Macacine Herpesvirus 1

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Julia K Hilliard - One of the best experts on this subject based on the ideXlab platform.

  • Identification of unique B virus (Macacine Herpesvirus 1) epitopes of zoonotic and macaque isolates using monoclonal antibodies.
    PloS one, 2017
    Co-Authors: David Katz, Irina Patrusheva, Mugdha Vasireddi, Wei Shi, Manjunath S. Gowda, Hyuk-kyu Seoh, Martin J. Wildes, Chadi Filfili, Julia K Hilliard
    Abstract:

    Our overall aim is to develop epitope-based assays for accurate differential diagnosis of B virus zoonotic infections in humans. Antibodies to cross-reacting epitopes on human-simplexviruses continue to confound the interpretation of current assays where abundant antibodies exist from previous infections with HSV types 1 and 2. To find B virus-specific epitopes we cloned ten monoclonal antibodies (mAbs) from the hybridomas we produced. Our unique collection of rare human sera from symptomatic and asymptomatic patients infected with B virus was key to the evaluation and identification of the mAbs as reagents in competition ELISAs (mAb-CE). The analysis of the ten mAbs revealed that the target proteins for six mAbs was glycoprotein B of which two are reactive to simian simplexviruses and not to human simplexviruses. Two mAbs reacted specifically with B virus glycoprotein D, and two other mAbs were specific to VP13/14 and gE-gI complex respectively. The mAbs specific to VP13/14 and gE-gI are strain specific reacting with B virus isolates from rhesus and Japanese macaques and not with isolates from cynomolgus and pigtail macaques. The mAb-CE revealed that a high proportion of naturally B virus infected rhesus macaques and two symptomatic humans possess antibodies to epitopes of VP13/14 protein and on the gE-gI complex. The majority of sera from B virus infected macaques and simplexvirus-infected humans competed with the less specific mAbs. These experiments produced a novel panel of mAbs that enabled B virus strain identification and confirmation of B virus infected macaques by the mAb-CE. For human sera the mAb-CE could be used only for selected cases due to the selective B virus strain-specificity of the mAbs against VP13/14 and gE/gI. To fully accomplish our aim to provide reagents for unequivocal differential diagnosis of zoonotic B virus infections, additional mAbs with a broader range of specificities is critical.

  • Regulation of PI3K/Akt dependent apoptotic markers during b virus infection of human and macaque fibroblasts.
    PloS one, 2017
    Co-Authors: Mugdha Vasireddi, Julia K Hilliard
    Abstract:

    B virus (Macacine Herpesvirus 1), a simplex virus endemic in macaques, causes encephalitis, encephalomyelitis, and death in 80% of untreated zoonotically infected humans with delayed or no treatment. Here we report a significant difference in PI3K/Akt-dependent apoptosis between B virus infected human and macaque dermal fibroblasts. Our data show that B virus infection in either human or macaque fibroblasts results in activation of Akt via PI3K and this activation does not require viral de novo protein synthesis. Inhibition of PI3K with LY294002 results in a significant reduction of viral titers in B virus infected macaque and human fibroblasts with only a modest difference in the reduction of virus titers between the two cell types. We, therefore, tested the hypothesis that B virus results in the phosphorylation of Akt (S473), which prevents apoptosis, enhancing virus replication in B virus infected macaque dermal fibroblasts. We observed markers of intrinsic apoptosis when PI3K activation of Akt was inhibited in B virus infected macaque cells, while, these apoptotic markers were absent in B virus infected human fibroblasts under the same conditions. From these data we suggest that PI3K activates Akt in B virus infected macaque and human fibroblasts, but this enhances virus replication in macaque fibroblast cells by blocking apoptosis.

  • B Virus (Macacine Herpesvirus 1) Divergence: Variations in Glycoprotein D from Clinical and Laboratory Isolates Diversify Virus Entry Strategies
    Journal of virology, 2016
    Co-Authors: Irina Patrusheva, Ludmila Perelygina, Ivan Y. Torshin, Julia C. Lecher, Julia K Hilliard
    Abstract:

    UNLABELLED B virus (Macacine Herpesvirus 1) can cause deadly zoonotic disease in humans. Molecular mechanisms of B virus cell entry are poorly understood for both macaques and humans. Here we investigated the abilities of clinical B virus isolates to use entry receptors of herpes simplex viruses (HSV). We showed that resistant B78H1 cells became susceptible to B virus clinical strains upon expression of either human nectin-2 or nectin-1. Antibody against glycoprotein D (gD) protected these nectin-bearing cells from B virus infection, and a gD-negative recombinant B virus failed to enter these cells, indicating that the nectin-mediated B virus entry depends on gD. We observed that the infectivity of B virus isolates with a single amino acid substitution (D122N) in the IgV-core of the gD ectodomain was impaired on nectin-1-bearing cells. Computational homology-based modeling of the B virus gD-nectin-1 complex revealed conformational differences between the structures of the gD-122N and gD-122D variants that affected the gD-nectin-1 protein-protein interface and binding affinity. Unlike HSV, B virus clinical strains were unable to use Herpesvirus entry mediator (HVEM) as a receptor, regardless of conservation of the gD amino acid residues essential for HSV-1 entry via HVEM. Based on the model of the B virus gD-HVEM interface, we predict that residues R7, R11, and G15 are largely responsible for the inability of B virus to utilize HVEM for entry. The ability of B virus to enter cells of a human host by using a combination of receptors distinct from those for HSV-1 or HSV-2 suggests a possible mechanism of enhanced neuropathogenicity associated with zoonotic infections. IMPORTANCE B virus causes brainstem destruction in infected humans in the absence of timely diagnosis and intervention. Nectins are cell adhesion molecules that are widely expressed in human tissues, including neurons and neuronal synapses. Here we report that human nectin-2 is a target receptor for B virus entry, in addition to the reported receptor human nectin-1. Similar to a B virus lab strain, B virus clinical strains can effectively use both nectin-1 and nectin-2 as cellular receptors for entry into human cells, but unlike HSV-1 and HSV-2, none of the clinical strains uses an HVEM-mediated entry pathway. Ultimately, these differences between B virus and HSV-1 and -2 may provide insight into the neuropathogenicity of B virus during zoonotic infections.

  • B Virus (Macacine Herpesvirus 1) Glycoprotein D Is Functional but Dispensable for Virus Entry into Macaque and Human Skin Cells
    Journal of Virology, 2015
    Co-Authors: Ludmila Perelygina, Irina Patrusheva, Mugdha Vasireddi, Nicole Brock, Julia K Hilliard
    Abstract:

    Glycoprotein D (gD) plays an essential role in cell entry of many simplexviruses. B virus (Macacine Herpesvirus 1) is closely related to herpes simplex virus 1 (HSV-1) and encodes gD, which shares more than 70% amino acid similarity with HSV-1 gD. Previously, we have demonstrated that B virus gD polyclonal antibodies were unable to neutralize B virus infectivity on epithelial cell lines, suggesting gD is not required for B virus entry into these cells. In the present study, we confirmed this finding by producing a B virus mutant, BV-ΔgDZ, in which the gD gene was replaced with a lacZ expression cassette. Recombinant plaques were selected on complementing VD60 cells expressing HSV-1 gD. Virions lacking gD were produced in Vero cells infected with BV-ΔgDZ. In contrast to HSV-1, B virus lacking gD was able to infect and form plaques on noncomplementing cell lines, including Vero, HEp-2, LLC-MK2, primary human and macaque dermal fibroblasts, and U373 human glioblastoma cells. The gD-negative BV-ΔgDZ also failed to enter entry-resistant murine B78H1 cells bearing a single gD receptor, human nectin-1, but gained the ability to enter when phenotypically supplemented with HSV-1 gD. Cell attachment and penetration rates, as well as the replication characteristics of BV-ΔgDZ in Vero cells, were almost identical to those of wild-type (wt) B virus. These observations indicate that B virus can utilize gD-independent cell entry and transmission mechanisms, in addition to generally used gD-dependent mechanisms. IMPORTANCE B virus is the only known simplexvirus that causes zoonotic infection, resulting in approximately 80% mortality in untreated humans or in lifelong persistence with the constant threat of reactivation in survivors. Here, we report that B virus lacking the gD envelope glycoprotein infects both human and monkey cells as efficiently as wild-type B virus. These data provide evidence for a novel mechanism(s) utilized by B virus to gain access to target cells. This mechanism is different from those used by its close relatives, HSV-1 and -2, where gD is a pivotal protein in the virus entry process. The possibility remains that unidentified receptors, specific for B virus, permit virus entry into target cells through gD-independent pathways. Understanding the molecular mechanisms of B virus entry may help in developing rational therapeutic strategies for the prevention and treatment of B virus infection in both macaques and humans.

  • please contact scholarworks@gsu.edu. B Virus Infection Activates p38 And JNK Pathways Differentially In Cells From Macaque Versus Human Hosts: Exploring Inflammation & Apoptosis
    2014
    Co-Authors: Rachael M. Farah-abraham, Under Direction, Julia K Hilliard
    Abstract:

    B virus (Macacine Herpesvirus 1), subfamily Alphaherpesvirinae, causes a fatal, neuro-‐ virulent infection in zoonotically infected humans. Macaques (Macaca sp.) serve as the natural host for B virus and they are frequently seropositive for B virus antibodies without showing any overt signs of disease. The global hypothesis of these studies is that B virus, a highly cytopathic virus in macaques, subverts the innate immune responses in the host (macaques) that has co-‐ evolved with it (the virus) differently than it does the foreign host (humans). The foreign host, frequently fails to produce neutralizing antibodies early after infection and this may be due to a dysregulation or inhibition of pathways known to play a role in the innate immune response which directs the adaptive defense responses. Current knowledge is that at least five major signaling pathways can be activated after a pathogen such as B virus enters a host cell (REF). These include the IRF3 pathway, the NFkB pathway, the NFAT pathway, and the MAPK path-‐ way. Early stimulation of one or more of these pathways leads to the induction of the proin-‐ flammatory response and subsequent induction of cytokines such as IL6, IL8 and IL10, and apoptosis. Cytokine induction and apoptosis play important roles in host-‐pathogen interactions

Gale G Galland - One of the best experts on this subject based on the ideXlab platform.

  • worker health and safety practices in research facilities using nonhuman primates north america
    Emerging Infectious Diseases, 2014
    Co-Authors: Emily W Lankau, Patricia V Turner, Robert J Mullan, Gale G Galland
    Abstract:

    To the Editor: Since 1975, federal quarantine regulations (1) have restricted nonhuman primate importation to scientific, educational, or exhibition purposes to limit risks for disease introduction (1,2). Infectious diseases resulting from importation of nonhuman primates need to be prevented to ensure that colonies of these animals are available for research and to protect persons working with them from exposure to established and emerging zoonotic diseases (2,3). Most imported nonhuman primates are bred for research and undergo standard screening and conditioning before shipment, which substantially reduce importation-associated health risks (4). However, many zoonotic agents can be difficult to exclude from even meticulously controlled breeding facilities (3,5). Nonhuman primates are commonly imported from regions with a high prevalence of potentially zoonotic diseases, such as tuberculosis and meliodosis, in humans and animals. Diagnosing tuberculosis in nonhuman primates can be difficult; inadvertent colony and human exposures can occur through undiagnosed cases (6). Similarly, Burkholderia pseudomallei, the causative agent of meliodosis, can be carried asymptomatically for extended periods before illness onset, posing a persistent exposure risk for persons working with imported nonhuman primates from regions to which meliodosis is endemic (7). Finally, nonhuman primates are host to potentially zoonotic viruses, such as simian foamy virus, which has unknown pathogenic potential in infected persons (8), and Macacine Herpesvirus 1, which causes severe, often fatal, neurologic disease in humans exposed to macaques with asymptomatic infection (9). Quarantine and testing of imported nonhuman primates, rigorous hygiene at research facilities, and strict personal protection equipment (PPE) standards are important to protect the health of nonhuman primate colonies and persons working with the animals (4). Importers must register with the Centers for Disease Control and Prevention (CDC) and implement disease control measures, including a 31-day quarantine for newly arrived animals (1). Specific PPE is mandated for quarantine facility staff, but individual facilities determine PPE standards after the animals are released from CDC-mandated quarantine (4). To better understand occupational health and safety practices at facilities housing nonhuman primates, in December 2012, the Association of Primate Veterinarians, with technical support from CDC, surveyed primate veterinarians in North America about animal handling practices and PPE standards at their institutions. The Association of Primate Veterinarians received completed surveys and removed identifying information before providing data to CDC for analysis. CDC and the University of Guelph (Guelph, ON, Canada) determined that the survey did not qualify as human subjects research. Information collected applied to the institution, not the individual respondent. Respondents were informed that participation was voluntary and anonymous, refusal carried no repercussions, and results would be presented in aggregate. Of 149 facilities, 7 (5%) indicated they were not currently housing nonhuman primates, and 26 (17%) provided completed surveys. Most responding facilities were university or private/contract research facilities (16 [62%] and 5 [19%] facilities, respectively). Most (18 [69%]) facilities maintained <500 nonhuman primates, primarily rhesus or cynomolgus macaques. Nineteen (73%) facilities acquired imported nonhuman primates during 2010–2012. During this period, 47,876 nonhuman primates were imported, of which 90% were cynomolgus macaques. Fewer nonhuman primates were acquired from domestic sources (1,877 animals; see also [10]). In a free-text field, we asked about quarantine and testing policies for newly acquired nonhuman primates. Most facilities reported applying standard health requirements to newly acquired animals, regardless of source, and requiring additional quarantine periods before moving new animals into the facility population or assigning them to a study. The number of staff working near nonhuman primates differed among facilities. Ten (38%) facilities reported that >30 staff members handle or work in close proximity to nonhuman primates for cleaning or observation each day (Table). All facilities required PPE for routine handling of animals, including use of surgical masks or N95 respirators; goggles, safety glasses, or full-face shields; specialized clothing (e.g., laboratory coat, scrubs, or coveralls); gloves; and either shoe covers, reusable boots, or facility-designated shoes (Table). Table Health and safety practices reported by 26 research facilities that use nonhuman primates, North America, December 2012* Twenty-one (81%) facilities reported routinely handling conscious nonhuman primates by using specialized safety equipment (e.g., pole and collar or restraint chair). Four (15%) facilities reported manually capturing conscious animals (“hand-catching”); 2 (8%) facilities performed intrapalpebral tuberculin skin tests on conscious animals (Table). All facilities reported routinely performing postmortem examinations. Five facilities specified that complete necropsies were performed only on animals found dead or euthanized because of illness or injury; for animals euthanized at study completion, targeted tissue specimens were collected to fulfill research objectives (Table). These results suggest that responding facilities generally maintained high standards for health and safety and are aware of disease risks. However, this survey has limitations for assessing the effectiveness of risk mitigation policies. Although a variety of facilities responded, response biases cannot be excluded. Additionally, these results summarize occupational health and safety standards on record but cannot address compliance or employee attitudes toward health and safety concerns in working with nonhuman primates. Facilities maintaining nonhuman primates need to strive for strict enforcement of occupational health and safety requirements; consider requiring regular continuing education about human health risks associated with working closely with animals; and consider the degree of risk pertaining to specific activities, particularly those generating infectious aerosols.

  • Worker health and safety practices in research facilities using nonhuman primates, North America.
    Emerging infectious diseases, 2014
    Co-Authors: Emily W Lankau, Patricia V Turner, Robert J Mullan, Gale G Galland
    Abstract:

