PIM1-HDAC2 axis modulates intestinal homeostasis through epigenetic modification.

The mucosal barrier is crucial for intestinal homeostasis, and goblet cells are essential for maintaining the mucosal barrier integrity. The proviral integration site for Moloney murine leukemia virus-1 (PIM1) kinase regulates multiple cellular functions, but its role in intestinal homeostasis during colitis is unknown. Here, we demonstrate that PIM1 is prominently elevated in the colonic epithelia of both ulcerative colitis patients and murine models, in the presence of intestinal microbiota. Epithelial PIM1 leads to decreased goblet cells, thus impairing resistance to colitis and colitis-associated colorectal cancer (CAC) in mice. Mechanistically, PIM1 modulates goblet cell differentiation through the Wnt and Notch signaling pathways. Interestingly, PIM1 interacts with histone deacetylase 2 (HDAC2) and downregulates its level via phosphorylation, thereby altering the epigenetic profiles of Wnt signaling pathway genes. Collectively, these findings investigate the unknown function of the PIM1-HDAC2 axis in goblet cell differentiation and ulcerative colitis/CAC pathogenesis, which points to the potential for PIM1-targeted therapies of ulcerative colitis and CAC.

Multi-step strategies for synergistic treatment of urinary tract infections based on D-xylose-decorated antimicrobial peptide carbon dots.

Urinary tract infections (UTIs) caused by Uropathogenic Escherichia coli (UPEC), often reoccur due to the formation of intracellular bacterial colonies (IBCs) and antibiotic resistance. Given the significance of YadC for UPEC infection in our previous study, we developed D-xylose-decorated ɛ-poly-L-lysine (εPL)-based carbon dots (D-xyl@εPLCDs) that can be traced, and employed multi-step approaches to elucidate the functional roles of D-xyl@εPLCDs in UPEC infection. Compared to undecorated particles, D-xyl@εPLCDs demonstrate YadC-dependent bacterial targeting and exhibit enhanced bactericidal activities both intracellularly and extracellularly. Moreover, pre-treatment of D-xyl@εPLCDs before infection blocked the subsequent adhesion and invasion of UPEC to bladder epithelial cells 5637. Increase of ROS production and innate immune responses were observed in bladder epithelial cells 5637 treated with D-xyl@εPLCDs. In addition, treatment of D-xyl@εPLCDs post-infection facilitated clearance of UPEC in the bladders of the UTI mouse model, and reduced ultimate number of neutrophils, macrophages and inflammatory responses raised by invaded bacteria. Collectively, we presented a comprehensive evaluating system to show that D-xyl@εPLCDs exhibits superior bactericidal effects against UPEC, making them a promising candidate for drug development in clinical UTI therapeutics.

Bacterial indole-3-lactic acid affects epithelium-macrophage crosstalk to regulate intestinal homeostasis.

Tryptophan and its derivatives perform a variety of biological functions; however, the role and specific mechanism of many tryptophan derivatives in intestinal inflammation remain largely unclear. Here, we identified that an Escherichia coli strain (Ec-TMU) isolated from the feces of tinidazole-treated individuals, and indole-3-lactic acid (ILA) in its supernatant, decreased the susceptibility of mice to dextran sulfate sodium-induced colitis. Ec-TMU and ILA contribute to the relief of colitis by inhibiting the production of epithelial CCL2/7, thereby reducing the accumulation of inflammatory macrophages in vitro and in vivo. Mechanistically, ILA downregulates glycolysis, NF-κB, and HIF signaling pathways via the aryl hydrocarbon receptor, resulting in decreased CCL2/7 production in epithelial cells. Clinical evidence suggests that the fecal ILA level is negatively correlated with the progression indicator of inflammatory bowel diseases. These results demonstrate that ILA has the potential to regulate intestinal homeostasis by modulating epithelium-macrophage interactions.

HSPA8 regulates anti-bacterial autophagy through liquid-liquid phase separation.

