• 대한바이러스학회지
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• 제26권1호
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• pp.67-76
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• 1996

The methods that make Hantavirus grow consist of inoculation into the experimental animals and cultured cells. The cultured cells, such as Vero-E6 and A549 cells, have been usually used for isolation of the virus and the animals, such as mice and rats, are used for large scale preparation of the virus so far. Furthermore, the cell can be used to maintain the virus and assay the infectivity and the animals can be used for the experiment of viral pathogenicity and challenge for assessment of vaccine. Apodemus mice, the own natural host of the virus, has been used for challenge test of Hantaan virus. However it has been pointed out to difficult handling and breeding the animal in laboratory. Therefore, we attempted to establish a new animal model for challenge test at the time of isolation of Maaji virus which is a new hantavirus similar but distinct to Hantaan virus. In suckling hamster, the titer of Maaji virus and the lethality to mice of the virus were increased gradually in the titer and lethality through passage by intracerebral (IC) inoculation. We tried to re-adapt this brain virus to lung of weanling hamster. The brain passaged virus was inoculated into weanling hamster intramuscularly. Again, the titer of the virus in lung was also increased by continuous passage of this virus. This facts could regarded as adaptation to new environment in which the virus proliferates. To identity the virus passaged in hamster with Maaji virus, both of the virus passaged in hamster brain and lung were compared with Maaji virus (MAA-I) and Hantaan virus (HTN 76-118) by means of restriction fragment length polymorphism (RFLP) and slingle strand conformation polymophism (SSCP). As a result, we conclude that Maaji virus could be adapted successfully to weanling hamster through this passage strategy. Utilizing this adapted Maaji virus strain, hamster model is able to be used for challenge test in hantaviral vaccinology and further experiments utilizing hamster system as a rather available and convenient lab animal are expected.

• Journal of Microbiology
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• 제43권6호
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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.

• Fisheries and Aquatic Sciences
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• 제21권3호
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• pp.5.1-5.8
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• 2018

Stimulator of interferon gene (STING) is induced by various inflammatory agents, such as lipopolysaccharide and microbial pathogens, including virus and bacteria. In this study, we obtained a full-length cDNA of a STING homolog from olive flounder using rapid amplification of cDNA ends PCR technique. The full-length cDNA of Paralichthys olivaceus STING (PoSTING) was 1442 bp in length and contained a 1209-bp open reading frame that translated into 402 amino acids. The theoretical molecular mass of the predicted protein sequence was 45.09 kDa. In the PoSTING protein, three transmembrane domains and the STING superfamily domain were identified as characteristic features. Quantitative real-time PCR revealed that PoSTING expressed in all the tissues analyzed, but showed the highest level in the spleen. Temporal expression analysis examined the significantly upregulated expression of PoSTING mRNA after viral hemorrhagic septicemia virus (VHSV) stimulation. In contrast, no significant changes in the PoSTING expression were detected in Edwardsiella tarda-challenged group compared to the un-injected control. The expression of P. olivaceus type I interferon (PoIFN-I) was also highly upregulated upon VHSV challenge. These results suggest that STING might be involved in the essential immune defense against viral infection together with the activation of IFN-I in olive flounder.

• BMB Reports
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• 제33권6호
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• pp.483-492
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• 2000

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• Journal of Veterinary Science
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• 제22권3호
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• pp.38.1-38.17
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• 2021

Background: The feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine, prepared from viruses grown in the Crandell-Rees feline kidney cell line, can induce antibodies to cross-react with feline kidney tissues. Objectives: This study surveyed the prevalence of autoantibodies to feline kidney tissues and their association with the frequency of FVRCP vaccination. Methods: Serum samples and kidneys were collected from 156 live and 26 cadaveric cats. Antibodies that bind to kidney tissues and antibodies to the FVRCP antigen were determined by enzyme-linked immunosorbent assay (ELISA), and kidney-bound antibody patterns were investigated by examining immunofluorescence. Proteins recognized by antibodies were identified by Western blot analysis. Results: The prevalences of autoantibodies that bind to kidney tissues in cats were 41% and 13% by ELISA and immunofluorescence, respectively. Kidney-bound antibodies were observed at interstitial cells, apical border, and cytoplasm of proximal and distal tubules; the antibodies were bound to proteins with molecular weights of 40, 47, 38, and 20 kDa. There was no direct link between vaccination and anti-kidney antibodies, but positive antibodies to kidney tissues were significantly associated with the anti-FVRCP antibody. The odds ratio or association in finding the autoantibody in cats with the antibody to FVRCP was 2.8 times higher than that in cats without the antibody to FVRCP. Conclusions: These preliminary results demonstrate an association between anti-FVRCP and anti-cat kidney tissues. However, an increase in the risk of inducing kidney-bound antibodies by repeat vaccinations could not be shown directly. It will be interesting to expand the sample size and follow-up on whether these autoantibodies can lead to kidney function impairment.

