• Journal of Veterinary Science
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• 제22권1호
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• pp.8.1-8.11
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• 2021

Background: Porcine circovirus type 2 (PCV2) is an important infectious pathogen implicated in porcine circovirus-associated diseases (PCVAD), which has caused significant economic losses in the pig industry worldwide. Objectives: A suitable viral vector-mediated gene transfer platform for the expression of the capsid protein (Cap) is an attractive strategy. Methods: In the present study, a recombinant adeno-associated virus 8 (rAAV8) vector was constructed to encode Cap (Cap-rAAV) in vitro and in vivo after gene transfer. Results: The obtained results showed that Cap could be expressed in HEK293T cells and BABL/c mice. The results of lymphocytes proliferative, as well as immunoglobulin G (IgG) 2a and interferon-γ showed strong cellular immune responses induced by Cap-rAAV. The enzyme-linked immunosorbent assay titers obtained and the IgG1 and interleukin-4 levels showed that humoral immune responses were also induced by Cap-rAAV. Altogether, these results demonstrated that the rAAV8 vaccine Cap-rAAV can induce strong cellular and humoral immune responses, indicating a potential rAAV8 vaccine against PCV2. Conclusions: The injection of rAAV8 encoding PCV2 Cap genes into muscle tissue can ensure long-term, continuous, and systemic expression.

• 대한바이러스학회지
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• 제27권2호
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• pp.143-149
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• 1997

Most of the current licenced hepatitis B vaccines are being produced by recombinant DNA technology in large fermentation cultures of Saccharomyces cerevisiae of yeast cells which carry the gene coded for hepatitis B virus surface antigen. These vaccines are proved very effective clinically and the immunogenicity of vaccines could be maintained for a long time under refrigeration. To develope the stabilizer that could increase the stability of hepatitis B virus vaccine which could be stored for a long period at room temperature or higher conditions, glucose, lactose and sucrose solutions in phosphate buffered saline were added into hepatitis B vaccine respectively to make 2.5%, 5%, 7.5% and 10% final concentration in vaccines. These sugar-vaccine mixtures were stored at room temperature for one month, two months and three months respectively and then inoculated into ICR mice intramuscularly. On the fourteenth day after inoculation, mice were bled and sera were tested for the evaluation of efficacies of vaccines. The results showed that 5% glucose, 7.5% lactose and sucrose increased the stability of vaccines in some degree and this method could be applied for the production of other viral vaccines and bacterial vaccines.

• 대한수의학회지
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• 제2권1호
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• pp.27-36
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• 1962

필자(筆者)는 한국(韓國)에 특(特)히 많이 유행(流行)하고 있는 가계(家鷄)의 유행병(流行病)인 Newcastle discase에 착안(着眼)하였다. 예방방법(豫防方法)으로서 종래(從來) 사용(使用)된 Forinalin-inactivated vaccine은 수산화(水酸化) "알미늄 젤"을 부가(賦加)하므로서 근육내주사방법(筋肉內注射方法)으로 사용(使用)할수 있게되어 사용방법(使用方法)이 비교적(比較的) 편리(便利)하게 되었으나 그 면역효과(免疫效果)에 대(對)하여서는 아직 이론(異論)이 있으므로 Hitcher등(等)이 제창(提唱)한 Attenuated Vaccine에 대(對)하여 더 흥미(興味)를 가지게 되었다. 특(特)히 이와같은 생독(生毒) Vaccine은 닭의 age에 차별(差別)없이 독력(毒力)이 거의 없을뿐만 아니라 경비적(經鼻的)으로 대단(大端)히 간이(簡易)한 방법(方法)으로도 투여(投與)할수 있고 고도(高度)의 면역(免疫)을 장기간(長期間) 부여(賦與)할수 있는 점(點)을 들어 기(其) 실용성(實用性)을 주목(注目)하게 되었다. 그러나 vaccine투여방법(投與方法)은 기면역율(其免疫率)과 vaccine투여(投與)로 인(因)한 손실(損失)을 좌우(左右)하게 되므로 중요(重要)한 연구과제(硏究課題)임을 인정(認定)하고 급수법(給水法)을 채택(採擇)하여 일정량(一定量)의 vaccine을 투여(投與)한후(後) 면역항체(免疫抗體)의 출현(出現)을 혈구응집조지항체(血球凝集阻止抗體)와 강독방어능력(强毒防禦能力)을 관찰(觀察)하여 구체적방법(具體的方法)을 검토(檢討)하려 하였다. 먼저 생독(生毒)vaccine으로 사용(使用)한 Lasota strain과 사독(死毒)으로 사용(使用)한 서울주(株)를 각각 $10^{-1}$에서 $10^{-11}$까지 희석(稀釋)하여 9일란(日卵)의 Allantoic Cavity로 접종(接種)하여 60시간(時間)이 경과(經過)한 후(後) Allantoic fluia를 채취(採取)하여 비교(比較)한 결과(結果) 서울주(株)는 $10^8MLD/ml$, $10^{9.6}LD_{50}/ml$였고 Lasota strain은 $10^9MID/ml$, $10^{10.5}ID_{50}/ml$였다. Formalin-mactivated Vaccine을 만들기 위(爲)하여 서울주(株)로 감염치사(感染致死)된 계태아(鷄胎兒), 양뇨막(羊尿膜), 요수등(尿水等)의 혼합유제(混合乳劑)에 Formaldehy를 1% 가(加)하여 48시간후(時間後)에 4배(四倍)의 생리식염수(生理食鹽水)를 가(加)하고 이 혼합물(混合物) 100ml에 4% "알미늄 젤"을 20ml.의 비(比)로 가(加)하여 Inactivation Test (9일(日)에 난접종(卵接種)) 및 Sterility Test(Thioglycollate Medium에 접종(接種))를 한 결과(結果) 완전(完全)히 inactivation 되었고 무균상태(無菌狀態)임을 인정(認定)한 후(後) 사용(使用)하였다. Attenuated Vaccine을 만들기 위(爲)하여는 Lasota strain을 9일란(日卵)의 Allantaic carity로 접종(接種)하여 3일간(日間) 발육(發育)케 한 후(後) Allantoic fluid를 채취(採取)하여 Allantoic fluid 100ml.에 탈지유(脫脂乳) 4ml.의 비(比)로 가(加)하여 2.5ml.식(式) 분주(分注)한 후(後) 냉동건조(冷凍乾燥)하여 $4^{\circ}C$에 보관(保管)하였다. 오개월팔일령(五個月八日齡)의 백색(白色)레구흥 200수(首)를 100수식(首式) 2군(二群)으로 나누어 각각(各各) 사독(死毒) Vaccine (1ml.I.M.) 및 생독(生毒) Vaccine (1 : 400, 10ml 경구(經口))을 투여(投與)한 후(後) 105일간(日間) 관찰(觀察)한 결과(結果) : 1. 사독(死毒) Vaccine은 HI항체(抗體)나 감염방어작용(感染防禦作用)이 105일간(日間)의 실험기간 동안 모두 고도(高度)로 유지(維持)되었다. 2. 생독(生毒) Vaccine은 감염방어작용(感染防禦作用)은 완전(完全)히 유지(維持)되었으나 HI항체(抗體)는 75일(日)까지 고도(高度)로 유지(維持)되었으나 75일후(日後)에는 약간(若干) 저하(低下)되었다. 3. 급수(給水)에 타서 경구적(經口的)으로 투여(投與)하는 생독예방접종(生毒豫防接種)은 면역기간(免疫期間)이 사독예방접종(死毒豫防接種)에 비(比)해 약간 짧은듯 싶지만 그접종방법(接種方法)이 대단히 간단하며 시간(時間)도 짧게 걸린다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.116-117
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• 2002

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.997-1006
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• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.849-859
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• 2018

Herpes simplex virus type 2 (HSV-2) infection has been a public health concern worldwide. It is the leading cause of genital herpes and a contributing factor to cervical cancer and human immunodeficiency virus (HIV) infection. No vaccine is available yet for the treatment of HSV-2 infection, and routinely used synthetic nucleoside analogs have led to the emergence of drug resistance. The small molecule $Retro-2^{cycl}$ has been reported to be active against several pathogens by acting on intracellular vesicle transport, which also participates in the HSV-2 lifecycle. Here, we showed that Retro-2.1, which is an optimized, more potent derivative of $Retro-2^{cycl}$, could inhibit HSV-2 infection, with 50% inhibitory concentrations of $5.58{\mu}M$ and $6.35{\mu}M$ in cytopathic effect inhibition and plaque reduction assays, respectively. The cytotoxicity of Retro-2.1 was relatively low, with a 50% cytotoxicity concentration of $116.5{\mu}M$. We also preliminarily identified that Retro-2.1 exerted the antiviral effect against HSV-2 by a dual mechanism of action on virus entry and late stages of infection. Therefore, our study for the first time demonstrated Retro-2.1 as an effective antiviral agent against HSV-2 in vitro with targets distinct from those of nucleoside analogs.

