Equine Herpesvirus Research Articles

Acute interstitial pneumonia in foals: A severe, multifactorial syndrome with lung tissue recovery in surviving foals.

Acute interstitial pneumonia in foals has been sparsely described in literature, and the individual authors disagree on the underlying aetiology. Histopathological follow-up from surviving foals is not available. Description of clinical and histopathological findings in the course of acute interstitial pneumonia and in recovery. Investigating the aetiology and possible triggering factors of acute interstitial pneumonia. Case series. Post-mortem examination of nine affected foals; seven died during the acute phase, and two had recovered from acute interstitial pneumonia. Data from clinical examinations on the day of death were recorded for all foals. Complete necropsy, special histological staining, virological and microbiological examinations were performed. Seven foals died during the acute phase with severe respiratory distress, fever and increased numbers of comet tail artefacts in lung ultrasound. In post-mortem examination, a wide variety of possible triggering factors was identified. Microbiology revealed Escherichia coli, Rhodococcus equi and Klebsiella pneumoniae as the most common bacterial pathogens. Equine herpesvirus 2 was detected in all foals by PCR. Those with high viral loads also displayed histopathological changes suggestive of viral infections. Pneumocystis carinii was detected in all acutely affected foals. Histopathological changes in lung parenchyma clearly differed between the foals that had recovered from acute interstitial pneumonia and those dying in the acute phase. While lungs in acute phase featured marked parenchymal collapse and necrosis, the recovered foals revealed nearly normal ventilated lung parenchyma and alveolar structure. Small number of cases. Because all foals are from the same breeding farm with endemic occurrence of pathogens, it is not certain whether the results of this study can be transferred to other foals without restrictions. Acute interstitial pneumonia seems to be based on a multifactorial aetiology. Lungs from foals that have survived acute interstitial pneumonia appear to be able to regenerate completely, leaving no permanent changes.

Identification of a New Equid Herpesvirus 1 DNA Polymerase (ORF30) Genotype with the Isolation of a C2254/H752 Strain in French Horses Showing no Major Impact on the Strain Behaviour.

Equid herpesvirus 1 is one of the most common viral pathogens in the horse population and is associated with respiratory disease, abortion and still-birth, neonatal death and neurological disease. A single point mutation in the DNA polymerase gene (ORF30: A2254G, N752D) has been widely associated with neuropathogenicity of strains, although this association has not been exclusive. This study describes the fortuitous isolation of a strain carrying a new genotype C2254 (H752) from an outbreak in France that lasted several weeks in 2018 and involved 82 horses, two of which showed neurological signs of disease. The strain was characterised as UL clade 10 using the equid herpesvirus 1 (EHV-1) multi-locus sequence typing (MLST) classification but has not been identified or isolated since 2018. The retrospective screening of EHV-1 strains collected between 2016 and 2018 did not reveal the presence of the C2254 mutation. When cultured in vitro, the C2254 EHV-1 strain induced a typical EHV-1 syncytium and cytopathic effect but no significant difference was observed when compared with A2254 and G2254 EHV-1 strains. An experimental infection was carried out on four Welsh mountain ponies to confirm the infectious nature of the C2254 strain. A rapid onset of marked respiratory disease lasting at least 2 weeks, with significant virus shedding and cell-associated viraemia, was observed. Finally, an in vitro antiviral assay using impedance measurement and viral load quantification was performed with three antiviral molecules (ganciclovir (GCV), aciclovir (ACV) and aphidicolin (APD)) on the newly isolated C2254 strain and two other A/G2254 field strains. The three strains showed similar sensitivity to ganciclovir and aphidicolin but both C2254 and A2254 strains were more sensitive to aciclovir than the G2254 strain, based on viral load measurement.

