• IMMUNE NETWORK
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• 제16권5호
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• pp.311-315
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• 2016

A pandemic influenza A (H1N1) virus strain was isolated from a pig farm in Korea in December 2009. The strain was propagated in and isolated from both the Madin-Darby canine kidney cell line and embryonated eggs. The partial and complete sequences of the strain were identical to those of A/California/04/2009, with >99% sequence similarity in the HA, NA, M, NS, NP, PA, PB1, and PB2 genes. The isolated strain was inactivated and used to prepare a swine influenza vaccine. This trial vaccine, containing the new isolate that has high sequence similarity with the pandemic influenza A (H1N1) virus, resulted in seroconversion in Guinea pigs and piglets. This strain could therefore be a potential vaccine candidate for swine influenza control in commercial farms.

• Pediatric Infection and Vaccine
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• 제13권2호
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• pp.163-173
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• 2006

목 적 : 근래에 영아 백신의 숫자가 증가하면서 동시에 접종하는 혼합백신의 개발이 증가하고 있다. 본 연구에서는 우리나라 영아에서 b형 H. influenzae에 대한 단백 결합 백신(PRP-OMP)과 B형 간염 백신의 혼합백신인 $Comvax^{TM}$(Merck & Co.)의 면역원성 및 안전성을 평가하고자 하였다. 대상 및 방법 : 생후 직후 B형 간염 백신(유박스 $B^{TM}$ 또는 헤파박스-진$^{TM}$)을 접종받은 영아를 대상으로 $Comvax^{TM}$를 단독으로 또는 DTaP, IPV, Prevenar 등과 2, 4개월에 접종한 후, 2회 접종 전(4개월 연령) 및 2회 접종 2개월 후(6개월 연령)에 혈청 내 항 PRP 항체가와 anti-HBs를 효소면역법과 chemiluminescent microparticle immunoassay로 측정하였다. 결 과 : 총 65명(남아 35명)이 포함되었으며, 58명이 계획된 접종을 완료하였다. 1회 접종 후 항PRP 항체가의 기하 평균치는 1.96 ${\mu}g/mL$(95% CI 1.38~2.78), 2회 접종 후의 기하평균치는 10.02 ${\mu}g/mL$(95% CI 7.04~14.26)이었다. 항체가가 1.0 ${\mu}g/mL$ 이상인 비율은 1회 접종 후 63.2%(95% CI 53.75~72.65), 2회 접종 후 96.6%(95% CI 93.05~100)이었다. 한편 anti-HBs가 10 mIU/mL 이상인 영아의 비율은 4개월에 73.7%(95% CI 65.07~82.33), 6개월에 94.8%(95% CI 90.45~99.15)이었으며, 기하 평균치는 4개월에 38.32 mIU/mL(95% CI 22.42~65.51), 6개월에 101.17 mIU/mL(95% CI; 65.94~155.25)이었다. 백신 접종 후 발생한 이상반응 중 전신 증세로서 보챔(24.8%)이 가장 많았고 졸음(19.2%), 식욕부진(19.2%) 그리고 발열(7.2%) 등이 있었다. 국소 증세로는 동통이 25.6%, 발적이 19.2%, 부종이 4.8%에서 관찰되었다. 이러한 이상반응은 대부분 경증에 속하는 것으로 모두 회복되었다. 결 론 : $Comvax^{TM}$는 우리나라 영아에서 안전하고, 매우 우수한 항-PRP 항체 반응을 보였으며, 본 연구에서 B형 간염에 대해서는 아직 표준화되지 않은 방법으로 접종되었으나 비교적 우수한 항체 반응을 보여 B형 간염에 대해서도 우수한 면역원성을 지녔을 것으로 추측된다.