• Pediatric Infection and Vaccine
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• 제15권1호
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• pp.12-19
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• 2008

Varicella is a highly infectious disease caused by the varicella zoster virus. The varicella vaccine was developed by Michiaki Takahashi in Japan in 1974. Despite the worldwide distribution of efficient vaccines, varicella vaccination policy is extremely variable from country to country. Although varicella vaccine is not currently recommended for universal vaccination in Japan, most countries throughout Europe, and developing countries, it had been introduced into Korea in 1988 and 20 years have elapsed since its use. Currently, varicella vaccine has been most extensively used in the United States where routine 2-dose vaccination program has been recently implemented for children. Recent 2-dose schedule in the United States and the availability of combination measles-rubella-varicella vaccines may lead to future varicella vaccination policy changes in many countries. With this background, this article summarizes the current status of varicella vaccination policies worldwide and presents provisional updated recommendation of varicella vaccination in Korea.

• EDISON SW 활용 경진대회 논문집
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• 제5회(2016년)
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• pp.86-89
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• 2016

The Zika virus, which is a member of the flavivirus genus, poses a serious threat to humanity because there is no vaccine or cure. Zika is suspected to cause microcephaly, and it is rapidly spreading throughout parts of Brazil. Surprisingly, there are no known protein structures for the virus which are essential for drug and vaccine development. This paper investigates the Zika virus's nonstructural proteins with template-based modeling by using GalaxyTBM/Refine/SC. GalaxyDock was used to examine the effectiveness of various known serine protease inhibitors in inhibiting the Zika viral protease. In testing five inhibitors, Kunitz soybean trypsin inhibitor showed the strongest binding affinity (-10.082 kcal/mol). This paper provides a rudimentary foundation for further drug discovery research regarding the Zika virus.

• 대한미생물학회지
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• 제16권1호
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• pp.83-89
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• 1981

Japanese encephalitis has been prevalent for long time in the Far East and many patients have been reported in both South East and Mid-West Asia recently. Recently, vaccine was used in prevention of this viral disease of man which was derived from formalin inactivated virus inoculated into mouse brain, but live attenuated active vaccine for human is not developed yet. Author inoculated Japanese encephalitis virus into several cell culture strains for development of live attenuated encephalitis virus strain and the results were as follows: 1. Japanese encephalitis virus was inactivated rapidly in cell free medium at $36^{\circ}C$ and totally inactivated by 72 hours. 2. In growth curve of Japanese encephalitis virus in HeLa cell cultures, maximal multiplication of the virus was occured at 4th day and virus multiplication was continued for at least 12 days. 3. After succeeding passage of the virus in HeLa cell cultures and human esophagus epithelial cell cultures, infectivity of virus for mice was disappeared from 2nd passage in HeLa cell cultures and 3rd passage in esophagus epithelial cell cultures. 4. In inoculation to monkey kidney epithelial cells and chick embryo cell cultures, infectivity of the virus for mice was continued after 10th passages.

• Animal cells and systems
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• 제3권3호
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• pp.337-341
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• 1999

Hepatitis C virus (HCV) is a positive strand RNA virus of the Flaviviridae family and the major cause of post-transfusion non-A, non-B hepatitis. Vaccine development for HCV is essential but has been slowed by poor understanding of the type of immunity that naturally terminates HCV infection. The DNA-based immunization technique offers the potential advantage of including cellular immune responses against conserved internal proteins of a virus, as well as the generation of antibodies to viral surface proteins. Here, we demonstrate that cell lines expressing the HCV core and/or NS3 proteins can induce a specific CTL response in mice, and these results suggest a possibility that the HCV core and NS3 DNA can be used to induce CTL activity against the antigen in mice and can be further developed as a therapeutic and preventive DNA vaccine.