• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.39-47
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• 1997

A large number of viruses belonging to Genus Hantavirus in Family Bunyaviridae are etiologic agents for hemorrhagic fever with renal syndrome (HFRS), or hantavirus pulmonary syndrome (HPS). Hantaan (HTN), Seoul (SED), Belgrade (BEL), Puumala (PUU) serotype viruses are well known causative agents for HFRS in Eurasian continent. Among those viruses Hantaan and Seoul serotypes are well known to cause HFRS in Korea, but there are some sporadic incidence by other than Hantaan or Seoul viruses. Recently we have developed the combined Hantaan-Puumala virus vaccine to prevent world-wide occuring HFRS. This combined vaccine is formalin inactivated, suckling mouse and suckling hamster brain extracts for Hantaan and Puumala viruses, respectively. Protein contents of this purified candidate vaccine is $27\;{\mu}g/ml$, which contains 1,024 ELISA antigen units to each virus, but content of myelin basic protein which is causing experimental allergic encephalomyelitis is less than 0.1 ng/ml. Thirty hamsters were given twice at one month interval intra-muscularly and bled on 30 days after each vaccination from retro-orbital sinus vein. Antibody titers were tested against 5 major serotype viruses, Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses by IFA and PRNT. The mean IF antibody titers on 30 days after primary shot were 78.4, 68.8, 68.8, 37.9, and 15.6; mean neutralizing antibody titers were 65.4, 12, 6.1, 65.6 and 0.5 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively. The mean IF antibody titers on 30 days after booster shot were 686.9, 567.5, 550.4, 516.3, and 430.9; and neutralizing antibody titers were 710.8, 41.9, 24.3, 409.9, and 1.6 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively.

• Journal of Microbiology
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• v.45 no.1
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• pp.41-47
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• 2007

The Hantaan virus (HTNV) is an enveloped virus that is capable of inducing low pH-dependent cell fusion. We molecularly cloned the viral glycoprotein (GP) and nucleocapsid (NP) cDNA of HTNV and expressed them in Vero E6 cells under the control of a CMV promoter. The viral gene expression was assessed using an indirect immunofluorescence assay and immunoprecipitation. The transfected Vero E6 cells expressing GPs, but not those expressing NP, fused and formed a syncytium following exposure to a low pH. Monoclonal antibodies (MAbs) against envelope GPs inhibited cell fusion, whereas MAbs against NP did not. We also investigated the N-linked glycosylation of HTNV GPs and its role in cell fusion. The envelope GPs of HTNV are modified by N-linked glycosylation at five sites: four sites on G1 (N134, N235, N347, and N399) and one site on G2 (N928). Site-directed mutagenesis was used to construct eight GP gene mutants, including five single N-glycosylation site mutants and three double-site mutants, which were then expressed in Vero E6 cells. The oligosaccharide chain on residue N928 of G2 was found to be crucial for cell fusion after exposure to a low pH. These results suggest that G2 is likely to be the fusion protein of HTNV.

• The Journal of Korean Society of Virology
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• v.28 no.4
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• pp.327-335
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• 1998

Hantaan (HTN) and Seoul (SEO) viruses, murid rodent-borne hantaviruses, are known to causes hemorrhagic fever with renal syndrome (HFRS) in Korea. To determine the genomic diversity and molecular phylogeny of HTN and SEO viruses found in Korea, we amplified for part of M and S genomic segments of hantaviruses from sera of HFRS patients and lung tissues of hantavirus seropositive striped-field mice. Both M and S segment of 16 HTN and 2 SEO viruses were amplified by nested reverse transcription-polymerase chain reaction. Based on 324 nucleotides in the M genomic segment, the HTN and SEO strains showed $93.8{\sim}100%$ and $99.1{\sim}99.4%$ homologies, respectively. Similarly, based on 230 nucleotides in the S genomic segment, HTN and SEO strains showed $90.9{\sim}100%$ and 100% homologies, respectively. Phylogenetic analysis of M and S segments indicated that HTN strains could be divided into at least two main groups in M and S trees and the sequence differences detected among the Sand M genomic segments of HTN viruses are consistent with reassortment having taken place between HTN virus strains.