HSPA8 (heat shock protein family A (Hsp70) member 8) plays a significant role in the autophagic degradation of proteins, however, its effect on protein stabilization and anti-bacterial autophagy remains unknown. Here, it is discovered that HSPA8, as a binding partner of RHOB and BECN1, induce autophagy for intracellular bacteria clearance. Using its NBD and LID domains, HSPA8 physically binds to RHOB residues 1-42 and 89-118 as well as to BECN1 ECD domain, preventing RHOB and BECN1 degradation. Intriguingly, HSPA8 contains predicted intrinsically disordered regions (IDRs), and drives liquid-liquid phase separation (LLPS) to concentrate RHOB and BECN1 into HSPA8-formed liquid-phase droplets, resulting in improved RHOB and BECN1 interactions. Our study reveals a novel role and mechanism of HSPA8 in modulating anti-bacterial autophagy, and highlights the effect of LLPS-related HSPA8-RHOB-BECN1 complex on enhancing protein interaction and stabilization, which improves the understanding of autophagy-mediated defense against bacteria.

Nasal delivery of an immunotherapeutic vaccine in thermosensitive hydrogel against allergic asthma.

Asthma poses a significant threat to public health, with an estimated burden of over 334 million people worldwide. Available treatments are often inadequate. We developed a thermo-sensitive hydrogel vaccine containing allergen and FK506 that induced immune tolerance via intranasal administration to treat experimental allergic asthma. The hydrogel delivery system was formulated based on Poloxamer 407 (P407), Carbopol 974P NF, and Polyoxyl 15 hydroxystearate (Kolliphor HS15, HS15). It flowed freely at room temperature and rapidly formed a hydrogel in the nasal cavity once the temperature rose over 33 °C. Ovalbumin and FK506 were slowly released from the hydrogel form and their mucosal residence time was significantly prolonged compared to the liquid formulation. In both an OVA-induced asthma model and an HDM-induced asthma model, the vaccines formulated in hydrogel gave lower levels of eosinophilic inflammation, and airway remodeling. The reduction of lung function was ameliorated, and Foxp3-expressing CD4 + Treg cells were significantly higher. The frequency of Foxp3 + Tregs in lung-draining lymph nodes (dLNs) was correlated with the amelioration. Depletion of Foxp3 + Treg cells abolished the beneficial effects of the allergen/FK506 hydrogel vaccinations. Thus, the allergen/FK506 hydrogel formulation has the potential to be a delivery system for therapeutic allergy vaccines to induce immune tolerance.

Distribution and accumulation of cadmium in soil under wheat-cultivation system and human health risk assessment in coal mining area of China.

The soil contamination caused by the discharge of cadmium (Cd) from coal mining activities has aroused continuous attention due to the detrimental effects on the human health. This study aimed to investigate the characteristics on distribution of Cd in soils and its accumulation in wheat grains under wheat-cultivation system, and further assess the human health risks to adults and children. 58 soils and wheat samples in pairs from Linhuan coal mining area, Anhui Province were collected and analyzed. Results showed that the concentrations of Cd in 17.24% of soil samples exceeded the limit value established by the Ministry of Ecology and Environment. The ordinary kriging interpolation displayed that the spatial variability of Cd concentrations in soils was mainly influenced by coal mining activities. The transfer capacity of Cd from soils to wheat roots was greater than that from the wheat roots to grains. Multiple linear regression model clarified that soil pH and exchangeable Cd fraction in soils were the critical factors affecting the Cd accumulation in wheat grains. The carcinogenic risk of Cd levels in our studied wheat grains was a concern but still within the acceptable range, while their non-carcinogenic hazard was negligible for adults and children. The calculation results were in accord with the uncertainty analysis conclusion based on Monte Carlo simulation. The study was expected to promote the source management and control strategy of reducing tailing discharge, and providing scientific references for current soil remediation and land degradation prevention.

