• Title/Summary/Keyword: Vaccine production

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Interaction between host cell proteins and open reading frames of porcine circovirus type 2

  • Si-Won Park;In-Byung Park;Seok-Jin Kang;Joonbeom Bae;Taehoon Chun
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.698-719
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    • 2023
  • Postweaning multisystemic wasting syndrome (PMWS) is caused by a systemic inflammation after porcine circovirus type 2 (PCV2) infection. It was one of the most economically important pathogens affecting pig production worldwide before PCV2 vaccine was first introduced in 2006. After the development of a vaccine against PCV2a type, pig farms gradually restored enormous economic losses from PMWS. However, vaccine against PCV2a type could not be fully effective against several different PCV2 genotypes (PCV2b - PCV2h). In addition, PCV2a vaccine itself could generate antigenic drift of PCV2 capsid. Therefore, PCV2 infection still threats pig industry worldwide. PCV2 infection was initially found in local tissues including reproductive, respiratory, and digestive tracks. However, PCV2 infection often leads to a systemic inflammation which can cause severe immunosuppression by depleting peripheral lymphocytes in secondary lymphoid tissues. Subsequently, a secondary infection with other microorganisms can cause PMWS. Eleven putative open reading frames (ORFs) have been predicted to encode PCV2 genome. Among them, gene products of six ORFs from ORF1 to ORF6 have been identified and characterized to estimate its functional role during PCV2 infection. Acquiring knowledge about the specific interaction between each PCV2 ORF protein and host protein might be a key to develop preventive or therapeutic tools to control PCV2 infection. In this article, we reviewed current understanding of how each ORF of PCV2 manipulates host cell signaling related to immune suppression caused by PCV2.

Immunomodulatory Properties of Lactobacillus plantarum NC8 Expressing an Anti-CD11c Single-Chain Fv Fragment

  • Liu, Jing;Yang, Guilian;Gao, Xing;Zhang, Zan;Liu, Yang;Yang, Xin;Shi, Chunwei;Liu, Qiong;Jiang, Yanlong;Wang, Chunfeng
    • Journal of Microbiology and Biotechnology
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    • v.29 no.1
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    • pp.160-170
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    • 2019
  • The lactic acid bacteria species Lactobacillus plantarum (L. plantarum) has been used extensively for vaccine delivery. Considering to the critical role of dendritic cells in stimulating host immune response, in this study, we constructed a novel CD11c-targeting L. plantarum strain with surface-displayed variable fragments of anti-CD11c, single-chain antibody (scFv-CD11c). The newly designed L. plantarum strain, named 409-aCD11c, could adhere and invade more efficiently to bone marrow-derived DCs (BMDCs) in vitro due to the specific interaction between scFv-CD11c and CD11c located on the surface of BMDCs. After incubation with BMDCs, the 409-aCD11c strain harboring a eukaryotic vector pValac-GFP could lead to more efficient expression of GFP compared with wild-type strains shown by flow cytometry analysis, indicating the enhanced translocation of pValac-GFP from L. plantarum to BMDCs. Similar results were also observed in an in vivo study, which showed that oral administration resulted in efficient expression of GFP in both Peyer's patches (PP) and mesenteric lymph nodes (MLNs) within 7 days after the last administration. In addition, the CD11c-targeting strain significantly promoted the differentiation and maturation of DCs, the differentiation of $IL-4^+$ and $IL-17A^+$ T helper (Th) cells in MLNs, as well as production of $B220^+$ $IgA^+$ B cells in the PP. In conclusion, this study developed a novel DC-targeting L. plantarum strain which could increase the ability to deliver eukaryotic expression plasmid to host cells, indicating a promising approach for vaccine study.

Reversibility and safety of KISS1 metastasis suppressor gene vaccine in immunocastration of ram lambs

  • Han, Yan-Guo;Liu, Gui-Qiong;Jiang, Xun-Ping;Xiang, Xing-Long;Huang, Yong-Fu;Nie, Bin;Zhao, Jia-Yu;Nabeel, Ijaz;Tesema, Birhanu
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.835-841
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    • 2018
  • Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results: The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS (KISS1-hepatitis B surface antigen S) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction. Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.

Evaluation on Immunogenicity and Safety of Avian Influenza Isolate(ADL0401) as a Candidate for the Killed Vaccine against tow-Pathogenic Avian Influenza (약병원성 조류인플루엔자 사독백신개발을 위한 후보주(ADL0401)의 면역 원성 및 안전성 평가)

  • Lee J. S.;Ha D. H.;Kim J. E.;Ha B. D.;Mo I. P.
    • Korean Journal of Poultry Science
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    • v.32 no.2
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    • pp.113-123
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    • 2005
  • Avian influenza (AI) virus (AIV) is distributed worldwide and it has been isolated from various species of wild and domestic birds. AI transfers with high speed and shows diverse pathogenicity syndroms. In Korea, several low Pathogenic AIV, H9N2, have been isolated from the commercial farms with severe decrease of egg production and mortality resulted in severe economic loss since 1996. Therefore, it has been requested to develop AI vaccines to prevent clinical signs and economic losses from the field infection of AIV. To develop a killed vaccine that efficiently prevents low pathogenic AIV (H9N2), evaluation on the pathogenicity and selection of an inactivator for H9N2 is taking place and is being tested safety and immunogenicity of vaccine produced. Based on the pathogenicity test and viral reisolation test, the ADL0401 isolate is the characteristic low pathogenic AIVs and has fairly similar biologic functions compared with MS96 which is the official low pathogenic AIV (H9N2) and one of the predominant AIV isolated from poultry farms in Korea. In antigenicity tests, the ADL0401 and MS96 virus have no significant antigenic difference. In inactivation tests, the ADL0401 isolates can be easily inactivated with $0.1\%$ Formalin at $37^{\circ}C$ within 1 hour with a little decrease of HA titer. The vaccine developed in the present report has no harmful effect on bird and forms good immune capability. Therefore, the isolates, ADL0401 can be used for a killed vaccine which can reduce the clinical signs and viral shedding in the birds infected with H9N2 low pathogenic AIVs.

Development of Transgenic Carrot Oral Vaccine to Protect against Diarrhea of Piglets (자돈 설사병 방지를 위한 경구백신용 형질전환 당근 개발)

  • 이영선;황철호
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.287-293
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    • 2002
  • We are trying to develop a transgenic carrot with aims of production and delivery of oral vaccine against microbial enteropathogen using a K88ac pilin gene. A K88ac antigen (pilin) gene was isolated by PCR from the K88ac genomic DNA. The pilin gene was constructed in pGA748 and introduced via Agrobacterium tumefaciens to the explants of carrot hypocotyl and then 494 transgenic lines were established. The amounts of the K88ac antigen produced in each of the cell lines were determined by western and two elite cell lines (M1-17, Y14-1) were selected based on higher levels of expression of the antigens as well as rate of cell growth and efficiency of embryogenesis. In order to test an immunization induced by oral administration of the transgenic carrot, serum of the mice fed with the carrot vaccine were tested in ELISA. It tumed out that the mice fed with 3 g of transgenic carrot showed a similar level of antibody compared to those applied with 10 $\mu\textrm{g}$ of the purified recombinant pilin protein. Besides, various clinical responses were measured after challenging with ETEC K88ac strain to the piglets experiencing an oral immunization with the transgenic carrot. The piglets fed with carrot vaccine showed a lower level of diarrhea in fecal score compared to those fed with non-transgenic carrot. A higher level of increase in weight of the piglets fed with the transgenic carrot vaccine was observed comparing to those fed with non-transgenic carrot as control.

OPTIMIZATION OF CULTURE CONDITIONS FOR PRODUCTION OF PNEUMOCOCCAL CAPSULAR POLYSACCHARIDE TYPE I

  • Kim, S.N.;K.K. Min;Kim, S.H.;Park, I.H.;Lee, S.H.;S.N. Pyo;D.K. Rhee
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.186-186
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    • 1996
  • Streptococcus pneumoniae (pneumococcus), the most common cause of bacterial pneumonia, has an ample polysaccharide(PS) capsule that is highly antigenic and is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 1 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based media were found to support luxuriant growth of pneumococcus type 1 at the same level. Because BHI medium is rather expensive and more complex than the Casitone based media, the Casitone based media was used to study optimization of the culture condition. The phase of growth which accomodated maximum PS production was logarithmic phase. Concentrations of glucose greater than 0.2% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplemention of various concentrations of metal ion (with the exception of calcium ion) had adverse effects on growth and PS production. On the other hand, low level aeration was beneficial for increased PS production. Addition of 3 mg/I concentration of methionine, phenylalanine, and threonine were found to enhance growth and PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

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Evaluation of Optimal Condition for Recombinant Bacterial Ghost Vaccine Production with Four Different Antigens of Streptococcus iniae-enolase, GAPDH, sagA, piaA (연쇄구균증 항원-enolase, GAPDH, sagA, piaA에 대한 재조합 고스트 박테리아 백신의 생산 최적화)

  • Ra, Chae-Hun;Kim, Yeong-Jin;Son, Chang-Woo;Jung, Dae-Young;Kim, Sung-Koo
    • Journal of Life Science
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    • v.19 no.7
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    • pp.845-851
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    • 2009
  • A vector harboring double cassettes; a heterologous gene expression cassette of pHCE-InaN-antigen and a ghost formation cassette of pAPR-cI-E lysis 37 SDM was constructed and introduced to E. coli DH5a. For the production of a bacterial ghost vaccine, bacterial ghosts from E. coli / Streptococcus iniae with four different types of antigens - enolase, GAPDH, sagA and piaA - were produced by the optimization of fermentation parameters such as a glucose concentration of 1 g/l, agitation of 300 rpm and aeration of 1 vvm. Efficiency of ghost bacteria formation was evaluated with cultures of OD$_{600}$=1.0, 2.0 and 3.0. The efficiency of the ghost bacteria formation was 99.54, 99.67, 99.99 and 99.99% with inductions at OD$_{600}$=3.0, 1.0, 2.0 and 1.0 for E. coli/S. iniae antigens enolase, piaA, GAPDH and sagA, respectively. Ghost bacteria as a vaccine was harvested by centrifugation. The antigen protein expressions were analyzed by SDS-PAGE and western blot analysis, and the molecular weights of the enolase, piaA, GAPDH and sagA were 78, 26, 67 and 26 kDa, respectively. The molecular weights of the expressed antigens were consistent with theoretical sizes obtained from the amino acid sequences.

Optimization of Culture Conditions for Production of Pneumococcal Capsular Polysaccharide Type IV

  • Kim, S.N.;Min, K.K.;Choi, I.H.;Kim, S.W.;Pyo, S.N.;Rhee, D.K.
    • Archives of Pharmacal Research
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    • v.19 no.3
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    • pp.173-177
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    • 1996
  • The Pneumococcus, Streptococcus pneumoniae, has an ample polysaccharide (PS) capsule that is highly antigenic and is the main virulence factor of the organism. The capsular PS is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 4 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based medium were found to support luxuriant growth of pneumococcus type 4 at the same level. Therefore in this study, the Casitone based medium was used to study optimization of the culture condition because of BHI broth's high cost and complex nature. The phase of growth which accomodated maximum PS production was exponential phase. Concentrations of glucose greater than 0.8% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplementation of various concentrations of metal ion (with the exception of calcium, copper, and magnesium ions) had adverse effects on growth and PS production. On the other hand, low level aeration and supplementation of 3 mg/l concentration of asparagine, phenylalanine, or threonine were beneficial for increased PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

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Opportunities and Challenges for Application of Poultry Science and Technology into the 21st Century

  • Sheldon, B.L.
    • Korean Journal of Poultry Science
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    • v.20 no.3
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    • pp.161-170
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    • 1993
  • Prospects are briefly reviewed for further advances in current poultry industry technology in the foreseeable future. It is concluded that in the most advanced industries progress should continue at a similar rate to the recent past in conventional genetics and breeding, nutrition and disease control. Significant benefits will also follow in the short-term from the application of molecular biotechnology to disease diagnosis and vaccine production. Technical advances now make it possible to produce transgenic chickens at acceptable success rates but applications of this technology to poultry breeding will not become significant till we have sufficient knowledge of the poultry genome, and especially the genes involved in production performance. For the undeveloped and less advanced industries it is argued that the level of advanced technologies to be implemented in those countries should be decided largely on market forces, informed by objective assessment of the diverse options available. The need for urgent international action on conservation of poultry genetic resources is also stressed.

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Optimal culture conditions for production of Escherichia coli Adhesin protein coupled to Escherichia coli Heat Labile Enterotoxin A2B in Escherichia coli TB1.

  • Lee, Yong-Hwa;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.226.2-226.2
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    • 2003
  • The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically linked to the Itxa2b gene, which was then cloned into the pMAL -p2E expression vector. The chimaeric construction of pMALfimH/Itxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

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