• Journal of Microbiology and Biotechnology
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• 제21권4호
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• pp.405-411
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• 2011

Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy ${\beta}$-glucan (SBG), a ${\beta}$-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 $log_2$ higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with ${\beta}$-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 $log_2$ following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.

• Journal of Microbiology and Biotechnology
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• 제28권1호
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• pp.157-164
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• 2018

Francisella tularensis (FT), a highly infectious pathogen, is considered to be a potential biological weapon owing to the current lack of a human vaccine against it. Tul4 and FopA, both outer membrane proteins of FT, play an important role in the bacterium's immunogenicity. In the present study, we evaluated the immune response of mice - humanized with human CD34+ cells (hu-mice) - to a cocktail of recombinant Tul4 and FopA (rTul4 and rFopA), which were codon-optimized and expressed in Escherichia coli. Not only did the cocktail-immunized hu-mice produce a significant human immunoglobulin response, they also exhibited prolonged survival against an attenuated live vaccine strain as well as human T cells in the spleen. These results suggest that the cocktail of rTul4 and rFopA had successfully induced an immune response in the hu-mice, demonstrating the potential of this mouse model for use in the evaluation of FT vaccine candidates.

• Clinical and Experimental Vaccine Research
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• 제12권2호
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• pp.156-171
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• 2023

Purpose: The development of vaccines that confer protection against multiple avian influenza A (AIA) virus strains is necessary to prevent the emergence of highly infectious strains that may result in more severe outbreaks. Thus, this study applied reverse vaccinology approach in strategically constructing messenger RNA (mRNA) vaccine construct against avian influenza A (mVAIA) to induce cross-protection while targeting diverse AIA virulence factors. Materials and Methods: Immunoinformatics tools and databases were utilized to identify conserved experimentally validated AIA epitopes. CD8⁺ epitopes were docked with dominant chicken major histocompatibility complexes (MHCs) to evaluate complex formation. Conserved epitopes were adjoined in the optimized mVAIA sequence for efficient expression in Gallus gallus. Signal sequence for targeted secretory expression was included. Physicochemical properties, antigenicity, toxicity, and potential cross-reactivity were assessed. The tertiary structure of its protein sequence was modeled and validated in silico to investigate the accessibility of adjoined B-cell epitope. Potential immune responses were also simulated in C-ImmSim. Results: Eighteen experimentally validated epitopes were found conserved (Shannon index <2.0) in the study. These include one B-cell (SLLTEVETPIRNEWGCR) and 17 CD8⁺ epitopes, adjoined in a single mRNA construct. The CD8⁺ epitopes docked favorably with MHC peptidebinding groove, which were further supported by the acceptable ∆G_bind (-28.45 to -40.59 kJ/mol) and Kd (<1.00) values. The incorporated Sec/SPI (secretory/signal peptidase I) cleavage site was also recognized with a high probability (0.964814). Adjoined B-cell epitope was found within the disordered and accessible regions of the vaccine. Immune simulation results projected cytokine production, lymphocyte activation, and memory cell generation after the 1st dose of mVAIA. Conclusion: Results suggest that mVAIA possesses stability, safety, and immunogenicity. In vitro and in vivo confirmation in subsequent studies are anticipated.

• Clinical and Experimental Pediatrics
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• 제48권9호
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• pp.960-968
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• 2005

목 적 : 본 연구는 한국인 소아를 대상으로 MMR 약독화 생백신인 프리오릭스주를 단독 접종한 후 공개, 다 기관, 비비교 시험으로 프리오릭스주의 안전성과 면역성 평가를 목적으로 수행되었다. 방 법 : 2002년 7월부터 2003년 2월까지 한일병원, 경희대학교 부속병원, 가톨릭대학교 성바오로병원, 고려대의과대학부속 안산병원 등 4개의 병원에 예방 접종을 위해 내원한 12-15개월 또는 4-6세의 건강한 남녀 소아 252명을 대상으로 하였으며, 이중에서 임상시험계획서의 모든 기준을 충족하는 피험자를 최종 분석대상군에 포함시켰다. 접종 후 이상 반응 및 중대한 이상 반응에 대하여 42일간의 추적관찰을 통하여 증례기록서의 이상 반응란에 기록하였다. 각 피험자들은 시험백신 투여 직전과 투여 후 42일 후 혈액검체를 채취하여 벨기에 GlaxoSmithKline Biologicals 실험실에서 면역분석법을 통하여 항체가를 측정하였다. 결 과 : 등록된 피험자 총 252명 중 최종적으로 12-15개월군에서 103명, 4-6세군에서 96명으로 총 199명이 분석대상으로 선정되었다. 본 임상시험 기간 동안 발생한 이상반응 중 시험약물과 관련 있는 국소 반응은 10.1% 전신 반응은 6.5%에 불과하였으며, 우려하였던 이상 반응, 예를 들면 무균성 뇌막염, 열성 경련 등은 발생하지 않았으며, 시험백신과 관련된 중대한 이상 반응은 없었다. 유효성 분석에서, 항체 음성인 피험자들의 혈청전환율은 홍역인 경우 99%, 유행성 이하선염인 경우 93%, 풍진인 경우 100%이었다. 시험백신 투여 전 항체 음성인 피험자군의 혈청전환시 항체가의 기하평균은, 홍역의 경우 12-15개월에 속하는 피험자군에서는 $383{\pm}8.6mIU/mL$, 4-6세에 속하는 피험자군에서는 $188{\pm}6.2mIU/mL$, 유행성 이하선염의 경우 12-15개월에 속하는 피험자군에서는 $95{\pm}6.3U/mL$, 4-6세에 속하는 피험자군에서는 $247{\pm}3.8U/mL$, 풍진의 경우 12-15개월에 속하는 피험자군에서는 $94.5IU/m{\pm}$, 4-6세에 속하는 피험자군에서는 $16{\pm}8.9IU/mL$을 보였다. 결 론 : 12-15개월 또는 4-6세의 건강한 소아를 대상으로 프리오릭스주를 접종하였을 때, 이와 관련된 이상 반응은 대부분의 백신 접종 후 나타날 수 있는 일반 반응에 한정된 것으로 나타나 한국인 소아에게서도 안전하게 사용될 수 있으며 또한 매우 높은 항체양전율 및 항체가의 기하평균치를 보여 우수한 면역효과를 기대할 수 있는 백신으로 사료된다.

• 대한수의학회지
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• 제39권4호
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• pp.743-750
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• 1999

Bovine viral diarrhea viruses (BVDVs) were isolated from cattle with respiratory and diarrhea signs as well as persistently infected cattle. These isolates were analysed serologically to characterize serogroups and to compare serological relationship with reference viruses of type I and II. Most isolates from calf diarrheal cases and persistently infected individuals showed a significant difference in cross-neutralization test with the viruses isolated from nasal discharges showing severe respiratory signs. Serologically most of the commercial vaccine strains could be classified into classical BVDV (type I) such as NADL strain. This serological difference among BVDV isolates suggested the need for new vaccines to protect cattle from both respiratory and enteric BVDV infections in field. The immunogenicity of BVDVs which showed a good propagation capability in MDBK cells and high rates of neutralizing activity (isolate : KD26-1, PHG, B5 and 95002) against all viruses used in this study, was confirmed in guinea pig when treated as single or combined groups.

• Archives of Pharmacal Research
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• 제21권5호
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• pp.543-548
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• 1998

We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross-immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. from the measurement of $ED_{50}$ after formalin- or heat-inactivation, both r-HBsAg and p-HBsAg and p-HBsAg showed $ED_{50}$ of 0.2-0.3 in formalin-inactivaton, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat-inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with $AIPO_{4}$ adjuvant. the mutagenicity of r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity.

