Rats의 한탄바이러스 및 서울바이러스에 대한 중화항체 형성

이평우,최순옥,이호왕 대한바이러스학회 1988 Journal of Bacteriology and Virology Vol.18 No.1

The production of neutralizing antibodies against Hantaan and Seoul viruses in sera from rats were investigated using plaque reduction neutralization test. Rats were bled at one week interval for 3 months after intramuscular inoculation of Han- taan and Seoul viruses and sera were used for titration of neutralizing antibodies against Hantaan and Seoul virus, respectively by plaque reduction neutralization method employing immunoperoxidase staining in Vero E6 cell cultures. In rats inoculated with Hantaan virus, titers of plaque reduction neutralizing against Hantaan virus increased significantly from 320 at one week to 2,560 at 3-4 weeks and persisted for 60 days. In contrast, neutralizing antibody titers against Seoul virus were much lower than those of Hantaan virus and antibody titer was 160, although it persisted for 2 months. In rats inoculated with Seoul virus, titers of neutralizing antibody against Seoul virus were shown similar values to that of Hantaan virus after Hantaan virus inoculation and maximum titer was 10,240 at 3-5 weeks but neutralizing antibody titers against Hantaan virus were very low and peak value was 160-320. In this study using a rat model, it was observed that neutralizing antibodies against Hantaan and Seoul viruses were specific and significantly different depend on inoculation of vurus and it was easy to differentiate Hantaan virus infection from Seoul virus infection. This study has also demonstrated that the formation of cross neutralizing antibody between Hantaan and Seoul virus after inoculation of the virus into rats for the first time.

일본 뇌염바이러스 감염에 대한 마크로파지의 역할

이종훈,이연태,김금용,임병욱 대한바이러스학회 1977 Journal of Bacteriology and Virology Vol.7 No.1

Macrophage, scavenger cell against foreign materials, may play an important role cn hcst defense mechanism, maintenance of homeostasis function, and various immune resporses. However, it is not certain wheather or not those function of macrophages derived from patients who were treated with immuncsuppressive drugs may be kept well as in normal persens. To investigate the effect of immuncsuppressive drugs (prednisolone, cyclophcsphamide and vincristine sulfate) on mcuse peritoneal macrophages against Japanese encephalitis virus(JE virus) infecticn, each drug was dissolved and adjusted with physiological saline at the proper concentration(prednisolone 20mg/kg B.W./day, 200mg/kg B.W./day; cyclcsphamide 10mg/kg B.W./day, 100mg/kg B.W./day; vincristine sulfate 0.05mg/kg B.W. /day, 0.5mg/kg B.W./day). Then, were injected intraperitoneally with each drug solution for three corsecutive days. In 24 hours after last three groups of mice injection, peritoneal macrophages were isolated. And then macrophage layers were formed both the inner surface of Leightcn tube and on the coversligs of Leighten tube. Thereafter, JE virus was inoculated on macrophage monolayer ar.d cultured at 37 C. At apprcpriate time interval, macrophage cultvres were sempled. JE virus antigrs retained in the macrophage were detected by immuncfluorescence technique, and viral infectivity was measured by the plaque-forming methad on the monolayer of chick embryo fibroblasts. Peritoneal macrophages obtained from untreated nomal mice were used as the control. The results were summarized as follows: 1. On the cultures of normal peritoneal macrophage phagccytized Japarese cncephslitis virus, mezximal rate of macrophages retaining JE virus antigens was reachcd at 6 hours after virus adsorption. Thereafter, the rate of immunofluorescent cells was rapidly decreased, but low rate was still observed at 96 hours after virus inoculation. In contrast, on the cultures of peritoneal macrophages obtained from mice that were pretreated with immunosuppressive drug, such as prednisolone, cyclophosphamide and vincristine sulfate, the rate of macrophage retaining JE virus antigens was higher than that of untreated control macrophages. Duration necessary for reaching to the maximal rate was required approxinately 2-6 hours more than that of the control macrophages.2. Within 24 hours after the virus inoculation, virus infectivity was rapidly inactivated in cell-free medium, but inact!vation of virus infectivity in tbe macrophage cultures was more rapid than that of cell-free medium. However, virus infectivity recovered from macrophage cultures, even though low in titer, was continuously maintained throughout 96 hours. Recovery rate of the infectivity, which was observed from macrophage cultures from immunosuppressant pretreated mice, was always bigher than that of the untreated control mcrophage cultures.

한탄바이러스 및 서울바이러스에 대한 단세포군 항체생산과 그 특성분석

반상자,이호자,박순희,조해월 대한바이러스학회 1995 Journal of Bacteriology and Virology Vol.25 No.1

To study the antigenic characteristics and differentiation serotypes of Hantavirus, the produced hybridoma secreting monoclonal antibodies(Mabs) against Hantaan virus, 76/118 strain and Seoul virus, 80/39 strain were produced by fusion of mouse myeloma cell, Sp2/0-Ag14 with spleen cells isolated from Balb/c mice immunized with Hantaan virus or Seoul virus. From the result in this study, the Mabs against Hantaan virus were screened by indirect immunofluorescent antibody techinque(IFA). 587 hybridoma cells were produced out of 768 wells and the fusion frequency was 76%. Seven among 587 hybridoma cells produced Mabs against Hantaan virus continuously. Isotype analysis of Mabs against Hantaan virus revealed that all these Mabs belong to the IgG2a subclass. All 7 Mabs reacted to nucleocapsid (N) proteins (MW.49-50Kd) of Hantaan virus by immunoblot assay. Mabs against Seoul virus were screened, 985 hybridoma cells were produced out of 1152 and the fusion frequency was 86%. Eight arnong 985 hybridoma cells produced Mabs against Seoul virus continuously. Isotype analysis of these Mabs against Seoul virus revealed the IgM and IgG2a subclass. These 8 Mabs reacted to nucleocapsid (N) proteins (MW.49-50Kd) of Seoul virus as demonstrated by immunoblot. Interestingly, these Mabs had neutralizing activity determined by PRNT analysis. Therefore it is suspected that the antigenic sites on N protein may be involved in the virus neutralization. The serological reactivity of 7 Mabs against Hantaan virus with Hantaan virus were strong, although Seoul, Puumala and prospect hill virus were not reacted to these Mabs. 8 Mabs against Seoul virus, reacted to Seoul virus strongly. Hantaan and Puumala virus reacted slightly to these Mabs but Prospect hill virus did not. To comparative study of sensitivity test between IFA and double sandwich ELISA method for human sera using ELISA of which these Mabs coated were tested, the ELISA method was more sensitive than IFA. These Mabs react.ed specifically to Hantaan and Seoul virus. There are antigenic relationships among Hantaviruses could be analysed by using these Mabs.

Our Hantaan Virus Became a New Family, Hantaviridae in the Classification of Order Bunyavirales. It will Remain as a History of Virology

Ho Wang Lee,Jin Won Song 대한바이러스학회 2019 Journal of Bacteriology and Virology Vol.49 No.2

한국형 출혈열 VII . 등줄쥐에서의 한국형 출혈열 병원체 한탄바이러스의 전퍼경로

이호왕,이평우,백락주,송철근,성인화,김정혜 대한바이러스학회 1982 Journal of Bacteriology and Virology Vol.12 No.1