Outbreak of equid herpesvirus 1 abortions at the Arabian stud in Poland

BackgroundEquid herpesvirus 1 (EHV-1) infections are endemic worldwide, including Poland. Many are subclinical, but some are associated with respiratory disease, abortion, neonatal foal death, or neurological disease. We describe an outbreak of abortions in Arabian mares at a well-managed State stud farm in Poland.Case presentationEight of 30 pregnant mares aborted and one gave birth to a weak foal that died within 72 h after birth. EHV-1 was isolated from all fetuses as well as from the diseased foal. All viruses belonged to the N752 variant based on the predicted open reading frame (ORF) 30 amino acid sequence. All were identical to each other and to previous EHV-1 viruses from the same stud based on the ORF68 sequence analysis. The outbreak coincided with the lapse in the routine yearly EHV-1/4 vaccinations of the mares.ConclusionsMultiple abortion due to EHV-1 infection can occur in well-managed groups of horses. Reactivation of latent EHV-1 in one of the resident mares followed by a horizontal spread was considered the most likely explanation for the outbreak. Routine vaccination is an important part of a herd-heath program.

A Pilot Serosurvey for Selected Pathogens in Feral Donkeys (Equus asinus).

Simple SummaryThis study aimed to assess the pathogen exposure status of recently captured feral donkeys from Death Valley National Park, California. Assays to detect the presence of antibodies to equine herpesvirus 1, equine influenza virus, West Nile virus, and Borrelia burgdorferi (the causative agent of Lyme disease) were performed on serum samples from these feral donkeys. The results indicate that this population is mostly naïve and likely susceptible to these common equid pathogens upon removal from the wild. Recent removal and relocation of feral donkeys from vast public lands to more concentrated holding pens, training facilities, and offsite adoption locations raises several health and welfare concerns. Very little is known regarding the common equid pathogens that are circulating within the feral donkey population in and around Death Valley National Park, California, USA. The aim of this study was to utilize serologic assays to assess previous exposure of these donkeys to equine herpesvirus 1 (EHV-1), equine influenza (EIV), West Nile virus (WNV), and Borrelia burgdorferi (the causative agent of Lyme disease). The results of this study indicate that this feral equid population is mostly naïve and likely susceptible to these common equid pathogens upon removal from the wild.

Equid Herpesvirus-1 Exploits the Extracellular Matrix of Mononuclear Cells to Ensure Transport to Target Cells

SummaryMononuclear cells are the first line of defense against microbial infection. Yet, several viruses have evolved different mechanisms to overcome host defenses to ensure their spread. Here, we show unique mechanisms of how equid herpesvirus-1 manipulates peripheral blood mononuclear cells (PBMC) to travel further in the body. (1) “PBMC-hitching”: at the initial contact, herpesviruses lurk in the extracellular matrix (ECM) of PBMC without entering the cells. The virus exploits the components of the ECM to bind, transport, and then egress to infect other cells. (2) “Intracellular delivery”: transendothelial migration is a physiological mechanism where mononuclear cells can transmigrate through the endothelial cells. The virus was intangible and probably did not interfere with such a mechanism where the infected PBMC can probably deliver the virus inside the endothelium. (3) “Classical-fusion”: this process is well mastered by herpesviruses due to a set of envelope glycoproteins that facilitate cell-cell fusion and virus spread.

Correlation of Immunomodulatory Cytokine Expression with Histopathological Changes and Viral Antigen in a Hamster Model of Equine Herpesvirus-9 Encephalitis

A group of hamsters (n=25) was intranasally infected with equine herpesvirus-9 (EHV-9) and mRNA transcription levels of several proinflammatory (IFN-γ, TNF-α and IL-6) and anti-inflammatory (IL-4, IL-10 and TGF-β) cytokines were investigated in brain tissue using RT-qPCR. These levels were correlated with the severity of sequential histopathological changes and intensity of immunohistochemical labelling of virus antigen in brain. Early and progressive upregulation of all the proinflammatory and anti-inflammatory cytokines investigated (P<0.05) was correlated with increasing severity of encephalitis and viral antigen expression from 2 days post infection (dpi) with a peak at 4-5 dpi (P<0.05).