    To the Editor: Since 1975, federal quarantine regulations (1) have restricted nonhuman primate importation to scientific, educational, or exhibition purposes to limit risks for disease introduction (1,2). Infectious diseases resulting from importation of nonhuman primates need to be prevented to ensure that colonies of these animals are available for research and to protect persons working with them from exposure to established and emerging zoonotic diseases (2,3). Most imported nonhuman primates are bred for research and undergo standard screening and conditioning before shipment, which substantially reduce importation-associated health risks (4). However, many zoonotic agents can be difficult to exclude from even meticulously controlled breeding facilities (3,5). Nonhuman primates are commonly imported from regions with a high prevalence of potentially zoonotic diseases, such as tuberculosis and meliodosis, in humans and animals. Diagnosing tuberculosis in nonhuman primates can be difficult; inadvertent colony and human exposures can occur through undiagnosed cases (6). Similarly, Burkholderia pseudomallei, the causative agent of meliodosis, can be carried asymptomatically for extended periods before illness onset, posing a persistent exposure risk for persons working with imported nonhuman primates from regions to which meliodosis is endemic (7). Finally, nonhuman primates are host to potentially zoonotic viruses, such as simian foamy virus, which has unknown pathogenic potential in infected persons (8), and Macacine Herpesvirus 1, which causes severe, often fatal, neurologic disease in humans exposed to macaques with asymptomatic infection (9). Quarantine and testing of imported nonhuman primates, rigorous hygiene at research facilities, and strict personal protection equipment (PPE) standards are important to protect the health of nonhuman primate colonies and persons working with the animals (4). Importers must register with the Centers for Disease Control and Prevention (CDC) and implement disease control measures, including a 31-day quarantine for newly arrived animals (1). Specific PPE is mandated for quarantine facility staff, but individual facilities determine PPE standards after the animals are released from CDC-mandated quarantine (4). To better understand occupational health and safety practices at facilities housing nonhuman primates, in December 2012, the Association of Primate Veterinarians, with technical support from CDC, surveyed primate veterinarians in North America about animal handling practices and PPE standards at their institutions. The Association of Primate Veterinarians received completed surveys and removed identifying information before providing data to CDC for analysis. CDC and the University of Guelph (Guelph, ON, Canada) determined that the survey did not qualify as human subjects research. Information collected applied to the institution, not the individual respondent. Respondents were informed that participation was voluntary and anonymous, refusal carried no repercussions, and results would be presented in aggregate. Of 149 facilities, 7 (5%) indicated they were not currently housing nonhuman primates, and 26 (17%) provided completed surveys. Most responding facilities were university or private/contract research facilities (16 [62%] and 5 [19%] facilities, respectively). Most (18 [69%]) facilities maintained 30 staff members handle or work in close proximity to nonhuman primates for cleaning or observation each day (Table). All facilities required PPE for routine handling of animals, including use of surgical masks or N95 respirators; goggles, safety glasses, or full-face shields; specialized clothing (e.g., laboratory coat, scrubs, or coveralls); gloves; and either shoe covers, reusable boots, or facility-designated shoes (Table). Table Health and safety practices reported by 26 research facilities that use nonhuman primates, North America, December 2012* Twenty-one (81%) facilities reported routinely handling conscious nonhuman primates by using specialized safety equipment (e.g., pole and collar or restraint chair). Four (15%) facilities reported manually capturing conscious animals (“hand-catching”); 2 (8%) facilities performed intrapalpebral tuberculin skin tests on conscious animals (Table). All facilities reported routinely performing postmortem examinations. Five facilities specified that complete necropsies were performed only on animals found dead or euthanized because of illness or injury; for animals euthanized at study completion, targeted tissue specimens were collected to fulfill research objectives (Table). These results suggest that responding facilities generally maintained high standards for health and safety and are aware of disease risks. However, this survey has limitations for assessing the effectiveness of risk mitigation policies. Although a variety of facilities responded, response biases cannot be excluded. Additionally, these results summarize occupational health and safety standards on record but cannot address compliance or employee attitudes toward health and safety concerns in working with nonhuman primates. Facilities maintaining nonhuman primates need to strive for strict enforcement of occupational health and safety requirements; consider requiring regular continuing education about human health risks associated with working closely with animals; and consider the degree of risk pertaining to specific activities, particularly those generating infectious aerosols.

Emily W Lankau - One of the best experts on this subject based on the ideXlab platform.

  • worker health and safety practices in research facilities using nonhuman primates north america
    Emerging Infectious Diseases, 2014
    Co-Authors: Emily W Lankau, Patricia V Turner, Robert J Mullan, Gale G Galland
    Abstract:

