• Journal of Veterinary Science
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• 제25권1호
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• pp.4.1-4.14
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• 2024

Background: Lawsonia intracellularis is the causative agent of proliferative enteropathy and is associated with several outbreaks, causing substantial economic loss to the porcine industry. Objectives: In this study, we focused on demonstrating the protective effect in the mouse model through the immunological bases of two vaccine strains against porcine proliferative enteritis. Methods: We used live-attenuated Salmonella Typhimurium (ST) secreting two selected immunogenic LI antigens (Lawsonia autotransporter A epitopes and flagellin [FliC]-peptidoglycan-associated lipoprotein-FliC) as the vaccine carrier. The constructs were cloned into a Salmonella expression vector (pJHL65) and transformed into the ST strain (JOL912). The expression of immunogenic proteins within Salmonella was evaluated via immunoblotting. Results: Immunizing BALB/c mice orally and subcutaneously induced high levels of LI-specific systemic immunoglobulin G and mucosal secretory immunoglobulin A. In immunized mice, there was significant upregulation of interferon-γ and interleukin-4 cytokine mRNA and an increase in the subpopulations of cluster of differentiation (CD) 4⁺ and CD 8⁺ T lymphocytes upon splenocytes re-stimulation with LI antigens. We observed significant protection in C57BL/6 mice against challenge with 10^6.9 times the median tissue culture infectious dose of LI or 2 × 10⁹ colony-forming units of the virulent ST strain. Immunizing mice with either individual vaccine strains or co-mixture inhibited bacterial proliferation, with a marked reduction in the percentage of mice shedding Lawsonia in their feces. Conclusions: Salmonella-mediated LI gene delivery induces robust humoral and cellular immune reactions, leading to significant protection against LI and salmonellosis.

• Journal of Animal Science and Technology
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• 제57권4호
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• pp.10.1-10.7
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• 2015

Control and prevention of foot and mouth disease (FMD) by vaccination remains unsatisfactory in endemic countries. Indeed, consistent and new FMD epidemics in previously disease-free countries have precipitated the need for a worldwide control strategy. Outbreaks in vaccinated animals require that a new and safe vaccine be developed against foot and mouth virus (FMDV). FMDV can be eradicated worldwide based on previous scientific information about its spread using existing and modern control strategies.

• Pediatric Infection and Vaccine
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• 제18권1호
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• pp.91-96
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• 2011

BCG는 M. bovis로부터 유래된 생백신으로 5-6세 미만의 소아 결핵 특히 결핵성 수막염이나 속립성 결핵과 같은 중증 결핵의 예방 효과가 인정되고 있다. BCG 접종 후 이상반응은 국소 궤양 형성, 국소 림프절염이 가장 흔하며, 드물지만 골수염, 파종성 감염과 같은 치명적인 합병증도 있다. 파종성 BCG 감염은 주로 면역저하 환자에게서 흔하게 관찰되며 면역 저하가 없는 소아에게서 발생한 파종성 BCG 감염은 국내 문헌상 보고가 없었다. 저자들은 13개월 여환이 BCG 접종 5개월 뒤 발생한 발열, 피부 결절 및 다발성 림프절 종대를 주소로 내원하여, 백 신 접종 부위 외에 2군데에서 M. bovis BCG를 확인한 후 파종성 BCG 감염으로 진단된 1례를 경험하였다. 환아는 isoniazid와 rifampin으로 치료받았으며 약 2주 간의 입원 치료 후 임상 증상이 호전되어 퇴원하였다. 3주뒤 외래 추적 관찰에서 역설적 반응으로 생각되는 림프 절 종대와 피부 결절이 진행하였으나 동일 약제를 유지하면서도 다시 임상 증상이 호전되었고 환아의 면역학적 검사상 이상 소견이 관찰되지 않았기에 이를 보고하는 바이다.

• 대한두경부종양학회지
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• 제19권1호
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• pp.25-33
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• 2003

Background and Objectives: The Herpes Simplex type 2 Defective Infectious Single Cycle virus (DISC virus) is attenuated virus originally produced as viral vaccines but are also efficient gene transfer vehicle. The main goals of this study were to examine the efficiencies of the gene transfer using DISC vectors for various head and neck squamous cell carcinoma cell lines and to evaluate the efficacy of vaccination with DISC virus carrying a immunomodulatory genes (GM-CSF) as cancer therapy in a organotopic oral cavity squamous cell cancer model. Materials and Methods : We determinated the gene transfer efficiency of DISC virus by x-gal stain method and proved gene and protein expression of DISC-GMCSF transfected SCCVII cells by RT-PCR and ELISA method. Also we evaluated the ex vivo vaccination effects of SCCVII/GMCSF (DISC-GMCSF transfected SCCVII vaccine) vaccine on preventing the recurrence of micro-residual tumor. After the vaccination of SCCVII/GMCSF, specific cytotoxic T-cell responses was evaluated by CTL assay. Results: At an MOI of 10 DISC virus showed 64-88% of transfection rates in various head and neck squamous cancer cell lines. SCCVII cells transduced by DISC virus vector (MOI=10) carrying the GM-CSF gene, produced 4.5 nanogram quantities of GM-CSF per $10^6$ cells. In vivo vaccination using tumor cells transduced ex vivo with DISC-GMCSF resulted in better protection rate against subsequent tumor recurrence in organotopic oral cavity cancer model. Although tumor free survival rate was not statistically significantly increased in vaccination group (p=0.078), tumor specific cytotocic T-cell responses were significantly increased in SCCVII/GMCSF vaccination group. Conclusion: These data demonstrate that; 1) The DISC virus vector is capable of efficient gene transfer to various head and neck squamous cancer cell lines, 2) GM-CSF secreting genetically modified tumor vaccine (SCCVII/GMCSF) efficiently protected against tumor recurrence in organotopic micro-residual oral cavity cancer model and produced tumor specific cytotoxic T-cell response. DISC virus-mediated, cytokine gene transfer may prove to be useful as a clinical therapy for head and neck cancers.

• 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.

• Clinical and Experimental Pediatrics
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• 제51권4호
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• pp.377-382
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• 2008

목 적 : 수두에 대한 개체 면역력을 평가하는데 있어서 약독화 수두백신을 불활성화 시킨 후 희석시켜 만든 용액을 이용한 피부시험이 유용한지를 알아보고자 하였다. 방 법 : 2005년도 7월에서 8월까지 한림대의료원 강동성심병원 의료진 19명과 입원한 소아 22명을 포함한 총 41명(남자 22명, 여자 19명, 만 1-32세)을 대상으로 하였다. 수두 감염의 과거력, 수두 예방접종 여부에 대한 사항은 설문지를 통해 이루어졌고 피부시험을 위한 용액은 Oka 균주의 약독화 수두백신을 상온에 10일간 방치하여 불활성화 시킨 후, 이를 다시 생리식염수로 50배 희석하여 사용하였다. 이 용액을 우측 상완 전박부에 피내 주사하였고 대조액으로 반대편 상완 전박부에 생리식염수액을 피내 주사하였다. 48시간 이후 경결의 직경 5 mm 이상일 때 양성으로 판독하였다. 또한 효소면역분석법을 이용하여 혈청 VZV 특이 IgG 항체가를 측정하여 피부시험 결과와 비교 분석하였다. 결 과 : 효소면역분석법을 이용한 항체 검사에 대한 성인에서의 수두 피부시험의 민감도는 94.7%이고 양성 예측도는 100%였다. 동일한 판정 기준을 소아에 적용하였을 때 양성 예측도와 특이도는 100%로 높았으나 민감도와 음성 예측도는 각각 50%, 30.7%로 낮았다. 소아에서의 VZV 피부반응도는 성인에 비해 낮았다. 결 론 : 약독화 수두백신을 불활성화, 희석시켜 만든 용액을 이용한 피부시험은 수두에 대한 면역력을 예측하는데 있어, 간편하고 안전한 저가의 유용한 검사이다. 그러나 피부시험에서 양성판정기준을 결정할 때 조사목적, 대상연령 등을 고려하여 신중히 판단하여야 할 것이며, 이에 대해서는 향후 더 많은 대상에서 추가 연구가 이루어져야 할 것으로 사료된다.

• 대한수의학회지
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• 제53권1호
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• pp.29-35
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• 2013

To develop a live vaccine candidate using an attenuated strain of Salmonella Typhimurium (ST), biochemical properties, plasmid profile, PFGE patterns and pathogenic analysis of the ST isolate were carried out after sequential passage of the ST isolate in porcine neutrophils. By the passage, the ability of the neutrophil-adapted isolate to utilize d-xylose was lost, while the ability of the strain to ferment trehalose was delayed after 2 or more days of the culture. Also, changes including deletion of the gene fragments were observed in PFGE analysis of the neutrophil-adapted isolates. Two plasmids, 105kb and 50kb, were cured in the strain passaged over 15 times in porcine neutrophils. The 50% of lethal dose ($LD_{50}$) of the parent strain was changed from $1{\times}10^5\;LD_{50}$ to $6{\times}10^6\;LD_{50}$ by the passage in intraperitoneal injection of the strains into mice. These results suggested that bacterial genotypic and phenotypic responses might be globally altered depending on the inside environment of neutrophils.

• Journal of Veterinary Science
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• 제24권5호
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• pp.54.1-54.13
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• 2023

Background: Porcine reproductive and respiratory syndrome virus (PRRSV) vaccines do not provide full cross-protection, mainly due to the virus genetic variability. Despite this, vaccines based on modified-live PRRSV (PRRSV-MLV) reduce the disease impact. Objectives: To assess the efficacy of two commercial vaccines-one based on PRRSV1 (PRRSV1-MLV) and another on PRRSV2 (PRRSV2-MLV)-against a Japanese PRRSV2 field strain. Methods: Two groups of three-week-old piglets were vaccinated (G1: PRRSV1-MLV; G2: PRRSV2-MLV) and two were kept as non-vaccinated (INF and CTRL). One month later, G1, G2, and INF were challenged with a PRRSV2 field strain. Results: After the challenge, clinical signs were only observed in INF. Moreover, the highest rectal temperatures and values for the area under the curve (AUC) were observed in INF. Regarding viral detection, both AUC and the proportion of positive samples in blood were higher in INF. In G1, viremic animals never reached 100%. At necropsy (21 d after the challenge), differences for titers among groups were only found in tonsils (G1 < G2 and INF). One animal (belonging to G1) was negative in all tissues. Regarding humoral responses, G1 and G2 seroconverted after vaccination, as detected in the corresponding enzyme-linked immunosorbent assay. Specific neutralizing antibodies (NA) against PRRSV1-MLV were already detected at 14 d after vaccination in G1, showing a significant booster after the challenge, while PRRSV2-MLV NA were detected in G2 at the end of the experiment. Conclusions: Despite genetic differences, PRRSV1-MLV has been demonstrated to confer partial protection against a Japanese PRRSV2 strain, at least as good as PRRSV2-MLV.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.330-338
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• 2019

Chronic infection with intracellular Brucella abortus (B. abortus) in livestock remains as a major problem worldwide. Thus, the search for an ideal vaccine is still ongoing. In this study, we evaluated the protective efficacy of a combination of B. abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12) and malate dehydrogenase (rMDH), cloned and expressed into a pMal vector system and $DH5{\alpha}$, respectively, and further purified and applied intraperitoneally into BALB/c mice. After first immunization and two boosters, mice were infected intraperitoneally (IP) with $5{\times}10^4CFU$ of virulent B. abortus 544. Spleens were harvested and bacterial loads were evaluated at two weeks post-infection. Results revealed that this combination showed significant reduction in bacterial colonization in the spleen with a log protection unit of 1.31, which is comparable to the average protection conferred by the widely used live attenuated vaccine RB51. Cytokine analysis exhibited enhancement of cell-mediated immune response as IFN-${\gamma}$ is significantly elevated while IL-10, which is considered beneficial to the pathogen's survival, was reduced compared to control group. Furthermore, both titers of IgG1 and IgG2a were significantly elevated at three and four-week time points from first immunization. In summary, our in vivo data revealed that vaccination with a combination of five different proteins conferred a heightened host response to Brucella infection through cell-mediated immunity which is desirable in the control of intracellular pathogens. Thus, this combination might be considered for further improvement as a potential candidate vaccine against Brucella infection.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.77-77
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• 1995

Human immunodeficiency virus type 1 (HIV-1) contains several nonstructural genes which are required for the viral replication and disease pathogenesis. Among them, tat and nef genes encode an essential transactivator of HIV-1 LTR and a pluripotent protein which seems to be essential for the in vivo but not in vitro viral replication, respectively. We constructed two tat and n of defective HIV-1 and tested for their ability to replicate in several T cells. The defective viruses did not replicate in CD4$\^$+/ T cells, but rescued in the recombinant Jurkat-tat cell which also contains tat gene. The replication of tat and nef defective HIV-1 which expresses chloramphenicol acetyltransferase(CAT) gene was easily detected by a sensitive CAT assay. No revertant was identified during the passages of the mutant viruses for more than two months in Jurkat-tat cells. tat and n of defective HIV-1 could be used instead of wild type viruse for several purposes such as inhibitor screening and development of attenuated AIDS vaccine.