A Satellite Symposium of the Joint Symposium between Seoul National University and Hokkaido University

Veterinary Medicine Today 2006 -ZoonosisProgram

15:00-15:15 Opening Remarks Takashi Umemura Dean of Graduate School of Veterinary Medicine, Hokkaido University Yong-Soon Lee Department of Veterinary Public Health, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI), Seoul National University

Session 1

Research on Prion Diseases Chairman: Motohiro Horiuchi Department of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University

15:15-15:35 Identification of biomarkers in the prion-infected mouse Han Sang Yoo Department of Infectious Diseases, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI), Seoul National University

15:35-15:55 Application of Real-time Immuno-PCR Method for Prion Protein in Meat I.B. Yim and H.J. Woo Department of Immunology, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI), Seoul National University 15:55-16:15 Propagation and inhibition of PrPSc formation in vitro and in vivo Motohiro Horiuchi Department of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University

16:15-16:30

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Break time

Session 2 Research on other Zoonoses Chairman: Misao Onuma Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University

16:30-16:50 Clostridium Infections in Captive Wild Animals at the Seoul Grand Park Zoo in Korea Nam-Shik Shin Department of Wild Animal Diseases, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI), Seoul National University

16:50-17:10 Leishmaniasis Ken Katakura Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University

17:10-17:30 Low Pathogenic Avian Influenza by H9N2 subtype virus in Korea Hyuk-Joon Kwon, Sun-Hee Cho and Sun-Joong Kim Laboratory of Influenzavirus, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI), Seoul National University

17:30-17:50 Efficacy of intracerebral immunization against neurotropic viruses in mice Jae-Ho Shin Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University

19:00- Social reception

Sapporo Beer Garden

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Profile Han Sang Yoo Associate Professor Department of Infectious Diseases, College of Veterinary Medicine, KRF Zoonotic Disease Priority Research Institute and BK 21 for Veterinary Science, Seoul National University San 56-1, Sillim-dong, Gwanak-Gu, Seoul 151-742, KOREA

ACADEMIC DEGREES: DVM MS

1982 1984

Ph.D.

1995

Seoul National University, College of Veterinary Medicine The Graduate School, Seoul National University (Major : Veterinary Microbiology) University of Minnesota (Major : Veterinary Microbiology), USA

PROFESSIONAL APPOINTMENTS: 1997.9 - Present

Associate Professor, Assistant Professor, Instructor College of Veterinary Medicine, Seoul National University 2005. 1- 2006.2 Visiting Professor Department of Animal Health and Biomedical Sciences The School of Veterinary Medicine University of Wisconsin-Madison, USA 2001.7 - Present Adjunct Senior Veterinary Researcher National Veterinary Research and Quarantine Service, Korea 1999. 11- 2001. 10 Editor-in-Chief and Secretary for Academic Affairs The Korean Association for Veterinary Science 1984. 4- 1997.8 Veterinary Researcher National Veterinary Research and Quarantine Service, Korea

RESEARCH INTERESTS: Biological and Pathogenic Mechanism of Zoonotic Diseases - Biological characterization of prion-infected cell lines - Isolation and identification of virulence factor(s) in Brucella spp. Development of Needle-Free Delivery System to Control Respiratory Diseases in Pigs - Development of oral vaccine based on yeast, plant and lactic acid bacteria - Development of spray vaccine using chitosan nanoparticle to control atrophic rhinitis Establish the Monitoring System in the Pig Facilities for the Production of Bio-organs - Development of bio-monitoring system and guideline in pig facilities to produce the Bio-organ

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Identification of biomarkers in the prion-infected mouse Han Sang Yoo Professor Department of Infectious Diseases, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI, KRF Priority Research Institute), and BK 21 for Veterinary Science, Seoul National University, San 56-1, Sillim-dong, Gwanak-Gu, Seoul 151-742, KOREA