• 한국동물위생학회지
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• 제32권3호
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• pp.201-207
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• 2009

Porcine transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV) and rotaviruses are considered as the most important causative agents of diarrhea in piglets. The study established 3 method vaccination programs to prevent PEDV. A (LL)group inoculated twice vaccinations on 2-weeks-interval during the late term of pregnant sows with PEDV live vaccine. The B (LKK) group was applied that one time single PEDV live vaccine at the pre-mate followed by the TGEV PEDV combined inactivated vaccine (twice vaccination on 2-weeks interval at the third-trimester). C (KK) group was applied to sow which inoculated twice vaccination on 2-weeks-interval during the late term of pregnant sows with by the TGEV, PEDV combined inactivated vaccine. As the result of SN test on sows in the pig farm before vaccination, antibody titers was showed 9/45 (20.0%). By comparison with the serum neutralizing antibody titers against PEDV of the vaccination programs after PEDV of the vaccination, A group and B group vaccination method was higher than those of C group in sows. In the piglets up to 2 weeks of age, A group was showed antibody titers of 17/22 (81.8%) that showed 2-128, and B group was showed antibody titers of 30/37 (81.1%) that showed 2-512, and C group was showed antibody titers of 14/28 (50.0%) that showed 2-32. On the other hand, PEDV antibody titers were tested for the survey. As the results of SN test, Aujeszky's disease survey in 54 pig farms from november 2005 to august 2006, antibody titers of 47/286 (16.4%) showed above 2. Five breeding farms were antibody titers of 38/77 (49.4%), Wanggung zone farms antibody titers of 59/85 (69.4%). In pigs farms vaccinated the first of twice PEDV live vaccine, and after 6 month, the second of twice TGEV PEDV combined inactivated vaccine (LLKK, 256-1024 titer) method was higher than those of vaccinated twice the early term of pregnant, and twice the late term of pregnant sows of PEDV live vaccine (LLLL, 32 titer).

• 대한수의학회지
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• 제41권2호
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1613-1619
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• 2016

Francisella tularensis is a highly virulent pathogen of humans and other mammals. Moreover, F. tularensis has been designated a category A biothreat agent, and there is growing interest in the development of a protective vaccine. In the present study, we determine the in vitro and in vivo immune responses of a subunit vaccine composed of recombinant peptides Tul4 and FopA from epitopes of the F. tularensis outer membrane proteins. The recombinant peptides with adjuvant CpG induced robust immunophenotypic change of dendritic cell (DC) maturation and secretion of inflammatory cytokines (IL-6, IL-12). In addition, the matured DCs enabled ex vivo proliferation of naive splenocytes in a mixed lymphocyte reaction. Lastly, we determined the in vivo immune response by assessment of antibody production in C57BL/6 mice. Total IgG levels were produced after immunization and peaked in 6 weeks, and moreover, Tul4-specific IgG was confirmed in the mice receiving peptides with or without CpG. Based on these results, we concluded that the recombinant peptides Tul4 and FopA have immunogenicity and could be a safe subunit vaccine candidate approach against F. tularensis.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.2135-2145
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• 2015

VP7, an outer capsid protein of grass carp reovirus (GCRV), was expressed and displayed on the surface of Saccharomyces cerevisiae for developing an efficient vaccine against hemorrhagic disease of grass carp. The result of flow cytometry analysis indicated that protein VP7 could be displayed on the surface of yeast cells after inducing with galactose. The expression of VP7 was confirmed by western blot analysis and further visualized with confocal microscopy. The specific antibodies against VP7 generated from mice were detectable from all immune groups except the control group, which was immunized with untransformed yeast cells. The displaying VP7 on glycosylation-deficient strain EBYΔMnn9 was detected to induce a relatively low level of specific antibody amongst the three strains. However, the antiserum of EBYΔM9-VP7 showed relative high capacity to neutralize GCRV. Further neutralization testing assays indicated that the neutralizing ability of antiserum of the EBYΔM9-VP7 group appeared concentration dependent, and could be up to 66.7% when the antiserum was diluted to 1:50. This result indicates that appropriate gene modification of glycosylation in a yeast strain has essential effect on the immunogenicity of a yeast-based vaccine.