• International Journal of Oral Biology
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• 제42권2호
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• pp.63-70
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• 2017

Selecting an appropriate antigen with optimal immunogenicity and physicochemical properties is a pivotal factor to develop a protein based subunit vaccine. Despite rapid progress in modern molecular cloning and recombinant protein technology, there remains a huge challenge for purifying and using protein antigens rich in hydrophobic domains, such as membrane associated proteins. To overcome current limitations using hydrophobic proteins as vaccine antigens, we adopted in silico analyses which included bioinformatic prediction and sequence-based protein 3D structure modeling, to develop a novel periodontitis subunit vaccine against the outer membrane protein FomA of Fusobacterium nucleatum. To generate an optimal antigen candidate, we predicted hydrophilicity and B cell epitope parameter by querying to web-based databases, and designed a truncated FomA (tFomA) candidate with better solubility and preserved B cell epitopes. The truncated recombinant protein was engineered to expose epitopes on the surface through simulating amino acid sequence-based 3D folding in aqueous environment. The recombinant tFomA was further expressed and purified, and its immunological properties were evaluated. In the mice intranasal vaccination study, tFomA significantly induced antigen-specific IgG and sIgA responses in both systemic and oral-mucosal compartments, respectively. Our results testify that intelligent in silico designing of antigens provide amenable vaccine epitopes from hard-to-manufacture hydrophobic domain rich microbial antigens.

• 대한수의학회지
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• 제37권4호
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• pp.875-882
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• 1997

To produce the recombinant fibronectin-binding protein(FnBP) for development of subunit vaccine against Staphylococcus aureus. The fnbp gene was amplified from the chromosomal DNA of S aureus KNU 196 strain using the polymerase chain reaction, and cloned into pGEX-4T-2. Then, the recombinant FnBP fused with glutathione-S-transferase was produced in E coli, purified by affinity chromatography, and identified its antigenicity and immunogenicity by Western blot. The recombinant FnBP produced in this study is considered to have the same property of native FnBP purified from S aureus, and is expected to be useful as a candidate for S aureus subunit vaccine.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.280-287
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• 2015

Current influenza vaccines are produced in embryonated chicken eggs. However, egg-based vaccines have various problems. To address these problems, recombinant protein vaccines have been developed as new vaccine candidates. Unfortunately, recombinant proteins frequently encounter aggregation and low stability during their biogenesis. It has been previously demonstrated that recombinantly expressed proteins can be greatly stabilized with high solubility by fusing stabilizing peptide (SP) derived from the C-terminal acidic tail of human synuclein (ATS). To investigate whether SP fusion proteins can induce protective immunity in mice, we produced influenza HA and SP fusion protein using a baculovirus expression system. In in vitro tests, SP-fused recombinant HA1 (SP-rHA1) was shown to be more stable than recombinant HA1 (rHA1). Mice were immunized intramuscularly with baculovirus-expressed rHA1 protein or SP-rHA1 protein ($2{\mu}g/mouse$) formulated with aluminum hydroxide. Antibody responses were determined by ELISA and hemagglutination inhibition assay. We observed that SP-rHA1 immunization elicited HA-specific antibody responses that were comparable to rHA1 immunization. These results indicate that fusion of SP to rHA1 does not negatively affect the immunogenicity of the vaccine candidate. Therefore, it is possible to apply SP fusion technology to develop stable recombinant protein vaccines with high solubility.

• IMMUNE NETWORK
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• 제10권6호
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• pp.198-205
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• 2010

Background: A crucial limitation of DNA vaccines is its weak immunogenicity, especially in terms of eliciting antibody responses in non-human primates or humans; therefore, it is essential to enhance immune responses to vaccination for the development of successful DNA vaccines for humans. Methods: Here, we approached this issue by evaluating interleukin-7 (IL-7) as a genetic adjuvant in cynomolgus monkeys immunized with multigenic HCV DNA vaccine. Results: Codelivery of human IL-7 (hIL-7)-encoding DNA appeared to increase DNA vaccine-induced antibody responses specific for HCV E2 protein, which plays a critical role in protecting from HCV infection. HCV-specific T cell responses were also significantly enhanced by codelivery of hIL-7 DNA. Interestingly, the augmentation of T cell responses by codelivery of hIL-7 DNA was shown to be due to the enhancement of both the breadth and magnitude of immune responses against dominant and subdominant epitopes. Conclusion: Taken together, these findings suggest that the hIL-7-expressing plasmid serves as a promising vaccine adjuvant capable of eliciting enhanced vaccine-induced antibody and broad T cell responses.

• Journal of Veterinary Science
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• 제21권5호
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• pp.74.1-74.13
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• 2020

Background: The quality of a vaccine depends strongly on the effects of the adjuvants applied simultaneously with the antigen in the vaccine. The adjuvants enhance the protective effect of the vaccine against a viral challenge. Conversely, oil-type adjuvants leave oil residue inside the bodies of the injected animals that can produce a local reaction in the muscle. The long-term immunogenicity of mice after vaccination was examined. ISA206 or ISA15 oil adjuvants maintained the best immunity, protective capability, and safety among the oil adjuvants in the experimental group. Objectives: This study screened the adjuvant composites aimed at enhancing foot-and-mouth disease (FMD) immunity. The C-type lectin or toll-like receptor (TLR) agonist showed the most improved protection rate. Methods: Experimental vaccines were fabricated by mixing various known oil adjuvants and composites that can act as immunogenic adjuvants (gel, saponin, and other components) and examined the enhancement effect on the vaccine. Results: The water in oil (W/O) and water in oil in water (W/O/W) adjuvants showed better immune effects than the oil in water (O/W) adjuvants, which have a small volume of oil component. The W/O type left the largest amount of oil residue, followed by W/O/W and O/W types. In the mouse model, intramuscular inoculation showed a better protection rate than subcutaneous inoculation. Moreover, the protective effect was particularly weak in the case of inoculation in fatty tissue. The initial immune reaction and persistence of long-term immunity were also confirmed in an immune reaction on pigs. Conclusions: The new experimental vaccine with immunostimulants produces improved immune responses and safety in pigs than general oil-adjuvanted vaccines.

• Clinical and Experimental Vaccine Research
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• 제12권1호
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• pp.70-76
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• 2023

Purpose: Typhoid remains a major health problem, especially in the developing world. Furthermore, the emergence of multidrug-resistant and extensively drug-resistant strains of Salmonella typhi added a sense of urgency to develop more effective typhoid vaccines, one of which is bacterial ghosts (BGs), prepared by both genetic and chemical means. The chemical method includes incubation with numerous agents for a short time at their minimum inhibitory or minimum growth concentrations. This study included the preparation of BGs by a sponge-like reduced protocol (SLRP). Materials and Methods: Critical concentrations of sodium dodecyl sulfate, NaOH, and H₂O₂ were used. Moreover, high-quality BGs were visualized by scanning electron microscope (SEM). Subculturing was used to confirm the absence of vital cells. Besides, the concentrations of the released DNA and protein were estimated spectrophotometrically. In addition, the integrity of cells was proved by visualizing Gram-stained cells using a light microscope. Furthermore, a comparison between the immunogenicity and safety of the prepared vaccine and the available whole-cell killed vaccine was established. Results: Improved preparation of high-quality BGs of S. typhi, visualized by SEM, revealed punctured cells with intact outer shells. Moreover, the absence of vital cells was confirmed by subculturing. At the same time, the release of respective amounts of proteins and DNA is another evidence of BGs' production. Additionally, the challenge test provided evidence that the prepared BGs are immunogenic and have the same efficacy as the whole cell vaccine. Conclusion: The SLRP provided a simple, economical, and feasible method for BGs preparation.

• 대한수의학회지
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• 제10권1호
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• pp.5-10
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• 1970

Due to the fact that an inactivated anthrax vaccine may lark its immunogenicity and stability of immunogen a number of spore vaccines were exclusively used worldwide. In these studies a number of important factors were emphasized to achieve the following: selection of non or less allergic strain of anthrax bacillus, capsulation of bacteria. obtaining of non sporulating but vegetative organisms, adequate inactivation of B. anthraccis by means of formalin, adsorption of immunogen to aluminum hydroxide gel. Non or less allergic strains of anthrax bacillus which is inactivated with formalin was selected by a hyperimmunization and shock test in rabbits. Obtaining capsular material and vegetative immunogen, a virulent anthrax organisms were cultivated on sodium bicarbonate medium with of without adding of l-alanine in which B, anthracis grew luxuriantly without forming spores. Inactivation was carried out at $37^{\circ}C$ water bath for 3 days after the bacterial culture was mixed with formalin, in a final concentration of two per cent of formalin. Aluminum hydroxide gel was added to the mixture of anthrax and blackleg bacterin. Vaccines were injected guinea pig via subcutaneous or intramusoular route and challenged after three weeks and the possibilities of protection was tested. Throughout the studies. the above mentioned vaccines possibly protected the vaccinated guinea pigs more than 80 per cent compared to that of the controls. This experimental results strongly suggest that the vaccine may possibly applicable to bovine.

• Pediatric Infection and Vaccine
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• 제5권2호
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• pp.245-257
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• 1998

목 적 : 1983년부터 B형 간염 예방접종을 시행한 이래 국내에서 사용 중인 B형 간염 백신들의 항체 양전률에 대한 보고는 접종 일정, 접종 용량, 항체 검사 방법 및 항체 양성기준 등이 서로 상이하여 그 효과를 비교하기에 논란이 있어 왔다. 이에 저자들은 B형 간염 예방접종 후에 항체 양전률을 조사한 국내 문헌들에 대한 분석을 통해 접종되고 있는 B형 간염 백신의 효과를 맡아 보고자 하였다. 방 법 : 각과 의학회지 및 분과 학회지, 의과대학 학회지 등에서 B형 간염 백신을 접종하고 항체 양전율을 보고한 논문을 대상으로 하였다. 논문의 포함 기준은 검사 방법이 방사선 면역법 또는 효소결합 면역흡착 검사법이고 혈청 방어 항체가를 10mIU/mL 이상으로 하거나 기준 비율(sample ratio unit)를 10 ratio unit(RU) 이상으로 한 경우이며 이를 기준으로 항체 양전률을 구할 수 있는 경우로 하였다. 제외 기준은 접종 용량이 불분명하거나 불규칙한 경우, 항체 양전률이 없는 경우, 양성 기준 항체가 또는 기준 비율을 10mIU/mL 또는 10RU로 정하지 않았거나 변환할수 없는 경우, 접종 일정이 0-1-2개월 또는 0-1-6개월이 아닌 경우, 접종 백신의 기원이 명시되지 않은 경우로 하여, 52편 중 29편을 제외한 23편의 논문을 대상으로 항체 양전률을 분석하였다. 결 과 : 1) 접종 연령에 따른 항체 양전률의 가중 펑균은 Hepaccine(제일제당)의 경우 영아에서 85.1%, 소아에서 83.3%, 성인에서 62.7%로 영아와 소아에서 성인보다 항체 양전률이 높았다(P<0.01). Hepavax(녹십자)의 경우는 영아에서 84.7%, 소아에서 81.1%, 성인에서 90.8%로 소아에서 더 낮은 항체양전률을 나타내었다(P<0.01). 2) Hepavax(녹십자)를 0-1-6개월에 접종한 경우 항체 양전률의 가중 평균은 85.6%였고 0-1-2개월에 접종한 경우는 78.5%로 0-1-6개월에 접종한 경우가 항체 양전률이 더 높은 것으로 나타났다(P<0.01). 3) 영아 및 소아에서 Hepavax(녹십자)를 $5{\mu}g$ 접종한 경우와 $10{\mu}g$ 접종한 경우의 항체 양전률에는 차이가 없었다(P<0.38). 4) 성인에서 Hepaccine 접종 후 항체 양전률의 가중 평균은 62.7%. Hepavax는 90.8%였고 Engerix-B의 경우 94.8%로 Hepavax와 Engerix-B가 Hepaccine 보다 항체 양전률이 높게 나타났다(P<0.01). 결 론 : 본 조사를 동하여 만성 B형 간염 환자의 보유율이 높은 국내에서 원래 정해진 일정을 변경하는 방법은 항체 생성을 극대화하지 못하므로 주의할 필요가 있으며 백신의 면역원성을 높이기 위해서는 과거보다 더 효과적인 백신을 개발하고, 올바른 접종 방법을 지키도록 노력하여야 할 것으로 생각되었다.