    To the Editor: Since 1975, federal quarantine regulations (1) have restricted nonhuman primate importation to scientific, educational, or exhibition purposes to limit risks for disease introduction (1,2). Infectious diseases resulting from importation of nonhuman primates need to be prevented to ensure that colonies of these animals are available for research and to protect persons working with them from exposure to established and emerging zoonotic diseases (2,3). Most imported nonhuman primates are bred for research and undergo standard screening and conditioning before shipment, which substantially reduce importation-associated health risks (4). However, many zoonotic agents can be difficult to exclude from even meticulously controlled breeding facilities (3,5). Nonhuman primates are commonly imported from regions with a high prevalence of potentially zoonotic diseases, such as tuberculosis and meliodosis, in humans and animals. Diagnosing tuberculosis in nonhuman primates can be difficult; inadvertent colony and human exposures can occur through undiagnosed cases (6). Similarly, Burkholderia pseudomallei, the causative agent of meliodosis, can be carried asymptomatically for extended periods before illness onset, posing a persistent exposure risk for persons working with imported nonhuman primates from regions to which meliodosis is endemic (7). Finally, nonhuman primates are host to potentially zoonotic viruses, such as simian foamy virus, which has unknown pathogenic potential in infected persons (8), and Macacine Herpesvirus 1, which causes severe, often fatal, neurologic disease in humans exposed to macaques with asymptomatic infection (9). Quarantine and testing of imported nonhuman primates, rigorous hygiene at research facilities, and strict personal protection equipment (PPE) standards are important to protect the health of nonhuman primate colonies and persons working with the animals (4). Importers must register with the Centers for Disease Control and Prevention (CDC) and implement disease control measures, including a 31-day quarantine for newly arrived animals (1). Specific PPE is mandated for quarantine facility staff, but individual facilities determine PPE standards after the animals are released from CDC-mandated quarantine (4). To better understand occupational health and safety practices at facilities housing nonhuman primates, in December 2012, the Association of Primate Veterinarians, with technical support from CDC, surveyed primate veterinarians in North America about animal handling practices and PPE standards at their institutions. The Association of Primate Veterinarians received completed surveys and removed identifying information before providing data to CDC for analysis. CDC and the University of Guelph (Guelph, ON, Canada) determined that the survey did not qualify as human subjects research. Information collected applied to the institution, not the individual respondent. Respondents were informed that participation was voluntary and anonymous, refusal carried no repercussions, and results would be presented in aggregate. Of 149 facilities, 7 (5%) indicated they were not currently housing nonhuman primates, and 26 (17%) provided completed surveys. Most responding facilities were university or private/contract research facilities (16 [62%] and 5 [19%] facilities, respectively). Most (18 [69%]) facilities maintained <500 nonhuman primates, primarily rhesus or cynomolgus macaques. Nineteen (73%) facilities acquired imported nonhuman primates during 2010–2012. During this period, 47,876 nonhuman primates were imported, of which 90% were cynomolgus macaques. Fewer nonhuman primates were acquired from domestic sources (1,877 animals; see also [10]). In a free-text field, we asked about quarantine and testing policies for newly acquired nonhuman primates. Most facilities reported applying standard health requirements to newly acquired animals, regardless of source, and requiring additional quarantine periods before moving new animals into the facility population or assigning them to a study. The number of staff working near nonhuman primates differed among facilities. Ten (38%) facilities reported that >30 staff members handle or work in close proximity to nonhuman primates for cleaning or observation each day (Table). All facilities required PPE for routine handling of animals, including use of surgical masks or N95 respirators; goggles, safety glasses, or full-face shields; specialized clothing (e.g., laboratory coat, scrubs, or coveralls); gloves; and either shoe covers, reusable boots, or facility-designated shoes (Table). Table Health and safety practices reported by 26 research facilities that use nonhuman primates, North America, December 2012* Twenty-one (81%) facilities reported routinely handling conscious nonhuman primates by using specialized safety equipment (e.g., pole and collar or restraint chair). Four (15%) facilities reported manually capturing conscious animals (“hand-catching”); 2 (8%) facilities performed intrapalpebral tuberculin skin tests on conscious animals (Table). All facilities reported routinely performing postmortem examinations. Five facilities specified that complete necropsies were performed only on animals found dead or euthanized because of illness or injury; for animals euthanized at study completion, targeted tissue specimens were collected to fulfill research objectives (Table). These results suggest that responding facilities generally maintained high standards for health and safety and are aware of disease risks. However, this survey has limitations for assessing the effectiveness of risk mitigation policies. Although a variety of facilities responded, response biases cannot be excluded. Additionally, these results summarize occupational health and safety standards on record but cannot address compliance or employee attitudes toward health and safety concerns in working with nonhuman primates. Facilities maintaining nonhuman primates need to strive for strict enforcement of occupational health and safety requirements; consider requiring regular continuing education about human health risks associated with working closely with animals; and consider the degree of risk pertaining to specific activities, particularly those generating infectious aerosols.

  • Worker health and safety practices in research facilities using nonhuman primates, North America.
    Emerging infectious diseases, 2014
    Co-Authors: Emily W Lankau, Patricia V Turner, Robert J Mullan, Gale G Galland
    Abstract:

    To the Editor: Since 1975, federal quarantine regulations (1) have restricted nonhuman primate importation to scientific, educational, or exhibition purposes to limit risks for disease introduction (1,2). Infectious diseases resulting from importation of nonhuman primates need to be prevented to ensure that colonies of these animals are available for research and to protect persons working with them from exposure to established and emerging zoonotic diseases (2,3). Most imported nonhuman primates are bred for research and undergo standard screening and conditioning before shipment, which substantially reduce importation-associated health risks (4). However, many zoonotic agents can be difficult to exclude from even meticulously controlled breeding facilities (3,5). Nonhuman primates are commonly imported from regions with a high prevalence of potentially zoonotic diseases, such as tuberculosis and meliodosis, in humans and animals. Diagnosing tuberculosis in nonhuman primates can be difficult; inadvertent colony and human exposures can occur through undiagnosed cases (6). Similarly, Burkholderia pseudomallei, the causative agent of meliodosis, can be carried asymptomatically for extended periods before illness onset, posing a persistent exposure risk for persons working with imported nonhuman primates from regions to which meliodosis is endemic (7). Finally, nonhuman primates are host to potentially zoonotic viruses, such as simian foamy virus, which has unknown pathogenic potential in infected persons (8), and Macacine Herpesvirus 1, which causes severe, often fatal, neurologic disease in humans exposed to macaques with asymptomatic infection (9). Quarantine and testing of imported nonhuman primates, rigorous hygiene at research facilities, and strict personal protection equipment (PPE) standards are important to protect the health of nonhuman primate colonies and persons working with the animals (4). Importers must register with the Centers for Disease Control and Prevention (CDC) and implement disease control measures, including a 31-day quarantine for newly arrived animals (1). Specific PPE is mandated for quarantine facility staff, but individual facilities determine PPE standards after the animals are released from CDC-mandated quarantine (4). To better understand occupational health and safety practices at facilities housing nonhuman primates, in December 2012, the Association of Primate Veterinarians, with technical support from CDC, surveyed primate veterinarians in North America about animal handling practices and PPE standards at their institutions. The Association of Primate Veterinarians received completed surveys and removed identifying information before providing data to CDC for analysis. CDC and the University of Guelph (Guelph, ON, Canada) determined that the survey did not qualify as human subjects research. Information collected applied to the institution, not the individual respondent. Respondents were informed that participation was voluntary and anonymous, refusal carried no repercussions, and results would be presented in aggregate. Of 149 facilities, 7 (5%) indicated they were not currently housing nonhuman primates, and 26 (17%) provided completed surveys. Most responding facilities were university or private/contract research facilities (16 [62%] and 5 [19%] facilities, respectively). Most (18 [69%]) facilities maintained 30 staff members handle or work in close proximity to nonhuman primates for cleaning or observation each day (Table). All facilities required PPE for routine handling of animals, including use of surgical masks or N95 respirators; goggles, safety glasses, or full-face shields; specialized clothing (e.g., laboratory coat, scrubs, or coveralls); gloves; and either shoe covers, reusable boots, or facility-designated shoes (Table). Table Health and safety practices reported by 26 research facilities that use nonhuman primates, North America, December 2012* Twenty-one (81%) facilities reported routinely handling conscious nonhuman primates by using specialized safety equipment (e.g., pole and collar or restraint chair). Four (15%) facilities reported manually capturing conscious animals (“hand-catching”); 2 (8%) facilities performed intrapalpebral tuberculin skin tests on conscious animals (Table). All facilities reported routinely performing postmortem examinations. Five facilities specified that complete necropsies were performed only on animals found dead or euthanized because of illness or injury; for animals euthanized at study completion, targeted tissue specimens were collected to fulfill research objectives (Table). These results suggest that responding facilities generally maintained high standards for health and safety and are aware of disease risks. However, this survey has limitations for assessing the effectiveness of risk mitigation policies. Although a variety of facilities responded, response biases cannot be excluded. Additionally, these results summarize occupational health and safety standards on record but cannot address compliance or employee attitudes toward health and safety concerns in working with nonhuman primates. Facilities maintaining nonhuman primates need to strive for strict enforcement of occupational health and safety requirements; consider requiring regular continuing education about human health risks associated with working closely with animals; and consider the degree of risk pertaining to specific activities, particularly those generating infectious aerosols.

R. Eberle - One of the best experts on this subject based on the ideXlab platform.

  • Role of the virion host shutoff protein in neurovirulence of monkey B virus (Macacine Herpesvirus 1)
    Virologica Sinica, 2014
    Co-Authors: Darla Black, Jerry Ritchey, Mark Payton, R. Eberle
    Abstract:

    Monkey B virus (Macacine Herpesvirus 1; BV) is noted for its extreme neurovirulence in humans. Since the vhs protein encoded by the UL41 gene has been shown to be a neurovirulence factor in the related human herpes simplex viruses, the role of the UL41 gene in BV neurovirulence was investigated. BV mutants were constructed that lacked the entire UL41 ORF (Δ41) or had the RNase active site mutated (Δ41A). Neither mutant shut off host protein synthesis, degraded β-actin mRNA, or prevented an IFN-β response, indicating that the vhs protein and its RNase activity are both necessary for these activities. Replication of both mutants in primary mouse cells was impaired and they exhibited a prolonged disease course in mice. Whereas Δ41 infected mice were euthanized for symptoms related to central nervous system (CNS) infection, Δ41A infected mice were euthanized primarily for symptoms of autonomic nervous system dysfunction. While neuroinvasiveness was not affected, lesions in the CNS were more limited in size, anatomical distribution, and severity than for wild-type virus. These results indicate that the vhs protein affects the general replicative efficiency of BV in vivo rather than being a specific neurovirulence factor critical for invasion of or preferential replication in the CNS.

  • Genome sequence of a pathogenic isolate of monkey B virus (species Macacine Herpesvirus 1)
    Archives of Virology, 2014
    Co-Authors: Kazutaka Ohsawa, Darla Black, Makiko Ohsawa, R. Eberle
    Abstract:

    The only genome sequence for monkey B virus (BV; species Macacine Herpesvirus 1 ) is that of an attenuated vaccine strain originally isolated from a rhesus monkey (BVrh). Here we report the genome sequence of a virulent BV strain isolated from a cynomolgus macaque (BVcy). The overall genome organization is the same, although sequence differences exist. The greatest sequence divergence is located in non-coding areas of the long and short repeat regions. Like BVrh, BVcy has duplicated Ori elements and lacks an ORF corresponding to the γ34.5 gene of herpes simplex virus. Nine of ten miRNAs and the majority of ORFs are conserved between BVrh and BVcy. The most divergent genes are several membrane-associated proteins and those encoding immediate early proteins.

