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Julio Reyesleyva - One of the best experts on this subject based on the ideXlab platform.

  • neuraminidase activity of Blue Eye Disease porcine rubulavirus specificity affinity and inhibition studies
    Research in Veterinary Science, 2017
    Co-Authors: G Santoslopez, Humberto Ramirezmendoza, Julio Reyesleyva, Maria Del Transito Borrazarguello, Veronica Vallejoruiz, Luis Marquezdominguez, Juan Carlos Floresalonso, Bernard Priem, Sebastien Fort, Irma Herreracamacho
    Abstract:

    Abstract Porcine rubulavirus (PorPV), also known as La Piedad Michoacan Virus (LPMV) causes encephalitis and reproductive failure in newborn and adult pigs, respectively. The hemagglutinin–neuraminidase (HN) glycoprotein is the most exposed and antigenic of the virus proteins. HN plays central roles in PorPV infection; i.e. , it recognizes sialic acid-containing cell receptors that mediate virus attachment and penetration; in addition, its neuraminidase (sialic acid releasing) activity has been proposed as a virulence factor. This work describes the purification and characterization of PorPV HN protein (isolate PAC1). The specificity of neuraminidase is restricted to sialyl(α2,3)lactose (3SL). HN showed typical Michaelis-Menten kinetics with fetuin as substrate (km = 0.029 μM, Vmax = 522.8 nmol min − 1  mg − 1 ). When 3SL was used as substrate, typical cooperative kinetics were found (S 50  = 0.15 μM, Vmax = 154.3 nmol min − 1  mg − 1 ). The influenza inhibitor zanamivir inhibited the PorPV neuraminidase with IC 50 of 0.24 μM. PorPV neuraminidase was activated by Ca 2 + and inhibited by nucleoside triphosphates with the level of inhibition depending on phosphorylation level. The present results open possibilities to study the role of neuraminidase in the pathogenicity of PorPV infection and its potential inhibitors.

  • production of an enzymatically active and immunogenic form of ectodomain of porcine rubulavirus hemagglutinin neuraminidase in the yeast pichia pastoris
    Journal of Biotechnology, 2016
    Co-Authors: Julio Reyesleyva, G Santoslopez, Sandra Cuevasromero, Nora Rosasmurrieta, Jose Luis Cerritenosanchez, Irma Herreracamacho
    Abstract:

    Blue-Eye Disease (BED) of swine is a viral Disease endemic in Mexico. The etiological agent is a paramyxovirus classified as Porcine rubulavirus (PoRV-LPMV), which exhibits in its envelope the hemagglutinin-neuraminidase (HN) glycoprotein, the most immunogenic and a major target for vaccine development. We report in this study the obtaining of ectodomain of PoRV HN (eHN) through the Pichia pastoris expression system. The expression vector (pPICZαB-HN) was integrated by displacement into the yeast chromosome and resulted in a Mut+ phenotype. Expressed eHN in the P. pastoris X33 strain was recovered from cell-free medium, featuring up to 67 nmol/min/mg after 6 days of expression. eHN was recognized by the serum of infected pigs with strains currently circulating in the Mexican Bajio region. eHN induces antibodies in mice after 28 days of immunization with specific recognition in ELISA test. These antibodies were able to inhibit >80% replication by viral neutralization assays in cell culture. These studies show the obtaining of a protein with similar characteristics to the native HN and which may be a candidate to propose a vaccine or to use the antigen in a serologic diagnostic test.

  • genetic and antigenic changes in porcine rubulavirus
    Canadian Journal of Veterinary Research-revue Canadienne De Recherche Veterinaire, 2012
    Co-Authors: Jose Ivan Sanchezbetancourt, Julio Reyesleyva, Maria Elena Trujillo, Susana E Mendoza, Rogelio A Alonso
    Abstract:

    Blue Eye Disease, caused by a porcine rubulavirus (PoRV), is an emergent viral swine Disease that has been endemic in Mexico since 1980. Atypical outbreaks were detected in 1990 and 2003. Growing and adult pigs presented neurological signs, mild neurological signs were observed in piglets, and severe reproductive problems were observed in adults. Amino acid sequence comparisons and phylogenetic analysis of the hemagglutinin-neuraminidase (HN) protein revealed genetically different lineages. We used cross-neutralization assays, with homologous and heterologous antisera, to determine the antigenic relatedness values for the PoRV isolates. We found antigenic changes among several strains and identified a highly divergent one, making up a new serogroup. It seems that genetically and antigenically different PoRV strains are circulating simultaneously in the swine population in the geographical region studied. The cross neutralization studies suggest that the HN is not the only antigenic determinant participating in the antigenic changes among the different PoRV strains.

  • molecular characterization of the hemagglutinin neuraminidase gene of porcine rubulavirus isolates associated with neurological disorders in fattening and adult pigs
    Research in Veterinary Science, 2008
    Co-Authors: Jose Ivan Sanchezbetancourt, Jesús Hernández, Humberto Ramirezmendoza, Julio Reyesleyva, G Santoslopez, Rogelio A Alonso, J M Doporto, S. Mendoza, Maria Elena Trujillo
    Abstract:

    Abstract “Blue Eye Disease” is a viral infection of swine endemic in Mexico, which produces fatal encephalitis accompanied by respiratory signs and corneal opacity in suckling piglets. An atypical Blue Eye Disease outbreak presented high rates of neurological signs in fattening and adult pigs from 2000 to 2003. In order to identify the basis of increased neurovirulence, the hemagglutinin-neuraminidase (HN) gene of several porcine rubulavirus isolates were sequenced and compared with that of La Piedad Michoacan virus and other isolates that did not produce neurological disorders in weaned pigs. Nine amino acid mutations distinguished the high neurovirulent PAC6–PAC9 viruses, whereas five mutations characterized the low neurovirulent PAC2 and PAC3 viruses. HN protein three-dimensional models showed that the main conformation and functional domains were preserved, although substitutions A223T and A291D occurred in PAC2 and PAC3 viruses, as well as A511K and E514K presented in PAC6–PAC9 viruses considerably modified the properties of the HN protein surface. The increased positive charge of the HN protein of PAC6–PAC9 viruses seems to be associated with their increased neurovirulence.

  • caracterizacion biologica de tres aislamientos naturales del rubulavirus porcino mexico
    Revista De Biologia Tropical, 2008
    Co-Authors: Maria Del Transito Borrazarguello, G Santoslopez, Veronica Vallejoruiz, Irma Herreracamacho, Julio Reyesleyva
    Abstract:

    Biological characterization of three natural isolates of the porcine rubulavirus (Mexico). Porcine rubulavirus (PoRV) produces a neurological and reproductive syndrome in pigs called the Blue-Eye Disease, known only from Mexico. Several isolates were grouped by the main symptoms presented during outbreaks: a) neurotropic in piglets, b) broadly neurotropic in piglets and gonadotropic in adults, and c) gonadotropic in adults. We studied some biological properties of three strains, which fall in one of each virus group: La Piedad Michoacan (LPM) and Produccion Animal Cerdos 1 (PAC1) and 3 (PAC3), respectively. The analyzed viral properties are mainly related with the trans-membrane hemagglutinin-neuraminidase (HN) and fusion (F) pro- teins, such as cytopathic effect, hemolysis, hemagglutinating (HA) and neuraminidase (NA) activities. In the infection assays PAC1 strain presented the highest fusogenicity level; however, the most cytolytic strain was PAC3. In addition, HA and NA activities and viral genome of PAC3 strain was detected in supernatants during cell infection earlier than in the other two strains, which shows that PAC3 virions release from the host cell earlier than LPM and PAC1. Experimental determination in purified viruses shows that PAC3 presented a higher HA and NA activities; however, PAC1 shows other interesting properties, such as a high thermostability of HN and differences about substrate profile respect to LPM and PAC3. Our data suggest that NA activity is associated with the virulence of RVP. Rev. Biol. Trop. 56 (2): 487-499. Epub 2008 June 30.

C. Mercado - One of the best experts on this subject based on the ideXlab platform.

  • Persistence of Escherichia coli, Salmonella choleraesuis, Aujeszky's Disease virus and Blue Eye Disease virus in ensilages based on the solid fraction of pig faeces.
    Journal of applied microbiology, 2001
    Co-Authors: R. Martínez‐gamba, P. Pradal‐roa, F.p. Castrejón, M. Herradora, E. Galvan, C. Mercado
    Abstract:

    Aims: This study was carried out to determine the survival time of Escherichia coli, Salmonella choleraesuis, Aujeszky’s Disease virus and Blue Eye Disease virus in ensilages based on the solid fraction of pig faeces. Methods and Results: The four micro-organisms were inoculated into microsilos based on the solid fraction of pig faeces, sorghum and molasses. They were left for 0, 7, 14, 28 and 56 days, after which the state of each microsilo was evaluated, and isolation of the inoculated agents was attempted. The four inoculated agents were isolated only on day 0 of ensilage. The viral agents were identified through the cytopathic effect and fluorescence. Conclusions, Significance and Impact of the Study: It is concluded that ensilages based on the solid fraction of pig faeces appear to reduce the risk of the transmission of the agents inoculated in this study and help to reduce the environmental impact by using the solid in animal feed.

Sergio Horacio Dueñas-jiménez - One of the best experts on this subject based on the ideXlab platform.

  • Blue Eye Disease porcine rubulavirus (PoRv) infects pig neurons and glial cells using sialo-glycoprotein as receptor
    Veterinary Journal, 2006
    Co-Authors: María Luisa Mendoza-magaña, Diana V. Godoy-martinez, Hugo Guerrero-cazares, Alejandra Rodriguez-peredo, Judith Marcela Duenas-jimenez, Sergio Horacio Dueñas-jiménez, Mario Alberto Ramírez-herrera
    Abstract:

    Abstract Pig neural cells express glycoproteins with sialylated N-linked oligosaccharide chains (SNOC) which are used by the porcine rubulavirus (PoRv) as receptors. Pig neuronal or glial cell cultures were employed to investigate (a) whether PoRv infects such cells using a molecule expressing SNOC, and (b) the role of viral envelope glycoproteins in establishing the infection. Enriched neuronal or glial cell cultures were exposed to PoRv and infection was detected immunocytochemically. Neuronal cultures prepared from neonatal pigs were treated enzymatically to eliminate sialic acid or N-linked oligosaccharide chains. Primary neural cultures were exposed to anti-HN or anti-F preincubated with PoRv to study the role of the viral glycoproteins. In enriched cultures, PoRv infected neurons and glial cells, and sialic acid expressed in N-linked oligosaccharide chains appeared to play a central role in infection. It was concluded that HN and F viral glycoproteins are required to infect neurons and glial cells.

  • Electrophysiological and morphological alterations in peripheral nerves by the pig paramyxovirus of Blue Eye Disease in neonatal pigs.
    Journal of veterinary medicine. B Infectious diseases and veterinary public health, 2001
    Co-Authors: Mario Alberto Ramírez-herrera, María Luisa Mendoza-magaña, Judith Marcela Duenas-jimenez, J. Mora-galindo, Sergio Horacio Dueñas-jiménez
    Abstract:

    The pig paramyxovirus of Blue Eye Disease (PPBED) produces central nervous system (CNS) damage leading to death in piglets. However, when PPBED was injected into the muscle and came into contact with hind limb peripheral nerves and was transported to the CNS, it did not cause death and could be a mechanism by which to induce protection. This study analyses whether PPBED causes electrophysiological and morphological alterations in infected hind limb peripheral nerves. It also studies, whether PPBED induces the onset of haemagglutination inhibitory antibodies (HIA) when it is transported to the spinal cord after medial gastrocnemius (MG) intramuscular injection. PPBED was detected by an immunohistochemical method and nerve morphology was studied using electron microscopy. The physiological status of the nerve was evaluated with electrophysiological techniques. The electrical threshold of the infected MG nerve increased four- or five fold compared to that in the ipsilateral lateral gastrocnemius or in the MG nerve on the control side. The infected nerve fibres underwent myelin sheet disarrangement and their internal fibre diameter decreased. PPBED induced the onset of HIA.

  • Pig Paramyxovirus of the Blue Eye Disease Binding to a 116 kDa Glycoprotein Expressed in Pig Neuronal Membranes
    Journal of veterinary medicine. B Infectious diseases and veterinary public health, 2001
    Co-Authors: María Luisa Mendoza-magaña, Judith Marcela Duenas-jimenez, Mario Alberto Ramírez-herrera, Sergio Horacio Dueñas-jiménez
    Abstract:

    UNLABELLED Pig paramyxovirus of the Blue Eye Disease (PPBED) is a novel member of the paramyxoviridac family which infects pigs. In neonatal pigs it causes neurological damage, whereas in adult pigs it affects the reproductive function. As PPBED damages the new-born pig central nervous system (CNS), it is important to study whether PPBED binds to the membrane proteins of all brain tissue, or selectively binds to neuronal tissue of the brain stem, olfactory bulb, hippocampus, cerebellum, frontal, temporal and parietal brain cortex. It is also important to establish whether it also infects neurones obtained from new-born, 60-day-old and adult pigs, and the role of carbohydrate residues in virus binding. The effect on virus binding of polyclonal antibodies against viral envelope proteins was also studied. Binding studies were performed using dot blot and virus overlay protein binding assays. PPBED was able to bind to membrane proteins from all brain regions, particularly to a protein band of approximately 116 kDa. Neuraminidase treatment of neuronal membrane proteins decreased virus binding; subsequent treatments with beta-galactosidase and manosidase did not increase virus binding inhibition. N-glycosidase F and trypsin also decreased virus binding, but not the O-glycanase. Antibodies against viral haemagglutinin-neuraminidase blocked virus binding more efficiently than antibodies against viral fusion protein. IN CONCLUSION (1) PPBFD is able to bind to pig neurones of all brain regions studied and at all ages analysed; (2) a 116 kDa membrane protein containing sialic acid residues with an N-linked oligosaccharide chain was specifically recognized; (3) PPBED haemagglutinin-neuraminidase protein seems to play a central role in neural receptor recognition.

Mario Alberto Ramírez-herrera - One of the best experts on this subject based on the ideXlab platform.

  • Blue Eye Disease porcine rubulavirus (PoRv) infects pig neurons and glial cells using sialo-glycoprotein as receptor
    Veterinary Journal, 2006
    Co-Authors: María Luisa Mendoza-magaña, Diana V. Godoy-martinez, Hugo Guerrero-cazares, Alejandra Rodriguez-peredo, Judith Marcela Duenas-jimenez, Sergio Horacio Dueñas-jiménez, Mario Alberto Ramírez-herrera
    Abstract:

    Abstract Pig neural cells express glycoproteins with sialylated N-linked oligosaccharide chains (SNOC) which are used by the porcine rubulavirus (PoRv) as receptors. Pig neuronal or glial cell cultures were employed to investigate (a) whether PoRv infects such cells using a molecule expressing SNOC, and (b) the role of viral envelope glycoproteins in establishing the infection. Enriched neuronal or glial cell cultures were exposed to PoRv and infection was detected immunocytochemically. Neuronal cultures prepared from neonatal pigs were treated enzymatically to eliminate sialic acid or N-linked oligosaccharide chains. Primary neural cultures were exposed to anti-HN or anti-F preincubated with PoRv to study the role of the viral glycoproteins. In enriched cultures, PoRv infected neurons and glial cells, and sialic acid expressed in N-linked oligosaccharide chains appeared to play a central role in infection. It was concluded that HN and F viral glycoproteins are required to infect neurons and glial cells.

  • Electrophysiological and morphological alterations in peripheral nerves by the pig paramyxovirus of Blue Eye Disease in neonatal pigs.
    Journal of veterinary medicine. B Infectious diseases and veterinary public health, 2001
    Co-Authors: Mario Alberto Ramírez-herrera, María Luisa Mendoza-magaña, Judith Marcela Duenas-jimenez, J. Mora-galindo, Sergio Horacio Dueñas-jiménez
    Abstract:

    The pig paramyxovirus of Blue Eye Disease (PPBED) produces central nervous system (CNS) damage leading to death in piglets. However, when PPBED was injected into the muscle and came into contact with hind limb peripheral nerves and was transported to the CNS, it did not cause death and could be a mechanism by which to induce protection. This study analyses whether PPBED causes electrophysiological and morphological alterations in infected hind limb peripheral nerves. It also studies, whether PPBED induces the onset of haemagglutination inhibitory antibodies (HIA) when it is transported to the spinal cord after medial gastrocnemius (MG) intramuscular injection. PPBED was detected by an immunohistochemical method and nerve morphology was studied using electron microscopy. The physiological status of the nerve was evaluated with electrophysiological techniques. The electrical threshold of the infected MG nerve increased four- or five fold compared to that in the ipsilateral lateral gastrocnemius or in the MG nerve on the control side. The infected nerve fibres underwent myelin sheet disarrangement and their internal fibre diameter decreased. PPBED induced the onset of HIA.

  • Pig Paramyxovirus of the Blue Eye Disease Binding to a 116 kDa Glycoprotein Expressed in Pig Neuronal Membranes
    Journal of veterinary medicine. B Infectious diseases and veterinary public health, 2001
    Co-Authors: María Luisa Mendoza-magaña, Judith Marcela Duenas-jimenez, Mario Alberto Ramírez-herrera, Sergio Horacio Dueñas-jiménez
    Abstract:

    UNLABELLED Pig paramyxovirus of the Blue Eye Disease (PPBED) is a novel member of the paramyxoviridac family which infects pigs. In neonatal pigs it causes neurological damage, whereas in adult pigs it affects the reproductive function. As PPBED damages the new-born pig central nervous system (CNS), it is important to study whether PPBED binds to the membrane proteins of all brain tissue, or selectively binds to neuronal tissue of the brain stem, olfactory bulb, hippocampus, cerebellum, frontal, temporal and parietal brain cortex. It is also important to establish whether it also infects neurones obtained from new-born, 60-day-old and adult pigs, and the role of carbohydrate residues in virus binding. The effect on virus binding of polyclonal antibodies against viral envelope proteins was also studied. Binding studies were performed using dot blot and virus overlay protein binding assays. PPBED was able to bind to membrane proteins from all brain regions, particularly to a protein band of approximately 116 kDa. Neuraminidase treatment of neuronal membrane proteins decreased virus binding; subsequent treatments with beta-galactosidase and manosidase did not increase virus binding inhibition. N-glycosidase F and trypsin also decreased virus binding, but not the O-glycanase. Antibodies against viral haemagglutinin-neuraminidase blocked virus binding more efficiently than antibodies against viral fusion protein. IN CONCLUSION (1) PPBFD is able to bind to pig neurones of all brain regions studied and at all ages analysed; (2) a 116 kDa membrane protein containing sialic acid residues with an N-linked oligosaccharide chain was specifically recognized; (3) PPBED haemagglutinin-neuraminidase protein seems to play a central role in neural receptor recognition.

Giuseppe Pisanelli - One of the best experts on this subject based on the ideXlab platform.

  • la piedad michoacan mexico virus v protein antagonizes type i interferon response by binding stat2 protein and preventing stats nuclear translocation
    Virus Research, 2016
    Co-Authors: Alan Belichavillanueva, Juliet Morrison, Bernardo Lozanodubernard, Felipa Castroperalta, Balaji Manicassamy, Giuseppe Pisanelli, Maudry Laurentrolle, Gerardo Iovane, Adolfo Garciasastre
    Abstract:

    La Piedad Michoacan Mexico Virus (LPMV) is a member of the Rubulavirus genus within the Paramyxoviridae family. LPMV is the etiologic agent of "Blue Eye Disease", causing a significant Disease burden in swine in Mexico with long-term implications for the agricultural industry. This virus mainly affects piglets and is characterized by meningoencephalitis and respiratory distress. It also affects adult pigs, causing reduced fertility and abortions in females, and orchitis and epididymitis in males. Viruses of the Paramyxoviridae family evade the innate immune response by targeting components of the interferon (IFN) signaling pathway. The V protein, expressed by most paramyxoviruses, is a well-characterized IFN signaling antagonist. Until now, there were no reports on the role of the LPMV-V protein in inhibiting the IFN response. In this study we demonstrate that LPMV-V protein antagonizes type I but not type II IFN signaling by binding STAT2, a component of the type I IFN cascade. Our results indicate that the last 18 amino acids of LPMV-V protein are required for binding to STAT2 in human and swine cells. While LPMV-V protein does not affect the protein levels of STAT1 or STAT2, it does prevent the IFN-induced phosphorylation and nuclear translocation of STAT1 and STAT2 thereby inhibiting cellular responses to IFN α/β.