• Toxicological Research
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• v.13 no.1_2
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• pp.157-160
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• 1997

The primary skin and eye irritancy 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. The KGCC-95VI was applied to the back skins of the New Zealand White rabbits. The rabbits were observed for 72 hours and did not exhibit erythema, eschar and edema. The eyes of the rabbits were exposed to the KGCC-95VI. The rabbits were observed for 7 days and did not exhibit any ocular findings on cornea, iris and conjuntivae. The KGCC-95VI is considered not to have the primary skin and eye toxicity in rabbits.

• Biomolecules & Therapeutics
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• v.5 no.2
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• pp.174-178
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• 1997

The immnunogenicity of the possible non-essential component of the combined vaccine (KGCC-957) for the prophylaxis against Japanese encephalitis and Hantaan virus infection recently developed by Korea Green Cross Corporation was investigated using the Hartley guinea pigs. The KGCC-95Vl was administered to the guinea pigs subcutaneously to sensitize the animals. The guinea pigs did not induce any anaphylactic immune responses which could be detectable by the active systemic anaphylaxis (ASA), the passive systemic anaphylaxis (PSA), and the passive cutaneous anaphylaxis (PCA) tests. The KGCC-95Vl is considered not to induce any anaphylactic immune responses except the prophylatic immune effects of the vaccine.

• The Journal of Korean Society of Virology
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• v.28 no.3
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• pp.287-293
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• 1998

Hantaan virus are widely distributed in rodents populations in Korea. Two antigenically distinct hantaviruses have been isolated from Apodemus agrarius in 1976 and Rattus norvegicus in 1980 in Korea. This study was designed to find the serological evidence of hantavirus infection among indigenous wild rodents captured in 7 Mountains located in Kangwon province of south Korea. A total 191 wild rodents of 3 species were trapped in Chumbong mountain, Kali mountain, Hansuk mounatin, Chachil peak, Bukam ridge, Kyebang mountain and Odae mountain in 1997. Serologic evidence for hantavirus infection were tested using hantavirus antigens by indirect immunofluorescent antibody technique (IFA). Among 85 Apodemus agrarius, 77 Apodemus peninsulae and 29 Eothenomys regulus; 8 A. agrarius (9.4%), 11 A. peninsulae (14.3%) and 4 E. regulus (13.8%) were immunofluorescent antibody positive against hantaan virus. IF antibody titers against Puumala virus of 3 E. regulus sera were higher than against hantaan virus. This data suggest that several antigenically distinct hantaviruses have been circulated in rodent populations in Korea.

• The Journal of Korean Society of Virology
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• v.29 no.3
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• pp.155-163
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• 1999

Hantaan virus (HTNV), the etiologic agent of hemorrhagic fever with renal syndrome (HFRS), belongs to the genus Hantavirus, and has three single negative stranded RNA genome segments. HTNV strain Howang isolated from the blood of severe case of Korean HFRS is more virulent than HTNV 76/118 and the M and S genome segments' nucleotide sequence of Howang strain showed 93.5% and 94% homology to each segment of HTNV 76/118. We have obtained 6533 nucleotides long sequence of the L genome segment of Howang strain using reverse transcriptase in conjunction with PCR amplification and compared to other hantaviruses. The messenger sense of the L segment contains one long single long open reading frame of 2151 amino acids, which encodes a deduced RNA dependent RNA polymerase of 246.4 kDa caculated molecular weight protein. The nucleotide sequence of the L segment of Howang strain shows 93%, 74%, 66%, 65% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus $H{\ddot{a}}lln{\ddot{a}}s$ B1 and Sin Nombre virus, respectively. The amino acid sequence of the L segment of Howang strain shows 99%, 85%, 68%, 68% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus $H{\ddot{a}}lln{\ddot{a}}s$ B1 and Sin Nombre virus, respectively.

• KSBB Journal
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• v.13 no.6
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• pp.649-655
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• 1998

Hantaviruses are rodent hosts-borne viruses belonging to the family Bunyaviridae, and are etiologic agents for two acute diseases, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome (HPS). There have been a lot of reports on prophylactic vaccines and diagnostics for the diseases, but most of viral antigens have been prepared by eukaryotic cell culture. Nucleocapsid proteins of Hantaviruses are known as the major viral antigens. Thereby, we prepared nucleocapsid genes of Hantaan virus and Seoul virus by RT-PCR and cloned into plasmid vectors, pET-3a and pKK223-3. Both genes were expressed in Escherichia coli with higher expression level of Seoul viral nucleocapsid protein compared to that of Hantaan in pET-3a. Hantaan viral gene was expressed much higher level in plasmid pET-3a that in pKK223-3. About 30% of expressed nucleocapsid protein was soluble and the rest was remained in insoluble fraction.