• The Journal of the Korean Society for Microbiology
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• v.22 no.1
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• pp.1-8
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• 1987

Hantaan virus(HV) 76-118 strain was inoculated into suckling ICR mice by intra-nasal route with an inoculum of $10LD_{50}$. Mortality was 65% at the 3rd week after inoculation, but declined to 35% at the 4th week. Infectivity was determined by the measuring immuno-fluorescent antibody in sera. The peak of infectivity was 80% at the 4'th week after inoculation. Viremia was reached peak level of $1.7{\times}10^4\;PFU/ml$ by day 10. Immunofluorescent antibody and neutralizing antibody appeared by 2 weeks and 15-17 days respectively, but achieved similar titer by 35 days. By using a monoclonal antibody to HV 76-118, viral antigens were initially detected in inguinal and axillary lymph node by 2 days. Viral antigens in bone marrow and lung were delayed much more than in those of lymph node. These were similar with those of intra-peritoneal and intra-muscular route. Immune complex against IgG, IgM and C3 appeared by 16 days, 14 days, and 18 days respectively. The pattern of immunofluorescence in the basement membrane of glomeruli was diffuse membranous. Spotted pattern was also observed in the tissue stained with anti-mouse C3 antibody. By 20 days, control tissue was also shown immune complex in the glomeruli.

• Toxicological Research
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• v.13 no.3
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• pp.281-291
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• 1997

The subacute toxicity of the combined vaccine (KGCC-95VI) for the prophylaxis against Japanese encephalitis and Hantaan virus infection, recently developed by Korea Green Cross Corporation, was investigated. KGCC-95VI was subcutaneously administered into the both sexes of New Zealand White rabbits at the dosage of 0, 10. 50 and 250 ml/kg body weight (20, 100 and 500 times the expected clincal dose) once a day for 30 days. There were no deaths and clinical findings during the experiment period. In both sexes. there were no statistically significant differences between the treated and control groups in urinalysis tests, hematological tests, blood chemistry tests and pathological examinations. The KGCC-95VI is considered not to have the subacute toxicity in the rabbits.

• The Journal of Korean Society of Virology
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• v.29 no.1
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• pp.1-9
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• 1999

Hantaan virus is widely distributed among rodent populations in Korea. Two antigenically distinct hantaviruses were isolated from Apodemus agrarius in 1976 and Rattus norvegicus in 1980 in Korea. This study was designed to investigate the serological evidence of hantavirus infection among indegenous wild rodents, which were captured in 11 mountains located in Kyunggi, Kangwon, Chungnam, Chunbug and Kyungnam province of South Korea. A total 252 wild rodents of 3 species were trapped from Myungsung Mt., Chumbong Mt., Kali Mt., Hansuk Mt., Chachil peak, Kyebang Mt., Odae Mt., Kyerong Mt., Kaya Mt., Togju Mtand Chiri Mtin 1997. Serologic test for hantavirus infection was performed using hantavirus antigens by indirect immunofluorescent antibody techniqueAmong 122 Apodemus agrarius, 88 Apodemus peninsulae and 42 Eothenomys regulus; 18 A. agrarius (14.8%), 12 A. peninsulae (13.6%) and 4 E. regulus (9.5%) were immunofluorescent antibody (IFA) positive against hantaan virus. IFA titers 3 Eothenomys regulus sera were higher against puumalavirus than hantaan virus. These data imply that above three species of rodent might be natural reservoirs of hantaviruses in Korea.

• Journal of the Korea Institute of Military Science and Technology
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• v.24 no.3
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• pp.348-355
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• 2021

Hantaan virus(HTNV) causes hemorrhagic fever with renal syndrome(HFRS) with a case fatality rate ranging from <1 to 15 % in human. Hantavax^Ⓡ is a vaccine against the Hantavirus, which has been conditionally approved by the Ministry of Food and Drug Safety(MFDS). However, only 50 % of volunteers had neutralizing antibodies 1 year following the boost. Effective antiviral treatments against HTNV infection are limited. Hantaviruses generally cause asymptomatic infection in adult mice. On the other hand, infection of suckling and newborn mice with hantaviruses causes lethal neurological diesease or persistant infection, which is different from the disease in humans. The development of vaccines and antiviral strategies for HTNV has been partly hampered by the lack of an efficient lethal mouse model to evaluate the efficacy of the candidate vaccines or antivirals. In this report, we established a lethal mouse model for HTNV, which may facilitate in vivo studies on the evaluation of candidate drugs against HTNV. The median lethal dose value of HTNV was calculated by probit analysis of deaths occurring within two weeks. Five groups of ten ICR mice were injected intracranially with serial 2-fold dilutions (from 50 to 3.125 PFU/head) of HTNV. Mice injected with HTNV began to die at 8 days post-infection. The lethal dose required to kill 50 % of the mice (LD₅₀) was calculated to be 2.365 PFU/head.