Chromium in Chinese coals: geochemistry and environmental impacts associated with coal-fired power plants.

Chromium (Cr), one of the prime hazardous trace elements in coals, may engender adverse effects on eco-environment and threaten human health during utilization of coal. Based on the samples obtained in our laboratory and published literature, the abundance and modes of occurrence of Cr in Chinese coals, and the environmental impacts associated with coal-fired power plants (CFPPs) were elucidated in this study. With a total of 1397 sets of data, the mean concentration of Cr in Chinese coals was calculated as 21.33 µg/g by the "reserve-concentration" weighted calculation method. Spatially, the average Cr contents increased gradually from North China to South China. Temporally, coals from T3, E-N and P2 were relatively enriched in Cr compared to the other geological time. The Cr concentration in coal varied with different coal ranks. The geological factors accounted for Cr enrichment in coals could be divided into the primary, secondary and epigenetic processes. Higher percentages of organically Cr occurred in low-rank coals, while inorganically associated Cr was mainly found in clay minerals. After coal combustion, most of Cr was enriched in solid wastes (e.g., fly ash and bottom ash). The leaching of Cr from solid wastes in the rainy season (especially acid rain) needs to be a concern for CFPPs. It was estimated that the atmospheric emission of Cr from CFPPs increased annually from 2015 to 2019 and reached approximately 159 tons in 2019.

Speciation, bioaccessibility and human health risk assessment of chromium in solid wastes from an ultra-low emission coal-fired power plant, China.

Chromium (Cr) in solid wastes from ultra-low emission (ULE) coal-fired power plants (CFPPs) could engender adverse effects on environment and human health. Hence, solid waste samples containing bottom ash, fly ash, gypsum and sludge were collected from a typical ULE CFPP in China to study the distribution, speciation, bioaccessibility and human health risk of Cr. The results showed that Cr was depleted in gypsum, whereas significantly enriched in bottom ash, fly ash and sludge comparing with feed coal. The ratios of Cr(VI) to total Cr in solid wastes were relatively low, but the increase of flow fractions in Cr chemical binding forms implied the deterioration of environmental stability. Based on the in vitro simulated digestion methods of solubility bioavailability research consortium (SBRC) and physiologically based extraction test (PBET), the bioaccessibility of Cr in the gastric and intestinal phases reached the highest values in either gypsum or sludge. After incorporating bioaccessibility in human health risk assessment, the carcinogenic risk (CR) within acceptable limits of Cr in solid wastes to adults and children was concluded, with the non-carcinogenic hazard quotient (HQ) was all within the safety threshold. The Monte Carlo model was applied to evaluate the uncertainty analysis of human health risk assessment at 5% and 95% confidence interval, and the fitting results were consistent with the calculation results of the carcinogenic and non-carcinogenic risk for adults and children. This study is expected to provide insights for the integration of bioaccessibility into the health risk assessment of Cr in solid wastes from ULE CFPPs, thus is conducive to the disposal of solid wastes and human health protection.

RhoB affects colitis through modulating cell signaling and intestinal microbiome.

BACKGROUND: The pathogenesis of inflammatory bowel diseases (IBD) is multifactorial, and diagnostic and treatment strategies for IBD remain to be developed. RhoB regulates multiple cell functions; however, its role in colitis is unexplored. RESULTS: Here, we found RhoB was dramatically increased in colon tissues of ulcerative colitis (UC) patients and mice with DSS-induced colitis. Compared with wild type mice, RhoB+/- and RhoB-/- mice developed milder DSS-induced colitis and increased goblet cell numbers and IEC proliferation. Decreased RhoB promoted goblet cell differentiation and epithelial regeneration through inhibiting Wnt signaling pathway and activating p38 MAPK signaling pathway. Moreover, increased SCFA-producing bacteria and SCFA concentrations were detected in intestinal microbiome of both RhoB+/- and RhoB-/- mice and upregulated SCFA receptor expression was also observed. CONCLUSIONS: Taken together, a higher level of RhoB is associated with UC, which also contributes to UC development through modulating cell signaling and altering intestinal bacterial composition and metabolites. These observations suggest that RhoB has potential as a biomarker and a treatment target for UC. Video Abstract.