Epidemic hemorrhagic fever with renal syndrome was recognized for the first time in 1951 among UN troops in Korea. Since that time it has been known as Korean hemorrhagic fever(KHF) and remained endemic near the demilitarized zone between South and North Korea. In recent years it appears to have spread slowly in a southwesterly direction and about 800 hospitalized cases are clinically diagnosed each year. In l976 Lee and Lee succeeded in demonstrating an antigen in the lungs of the striped field mouse, Apodemus agrarius, which gave specific immunofluorescent reactions with sera from patients convalescent from KHF for the first time. The natural reservoir host of Korean hemorrhagic fever is Apodemus agrarius coreae in the rural endemic areas in Korea however, mode of transmission of Hantaan virus in Apodemus mice is not known to date yet. This is the first report on demonstration of the mode of transmission of the virus experimentally in Apodemus agrarius. 1. Mice inoculated by the intramuscular route experienced viremia for about 5 days beginning on day 7. After 3 weeks, immunofluorescent and neutralizing antibodies were present and no mouse ever developed signs of acute illness. 2. Virus was recovered from lung, kidney, salivary gland, and liver and virus excretion in urine, saliva, and feces occurred from about day 10 through day 640 (urine) postinoculation. Antigen, but not infectious virus, was persistent in lung tissue for as long as 1 year. 3. Horizontal contact infection occurred among cage-mates regardless of sexual pairing up to 640 days after infection and no evidence for participation of ectoparasitic arthropods in such transmission was obtained.

한국인에서 E형 간염 바이러스의 항체 유병률에 관한 조사

신학균,윤재득,정연호,김문보,서순덕,김정서 대한바이러스학회 1992 Journal of Bacteriology and Virology Vol.22 No.2

Serosurvey of hepatitis E virus (HEV) infection, known as enterically transmitted non-A, non-B hepatitis (ET-NANBH), was done in Korea between May through August, 1991 using anti HEV EIA method developed by Genelabs Inc. in 1990. The coated polypeptide antigens were expressed in Baculovirus vector from the specific HEV cDNA clones derived from two patients infected with HEV, one from Burma (antigen A) and the other from Mexico (antigen B). The 1,079 examinees was arbitrarily grouped into risk (406) and non-risk (673) group according to the serologic markers of hepatitis B virus (HBsAg, HBsAb, and HBcAb), anti-hepatitis C virus (HCV), and alanine transarninase (ALT) value but not to the risk of HEV. The overall seroprevalences of HEV in non-risk and risk group were 10.4 %, and 8.9%, respectively. Among the non-risk subgroups, the prevalence of anti-HEV was about 10 % regard- less of the result of HBsAb or HBcAb. There was no sexual and regional difference. The peak prevalence was shown at the age between 30 and 39. In the risk groups, the highest prevalence was 15% shown in the anti-HCV positive group and the lowest was the 5.9% of HBsAg positive group. There was a little higher prevalence rate in male group but there was no significant statistical difference between male and female. Anti-HEV posit,ive rates of normal group were relatively higher than the expected risk group and these results strongly suggested the possibility of existence of HEV infection in Korea, particularly in rural areas where the tap water supply facilities are not enough.

발육란뇨막 (發育卵尿膜) - 卵嗀片 ( allantois - on - shell )을 使用한 인풀루엔자 바이러스 감염가측정법의 (感染價測定法) 실용성검토 (實用性檢討)

윤승태(尹承泰),양용태(梁容泰),최철순(崔哲淳) 대한바이러스학회 1977 Journal of Bacteriology and Virology Vol.7 No.1

A study was made to confirm both the validity and the applicablitity of influenza virus infectivity titration by the allatois-on-shell technique, originally devised by Far.ekas de St. Groth and White in 1958. Furthermore, a series of experiments was conducted to improve the infectivity titers of influenza virus type A, PR 8 strain by this method. The results are summarized as follows. Among the embryonated eggs tested, the 13-day old eggs were best suitable for successful deembryonation and for the preparation of allantois-on-shell pieces, and at the same time provided the highest infectivity titers. 2. Shaking incubation for 72 hours at 37'C resulted in higher infectivity titers than those of 48 hours of incubation. 3. Addition of mycostatin (20 units per ml) to Star.dard Medium effectively controlled the growth of fungi during the shaking incubation without any inhibitory effect on the virus titers. 4. Allocation of (virus) adsorption time (37C, 1 hr) prior to shakirg incubation of virus-inoculated allantois-on-shell pieces appeared to improve the results of infectivity titration. 5. Ambient temperature (room temperature or cold room) at which preparation of allantois-on-shell pieces, virus inoculation and setting up of the test were conducted did effect very little the infectivity titers when the (virus) adsorption period (37C, l hr) was applied before the shaking incubation. 6. A significant drop in the virus titers was observed when the infectivity titrations were carried out with sodiun bicarbonate-added Standard Medium. 7. Infectivity titers obtained by the allantois-on-shell technique were approximately 25 times lower than those of egg infectivity titrations.