Comparative Study of the Pathogenesis of Rhinopneumonitis Induced by Intranasal Inoculation of Hamsters with Equine Herpesvirus-9, Equine Herpesvirus-1 strain Ab4p and Zebra-borne Equine Herpesvirus-1

Equine herpesvirus-9 (EHV-9), equine herpesvirus-1 (EHV-1) and zebra-borne EHV-1 are members of the family Herpesviridae and cause encephalitis and rhinopneumonitis in a range of animal species. The aim of this study was to characterize and compare the rhinopneumonitis induced by experimental intranasal inoculation of groups of hamsters with EHV-9, EHV-1 strain Ab4p or zebra-borne EHV-1 viruses. Animals inoculated with EHV-9 had earlier and more severe neurological and respiratory signs than those inoculated with EHV-1 strain Ab4p or zebra-borne EHV-1. At 4-5 days post inoculation (dpi), hamsters inoculated with EHV-9 had significantly increased expression of open reading fame (ORF) 30, the viral gene encoding the DNA polymerase, in lung tissue. ORF 30 expression at these time points was higher in the hamsters infected with EHV-9 than in those inoculated with the other two viruses. Severe, mild or very mild rhinitis was seen in animals inoculated with EHV-1 strain Ab4p, EHV-9 and zebra-borne EHV-1, respectively. Viral antigen was detected in olfactory receptor neurons, inflammatory cells and desquamated epithelial cells in animals in all groups until 5 dpi. Tracheitis was also seen in all three virus-infected groups with viral antigen detected in tracheal epithelium. Inoculated hamsters developed interstitial pneumonia of increasing severity over the course of the experiment. Bronchopneumonia and vasculitis were also seen in all three infected groups. These results confirm that, in addition to their neurotropism, EHV-9 and zebra-borne EHV-1 are pneumotropic viruses. EHV-1 strain Ab4p caused more severe upper respiratory tract disease, but no significant differences were detected in the severity of pneumonia induced by each virus.

Equid Herpesvirus 8 Isolated From an Adult Donkey in Israel

We report the isolation of Equid herpesvirus 8 from a rescued donkey that suffered severe postcastration complications. Despite intensive treatment, the donkey deteriorated and was euthanized. Postmortem virologic analysis revealed the isolation of a herpesvirus that is closely related to herpesviruses reported from donkeys and horses in Australia, China, and Ireland, causing respiratory disease in donkeys and abortion in mares. To our knowledge, this is the first report of this equid herpesvirus in Israel. The potential significance of this herpesvirus to the equid population in Israel needs further investigation.

Identification of antiviral compounds against equid herpesvirus-1 using real-time cell assay screening: Efficacy of decitabine and valganciclovir alone or in combination

Equid herpesvirus-1 infections cause respiratory, neurological and reproductive syndromes. Despite preventive treatments with vaccines, resurgence of EHV-1 infection still constitutes a major threat to equine industry. However, no antiviral compound is available to treat infected horses. In this study, 2891 compounds were screened against EHV-1 using impedance measurement. 22 compounds have been found to be effective in vitro against EHV-1. Valganciclovir, ganciclovir, decitabine, aphidicolin, idoxuridine and pritelivir (BAY 57–1293) are the most effective compounds identified, and their antiviral potency was further assessed on E. Derm, RK13 and EEK cells and against 3 different field strains of EHV-1 (ORF30 2254 A/G/C). We also provide evidences of synergistic interactions between valganciclovir and decitabine in our in vitro antiviral assay as determined by MacSynergy II, isobologramm and Chou-Talalay methods. Finally, we showed that deoxycytidine reverts the antiviral effect of decitabine, thus supporting some competition at the level of nucleoside phosphorylation by deoxycytidine kinase and/or DNA synthesis. Deoxycitidine analogues, like decitabine, is a family of compounds identified for the first time with promising antiviral efficacy against herpesviruses.

Equine Herpesvirus Type 1 Modulates Cytokine and Chemokine Profiles of Mononuclear Cells for Efficient Dissemination to Target Organs.

Equine herpesvirus type 1 (EHV-1) causes encephalomyelopathy and abortion, for which cell-associated viremia and subsequent virus transfer to and replication in endothelial cells (EC) are responsible and prerequisites. Viral and cellular molecules responsible for efficient cell-to-cell spread of EHV-1 between peripheral blood mononuclear cells (PBMC) and EC remain unclear. We have generated EHV-1 mutants lacking ORF1, ORF2, and ORF17 genes, either individually or in combination. Mutant viruses were analyzed for their replication properties in cultured equine dermal cells, PBMC infection efficiency, virus-induced changes in the PBMC proteome, and cytokine and chemokine expression profiles. ORF1, ORF2, and ORF17 are not essential for virus replication, but ORF17 deletion resulted in a significant reduction in plaque size. Deletion of ORF2 and ORF17 gene significantly reduced cell-to-cell virus transfer from virus-infected PBMC to EC. EHV-1 infection of PBMC resulted in upregulation of several pathways such as Ras signaling, oxidative phosphorylation, platelet activation and leukocyte transendothelial migration. In contrast, chemokine signaling, RNA degradation and apoptotic pathways were downregulated. Deletion of ORF1, ORF2 and ORF17 modulated chemokine signaling and MAPK pathways in infected PBMC, which may explain the impairment of virus spread between PBMC and EC. The proteomic results were further confirmed by chemokine assays, which showed that virus infection dramatically reduced the cytokine/chemokine release in infected PBMC. This study uncovers cellular proteins and pathways influenced by EHV-1 after PBMC infection and provide an important resource for EHV-1 pathogenesis. EHV-1-immunomodulatory genes could be potential targets for the development of live attenuated vaccines or therapeutics against virus infection.

Viral Load and Cell Tropism During Early Latent Equid Herpesvirus 1 Infection Differ Over Time in Lymphoid and Neural Tissue Samples From Experimentally Infected Horses.

Upper respiratory tract infections with Equid Herpesvirus 1 (EHV-1) typically result in a peripheral blood mononuclear cell-associated viremia, which can lead to vasculopathy in the central nervous system. Primary EHV-1 infection also likely establishes latency in trigeminal ganglia (TG) via retrograde axonal transport and in respiratory tract-associated lymphatic tissue. However, latency establishment and reactivation are poorly understood. To characterize the pathogenesis of EHV-1 latency establishment and maintenance, two separate groups of yearling horses were experimentally infected intranasally with EHV-1, strain Ab4, and euthanized 30 days post infection (dpi), (n = 9) and 70 dpi (n = 6). During necropsy, TG, sympathetic trunk (ST), retropharyngeal and mesenteric lymph nodes (RLn, MesLn) and kidney samples were collected. Viral DNA was detected by quantitative PCR (qPCR) in TG, ST, RLn, and MesLn samples in horses 30 and 70 dpi. The number of positive TG, RLn and MesLn samples was reduced when comparing horses 30 and 70 dpi and the viral copy number in TG and RLn significantly declined from 30 to 70 dpi. EHV-1 late gene glycoprotein B reverse transcriptase PCR and IHC results for viral protein were consistently negative, thus lytic replication was excluded in the present study. Mild inflammation could be detected in all neural tissue samples and inflammatory infiltrates mainly consisted of CD3+ T-lymphocytes (T-cells), frequently localized in close proximity to neuronal cell bodies. To identify latently infected cell types, in situ hybridization (ISH, RNAScope®) detecting viral DNA was used on selected qPCR- positive neural tissue sections. In ganglia 30 dpi, EHV-1 ISH signal was located in the neurons of TG and ST, but also in non-neuronal support or interstitial cells surrounding the neuron. In contrast, distinct EHV-1 signal could only be observed in neurons of TG 70 dpi. Overall, detection of latent EHV-1 in abdominal tissue samples and non-neuronal cell localization suggests, that EHV-1 uses T-cells during viremia as alternative route toward latency locations in addition to retrograde neuronal transport. We therefore hypothesize that EHV-1 follows the same latency pathways as its close relative human pathogen Varicella Zoster Virus.

