• Korean Journal of Veterinary Research
• /
• v.57 no.4
• /
• pp.249-252
• /
• 2017

Rabies virus (RABV) causes a neurological disease in warm-blooded animals that is nearly always fatal. In this study, we analyzed the matrix (M) genes in 10 Korean street RABV strains isolated from two Provinces during 2011-2013. The M genes in these 10 Korean strains were highly conserved during 1999-2013. Phylogenetic analysis revealed they were closely related to the M genes of RABVs isolated in northeastern China. Specific amino acid substitutions were identified in the KRVB1206, KRVF1301, and BV9901PJ strains. However, functional domains, including those involved in virus production and pathogenicity, were conserved in all 10 strains.

• Korean Journal of Veterinary Research
• /
• v.52 no.1
• /
• pp.9-18
• /
• 2012

Rabies is a major zoonotic disease that causes approximately 55,000 human deaths worldwide on an annual basis. The nucleocapsid protein and glycoprotein genes of the Korean rabies virus (RABV) have been subjected to molecular and phylogenetic analyses. Although the phosphoprotein (P) has several important functions in viral infection and pathogenicity, the genetic characterizations of the P of Korean RABV isolates have not yet been established. In the present study, we conducted genetic analyses of P genes of 24 RABV isolates circulating in the Republic of Korea (hereafter, Korea) from 2008 to 2011. This study revealed that the P genes of Korean RABVs are genetically similar to those of RABV strains of lyssavirus genotype I including V739 (dogs, Korea), NNV-RAB-H (humans, India), NeiMeng925 (raccoon dogs, China), and RU9.RD (raccoon dogs, Russia). Among Korean isolates, the RABV P genes showed low variability in the variable domains among Korean isolates; they had specific consensus sequences and amino acid substitutions capable of identifying geographic characteristics and retained specific sequences thought to be important for viral function. These results provide important genetic characteristics and epidemiological information pertaining to the P gene of the Korean RABV.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.4
• /
• pp.718-724
• /
• 2017

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.4
• /
• pp.366-373
• /
• 2011

We characterized the proteomes of murine N2a cells following infection with three rabies virus (RV) strains, characterized by distinct virulence phenotypes (i.e., virulent BD06, fixed CVS-11, and attenuated SRV9 strains), and identified 35 changes to protein expression using two-dimensional gel electrophoresis in whole-cell lysates. The annotated functions of these proteins are involved in various cytoskeletal, signal transduction, stress response, and metabolic processes. Specifically, a-enolase, prx-4, vimentin, cytokine-induced apoptosis inhibitor 1 (CIAPIN1) and prx-6 were significantly up-regulated, whereas Trx like-1 and galectin-1 were down-regulated following infection of N2a cells with all three rabies virus strains. However, comparing expressions of all 35 proteins affected between BD06-, CVS-11-, and SRV9-infected cells, specific changes in expression were also observed. The up-regulation of vimentin, CIAPIN1, prx-4, and 14-3-3 ${\theta}/{\delta}$, and down-regulation of NDPK-B and HSP-1 with CVS and SRV9 infection were ${\geq}2$ times greater than with BD06. Meanwhile, Zfp12 protein, splicing factor, and arginine/serine-rich 1 were unaltered in the cells infected with BD06 and CVS-11, but were up-regulated in the group infected with SRV9. The proteomic alterations described here may suggest that these changes to protein expression correlate with the rabies virus' adaptability and virulence in N2a cells, and hence provides new clues as to the response of N2a host cells to rabies virus infections, and may also aid in uncovering new pathways in these cells that are involved in rabies infections. Further characterization of the functions of the affected proteins may contribute to our understanding of the mechanisms of RV infection and pathogenesis.

• Microbiology and Biotechnology Letters
• /
• v.46 no.3
• /
• pp.269-276
• /
• 2018

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

• Korean Journal of Veterinary Research
• /
• v.50 no.2
• /
• pp.133-137
• /
• 2010

Rabies virus (family Rhabdoviridae, genus Lyssavirus, RV) is the causative agent of rabies in mammals. We conducted a sero-epidemiological survey for RV using sera from South Korean stray and companion dogs in the present study. A total of 533 canine serum samples were collected between February 2006 and December 2007 and were screened for rabies immunity with a neutralizing peroxidase linked assay. Both companion (49.1%) and stray (60.1%) dogs demonstrated RV seropositivity. Regional RV antibody prevalence was measured in the Jeju (87.5%), Gyeonggi (62%), Gyeongsang (59.1%), Jeonra (42%), Chungcheong (37.9%), and Gangwon (30.4%) provinces. Prevalence increased with age but did not exceed 80% in any age group. Stray and companion dogs had RV antibody prevalence values of 26.7% and 23.7%, respectively. Seroprevalence was significantly associated with age $({\chi}2\;=\;9.46;\;p\;=\;0.024)$ for companion dogs, although this association was not evident in stray dogs. There were no significant differences in age between stray and companion dogs and no gender differences in RV seroprevalence. Our results suggested that a widespread and reinforced vaccination program must be applied to Korean dogs.