혈구응집을 일으킨 한탄바이러스의 전자현미경 및 면역전자현미경적 소견

성인화,이호왕 대한바이러스학회 1988 Journal of Bacteriology and Virology Vol.18 No.2

Bunyaviridae-like morphology of Hantaan virus was reported by negative staining of supernatant fluids of A-549 cell cultures infected with strain 76-118 of Hantaan virus and plaque-purified Hantaan virus by several workers. In this study, Hantaan viruses in suspending medium could be observed without negative staining. Hantaan viruses adsorbed on the goose erythrocytes and hemagglutinated Hantaan viruses were observed by electronmicroscopy and immune electronmicroscopy using monoclonal, polyclonal antibodies against Hantaan virus and ProteinA-Gold reagents. Hantaan viruses were spherical to oval or pleomorphic, 80-160nm in size, and didnt cause hemolysis of goose erythrocytes or fusion of envelope of virus with cytoplasmic membrane of erythrocytes, but caused indentation of cytoplasmic membranes in some cases. Gold particles were attached to Hantaan virions which agglutinated goose erythrocytes. Any virus which has hemagglutinating ability, even in suspension, could be studied by electronmicroscopy and irnmune electronmicroscopy without negative staining.

강원도 산악지대에서 채집한 야생들쥐의 한타바이러스 감염에 대한 혈청학적 연구

송진원(Jin Won Song),송기준(Ki Joon Song),백락주(Luck Ju Baek),이용주(Yong Ju Lee),정기모(Ki Mo Jung),고은영(Eun Young Go),박광숙(PS Park) 대한바이러스학회 1998 Journal of Bacteriology and Virology Vol.28 No.3

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

면역효소법에 의한 한타바이러스 플라크의 분석과 그 응용에 관한 연구

이평우 대한바이러스학회 1988 Journal of Bacteriology and Virology Vol.18 No.2

The new method for formation of plaques of Hantaviruses was developed using the system of enzyme-labelled antibody and Vero-E6cells maintained with semi-solid rnedium. To establish the method, several preliminary tests were carried out and consecutive studies were included for evaluation and practical application. The obtained conclusions are as follows; 1. The method, immunoenzyme plaque assay, was constructed with 1) anti-Hantavirus antibody and peroxidase labelled anti-rat IgG for antisera, 2) 3, 3-diaminobenzidine tetrahydrochloride for substrate and 3) Vero-E6 monolayer maintained with semisolid methyl cellulose DMEM for virus propagation. 2. The optimal units of antibody and enzyme-labelled antibody were determined as 4-B unit in both. Otherwise, at the higher or lower dilutions, it was not satisfiable in the contrast with background and the intensity of plaque staining. 3. The optimal concentrations of methyl cellulose in culture medium to form the countable, well sized plaques were 0.4% and 0.6% for prospect Hill virus and Hantaan viurs, respectively. 4. The plaques formed by this mathod were proved as to be produced by at least one virus particle according to the result of virus dilution experiment. Also, the observation of plaque by light microscope revealed that the plaque is the group of cells bearing viral antigens which was deposited with reaction product produced histochemically. 5. To determine whether plaques are formed by Hantaan virus, the virus was mixed with either sera from the patients of KHF or sera of rats irnmunized with Hantaan virus. The positive neutralization was evident by remarked reduction of nurnbers of plaques. 6. The sensitivity, specificity and usefulness of the rnethod could be proved by being applied in differential diagnosis of HFRS, analysis of Hantaviral antigens, test for virus interference and massive screening of neutralizing antibody or virus titration.