of NM, the precise target and molecular mechanism underlying its anti-inflammatory activity in AP treatment remain largely unknown. METHODS: The protection of NM against pancreatic injury and inhibitory effect on the NOD-like receptor protein 3 (NLRP3) inflammasome activation were investigated in an experimental mouse model of AP. To decipher the molecular mechanism of NM, the effects of NM on nuclear factor kappa B (NF-κB) activity and NF-κB mediated NLRP3 inflammasome priming were examined in lipopolysaccharide (LPS)-primed THP-1 cells. Additionally, the potential of NM to block the activity of histone deacetylase 6 (HDAC6) and disrupt the association between HDAC6 and NLRP3 was also evaluated. RESULTS: NM significantly suppressed NLRP3 inflammasome activation in the pancreas, leading to a reduction in pancreatic inflammation and prevention of pancreatic injury during AP. NM was found to interact with HDAC6 and effectively inhibit its function. This property allowed NM to influence HDAC6-dependent NF-κB transcriptional activity, thereby blocking NF-κB-driven transcriptional priming of the NLRP3 inflammasome. Furthermore, NM exhibited the potential to interfere the association between HDAC6 and NLRP3, impeding HDAC6-mediated intracellular transport of NLRP3 and ultimately preventing NLRP3 inflammasome activation. CONCLUSIONS: Our current work has provided valuable insight into the molecular mechanism underlying the immunomodulatory effect of NM in the treatment of AP, highlighting its promising application in the prevention of NLRP3 inflammasome-associated inflammatory pathological damage.

Prevalence of sufficient physical activity among general adult population and sub-populations with chronic conditions or disability in the USA.

BACKGROUND: Recently, the World Health Organization (WHO) released an updated global guideline on physical activity and sedentary behavior, including recommendations for sub-populations living with chronic conditions or disabilities. We aimed to examine the prevalence of meeting the WHO recommendations among these sub-populations in the USA. METHODS: We conducted a cross-sectional study using data from the 2017 to 2018 cycle of the National Health and Nutrition Examination Survey (NHANES). RESULTS: We revealed variations in physical activity levels among individuals with chronic conditions and disability. US adults with diabetes, hypertension or disability had a lower prevalence of recommended physical activity levels than the general population. In addition, certain demographic groups such as being female, older and having lower socioeconomic status were associated with a lower likelihood of meeting the WHO recommendations on physical activity. CONCLUSIONS: Our findings underscore the importance of promoting physical activity levels among US adults, especially those with older age, low socioeconomic status, hypertension and disability.

A practical nomogram and risk stratification system for predicting survival outcomes in neuroblastoma patients: a SEER population-based study.

BACKGROUND: Neuroblastoma (NB) is a childhood malignancy with marked heterogeneity, resulting in highly variable outcomes among patients. This study aims to establish a novel nomogram and risk stratification system to predict the overall survival (OS) for patients with NB. METHODS: We analyzed neuroblastoma patients from the Surveillance, Epidemiology, and End Results (SEER) database between 2004 and 2015. The nomogram was constructed using independent risk factors for OS, identified through univariate and multivariate Cox regression analyses. The accuracy of this nomogram was evaluated with the concordance index, receiver operating characteristic curve, calibration curve, and decision curve analysis. In addition, we developed a risk stratification system based on the total score of each patient in the nomogram. RESULTS: A total of 2185 patients were randomly assigned to the training group and the testing group. Six risk factors, including age, chemotherapy, brain metastases, primary site, tumor stage, and tumor size, were identified in the training group. Using these factors, a nomogram was constructed to predict 1-, 3-, and 5-year OS of NB patients. This model exhibited superior accuracy in the training and testing groups, exceeding traditional tumor stage prediction. Subgroup analysis suggested worse prognosis for retroperitoneal origin in the intermediate-risk group and adrenal gland origin in the high-risk group compared to other sites. Additionally, the prognosis for high-risk patients significantly improved after surgery. We also developed a web application to make the nomogram more user-friendly in clinical practices. CONCLUSION: This nomogram demonstrates excellent accuracy and reliability, offering more precise personalized prognostic predictions to clinical patients.

Chitosan-based mussel-inspired hydrogel for rapid self-healing and high adhesion of tissue adhesion and wound dressings.

The hydrogel wound dressing with self-healing and adhesive property can provide better protection to the wound and prolong the service life of the material. Inspired by mussels, a high-adhesion, injectable, self-healing and antibacterial hydrogel was designed in this study. The lysine (Lys) and the catechol compound 3,4-dihydroxyphenylacetic acid (DOPAC) were grafted onto chitosan (CS). The presence of catechol group endows the hydrogel strong adhesion and antioxidation. In the experiment of wound healing in vitro, the hydrogel can adhere to the wound surface and promote wound heal. In addition, it has been proved the hydrogel has good antibacterial properties against S. aureus and E. coli. The treatment of CLD hydrogel, the degree of wound inflammation was significantly alleviated. The levels of TNF-α, IL-1ß, IL-6 and TGF-ß1 were reduced from 39.8379 %, 31.6768 %, 32.1015 % and 38.4911 % to 18.5931 %, 12.2275 %, 13.0524 % and 16.9959 %, respectively. And the levels of PDGFD and CD31 were increased from 35.6054 %, 21.7394 % to 51.8555 %, 43.9326 %, respectively. These results indicated that the CLD hydrogel has a good ability to promote angiogenesis, thickening of skin and epithelial structures.

Orf virus DNA prime-protein boost strategy is superior to adenovirus-based vaccination in mice and sheep.

Contagious ecthyma (Orf), an acute and highly contagious zoonosis, is prevalent worldwide. Orf is caused by Orf virus (ORFV), which mainly infects sheep/goats and humans. Therefore, effective and safe vaccination strategies for Orf prevention are needed. Although immunization with single-type Orf vaccines has been tested, heterologous prime-boost strategies still need to be studied. In the present study, ORFV B2L and F1L were selected as immunogens, based on which DNA, subunit and adenovirus vaccine candidates were generated. Of note, heterologous immunization strategies using DNA prime-protein boost and DNA prime-adenovirus boost in mice were performed, with single-type vaccines as controls. We have found that the DNA prime-protein boost strategy induces stronger humoral and cellular immune responses than DNA prime-adenovirus boost strategy in mice, which was confirmed by the changes in specific antibodies, lymphocyte proliferation and cytokine expression. Importantly, this observation was also confirmed when these heterologous immunization strategies were performed in sheep. In summary, by comparing the two immune strategies, we found that DNA prime-protein boost strategy can induce a better immune response, which provides a new attempt for exploring Orf immunization strategy.

Orf virus induces complete autophagy to promote viral replication via inhibition of AKT/mTOR and activation of the ERK1/2/mTOR signalling pathway in OFTu cells.

Orf virus (ORFV) is the causative agent of contagious ecthyma, which is an important zoonotic pathogen with a widespread distribution affecting sheep, goats and humans. Our previous research showed that autophagy can be induced in host cells by ORFV infection. However, the exact mechanism of ORFV-induced autophagy remains unknown. In this study, we investigated the underlying mechanisms of autophagy induced by ORFV in OFTu cells and the impact of autophagy on ORFV replication. By using specific autophagy inhibitors and activators, Western blotting, immunofluorescence and transmission electron microscopy imaging, we confirmed that ORFV infection triggered intracellular autophagosome accumulation and the activation of autophagic flux. Moreover, ORFV-induced autophagic activity was found to rely on an increase in the phosphorylation of tuberous sclerosis complex 2 (TSC2) and a decrease in the phosphorylation of mammalian target of rapamycin (mTOR), which is mediated by the suppression of the PI3K/AKT/mTOR signalling pathway and activation of the ERK1/2/mTOR signalling pathway. Furthermore, we investigated the role of mTOR-mediated autophagy during ORFV replication using pharmacological agents and demonstrated that ORFV-induced autophagy correlated positively with viral replication. Taken together, our data reveal the pathways of ORFV-induced autophagy and the impact of autophagy on ORFV replication, providing new insights into ORFV pathogenesis.

Speckle autocorrelation separation for multi-target scattering imaging.

Imaging through scattering media remains a big challenge in optics while the single-shot non-invasive speckle autocorrelation technique (SAT) is well-known as a promising way to handle it. However, it usually cannot recover a large-scale target or multiple isolated small ones due to the limited effective range of the optical memory effect (OME). In this paper, we propose a multi-target scattering imaging scheme by combining the traditional SA algorithm with a Deep Learning (DL) strategy. The basic idea is to extract each autocorrelation component of every target from the autocorrelation result of a mixed speckle using a suitable DL method. Once we get all the expected autocorrelation components, a typical phase retrieval algorithm (PRA) could be applied to reveal the shapes of all those corresponding small targets. In our experimental demonstration, up to five isolated targets are successfully recovered.

Natural flavonoid sinensetin inhibits cisplatin-induced pyroptosis and attenuates intestinal injury.

The demand of exploring strategies to enhance chemotherapy drug efficacy and alleviate adverse effects by using natural compounds is increasing. Sinensetin (SIN) is a kind of natural flavonoids with anti-inflammatory activities. However, its protective impact on chemotherapy-induced adverse effects has not been well demonstrated. Here, we found that SIN could inhibit Cisplatin-induced release of proinflammatory cellular contents and inflammatory cell death-pyroptosis. In addition, Cisplatin-induced activation of gasdermin E (GSDME), a critical mediator of chemotherapy-induced tissue injury, could also be reversed by SIN. Furthermore, SIN impaired Cisplatin-induced intracellular damages, including ROS release and DNA damages. Importantly, SIN was able to alleviate intestinal injury in Cisplatin-challenged mice, which was accompanied by the decrease of lytic cell death and immune cell infiltration. Of note, SIN administration did not reverse Cisplatin-caused tumor suppression in vivo. In conclusion, our result provides a potential application of SIN to reduce Cisplatin-caused adverse effects, without impairing its anti-tumor capacity.

Oncolytic Parapoxvirus induces Gasdermin E-mediated pyroptosis and activates antitumor immunity.

The advantage of oncolytic viruses (OV) in cancer therapy is their dual effect of directly killing tumours while prompting anti-tumour immune response. Oncolytic parapoxvirus ovis (ORFV) and other OVs are thought to induce apoptosis, but apoptosis, being the immunogenically inert compared to other types of cell death, does not explain the highly inflamed microenvironment in OV-challenged tumors. Here we show that ORFV and its recombinant therapeutic derivatives are able to trigger tumor cell pyroptosis via Gasdermin E (GSDME). This effect is especially prominent in GSDME-low tumor cells, in which ORFV-challenge pre-stabilizes GSDME by decreasing its ubiquitination and subsequently initiates pyroptosis. Consistently, GSDME depletion reduces the proportion of intratumoral cytotoxic T lymphocytes, pyroptotic cell death and the success of tumor ORFV virotherapy. In vivo, the OV preferentially accumulates in the tumour upon systemic delivery and elicits pyroptotic tumor killing. Consequentially, ORFV sensitizes immunologically 'cold' tumors to checkpoint blockade. This study thus highlights the critical role of GSDME-mediated pyroptosis in oncolytic ORFV-based antitumor immunity and identifies combinatorial cancer therapy strategies.