Enterobacter ludwigii protects DSS-induced colitis through choline-mediated immune tolerance.

Commensal intestinal bacteria play key roles in regulating host immune tolerance; however, bacterial strains and related metabolites directly involved in this regulation are largely unknown. Here, using a mouse model of dextran sulfate sodium (DSS)-induced colitis combined with different antibiotic treatment, Enterobacter ludwigii, abundant in microbiota of mice treated with metronidazole, is screened out to have prophylactic and therapeutic effects on DSS-induced colitis with or without the presence of complex intestinal bacteria. E. ludwigii is found to induce CD103+DC and regulatory T (Treg)-mediated immune tolerance for colitis remission using in vitro and in vivo experiments. Moreover, choline, one metabolite of E. ludwigii, is identified to increase dendritic cells' (DCs) immune tolerance to promote Treg differentiation. E. ludwigii is found to induce DCs' immune tolerance ability for Treg differentiation through choline and α7nAChR-mediated retinoic acid (RA) and transforming growth factor beta (TGF-ß) upregulation, resulting in protecting mice against DSS-induced colitis. This study suggests potential therapeutic approaches for inflammatory bowel diseases (IBDs).

Interactions of Bacterial Toxin CNF1 and Host JAK1/2 Driven by Liquid-Liquid Phase Separation Enhance Macrophage Polarization.

Urinary tract infections (UTIs) are a global public health concern, which is mainly caused by uropathogenic Escherichia coli (UPEC). Cytotoxic necrotizing factor 1 (CNF1) is a key UPEC toxin and regulates multiple host cellular processes through activating the Rho GTPases; however, the effect of CNF1 on macrophage polarization remains unknown. Here, we found that CNF1 promoted M1 macrophage polarization through regulating NF-κB and JAK-STAT1 signaling pathways in kidney at an early stage of acute UTIs. Notably, we identified CNF1 could directly interact with JAK1/2 through its domain without Rho GTPases activation, which induced JAK1/2 phosphorylation, subsequent STAT1 activation and M1 polarization. Moreover, CNF1 exhibited liquid-liquid phase separation (LLPS) to induce a CNF1-JAK1/2 complex, promoting macrophage reprogramming. These findings highlight the LLPS-dependent and Rho GTPase-independent effect of CNF1 as an adaptor on interfering with host cell signals. IMPORTANCE CNF1 is a key toxin secreted by UPEC, which induces inflammation during UPEC infections. CNF1 is well known to activate Rho GTPases to disturb host cell signaling pathways. Macrophage reprogramming plays important roles in inflammation; however, the effect of CNF1 on macrophage polarization is not reported. This study demonstrated the role and mechanism of CNF1 in promoting M1 macrophage polarization during UPEC-induced acute kidney infections. Importantly, we identified Rho GTPase-independent effect of CNF1 as an adaptor on interfering with host cell signals and demonstrated that CNF1 exhibited LLPS to drive its interaction with host proteins, which improve our understanding of the UPEC-host interactions and UTI pathogenesis.

Impact of ultra-low emission retrofitting on partitioning and emission behavior of chromium in a Chinese coal-fired power plant.

Due to its low vapor pressure, chromium (Cr) mostly emitted as fly ash particles (especially PM2.5) into environment in coal-fired power plants (CFPPs). The ultra-low emission (ULE) control technologies used in current CFPPs may be beneficial to reducing both the regular pollutants and hazardous trace elements (e.g., Cr), but the insight into the removal efficiency of Cr by different upgrading air pollution cleaning devices (APCDs) and the environmental stability of the Cr-bearing wastes produced from those APCDs in the ULE CFPPs has rarely reported. This study investigated and compared the distribution and emission characteristics of Cr in a Chinese CFPP before and after ULE, and the leaching behavior of Cr after ULE retrofitting in combustion byproducts was also revealed. The results showed that Cr was primarily captured in bottom and fly ashes (80.85%), followed by gypsum (0.02%) and sludge from wet electrostatic precipitator (WESP) (4.52 × 10-4%), with only 3.02 × 10-8% emitted into the atmosphere. Additional WESP had a large removal efficiency of Cr with the value of 92.04%, and the overall Cr removal efficiency of selective catalytic reduction (SCR) equipment, electrostatic precipitator (ESP), wet flue gas desulphurization (WFGD) system, and WESP equipped after ULE retrofitting was 99.99%. Notably, although the mass percentage of Cr in WESP sludge was negligible, the concentration of Cr in WESP sludge was 324.04 mg/kg. The leaching concentrations of Cr in combustion byproducts were in the descending order: fly ash > WESP sludge > bottom ash > gypsum. The atmospheric emission factor of Cr in the studied power plant was 1.08 mg/t coal, which was significantly lower than those of the CFPPs before ULE retrofitting. Therefore, the ULE retrofitting for CFPP was beneficial to reduce Cr emissions. More attention should be paid to the subsequent processing problem of solid combustion byproducts, especially the WESP sludge.

A Survey of Common Pathogens of Apis spp. in Wild Non-Cave Honeybees in Southwest China.

Honeybees are crucial pollinators with significant ecologic value. The decline of wild honeybee populations has been recognized and documented during recent decades. However, the health status of wild non-cave Apis spp., including giant and dwarf honeybees, remains generally unknown. We investigated eight common viruses and five bacterial or fungal pathogens in four wild non-cave honeybee species at 11 locations in Southwest China. As a result, Melissococcus plutonius, the pathogenic agent of European foulbrood, was detected in all the species, and the sequences were identical to the pathogen in managed cave honeybees. Only one virus, black queen cell virus (BQCV), was positive in one dwarf species, Apis florea, in our study. The positive BQCV infected three A. florea colonies in Guangxi Province, with distinct sequences from this virus reported in cave honeybees or in the same host in the nearby Yunnan Province. Although our results indicated a low pathogenic level of common diseases in the wild non-cave Apis spp. in Southwest China, the conservation of these wild pollinators is of importance in light of the noticeable decline in populations and the irreplaceable position of pollination.

The composition of bacteria in gut and beebread of stingless bees (Apidae: Meliponini) from tropics Yunnan, China.

Stingless bees are the main pollinators in tropical and subtropical regions. However, there are only a few studies on the structure and composition of bacteria in the gut and beebread of stingless bees, especially in China. To address this shortage of information, we characterized the microbiota of three common species of stingless bees (Lepidotrigona terminata, Lepidotrigona ventralis and Tetragonula pagdeni) and beebread samples of T. pagdeni. The results showed that the gut of stingless bees contained a set of dominant bacteria, including Acetobacter-like, Snodgrassella, Lactobacillus, Psychrobacter, Pseudomonas, Bifidobacterium and other species. The gut microbiota structures of the three stingless bees were different, and the abundances of bacterial species in the gut varied between communities of the same bee species. The reasons for this are manifold and may include food preference, age and genetic differences. In addition, the abundances of Lactobacillus, Carnimonas, Escherichia-Shigella, Acinetobacter and other species were high in the beebread of stingless bees. In conclusion, our findings reveal the bacteria composition and structure of the gut and beebread of stingless bees in China and deepen our understanding of the dominant bacteria of the gut and beebread of stingless bees.

An infection-induced RhoB-Beclin 1-Hsp90 complex enhances clearance of uropathogenic Escherichia coli.