• KSBB Journal
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• v.13 no.6
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• pp.662-668
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• 1998

Viruses belonging to the Hantavirus genus cause two acute severe illness in humans, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome(HPS). Among them, Hantaan virus is one of the most important viruses causing HFRS. Recombinant expression vectors, pKK-NP and pET-NP, with Hantaan viral nucleocapsid gene were constructed, and used to transform Eschericia coli BL21(DE3). Stability of the vectors in the host strain, and effects of some environmental conditions on the expression of nucleocapsid gene were studied. Expression vector, pKK-NP, was very unstable, and the expression level of nucleocapsid gene was very low compared to that of pET-NP. BL21(pET-NP) produced about 100 mg of N protein per liter of culture broth. Induction time did not show any significant difference on the expression level of nucleocapsid gen and cell growth. BL21(pET-NP) culture at 35$^{\circ}C$ showed a little higher expression level than at 30$^{\circ}C$ during growth phase, but reached to the same level at stationary phase. Total expression level was proportional to supplemented glucose concentration of media up to 0.5% along with cell growth, but expression level per unit cell mass was inversely proportional to glucose concentration and maximal when glucose was not supplemented at all.

• Toxicological Research
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• v.13 no.3
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• pp.275-279
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• 1997

The acute 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 administered to the Balb /c mice in two routes, orally and subcutaneously, and into the New Zealand White rabbits subcutaneously. $LD_{50}$ was not accessible as there were no deaths in the group treated even at a dose 800 times the expected clinical dose in both animal species. Between the treated and control groups there were no statistically significant differences in body weight changes and clinical signs during the 14-day observation period, and no pathological gross findings. Accordingly KGCC-95VI is considered not to have the acute toxicity in mice and rabbits.

• Toxicological Research
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• v.13 no.4
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• pp.353-357
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• 1997

The possibility of the allergic encephalomyelitis caused by the combined vaccine (KGCC95VI) for the prophylaxis against Japanese encephalitis and Hantaan virus infection, recently developed by Korea Green Cross Corporation, was investigated in the Hartley guinea pigs. The KGCC-95VI was administered to the guinea pigs subcutaneously to sensitize the animals three times at one month intervals. There were no clinical signs or gross pathological findings. There were no abnormal histopathological findings at cerebrums, cerebellums, brain stems and the spinal cords. The concentration of myelin basic protein was 1.10 ng/dose quantified by ELISA, which met the guide4ine of below 2 ng/ml/dose recommended by American Society of Health -System Pharmacists(AHPS) Drug Information. Accordingly, the KGCC-95VI is considered not to induce any allergic immune responses which may lead to the experimental allergic encephalomyelitis.

• The Journal of Korean Society of Virology
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• v.30 no.1
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• pp.11-18
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• 2000

Since $Hantavax^{TM}$, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after primary basic vaccination with $Hantavax^{TM}$. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees at one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result $Hantavax^{TM}$ has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.

• Journal of Microbiology
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• v.43 no.6
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• pp.537-545
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• 2005

Even though neutralizing antibodies against the Hantaan virus (HTNV) has been proven to be critical against viral infections, the cellular immune responses to HTNV are also assumed to be important for viral clearance. In this report, we have examined the cellular and humoral immune responses against the HTNV nucleocapsid protein (NP) elicited by virus infection or DNA vaccination. To examine the cellular immune response against HTNV NP, we used $H-2K^b$ restricted T-cell epitopes of NP. The NP-specific $CD8^+$ T cell response was analyzed using a $^{51}Cr-release$ assay, intracellular cytokine staining assay, enzyme-linked immunospot assay and tetramer binding assay in C57BL/6 mice infected with HTNV. Using these methods, we found that HTNV infection elicited a strong NP-specific $CD8^+$ T cell response at eight days after infection. We also found that several different methods to check the NP-specific $CD8^+$ T cell response showed a very high correlation among analysis. In the case of DNA vaccination by plasmid encoding nucleocapsid gene, the NP-specific antibody response was elicited $2\~4$ weeks after immunization and maximized at $6\~8$ weeks. NP-specific $CD8^+$ T cell response reached its peak 3 weeks after immunization. In a challenge test with the recombinant vaccinia virus expressing NP (rVV-HTNV-N), the rVV-HTNV-N titers in DNA vaccinated mice were decreased about 100-fold compared to the negative control mice.