Molecular characterization of equine herpesvirus-1 and asinine herpesvirus -5 isolated from aborted fetuses of arabian horses

Molecular characterization of equine herpesvirus-1 and asinine herpesvirus -5 isolated from aborted fetuses of arabian horses

Efficiency of alfalfa granulated feed additive in balanced ration of Ukrainian Riding Horse

We established positive influence of granulated alfalfa feed on the growth, development intensity, and sport qualities of the race horses. We registered the increase in live weight of young horses, which consumed the full granulated alfalfa feed by 8.0-9.8%. These animals had a more pronounced constitutional breed type. Thus, at the age of 18 months, they outclassed their peers in main linear dimensions: they had higher values of thoracic development (by 4.6%), higher thoracic diameter (by 4.5%), higher body length (by 5.7%). At the age of 24 months, these advantages were 6.4, 8.2, and 6.0% respectively. We found that the colts and fillies in the experimental group at the age of 12 months outperformed the same year horses from the control group by 1.1% by the index of format. Further, at the age of 18 and 24 months, the animals of the experimental group outperformed their counterparts from the control group by 0.2 and 0.4%, respectively. In terms of breast circumference index, the advantage of the exerimentary group animals at the age of 12 months was 0.1%, at the age of 18 months - 1.7%, at the age of 24 months - 1.8%, respectively. Similar results were recorded for the compactness index, where the advantage at the age of 12 months was 1.2%, at 18 months - 1.9%, at 24 months - 2.3% respectively. The effective use of environmentally friendly feed is confirmed by the results of hematological blood tests. It was found that for the vast majority of indicators they correspond to the physiological norm. This blood composition naturally characterizes changes in horses during the whole period of research. The received hematological profile is the basic characteristic of the state of health of horses in the research enterprise.

Time Course-Dependent Study on Equine Herpes Virus 9-Induced Abortion in Syrian Hamsters

Simple SummaryEquine herpesvirus 9 (EHV-9) is a virus belonging to the family of equine herpesviruses. EHV-9 has been isolated from natural infections of different wild and zoo animals. In addition, it has been associated with encephalitis and abortion in several animal species. However, the host range and pathogenesis of this virus are still unknown. Herein, we investigated the underlying pathogenesis of EHV-9-induced abortion in relation to the gestation period in either early or late trimester infection. We noticed that the late trimester infection of EHV-9 was associated with more severe death and both placental and fetal tissue localization of the virus. Also, early stage infection was accompanied by band necrotic changes within the placenta, which usually led to abortion.This study aimed to follow the time-course pathogenesis of EHV-9 abortion in early and late trimesters. Twenty-seven pregnant hamster dams were divided into three groups: (G1) control, (G2) EHV-9-inoculated on the 5th day (early trimester), and (G3) EHV-9-inoculated on the 10th day of gestation (late trimester). Dams were sacrificed at different time points during gestation and examined for viremia and viral DNA in different fetal and maternal tissues and pathological changes in fetal tissue, placenta, and cytokines. Animals in G3 showed a marked increase in the number of dead fetuses than those in G2. Histopathological findings of G2 showed early band coagulative necrosis of maternal spaces and stromal decidual cells. Necrotic changes were observed within the decidua basalis, spongiotrophoblast layer, and labyrinth. First, the virus was localized within mononuclear leukocytes in the decidua capsularis and basalis, and within the necrotic chorionic villi and cervical epithelium. G3 demonstrated degenerative changes within the chorionic villi and trophospongium. The virus antigen was observed within the chorionic villi, trophoblasts, mononuclear cells, and fetal tissues. In conclusion, EHV-9 induced abortion mostly occurs through necrosis of the chorionic villi and cannot cross through the capsular placenta in the early trimester but can through the developed decidual placentation.

Antibody Responses Against Equine Influenza Virus Induced by Concurrent and by Consecutive Use of an Inactivated Equine Influenza Virus Vaccine and a Modified Live Equine Herpesvirus Type 1 Vaccine in Thoroughbred Racehorses

An inactivated equine influenza virus (EIV) vaccine and a live equine herpesvirus type 1 (EHV-1) vaccine are usually administered concurrently to Thoroughbred racehorses in Japan. The objective of this study was to evaluate whether concurrent administration of an inactivated EIV vaccine and a live EHV-1 vaccine in Thoroughbred racehorses influences the antibody response against EIV. We compared the antibody response against EIV in horses administered both vaccines on the same day (Group A; n = 27) and the response in horses administered an inactivated EIV vaccine first and then a live EHV-1 vaccine 1–2 weeks later (Group B; n = 20). In both groups, geometric mean hemagglutination inhibition (HI) titers against A/equine/Ibaraki/1/2007 and A/equine/Yokohama/aq13/2010 increased significantly after EIV vaccination. However, the percentage of horses that showed a twofold increase or greater in HI titers against A/equine/Yokohama/aq13/2010 was significantly higher in Group B (75%) than in Group A (37%; P = .02). These results suggest that the concurrent use of an inactivated EIV vaccine and a live EHV-1 vaccine reduced the immune response against EIV to some extent, and it would be better to use these vaccines consecutively, especially for naïve horses or horses whose vaccination history is incomplete.

Characterization of high virulent strains of equine rhinopneumonitis virus

This article presents results of screening samples collected from horses during acute outbreaks of abortion and neurological disorders recorded in 2019-2020 in Russia. The signs of disease were characterized by severe hemorrhagic vasculitis. Massive hemorrhages were observed in organs of aborted fetuses and dead horses. Equine herpesvirus type one was isolated in cell culture. Testing to identify genes encoding glycoprotein B and polymerase catalytic subunit of certain viral isolates were carried out. Virus isolated from horses with signs of encephalomyelitis from the Northwest region had a high degree of phylogenetic relationship with the strain “89c25” of equine herpesvirus type one from Japan, 1989. The same virus was found in horses with signs of rhinopneumonia from the Ural region. The virus isolated after equine abortion from the North Caucasus region had a high degree of phylogenetic relationship with the strain “IT19m10” of equine herpesvirus type one from Italy, 2019. It was established that all viral isolates belong to non-neuropathogenic genotype - A2254.

Topical Ganciclovir Reduces Viral Excretion in Mares With Equine Coital Exanthema

Equid alphaherpesvirus 3 (EHV-3) is the etiological agent of equine coital exanthema (ECE). Because no vaccines or antiviral therapies are available, prevention consists of clinical examination of mares and stallions before mating or semen collection and resting from breeding activities when lesions are present. However, this methodology does not identify subclinically infected animals. Ganciclovir is the most potent compound known to reduce EHV-3 replication. This study aimed to evaluate the efficacy of topical ganciclovir application to reduce EHV-3 replication in experimentally infected mares. A pilot study, after a double-blind completely randomized design, was carried out. Twenty mares were randomly divided into five groups (three treated with ganciclovir with different regimen of doses, one treated with a placebo, and one nontreated). Mares were experimentally infected with EHV-3 on day 0. Rectal temperature, clinical signs, and lesions were recorded. Daily perineal and vaginal swabs were evaluated by quantitative polymerase chain reaction for virus detection. The antibody response was assessed by a virus neutralization test in serum samples collected weekly. Mares experimentally infected with EHV-3 and treated with ganciclovir twice a day for 13 days showed reduced levels and duration of viral excretion and less severe lesions. The viral excretion period was reduced from 18 to nine days compared with the untreated groups. We concluded that ganciclovir had an antiviral effect on EHV-3 replication when topically administered in mares showing clinical signs of ECE. Further trials should be performed to optimize the dose of the antiviral for a definitive formulation.

Environmental persistence of equid herpesvirus type-1.

Equid herpesvirus type 1 (EHV-1) is ubiquitous in equine populations causing respiratory disease, and complications including late-term abortion and neurological disease. Eradication of EHV-1 from housing environments that typically contain unsealed wood and porous bedding materials can be challenging. However, consideration should be given to take advantage of the viral envelope's susceptibility to environmental conditions. To determine environmental persistence of EHV-1 on materials and in environmental conditions commonly found in equine facilities. We hypothesised that environmental conditions and materials would limit environmental persistence of EHV-1 in horse housing environments. Experimental study. Standard inoculum of EHV-1 strain OH03 was applied to leather, polyester-cotton fabric, two bedding materials (pinewood shavings and wheat straw) and polystyrene (plastic), and placed under three different environmental conditions (4°C, indoors and outdoors). Virus titration and quantitative PCR (qPCR) were performed at six time points between 0 and 48hours and the number of plaque-forming units (PFUs) was determined. Viable EHV-1 was recovered up to 48hours from all material-environmental condition combinations, with persistence decreasing over time. In general, outdoor environment had the greatest impact, irrespective of material tested, followed by indoor environment and 4°C. On average, wood shavings had the greatest impact on persistence, followed by leather, straw, fabric and polystyrene. The inoculum used in this study was not in a milieu consistent with nasal secretions. As such, virus particles may have been more sensitive to the materials and/or environmental conditions evaluated. Environmental factors had variable effects on environmental persistence. Although there were significant reductions in PFUs within the first 3hours, irrespective of environment-material evaluated, viable virus was still recovered at 48hours likely representing a transmission risk. Barrier precautions should be used to prevent spread of EHV-1 from unrecognised environmental reservoirs.

Equine Alphaherpesviruses Require Activation of the Small GTPases Rac1 and Cdc42 for Intracellular Transport.

Viruses utilize host cell signaling to facilitate productive infection. Equine herpesvirus type 1 (EHV-1) has been shown to activate Ca2+ release and phospholipase C upon contact with α4β1 integrins on the cell surface. Signaling molecules, including small GTPases, have been shown to be activated downstream of Ca2+ release, and modulate virus entry, membrane remodeling and intracellular transport. In this study, we show that EHV-1 activates the small GTPases Rac1 and Cdc42 during infection. The activation of Rac1 and Cdc42 is necessary for virus-induced acetylation of tubulin, effective viral transport to the nucleus, and cell-to-cell spread. We also show that inhibitors of Rac1 and Cdc42 did not block virus entry, but inhibited overall virus infection. The Rac1 and Cdc42 signaling is presumably orthogonal to Ca2+ release, since Rac1 and Cdc42 inhibitors affected the infection of both EHV-1 and EHV-4, which do not bind to integrins.

Generation of Equine Herpesvirus type 1 glycoprotein pseudotyped lentiviral particles for use as a tool for tropism and diagnostic studies

Equine herpesviruses (EHVs) are enveloped DNA viruses infecting mainly members of the Equidae family and also members of other taxa. EHVs primarily causing respiratory disease, however EHV type 1 (EHV-1) can produce cases of a neurological disease, abortion and neonatal death, sometimes as regional outbreaks. Thus these viruses represent a welfare issue for the equine industry and scientific focus for researchers. EHV-1 presents a complex array of 12 glycoproteins on its surface envelope, but it is unclear which ones are important for virus cell entry and the role of each in host immune response. In order to investigate the contribution of these glycoproteins, pseudotype viruses (PVs) could provide a perfect study tool. In 2016, Rogalin &amp; Heldwein successfully generated the first functional herpesvirus pseudotype, bearing the four glycoproteins gB, gD, gH and gL from human Herpes simplex 1. Our study is the first to attempt pseudotyping of EHV-1. We have employed homologous glycoproteins of EHV-1 in lentivirus PV generation, using different mammalian cells (e.g. epithelial, dermal, CNS) as transduction targets. The glycoprotein sequences obtained from an EHV-1 strain isolated from organs of aborted foetus during a significant outbreak in Normandy (France) in 2010. Future work will focus on the development of a PV assay for detection of neutralising antibodies in naturally infected horses for diagnostics and for vaccine evaluation.