Transmissible spongiform encephalopathy (TSE) diseases caused by prion infection are a group of fatal neurodegenerative disorders such as BSE, CWD, Scrapie and CJD. These disorders are believed to occur through the accumulation of an abnormal pathogenic isoform (PrPSc) of cellular protein (PrPc). However, the molecular mechanism of disease pathology is not fully understood, yet. Also, all validated diagnostics of the diseases rely on the immunochemical detection of PrPSc, precluding diagnosis of presymptomatic prion infections. Although the differential expression of a number of transcripts in CNS tissue at late stage of the infection has been reported, a useful biomarker has not yet been identified. Ideally, these surrogate markers should be detectable (and differentially expressed) in easily accessible body fluids such as blood or urine. Also, assessment of the levels of surrogate markers in healthy individuals is crucial in order to define the normal range of expression in order to determine what represents abnormal levels. Changes in gene expression may occur as part of the primary pathogenesis of disease or secondary to other factors involved in disease progression. In order to find out appropriate surrogate biomarker(s), gene expression was comprehensively analyzed in different stages of prion-infected mice. Eight-week-old C57B1/6 mice were infected intraperitoneally with the Rocky Mountain Laboratory (RML) strain of mouse prion. At 108, 158 and 198 days post-inoculation, mice were sacrificed and gene expression profiles in the brain, PBMC and spleen were obtained using Affymetrix gene arrays. Of the 45,000 genes whose expression was measured, numbers of genes were up or down-regulated at different time points. The patterns were also different depending on the organs. Those genes were involved in multiple biological processes. Presence of pathogenic isoform of prion (PrPSc) in brains and spleens was detected by Western blot analysis with monoclonal antibody, 6H4. In the analysis, PrPSc in the spleen was detected earlier than that in brain. Also, expression of some important genes including glial fibrilliary acidic protein in microarray was validated by iqRT-PCR. Expression of the genes was organ and time specifically increased. Concordant trends in the expression levels yielded by microarrays and RTPCR were observed for all genes. The changes in expression in RML-infected mice vs uninfected mice observed by microarray analysis were generally lower than those derived by RTPCR. These results suggested that the genes which showed the increased expression profile, especially in PMBC might be used as biomarkers in the prion infected animals even though further analysis is required.

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Profile Hee-Jong Woo Professor Department of Immunology, College of Veterinary Medicine Seoul National University, Seoul, Republic of Korea #707, Bldg 85, Seoul national University, San 56-1, Shillim-dong, Gwanak-Gu, Seoul, Korea

ACADEMIC DEGREES: B.S.

1983

Ph.D. 1986

Department of Pharmacy, University of Tokyo (Immunochemistry /Cellular Immunology) Department of Pharmacy, University of Tokyo (Immunochemistry /Cellular Immunology)

PROFESSIONAL APPOINTMENTS: 2004.3-present 1998.10-2004.2 1994.10-1998.9 1992.8-1994.9 1992.51989.7-1992.4 1988.9-1989.6 1987.9-1989.6

College of Veterinary Medicine, Seoul National University, Professor College of Veterinary Medicine, Seoul National University, Associate Professor College of Veterinary Medicine, Seoul National University, Assistant Professor College of Veterinary Medicine, Seoul National University, Instructor Boston University, Medical School, Assistant Professor Harvard Medical School, Laboratory of Cancer Biology, Boston, Instructor The Wistar Institute, Philadelphia , Research fellow Univ. Pennsylvania, Medical School, Division of Molecular Diseases, Philadelphia, Research fellow

RESEARCH INTERESTS: Functional study of Galectin-3 (Molecular embryology and transmission) Apoptosis of T cells and nerve cells Interaction between nerve and immune cells Research of Prion expression Development of animal monoclonal antibodies Development of anticancer- natural substance

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Application of Real-time Immuno-PCR Method for Prion Protein in Meat I.B. Yim and H.J. Woo Professor Department of Immunology, College of Veterinary Medicine, Zoonotic Disease Institute (ZooDI, KRF Priority Research Institute), and BK 21 for Veterinary Science, Seoul National University, San 56-1, Sillim-dong, Gwanak-Gu, Seoul 151-742, KOREA

Prion diseases are fatal neurodegenerative diseases caused by a conformational change of prion protein (PrP). Recent reports have shown that PrP accumulates in the skeletal muscles of diseased animals and man. As the Western blot or ELISA, officially used to detect PrP, are not sufficient because of very low level of PrP in the skeletal muscles of animals for food, the novel approaches to improve current protocols for detecting PrP in meat. As a model system, we have focused on the murine system including related proteins, antibodies and cell lines for in vitro studies, as well as numerous strains of animals for in vivo investigations. Taking advantage of the system, we have expressed and purified recombinant mouse (recMo) PrP, and chosen several peptides from PrP of animals for immunizing goat to get polyclonal antibodies. Using recMo PrP and polyclonal antibodies, immuno-polymerase chain reaction (IPCR) was performed to improve the limit of detection by ELISA for PrP. The PCR was performed in realtime. Sensitivity of our results using IPCR can be compared with conventional detection by Western blot and ELISA. We are establishing this technique for detecting PrP in food including red meat.

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Profile Motohiro Horiuchi Professor Laboratory of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, JAPAN

ACADEMIC DEGREES: B.S. Ph.D.

1986 1994

Hokkaido University (Veterinary Medicine) Hokkaido University (Veterinary Medicine)

PROFESSIONAL APPOINTMENTS: 2003, 8 - present 1995, 6 - 2003, 7

1989, 1 - 1995, 6 1988, 4 - 1988, 12

Professor, Laboratory of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University Associate Professor, Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine (1997,7 - 1999, 7, Visiting fellow, Rocky Mountain Laboratories, National Institute of Health, USA) Research Associate, Department of Veterinary Public Health, Obihiro University of Agriculture and Veterinary Medicine Researcher, Nippon Roche, Co. Ltd.

RESEARCH INTERESTS: We are now investigating on the following topics on prion diseases: 1) Understanding the entity of the causative agent ