scholarly journals Preparation of Anti-Rabies Virus N Protein IgYs by DNA Immunization of Hens Using Different Types of Adjuvants

2021 ◽  
Author(s):  
Nanase Kubo ◽  
Mari Nishii ◽  
Satoshi Inoue ◽  
Akira Noguchi ◽  
Hajime Hatta
Keyword(s):  
Rabies Virus ◽  
Dna Immunization ◽  
N Protein ◽  
Different Types

Molecular characterisation of rabies virus detected in livestock animals in the southern part of Egypt during 2018 and 2019

2021 ◽  
Author(s):  
Serageldeen Sultan ◽  
Soheir Abdou Hussein Ahmed ◽  
Mohamed Wael Abdelazeem ◽  
Sabry Hassan
Keyword(s):  
Amino Acid ◽  
Rabies Virus ◽  
Histopathological Examination ◽  
Fluorescent Antibody ◽  
N Gene ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
N Protein ◽  
Geographical Regions ◽  
The One

AbstractBrain samples were collected from 33 animals of different species, including buffalo, cattle, dog, donkey, fox and wolf, that had been suspected to be infected by rabies virus (RABV) in different geographical regions of Aswan and Luxor governorates in Egypt. The samples were submitted for histopathological examination and the presence of the nucleic acid and antigens of RABV was tested by RT-PCR and indirect fluorescent antibody technique (IFAT), respectively. Sixteen samples were found positive by all the three examinations. Three samples were selected for further study from animals in which the highest virus loads were detected. The partial sequence of the RABV N gene was determined and analysed from the samples of a buffalo, a cow and a donkey. The viruses in the samples were found to share 95–98% and 95–97% nucleotide and amino acid sequence identities, respectively. In comparison to reference sequences, a few amino acid substitutions occurred in the N protein antigenic sites I and IV in the immunodominant epitopes of the viruses detected in the cow and the donkey but not in the one from the buffalo. The phylogenetic analysis revealed that the RABVs sequenced from the samples belonged to genotype 1, Africa-4 clade, and formed two distinct sub-clades within the Egyptian clade. These findings indicate the circulation of RABV among livestock animals in the southern part of Egypt and raise public health concerns. The amino acid changes detected in this work may contribute to the antigenic diversification of RABVs.

scholarly journals A surface plasmon resonance biosensor for direct detection of the rabies virus

2012 ◽  
Vol 81 (2) ◽  
pp. 107-111 ◽  
Author(s):  
Jing Xu ◽  
Jia-yu Wan ◽  
Song-tao Yang ◽  
Shou-feng Zhang ◽  
Na Xu ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Rabies Virus ◽  
Surface Plasmon ◽  
Specific Antibody ◽  
Plasmon Resonance ◽  
Fluorescent Antibody ◽  
Test Surface ◽  
N Protein ◽  
Surface Plasmon Resonance Biosensor ◽  
Biosensor Chip

A surface plasmon resonance biosensor chip was constructed for detection of rabies virus. For the construction of the biosensor chip, N protein specific antibody and N protein specific antibody combined with G protein specific antibody of rabies virus were linked on two different flow cells on one CM5 chip, respectively. The chip was tested for the detection of rabies virus antigens using the crude extract of rabies virus from infected BHK cell strain culture. Tenfold serial dilutions of SRV9 strain virus-infected cell cultures were tested by the biosensor chip to establish the detection limit. The limit detection was approximately 70 pg/ml of nucleoprotein and glycoprotein. The biosensor chip developed in this study was employed for the detection of rabies virus in five suspect infectious specimens of brain tissue from guinea pigs; the results were compared by fluorescent antibody test. Surface plasmon resonance biosensor chip could be a useful automatic tool for prompt detection of rabies virus infection.

scholarly journals Genetic Mapping of Rabies Virus in Dogs as a Basis for Disease Control

2016 ◽  
Vol 10 (1) ◽  
Author(s):  
Muharam Saepulloh ◽  
R. M. Abdul Adji
Keyword(s):  
Phylogenetic Analysis ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Phylogenetic Relationship ◽  
Rabies Virus ◽  
Virus Isolate ◽  
Protein Gene ◽  
Genetic Characteristics ◽  
Nucleoprotein Gene ◽  
N Protein

The purpose of this study was to determine the genetic characteristics of rabies virus based on phylogenetic relationship among rabies virus in various regions in Indonesia. The amino acid sequence of the nucleoprotein gene of rabies virus isolate from Banten (RV/Banten01/dog/2007),Makasar (RV/MKS-26/dog/2010), Bukit Tinggi (RV/BKT-52/dog/2009 and RV/BKT-58/dog/2009), Medan (RV/Medan27/dog/2007)andBali(RV/Bali-1/dog/2009;RV/Bali-2/dog/2009;RV/Bali-3/dog/2009),Indonesiawasdetermined.TheseisolatesshowedahighdegreeofhomologyamongIndonesianisolateswhichreached100%.Meanwhile,thelevelofhomologybetweenrabiesvirusisolatesfromcatsrabiesvirusisolatesfromdogsreached97%.ResultsofphylogeneticanalysisusingtheaminoacidsequencesoftheNgenesshowedthatallofIndonesianrabiesvirusisolateswerecloselyrelatedtorabiesvirusesfromChinathanthosefromThailand,Laos,Burma,andVietnamwhichgeograficallycloser to Indonesia. Data obtained from the phylogenetic analysis is expected to trace the source of rabies spread and thepossibility to create a vaccines which more suitable with rabies virus that spreads in Indonesia. Based on the phylogenetic relationship analysisusing the amino acid sequence of the rabies virus N protein gene showed that all of rabies virus isolated from Indonesian regions share a highhomology with others ranging from 97-100%..Key words: sequencing, rabies, nucleoprotein gene (N), homology

scholarly journals Structural relationship between nucleocapsid-binding activity of the rabies virus phosphoprotein (P) and exposure of epitope 402-13 located at the C terminus

2002 ◽  
Vol 83 (12) ◽  
pp. 3035-3043 ◽  
Author(s):  
Harufusa Toriumi ◽  
Yoshikazu Honda ◽  
Kinjiro Morimoto ◽  
Tadafumi S. Tochikura ◽  
Akihiko Kawai
Keyword(s):  
Rabies Virus ◽  
Structural Changes ◽  
Binding Activity ◽  
Free Form ◽  
Gene Products ◽  
Epitope Region ◽  
N Protein ◽  
P Gene ◽  
P Protein ◽  
P Proteins

The structural changes of the nominal phosphoprotein (P) of rabies virus using a monoclonal antibody, mAb #402-13, was investigated. This mAb recognized a linear epitope that was mapped roughly to a C-terminal region of the P protein, ranging from aa 256 to 297. The P gene products were detected by the mAb in immunoblot assays, the products of which were produced either in BHK-21 cells or in Escherichia coli cells. The mAb, however, detected very low levels of P gene products in immunoprecipitation assays. The mAb recognized the nucleocapsid (NC)-associated P proteins but recognized free P protein and free N–P complex produced in the infected cells much less efficiently. When the P proteins were released from the NC, however, they were no longer recognized by the mAb. Similar results were obtained from BHK-21 cells co-transfected with P and N cDNAs. Furthermore, studies with C-terminally truncated P protein mutants revealed that the NC-binding ability of the P protein was dependent on the presence of the C-terminal epitope region. From these results, it is thought that the 402-13 epitope region is concealed when the P protein is present in a free form or free N–P complex but is exposed when it is associated with the NC. The C-terminal epitope region seemed to be essential for the P protein to be associated with the NC but not for the formation of free N–P complexes with newly synthesized N protein.

DNA immunization protects nonhuman primates against rabies virus

1998 ◽  
Vol 4 (8) ◽  
pp. 949-952 ◽  
Author(s):  
Donald L. Lodmell ◽  
Nancy B. Ray ◽  
Michael J. Parnell ◽  
Larry C. Ewalt ◽  
Cathleen A. Hanlon ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Nonhuman Primates ◽  
Dna Immunization

scholarly journals Pathogenicity of Different Rabies Virus Variants Inversely Correlates with Apoptosis and Rabies Virus Glycoprotein Expression in Infected Primary Neuron Cultures

1999 ◽  
Vol 73 (1) ◽  
pp. 510-518 ◽  
Author(s):  
Kinjiro Morimoto ◽  
D. Craig Hooper ◽  
Sergei Spitsin ◽  
Hilary Koprowski ◽  
Bernhard Dietzschold
Keyword(s):  
Protein Expression ◽  
G Protein ◽  
Rabies Virus ◽  
Antigenic Site ◽  
Expression Levels ◽  
N Protein ◽  
Adult Mice ◽  
Glycoprotein G ◽  
Neuronal Processes ◽  
Nucleoprotein N

ABSTRACT The mouse-adapted rabies virus strain CVS-24 has stable variants, CVS-B2c and CVS-N2c, which differ greatly in their pathogenicity for normal adult mice and in their ability to infect nonneuronal cells. The glycoprotein (G protein), which has previously been implicated in rabies virus pathogenicity, shows substantial structural differences between these variants. Although prior studies have identified antigenic site III of the G protein as the major pathogenicity determinant, CVS-B2c and CVS-N2c do not vary at this site. The possibility that pathogenicity is inversely related to G protein expression levels is suggested by the finding that CVS-B2c, the less pathogenic variant, expresses at least fourfold-higher levels of G protein than CVS-N2c in infected neurons. Although there is some difference between CVS-B2c- and CVS-N2c-infected neurons in G protein mRNA expression levels, the differential expression of G protein appears to be largely determined by posttranslational mechanisms that affect G protein stability. Pulse-chase experiments indicated that the G protein of CVS-B2c is degraded more slowly than that of CVS-N2c. The accumulation of G protein correlated with the induction of programmed cell death in CVS-B2c-infected neurons. The extent of apoptosis was considerably lower in CVS-N2c-infected neurons, where G protein expression was minimal. While nucleoprotein (N protein) expression levels were similar in neurons infected with either variant, the transport of N protein into neuronal processes was strongly inhibited in CVS-B2c-infected cells. Thus, downregulation of G protein expression in neuronal cells evidently contributes to rabies virus pathogenesis by preventing apoptosis and the apparently associated failure of the axonal transport of N protein.

scholarly journals Definition of the immune evasion-replication interface of rabies virus P protein

2021 ◽  
Vol 17 (7) ◽  
pp. e1009729
Author(s):  
Jingyu Zhan ◽  
Angela R. Harrison ◽  
Stephanie Portelli ◽  
Thanh Binh Nguyen ◽  
Isshu Kojima ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Immune Evasion ◽  
Structural Data ◽  
Direct Analysis ◽  
Multifunctional Protein ◽  
N Protein ◽  
Binding Interface ◽  
P Protein ◽  
Definition Of ◽  
Nucleoprotein N

Rabies virus phosphoprotein (P protein) is a multifunctional protein that plays key roles in replication as the polymerase cofactor that binds to the complex of viral genomic RNA and the nucleoprotein (N protein), and in evading the innate immune response by binding to STAT transcription factors. These interactions are mediated by the C-terminal domain of P (PCTD). The colocation of these binding sites in the small globular PCTD raises the question of how these interactions underlying replication and immune evasion, central to viral infection, are coordinated and, potentially, coregulated. While direct data on the binding interface of the PCTD for STAT1 is available, the lack of direct structural data on the sites that bind N protein limits our understanding of this interaction hub. The PCTD was proposed to bind via two sites to a flexible loop of N protein (Npep) that is not visible in crystal structures, but no direct analysis of this interaction has been reported. Here we use Nuclear Magnetic Resonance, and molecular modelling to show N protein residues, Leu381, Asp383, Asp384 and phosphor-Ser389, are likely to bind to a ‘positive patch’ of the PCTD formed by Lys211, Lys214 and Arg260. Furthermore, in contrast to previous predictions we identify a single site of interaction on the PCTD by this Npep. Intriguingly, this site is proximal to the defined STAT1 binding site that includes Ile201 to Phe209. However, cell-based assays indicate that STAT1 and N protein do not compete for P protein. Thus, it appears that interactions critical to replication and immune evasion can occur simultaneously with the same molecules of P protein so that the binding of P protein to activated STAT1 can potentially occur without interrupting interactions involved in replication. These data suggest that replication complexes might be directly involved in STAT1 antagonism.

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