Host cells use several anti-bacterial pathways to defend against pathogens. Here, using a uropathogenic Escherichia coli (UPEC) infection model, we demonstrate that bacterial infection upregulates RhoB, which subsequently promotes intracellular bacteria clearance by inducing LC3 lipidation and autophagosome formation. RhoB binds with Beclin 1 through its residues at 118 to 140 and the Beclin 1 CCD domain, with RhoB Arg133 being the key binding residue. Binding of RhoB to Beclin 1 enhances the Hsp90-Beclin 1 interaction, preventing Beclin 1 degradation. RhoB also directly interacts with Hsp90, maintaining RhoB levels. UPEC infections increase RhoB, Beclin 1 and LC3 levels in bladder epithelium in vivo, whereas Beclin 1 and LC3 levels as well as UPEC clearance are substantially reduced in RhoB+/- and RhoB-/- mice upon infection. We conclude that when stimulated by UPEC infections, host cells promote UPEC clearance through the RhoB-Beclin 1-HSP90 complex, indicating RhoB may be a useful target when developing UPEC treatment strategies.

Characteristics of the Two Asian Bumblebee Species Bombus friseanus and Bombus breviceps (Hymenoptera: Apidae).

This study compared the food plants, life cycle, colony development, and mating behaviour of the two Asian bumblebee species Bombus friseanus and B. breviceps, which are very important pollinators for many wild flowers and crops in local ecosystems. Both species were shown to be highly polylectic. Differences were observed in their life cycles and colony development patterns. The colony foundation rate of the field-collected queens was high in both species, 95.5% in B. friseanus and 86.5% in B. breviceps. The intervals from colony initiation to colony sizes of 30, 60, and 80 workers and to the first male and gyne emergence were significantly shorter in B. friseanus than in B. breviceps (p < 0.01). The development period of the first batch of workers showed no significant difference between the two species (p > 0.05). Compared with B. friseanus, B. breviceps produced remarkably higher numbers of workers (135 ± 30 workers/colony in B. friseanus and 318 ± 123 workers/colony in B. breviceps) and males (199 ± 46 males/colony in B. friseanus and 355 ± 166 males/colony in B. breviceps) (p < 0.01), with notable variation was found among the colonies in both species. With no significant difference in the mating rate between these two species, the copulation duration of B. breviceps (1.54 ± 0.63 min) was strikingly shorter than that of B. friseanus (27.44 ± 11.16 min) (p < 0.001). This study highlights the characteristics of the two Asian bumblebee species and will aid further studies on their conservation and agricultural pollination use.

Alpha-hemolysin of uropathogenic Escherichia coli induces GM-CSF-mediated acute kidney injury.

Uropathogenic Escherichia coli (UPEC) is the leading cause of urinary tract infections (UTIs), inducing acute pyelonephritis and may result in permanent renal scarring and failure. Alpha-hemolysin (HlyA), a key UPEC toxin, causes serious tissue damage; however, the mechanism through which HlyA induces kidney injury remains unclear. In the present study, granulocyte-macrophage colony-stimulating factor (GM-CSF) secreted by renal epithelial cells was upregulated by HlyA in vitro and in vivo, which induced M1 macrophage accumulation in kidney, and ADAM10 was found involved in HlyA-induced GM-CSF. Macrophage elimination or GM-CSF neutralization protected against acute kidney injury in mice, and increased GM-CSF was detected in urine of patients infected by hlyA-positive UPEC. In addition, HlyA was found to promote UPEC invasion into renal epithelial cells by interacting with Nectin-2 in vitro. However, HlyA did not affect bacterial titers during acute kidney infections, and HlyA-induced invasion did not contribute to GM-CSF upregulation in vitro, which indicate that HlyA-induced GM-CSF is independent of bacteria invasion. The role of GM-CSF in HlyA-mediated kidney injury may lead to novel strategies to treat acute pyelonephritis.

Alteration behavior of mineral structure and hazardous elements during combustion of coal from a power plant at Huainan, Anhui, China.