• Korean Journal of Veterinary Service
• /
• v.37 no.4
• /
• pp.281-290
• /
• 2014

The purpose of this study was to determine the zoonotic diseases of stray dog and cat in Gwangju, Korea. We chose randomly 300 samples which is 265, dogs and 35, cats in the public animal shelter from March to August of 2013. The animals' blood biochemistry values measured, and the out of normal values were 49.7% GOT, 36.3% GPT, and 78.0% GGT. The tested items were Dirofilaria immitis, Toxoplasma gondii, Brucella canis, Rabies virus. The positive results were 10% Dirofilaria immitis, 6.3% Toxoplasma gondii (antibody detected), 10% Rabies (antibody detected) but 0.0% in B. canis. The stray animals' antibody antigen positivity take effect high from Mar.-May. Therefore, we propose that those diseases should be monitering and vaccinating in Korea.

• Korean Journal of Veterinary Service
• /
• v.18 no.1
• /
• pp.57-71
• /
• 1995

During the period of 2 years from 1993 to 1994, epidemiological investigation were carried out each time the outbreak of rabies suspected cases were informed and all specimens were transported to the Kangwon-do Veterinary Service Lab. At the Lab., autopsy and laboratory examinations were conducted as well. The results obtained as follows 1. The dogs and cattles were confirmed to be infected with rabies virus by routine histopathological findings, indirect flulrescent antibody technique, and mouth inoculation test. 2. The total numbers of rabies occured in dogs and cattles were 9, 5 heads, respectively. The regional distribution of total of 14 heads were 9 heads in Chulwon, 2 heads in Hwachun, 1 head Yangku, 2 heads in Inje and the highest number of outbreaks were orrured in Chulwon. 3. Most of dogs showed furious type and dumb type found in only one dog. 4. Cattles showed the symtoms of uneasiness feeling, excitement, disturbance howling, the change of voice, sexual accelaration, foreign body swatting and attacked to human and ani-mal and found to be died after two or four days. 5. In the seasonal outbreaks of rabies, 8 heads of the highest incidence were occured in win-ter followed by 4 heads in autumn, 1 head in spring and 1 head in summer. 6. The first outbreaks of rabies was reported in Chulwon and then transmitted into Hwachun, Yangku and Inje in turn. 7 The infectious source was supposed to be wild animals such as laccon dog, badger etc. living around the D. M. Z.

• Korean Journal of Veterinary Research
• /
• v.57 no.4
• /
• pp.209-214
• /
• 2017

Wild raccoon dogs (Nyctereutes procyonoides koreensis) may play a role transmitting several pathogens to humans and pet animals. Information concerning the incidence of rabies, canine distemper virus (CDV), canine parvovirus (CPV), canine adenovirus type 2 (CAdV-2), canine parainfluenza virus type 5 (CPIV-5), and canine herpesvirus (CHV) is needed in wild raccoon dogs. In total, 62 brain samples of raccoon dogs were examined for rabies virus (RABV) and CDV, and 49 lung samples were screened for CDV, CAdV-2, CPIV-5, and CHV. No RABV, CAdV-2, CPIV-5, or CHV was identified, but nine CDV antigens (8.1%, 9/111) were detected. Moreover, 174 serum samples from wild raccoon dogs were screened for antibodies against the five major viral pathogens. The overall sero-surveillance against CDV, CPV, CAdV-2, CPIV-5, and CHV in wild raccoon dogs was 60.3%, 52.9%, 59.8%, 23.6%, and 10.3%, respectively. Comparisons of the sero-surveillance of the five pathogens showed that raccoon dogs of Gyeonggi province have slightly higher sero-positive rates against CDV, CPV, and CHV than those of Gangwon province. These results indicate high incidences of CDV, CPV, and CAdV-2 in wild raccoon dogs of two Korean provinces and a latent risk of pathogen transmission to companion and domestic animals.

• Molecules and Cells
• /
• v.44 no.10
• /
• pp.770-779
• /
• 2021

Transgenic Arabidopsis thaliana expressing an anti-rabies monoclonal antibody (mAb), SO57, was obtained using Agrobacterium-mediated floral dip transformation. The endoplasmic reticulum (ER) retention signal Lys-Asp-Glu-Leu (KDEL) was tagged to the C-terminus of the anti-rabies mAb heavy chain to localize the mAb to the ER and enhance its accumulation. When the inaccurately folded proteins accumulated in the ER exceed its storage capacity, it results in stress that can affect plant development and growth. We generated T₁ transformants and obtained homozygous T₃ seeds from transgenic Arabidopsis to investigate the effect of KDEL on plant growth. The germination rate did not significantly differ between plants expressing mAb SO57 without KDEL (SO plant) and mAb SO57 with KDEL (SOK plant). The primary roots of SOK agar media grown plants were slightly shorter than those of SO plants. Transcriptomic analysis showed that expression of all 11 ER stress-related genes were not significantly changed in SOK plants relative to SO plants. SOK plants showed approximately three-fold higher mAb expression levels than those of SO plants. Consequently, the purified mAb amount per unit of SOK plant biomass was approximately three times higher than that of SO plants. A neutralization assay revealed that both plants exhibited efficient rapid fluorescent focus inhibition test values against the rabies virus relative to commercially available human rabies immunoglobulins. KDEL did not upregulate ER stress-related genes; therefore, the enhanced production of the mAb did not affect plant growth. Thus, KDEL fusion is recommended for enhancing mAb production in plant systems.