초록 ▼

• 기존의 cI857 / λ PR / 유전자 E 발현 시스템에 anti-sense ParaBAD 프로모터를 추가하여 효율이 우수한 고스트백신 시스템 개발.
• Salmonella Typhimurium (ST) 및 Salmonella Enteritidis (SE)의 교차방어가 가능한 SE 생균 백신 시스템 개발.
• PrP(106-126) 처리에 의한 autophagy의 활성화와 활성화된 autophagy에 의한 신경세포 사멸 유도 확인, 신경세포에 많이 존재하는 cellular prion protein (PrPc)이

• 기존의 cI857 / λ PR / 유전자 E 발현 시스템에 anti-sense ParaBAD 프로모터를 추가하여 효율이 우수한 고스트백신 시스템 개발.
• Salmonella Typhimurium (ST) 및 Salmonella Enteritidis (SE)의 교차방어가 가능한 SE 생균 백신 시스템 개발.
• PrP(106-126) 처리에 의한 autophagy의 활성화와 활성화된 autophagy에 의한 신경세포 사멸 유도 확인, 신경세포에 많이 존재하는 cellular prion protein (PrPc)이 방어요인 작용 확인.
• IFN-I signal pathway는 monocyte/수지상세포를 비롯한 NK 세포의 이주 및 분화에 관여함을 flow cytometric analys/confocal microscopy 등의 분석법으로 규명, CD11b+Ly-6Chi monocyte의 분화

(출처 : 보고서 요약서 3p)

Abstract ▼

□ Purpose
This research laboratory studies the control systems for interventions, preventions and treatments of various zoonotic diseases using an integrated modern molecular biological and new cellular technologic framework with an “one health” concept. Especially, the goal of the control system

□ Purpose
This research laboratory studies the control systems for interventions, preventions and treatments of various zoonotic diseases using an integrated modern molecular biological and new cellular technologic framework with an “one health” concept. Especially, the goal of the control system is to link the Brucellosis, Salmonellosis, prion diseases and emerging pathogen associated with global climate changes. This basic research laboratory (BRL) project promises most efficient and effective control strategies of zoonosis and cut the costs and risks of the newly emerging zoonotic diseases by protruding specific delivery system and/or molecules.

□ contents
Specific aim 1: Development of system to control food derived zoonotic diseases. We will develop Salmonella and/or Brucella live delivery or ghost vaccine induced by autolysis using Ghost cassette (Pm/xylS gene E-lysiscontrol) for prevention salmonellosis/brcellosis. Post immunization with these vaccines in various domestic animals, the characteristics of the induced immune responses will investigated. Serum IgG, mucosal IgA and cytokines will investigated post immunization with Salmonella and/or Brucella ive delivery or ghost vaccine. In addition, the efficacy of the Salmonella and/or Brucella ive delivery or ghost vaccine in the immunized animals will be evaluated via challenge with wild type Salmonella or Brucella strain.
Specific aim 2: Discovery of molecular and cellular control targets in prion disease based on autophagic regulation. We will identify the role of autophagy in the process of neuronal damage caused by specific prion protein (PrP106-126) in order to investigate the protective effect toward neuronal injury via regulatory mechanisms of autophagic activation control and autophagic stress on neuronal injuries. First, we will search for activating molecules of autophagy and verify its neuroprotective efficacy; second, we will investigate the effect and mechanisms of autophagic activities on neurotoxicity in prion disease; third, we will identify autophagy-related factors and neuronal injuries due to prion protein; fourth, we will assess the activity of autophagy via regulation of PrPc expression; finally, we will evaluate the expression of PrPc within a neuron cell through the regulation of autophagy activities. Overall, the ultimate goal of this research is to explore the molecular and cellular control targets in prion disease through autophagy regulation.
Specific aim 3: Meta-immunobiological study on newly emerging zoonotic diseases caused by global climate changes for the development of regulatory techniques. The main zoonotic diseases caused by global climate changes, such as flaviviral infection transmitted by mosquitos, pose an increasing threat to global health and welfare. The third specific aim includes meta-immunobiological study on such newly emerging zoonotic diseases to protrude regulatory molecules. In particular, this study will be focused on the role of IFN-I and TLR signal pathways in controlling the progression of acute inflammatory diseases (viral encephalitis and hemorrhage), and the role of CD4+Foxp3+ Tregs will be performed in elaborated infectious models.