The alteration behavior of minerals and hazardous elements during simulated combustion (100-1200 °C) of a raw coal collected from a power plant were studied. Thermogravimetric analysis indicated that there were mainly four alteration stages during coal combustion. The transformation behavior of mineral phases of raw coal, which were detected by X-ray polycrystalline diffraction (XRD) technique, mainly relied on the combustion temperature. A series of changes were derived from the intensities of mineral (e.g. clays) diffraction peaks when temperature surpassed 600 °C. Mineral phases tended to be simple and collapsed to amorphous glass when temperature reached up to 1200 °C. The characteristics of functional groups for raw coal and high-temperature (1200 °C) ash studied by Fourier transform infrared spectroscopy (FTIR) were in accordance with the result obtained from XRD analysis. The volatilization ratios of Co, Cr, Ni and V increased consistently with the increase of combustion temperature, suggesting these elements were gradually released from the organic matter and inorganic minerals of coal.

Induction of Cell-Cell Fusion by Ebola Virus Glycoprotein: Low pH Is Not a Trigger.

Ebola virus (EBOV) is a highly pathogenic filovirus that causes hemorrhagic fever in humans and animals. Currently, how EBOV fuses its envelope membrane within an endosomal membrane to cause infection is poorly understood. We successfully measure cell-cell fusion mediated by the EBOV fusion protein, GP, assayed by the transfer of both cytoplasmic and membrane dyes. A small molecule fusion inhibitor, a neutralizing antibody, as well as mutations in EBOV GP known to reduce viral infection, all greatly reduce fusion. By monitoring redistribution of small aqueous dyes between cells and by electrical capacitance measurements, we discovered that EBOV GP-mediated fusion pores do not readily enlarge-a marked difference from the behavior of other viral fusion proteins. EBOV GP must be cleaved by late endosome-resident cathepsins B or L in order to become fusion-competent. Cleavage of cell surface-expressed GP appears to occur in endosomes, as evidenced by the fusion block imposed by cathepsin inhibitors, agents that raise endosomal pH, or an inhibitor of anterograde trafficking. Treating effector cells with a recombinant soluble cathepsin B or thermolysin, which cleaves GP into an active form, increases the extent of fusion, suggesting that a fraction of surface-expressed GP is not cleaved. Whereas the rate of fusion is increased by a brief exposure to acidic pH, fusion does occur at neutral pH. Importantly, the extent of fusion is independent of external pH in experiments in which cathepsin activity is blocked and EBOV GP is cleaved by thermolysin. These results imply that low pH promotes fusion through the well-known pH-dependent activity of cathepsins; fusion induced by cleaved EBOV GP is a process that is fundamentally independent of pH. The cell-cell fusion system has revealed some previously unappreciated features of EBOV entry, which could not be readily elucidated in the context of endosomal entry.

Cell-cell contact promotes Ebola virus GP-mediated infection.

Ebola virus (EBOV) is a highly pathogenic filovirus that causes hemorrhagic fever in humans and animals. Here we provide evidence that cell-cell contact promotes infection mediated by the glycoprotein (GP) of EBOV. Interestingly, expression of EBOV GP alone, even in the absence of retroviral Gag-Pol, is sufficient to transfer a retroviral vector encoding Tet-off from cell to cell. Cell-to-cell infection mediated by EBOV GP is blocked by inhibitors of actin polymerization, but appears to be less sensitive to KZ52 neutralization. Treatment of co-cultured cells with cathepsin B/L inhibitors, or an entry inhibitor 3.47 that targets the receptor NPC1 for virus binding, also blocks cell-to-cell infection. Cell-cell contact also enhances spread of rVSV bearing GP in monocytes and macrophages, the primary targets of natural EBOV infection. Altogether, our study reveals that cell-cell contact promotes EBOV GP-mediated infection, and provides new insight into understanding EBOV spread and viral pathogenesis.