  • Papiine Herpesvirus 2 as a predictive model for drug sensitivity of Macacine Herpesvirus 1 (monkey B virus).
    Comparative medicine, 2014
    Co-Authors: Lauren A Brush, Darla H. Black, Lara K. Maxwell, Kimberly A Mccormack, George E. Wright, Jerry W. Ritchey, Mark E. Payton, R. Eberle
    Abstract:

    Monkey B virus (Macacine Herpesvirus 1; BV) is endemic in macaques. BV (a BSL4 agent) is the primary zoonotic concern for persons working with macaques in research, and human BV infections frequently are fatal. We assessed the use of a BSL2 baboon Herpesvirus (Papiine Herpesvirus 1; HVP2) for predicting the drug sensitivity of BV by comparing the sensitivity of the 2 viruses to 12 antiherpetic drugs. Plaque reduction assays showed that 4 drugs (HBPG, BVdU, PFA, and BrdU) were ineffective against both viruses. Of the 8 effective drugs, both viruses were most sensitive to TFT, whereas sensitivity to the remaining 7 drugs varied between BV and HVP2 as well as between strains of HVP2. In addition, the efficacy of 5 drugs (ACV, PCV, GCV, CDV, and EDU) was tested by using a murine model. ACV and EDU were completely ineffective against both HVP2 and BV, and high doses of PCV only delayed death by a few days. GCV and CDV both protected mice against death, and CDV also prevented the development of neurologic symptoms. When the initiation of drug therapy was delayed until after virus gained access to the CNS, both GCV and CDV were ineffective. The similarity of the drug sensitivities of HVP2 and BV in both models validates the use of HVP2 as a BSL2 level model that can be used to predict drug sensitivity of BV. The greater efficacy of CDV relative to GCV suggests the potential for use of CDV in the treatment of zoonotic BV infections.

  • Characterization of a spontaneous drug-resistant mutant of monkey B virus (Macacine Herpesvirus 1)
    Archives of Virology, 2009
    Co-Authors: Darla H. Black, Lara K. Maxwell, R. Eberle
    Abstract:

    Monkey B virus ( Macacine Herpesvirus 1; BV) is an α-Herpesvirus of macaques that causes serious infections in humans. A spontaneous mutant resistant to penciclovir (PCV) was isolated. Several genes were sequenced to identify mutations potentially responsible for PCV resistance. A single nucleotide deletion in the thymidine kinase (TK) gene was identified. To confirm its role in PCV resistance, several TK recombinants were constructed. A TK-deletion virus and a recombinant carrying the mutation were both resistant to PCV, while a revertant was PCV-sensitive. These results demonstrate that spontaneous drug-resistant mutants of BV do occur and that the BV TK is responsible for sensitivity to PCV.

Irina Patrusheva - One of the best experts on this subject based on the ideXlab platform.

  • Identification of unique B virus (Macacine Herpesvirus 1) epitopes of zoonotic and macaque isolates using monoclonal antibodies.
    PloS one, 2017
    Co-Authors: David Katz, Irina Patrusheva, Mugdha Vasireddi, Wei Shi, Manjunath S. Gowda, Hyuk-kyu Seoh, Martin J. Wildes, Chadi Filfili, Julia K Hilliard
    Abstract:

    Our overall aim is to develop epitope-based assays for accurate differential diagnosis of B virus zoonotic infections in humans. Antibodies to cross-reacting epitopes on human-simplexviruses continue to confound the interpretation of current assays where abundant antibodies exist from previous infections with HSV types 1 and 2. To find B virus-specific epitopes we cloned ten monoclonal antibodies (mAbs) from the hybridomas we produced. Our unique collection of rare human sera from symptomatic and asymptomatic patients infected with B virus was key to the evaluation and identification of the mAbs as reagents in competition ELISAs (mAb-CE). The analysis of the ten mAbs revealed that the target proteins for six mAbs was glycoprotein B of which two are reactive to simian simplexviruses and not to human simplexviruses. Two mAbs reacted specifically with B virus glycoprotein D, and two other mAbs were specific to VP13/14 and gE-gI complex respectively. The mAbs specific to VP13/14 and gE-gI are strain specific reacting with B virus isolates from rhesus and Japanese macaques and not with isolates from cynomolgus and pigtail macaques. The mAb-CE revealed that a high proportion of naturally B virus infected rhesus macaques and two symptomatic humans possess antibodies to epitopes of VP13/14 protein and on the gE-gI complex. The majority of sera from B virus infected macaques and simplexvirus-infected humans competed with the less specific mAbs. These experiments produced a novel panel of mAbs that enabled B virus strain identification and confirmation of B virus infected macaques by the mAb-CE. For human sera the mAb-CE could be used only for selected cases due to the selective B virus strain-specificity of the mAbs against VP13/14 and gE/gI. To fully accomplish our aim to provide reagents for unequivocal differential diagnosis of zoonotic B virus infections, additional mAbs with a broader range of specificities is critical.

  • B Virus (Macacine Herpesvirus 1) Divergence: Variations in Glycoprotein D from Clinical and Laboratory Isolates Diversify Virus Entry Strategies
    Journal of virology, 2016
    Co-Authors: Irina Patrusheva, Ludmila Perelygina, Ivan Y. Torshin, Julia C. Lecher, Julia K Hilliard
    Abstract:

    UNLABELLED B virus (Macacine Herpesvirus 1) can cause deadly zoonotic disease in humans. Molecular mechanisms of B virus cell entry are poorly understood for both macaques and humans. Here we investigated the abilities of clinical B virus isolates to use entry receptors of herpes simplex viruses (HSV). We showed that resistant B78H1 cells became susceptible to B virus clinical strains upon expression of either human nectin-2 or nectin-1. Antibody against glycoprotein D (gD) protected these nectin-bearing cells from B virus infection, and a gD-negative recombinant B virus failed to enter these cells, indicating that the nectin-mediated B virus entry depends on gD. We observed that the infectivity of B virus isolates with a single amino acid substitution (D122N) in the IgV-core of the gD ectodomain was impaired on nectin-1-bearing cells. Computational homology-based modeling of the B virus gD-nectin-1 complex revealed conformational differences between the structures of the gD-122N and gD-122D variants that affected the gD-nectin-1 protein-protein interface and binding affinity. Unlike HSV, B virus clinical strains were unable to use Herpesvirus entry mediator (HVEM) as a receptor, regardless of conservation of the gD amino acid residues essential for HSV-1 entry via HVEM. Based on the model of the B virus gD-HVEM interface, we predict that residues R7, R11, and G15 are largely responsible for the inability of B virus to utilize HVEM for entry. The ability of B virus to enter cells of a human host by using a combination of receptors distinct from those for HSV-1 or HSV-2 suggests a possible mechanism of enhanced neuropathogenicity associated with zoonotic infections. IMPORTANCE B virus causes brainstem destruction in infected humans in the absence of timely diagnosis and intervention. Nectins are cell adhesion molecules that are widely expressed in human tissues, including neurons and neuronal synapses. Here we report that human nectin-2 is a target receptor for B virus entry, in addition to the reported receptor human nectin-1. Similar to a B virus lab strain, B virus clinical strains can effectively use both nectin-1 and nectin-2 as cellular receptors for entry into human cells, but unlike HSV-1 and HSV-2, none of the clinical strains uses an HVEM-mediated entry pathway. Ultimately, these differences between B virus and HSV-1 and -2 may provide insight into the neuropathogenicity of B virus during zoonotic infections.

  • B Virus (Macacine Herpesvirus 1) Glycoprotein D Is Functional but Dispensable for Virus Entry into Macaque and Human Skin Cells
    Journal of Virology, 2015
    Co-Authors: Ludmila Perelygina, Irina Patrusheva, Mugdha Vasireddi, Nicole Brock, Julia K Hilliard
    Abstract:

    Glycoprotein D (gD) plays an essential role in cell entry of many simplexviruses. B virus (Macacine Herpesvirus 1) is closely related to herpes simplex virus 1 (HSV-1) and encodes gD, which shares more than 70% amino acid similarity with HSV-1 gD. Previously, we have demonstrated that B virus gD polyclonal antibodies were unable to neutralize B virus infectivity on epithelial cell lines, suggesting gD is not required for B virus entry into these cells. In the present study, we confirmed this finding by producing a B virus mutant, BV-ΔgDZ, in which the gD gene was replaced with a lacZ expression cassette. Recombinant plaques were selected on complementing VD60 cells expressing HSV-1 gD. Virions lacking gD were produced in Vero cells infected with BV-ΔgDZ. In contrast to HSV-1, B virus lacking gD was able to infect and form plaques on noncomplementing cell lines, including Vero, HEp-2, LLC-MK2, primary human and macaque dermal fibroblasts, and U373 human glioblastoma cells. The gD-negative BV-ΔgDZ also failed to enter entry-resistant murine B78H1 cells bearing a single gD receptor, human nectin-1, but gained the ability to enter when phenotypically supplemented with HSV-1 gD. Cell attachment and penetration rates, as well as the replication characteristics of BV-ΔgDZ in Vero cells, were almost identical to those of wild-type (wt) B virus. These observations indicate that B virus can utilize gD-independent cell entry and transmission mechanisms, in addition to generally used gD-dependent mechanisms. IMPORTANCE B virus is the only known simplexvirus that causes zoonotic infection, resulting in approximately 80% mortality in untreated humans or in lifelong persistence with the constant threat of reactivation in survivors. Here, we report that B virus lacking the gD envelope glycoprotein infects both human and monkey cells as efficiently as wild-type B virus. These data provide evidence for a novel mechanism(s) utilized by B virus to gain access to target cells. This mechanism is different from those used by its close relatives, HSV-1 and -2, where gD is a pivotal protein in the virus entry process. The possibility remains that unidentified receptors, specific for B virus, permit virus entry into target cells through gD-independent pathways. Understanding the molecular mechanisms of B virus entry may help in developing rational therapeutic strategies for the prevention and treatment of B virus infection in both macaques and humans.

  • An automated ELISA using recombinant antigens for serologic diagnosis of B virus infections in macaques.
    Comparative medicine, 2012
    Co-Authors: David Katz, Ludmila Perelygina, Irina Patrusheva, Wei Shi, Manjunath S. Gowda, Chadi Filfili, Peter W. Krug, John A. Ward, Julia K Hilliard
    Abstract:

    B virus (Macacine Herpesvirus 1) occurs naturally in macaques and can cause lethal zoonotic infections in humans. Detection of B virus (BV) antibodies in macaques is essential for the development of SPF breeding colonies and for diagnosing infection in macaques that are involved in human exposures. Traditionally, BV infections are monitored for presence of antibodies by ELISA (a screening assay) and western blot analysis (WBA; a confirmatory test). Both tests use lysates of infected cells as antigens. Because WBA often fails to confirm the presence of low-titer serum antibodies detected by ELISA, we examined a recombinant-based ELISA as a potential alternative confirmatory test. We compared a high-throughput ELISA using 384-well plates for simultaneous antibody screening against 4 BV-related, recombinant proteins with the standard ELISA and WBA. The recombinant ELISA results confirmed more ELISA-positive sera than did WBA. The superiority of the recombinant ELISA over WBA was particularly prominent for sera with low (