□ Developement results
Specific aim 1: Development of system to control food derived zoonotic diseases. A Salmonella Gallinarum (SG) with controlled expression of the bacteriophage PhiX174 lysis gene E or a live vaccine system were constructed to develop a novel, safe and immunogenic fowl typhoid (FT) vaccine. The safety and protective efficacy of the ghost and live attenuated vaccine was tested in chickens. Significant protection against the virulent challenge was observed in all immunized groups. A potent antigen-specific lymphocyte activation response along with significantly increased percentages of CD4+ and CD8+ T lymphocytes found in all immunized groups clearly indicate the induction of cellular immune responses. Live ST vaccine expressing BCSP31, Omp3b and SOD proteins of Brucella abortus was constructed. IP immunization with the mixture can induce immune responses, and can effectively protect mice against brucellosis. Brucella abortus ghost vaccine was generated using the GI24. The IP immunization with the ghost can effectively protect mice from infections of virulent Brucella abortus.
Specific aim 2: Discovery of molecular and cellular control targets in prion disease based on autophagic regulation. Prion protein-induced autophagy flux is involved in neuron cell death in prion disease and suggest that autophagy flux might play a critical role in neurodegenerative diseases including prion disease. Autophagy flux induced by prion protein is more activated in prpc expressing cells than in prpc silencing cells. Reduction of mitochondrial membrane potential, translocation of Bax, and cytochrome c release which induced by PrP (106–126) treatment were inhibited by b-catenin activation, which contributed to prevented PrP (106–126)-induced neuronal cell death. b-catenin activation by melatonin prevented PrP (106–126)-induced neuronal cell death through regulating anti-apoptotic proteins and mitochondrial pathways. These results also suggest the therapeutic value of Wnt/b-catenin signaling in prion-related disorders as influenced by melatonin.
Specific aim 3: Meta-immunobiological study on newly emerging zoonotic diseases caused by global climate changes for the development of regulatory techniques. IFN-I signal pathway played an essential role to control acute inflamamtion via regualting Ly-6C^hi monocytes and showed distinct upstream role in establishing early orchestrated environment to tissue resident and hematopoietic stem cells (published in PLoS Pathogens). TLR2/9 dual signal pathway showed regulatory function in differentiation of Ly-6C^hi monocytes and NK cells, and thereby regulate viral dissemination (submitted to PLoS Pathogens). Intriguingly, IFN-I pathway ablation altered viral tropism of neurotropic viruses, like inducing viral hemorrhage rather than viral encephalitis. Also, elaborated infectious models (Dereg.DKO) for studying on dengue hemorrhage fever (DHF) were stably established during this study.

□ Expected Contribution
▪Prevention of human infection from domestic animals via eradication of salmonellosis and brucellosis in domestic animals.
▪The data generated from this proposal will provide the information when new vaccine development willl be tried against domestic animals and human infections such as Camphylobacter, Clostridium as well as Listera infections, that are important in public health, and contribute to enhance the national economic.
▪Novel regulatory factors and genes which modulate the autophagy flux may be developed therapeutic targets or strategies against neurodegenerative diseases including prion-mediated neuronal damage.
▪The new crucial factors that regulate acute inflammatory disease caused by newly emerging virus like JEV and Zika virus may be able to enlarge the immunobiological study on therapeutic approaches.
▪The data generated from this proposal will provide new paradigm in development of therapeutics for treatment of various acute inflammatory diseases, such as Zika virus, MERS as well as influenza, that are important in public health, and contribute to enhance the national economic.

(출처 : SUMMARY 6p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 4
• SUMMARY ... 6
• 목차 ... 7
• 1. 연구개발 목표 및 내용 ... 8
• 가. 최종 목표 ... 8
• 나. 단계목표(당초 목표 및 수정·보완 목표) ... 13
• 다. 당초 목표의 수정·보완(중요 연구변경) 사유 ... 16
• 라. 1단계 연차별 연구목표 및 내용 ... 17
• 2. 연구 추진전략 및 방법 ... 21
• 3. 주요 연구개발 결과 ... 34
• 가. 목표달성도 ... 34
• 나. 대표적 연구업적 ... 258
• 다. 기타 연구성과 ... 259
• 4. 연구수행에 따른 문제점 및 개선방향 ... 259
• 5. 연구개발성과 현황 ... 260
• 6. 국가과학기술종합정보시스템에 등록한 연구시설·장비 현황 ... 284
• 7. 기타사항 ... 284
• 끝페이지 ... 285