• Journal of Yeungnam Medical Science
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• v.15 no.1
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• pp.55-66
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• 1998

Pathophysiological mechanism of hemorrhagic fever with renal syndrome (HFRS) is not fully understood. Major clinical findings of HFRS patients are widespread hemorrhage, acute renal failure and shock. Basic lesion is vascular injury with microvascular hemorrhage and relatively little inflammation. According to autopsy findings, renal medulla shows focal hemorrhage, tubular necrosis and interstitial mononuclear infiltrates. The predominant cell type in the renal and pulmonary interstitium is a fibroblast and it participates in the healing process at the injury site by secreting a large amount of extracellular matrix proteins. Cultured human lung fibroblasts and Mongolian gerbil fibroblasts were known to be good host cells for the hantaan virus. It is possible that not only the endothelial cell but also the fibroblast is a target of Hantaan virus and the fibroblast might be involved in the pathogenesis and the healing process in HFRS. Integrins are adhesion molecules, and act as receptors for many extracellular matrix proteins. Recently, there are many reports that cell surface integrins influence on some viral infections or reversely viruses influence on the expression of integrins. The ${\alpha}_5{\beta}_1$ integrin is a major receptor for the fibronectin which is an important extracellular matrix protein secreted by fibroblasts. In this study, the role of ${\alpha}_5{\beta}_1$ integrin in the infection of Hantaan virus was examined by using anti-${\alpha}_5{\beta}_1$, integrin, anti-${\alpha}_5$ integrin and anti-${\beta}_1$, integrin antibodies in chicken embryo fibroblasts (CEF) and Mongolian gerbil fibroblasts(MGF). The treatment of anti-${\alpha}_5{\beta}_1$, integrin antibody in CEF reduced the virion titers 26.8% and the amount of nucleocapsid N protein 32.6% when compared with control CEF. When MGF were treated with anti-${\alpha}_5$, anti-${\beta}_1$ and anti-${\alpha}_5{\beta}_1$ integrin antibodies, virion titers were reduced by 26.5%, 29.4% and 28.7% and the amount of nucleocapsid N protein were reduced by 65.2%, 59.7% and 72.6%. These results suggested that ${\alpha}_5{\beta}_1$ integrin might act as a receptor for the Hantaan virus or blocking of ${\alpha}_5{\beta}_1$ integrin influences on the viral replication in CEF and MGF. It is also possible that the blocking of only one subunit of integrin represents similar results in that of whole molecule.

• Biomedical Science Letters
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• v.6 no.2
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• pp.141-149
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• 2000

The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

• The Journal of Korean Society of Virology
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• v.29 no.4
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• pp.201-209
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• 1999

Results of the studies on the morphologic and molecular biologic characteristics of Hantaan virus (HTNV), one of the etiologic agents of Hemorrhagic fever with renal syndrome (HFRS), revealed that HTNV was a member of Family Bunyaviridae and its RNA divided into three segments. And the nucleotide sequences of these segments also were known and the differences in nucleotide sequences of HTNV from other members of genus Hantavirus were clearly evaluated. But the morphorgenesis, pathogenesis of HFRS and the replication time had not been clearly determined. In this study, to estimate the replication time of HTNV in Vero E-6 cells, Vero cells were infected with HTNV 76/118 strain, and cells were harvested from two hours post-infection up to 24 hours at two hours-intervals. Harvested cells were treated with ordinary techniques for electron microscopy and immune-electron microscopy. And then thin sections were observed under transmission electron microscope. HTNV particles were not found in the cytoplasm and in the extracellular space between $2{\sim}8$ hours after inoculation of virus, but virus particles were observed in extracellular space near the cell membrane of Vero-E6 cells 10 hours after infection. In immune electron microscopy, mature HTNV particles in extracellular spaces and immature virus labelled with gold particles in the cytoplasm of Vero E-6 cell 10 hours after infection of HTNV could be seen. This results suggest that the replication time of HTNV might be about 10 hours.