• IMMUNE NETWORK
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• 제21권1호
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• pp.8.1-8.17
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• 2021

The global crisis caused by the coronavirus disease 2019 (COVID-19) led to the most significant economic loss and human deaths after World War II. The pathogen causing this disease is a novel virus called the severe acute respiratory syndrome coronavirus 2 (SARSCoV-2). As of December 2020, there have been 80.2 million confirmed patients, and the mortality rate is known as 2.16% globally. A strategy to protect a host from SARS-CoV-2 is by suppressing intracellular viral replication or preventing viral entry. We focused on the spike glycoprotein that is responsible for the entry of SARS-CoV-2 into the host cell. Recently, the US Food and Drug Administration/EU Medicines Agency authorized a vaccine and antibody to treat COVID-19 patients by emergency use approval in the absence of long-term clinical trials. Both commercial and academic efforts to develop preventive and therapeutic agents continue all over the world. In this review, we present a perspective on current reports about the spike glycoprotein of SARS-CoV-2 as a therapeutic target.

• Pediatric Infection and Vaccine
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• 제5권1호
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• pp.96-103
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• 1998

목 적 : B형 간염 바이러스(hepatitis B virus; HBV) 보유 산모로부터 출생한 신생아의 수직감염 양상을 관찰하고 HBV 백신의 면역반응 및 방어효과를 평가하기 위하여 본 연구를 시행하였다. 대상 및 방법 : 1995년 7월부터 1996년 12월까지 18개월간 가톨릭대학교 성빈센트병원을 방문한 HBV 보유 산모와 이들로부터 분만된 신생아중 12개월간 추적관찰이 가능하였던 78례를 연구대상으로 하였다. 이들 HBV 보유 산모의 신생아에게 출생 직후부터 HBV 면역 글로불린(HBIG)과 가열 불활성화 혈장 백신을 접종한 후 4, 8, 12개월시에 검체를 채취하여 일반표지자의 양상을 면역효소법 및 방사면역법으로 측정한 후 HBV 보유 산모와 그 신생아의 각 표지자간의 관계 및 변화를 분석하였다. 결 과 : 1) 연구 대상 산모 중 HBV 보유자는 5.0%(106/2,117), HBeAg 양성 및 음성의 비율은 38.5%(37/96)와 61.5%(59/96)이었다. 2) HBV 보유 산모로부터 출생한 영아에 있어서 백신에 대한 anti-HBs 양전율은 4, 8, 12개월 시 각각 85.9%(67/78), 75.6%(59/78), 73.1%(57/78)로 유의한 차이가 있었으나(P<0.05), 산모의 HBeAg 유무와는 관련이 없었다. 형성된 anti-HBs의 기하 항체가는 4개월에 비하여 8개월, 12개월에 유의하게 증가하였다(P<0.05). HBV 백신과 HBIG 투여의 HBV 보유자 이행에 대한 12개월시의 방어 효과는 HBeAg 양성 및 음성 산모로부터 분만된 영아에서 각각 89.8%와 100%이었다. 3) 12개월의 관찰 기간동안 산모로부터 감염이 있었던 경우는 78명 중 5명으로 모두 HBeAg 양성 산모에서 분만된 영아이었고 이 중 3명에서 HBV 보유자로 이행되었다. 출생 직후 HBeAg 양성 및 음성 산모의 신생아에서 HBsAg 양성이었던 경우는 각각 4명으로 HBeAg 양성 산모의 신생아 4명 중 3명에서 감염이 발생하였다. 결 론 : 국내에서 개발된 백신의 하나인 가열 불활성화 B형 간염 혈장 백신의 항체 양전률은 85.9%이었으며, HBIG과 동시 투여하였을 때 12개월간의 HBV 보유자 이행에 대한 방어효과는 HBeAg 양성 산모에서 출생한 경우 89.8%인 반면 HBeAg 음성 산모에서 출생한 경우는 100%이었다.

• Pediatric Infection and Vaccine
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• 제25권1호
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• pp.35-44
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• 2018

목적: 인플루엔자 유행 예방에 대한 가장 효과적인 방법은 인플루엔자 백신이나 한국의 소아청소년을 대상으로 한 면역원성 및 안전성에 대한 자료가 많지 않다. 이에 본 연구는 국내에서 많이 사용되는 불활성화 3가 백신의 면역원성과 안전성을 확인하고자 하였다. 방법: 2008년 10월부터 12월까지 고려대학교 의료원 안산병원 소아청소년과에 인플루엔자 예방접종을 위해 내원한 건강한 소아청소년 중 임상시험 지원자 65명을 대상으로 하여 접종 전과 접종 후 30일째 혈구응집억제(hemagglutinin inhibition) 항체검사를 시행하고 접종 직후부터 접종 후 7일까지 국소반응과 전신반응을 포함한 이상반응을 관찰하여 기록하도록 하였다. 결과: 분할 인플루엔자 백신의 세 항원(H1N1, H3N2, B) 각각에 대한 접종 후 혈청보호율은 87.7%, 89.2%, 89.2% (${\geq}70%$)였으며 혈청전환율은 44.6%, 73.8%, 63.1% (${\geq}40%$), 혈청전환인자는 3.35, 7.18, 5.13 (>2.5)으로 Committee for Proprietary Medicinal Products (CPMP) 기준을 만족하였다. 전체 피험자 65명 중 48명(73.8%)이 백신 접종 후 이상반응을 보고하였으나 아나필락시스나 경련과 같은 심각한 이상반응은 없었다. 발적(29.2%), 동통(43.1%), 종창(41.5%)과 같은 국소반응을 보고한 피험자는 42명(64.6%)이었고, 권태(23.1%), 근육통(20.0%), 두통(10.8%), 관절통(10.8%), 오한(9.2%), 발열(7.7%) 등과 같은 전신반응을 보고한 피험자는 26명(40.0%)이었다. 결론: 6개월에서 18세까지의 소아청소년을 대상으로 한 불활성화 3가 백신의 면역원성과 안전성에 대한 연구 결과 H1N1, H3N2, B 항원 모두 CPMP 기준에 부합하여 적합한 면역원성이 있는 것으로 나타났으며, 국소반응 및 경미한 전신반응 이외에 심각한 이상반응은 보이지 않아 비교적 안전하다고 판단된다.

• Pediatric Infection and Vaccine
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• 제9권2호
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• pp.193-200
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• 2002

목 적 : 바이러스에 의한 급성 하기도염은 병원에 입원하게 되는 가장 흔한 질병으로 이를 진단하는데는 역학적 진단이 절대적으로 중요하다. 이에 저자들은 서로 다른 두 지역에서의 바이러스성 급성 하기도염의 역학조사를 시행함으로써 지역간의 차이를 알아보고자 하였다. 방 법 : 2000년 6월부터 2001년 6월까지 마산 파티마병원과 삼성서울병원에 급성 하기도염으로 입원한 환자 796명을 대상으로 하였으며, 바이러스 검출은 비인두 분비물을 통해 시행하였다. 두 병원간 원인 바이러스, 연령분포, 임상증상, 계절 유행양상 등을 비교하였다. 결 과 : 1) 바이러스가 배양된 환아는 208명(26.1%)였다. 바이러스 검출율은 삼성서울병원 21.9%, 마산 파티마병원 30.0%였다. 삼성서울병원에서의 연령분포는 2세 이하의 환아가 60.2%였으며 5세 이상의 환아는 12.8%이었는데 비해 마산 파티마병원에서는 2세 이하의 환아가 90.0%였으며 5세 이상의 환자는 0.8%였다(P<0.05). 2) 마산 파티마병원에서는 RSV의 검출율이 높았고(72.3%), 삼성서울병원에서는 adenovirus, influenza type A와 parainfluenza virus의 검출율이 상대적으로 높았다(P<0.05). 3) 임상양상을 비교해 보면 삼성서울병원에서는 기관기관지염과 폐렴이, 마산 파티마병원에서는 모세기관지염과 크룹이 더 많은 비율을 차지하고 있었다(P<0.05). 두 병원 모두 모세기관지염은 RSV, 크룹은 parainfluenza virus가 가장 많았다. 폐렴의 경우 삼성서울병원에서는 adenovirus, 마산 파티마병원은 RSV가 가장 많았고, 기관기관지염의 경우 삼성서울병원에서는 adenovirus, 마산 파티마병원은 influenza virus-type A가 가장 많았다. 4) 두 병원 모두 RSV는 가을과 겨울, parainfluenza virus는 봄, influenza virus는 겨울과 봄에 나타나는 양상을 보였고 adeonvirus는 연중 발생하는 양상을 보였다. 결 론 : 두 병원의 바이러스의 유행시기, 임상양상에 따른 원인바이러스의 분포결과는 유사하였다. 각 바이러스의 검출률과 임상상의 분포 등에서는 차이를 보였는데 이는 지역적 차이보다는 주로 대상연령군의 차이에 기인하는 것으로 생각된다. 우리나라에서 바이러스성 호흡기감염의 지역적 차이 유무를 확인하기 위해서는 비슷한 연령군을 대상으로 한 전국적이고 지속적인 역학조사가 이루어져야 할 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.591-599
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• 2008

A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV.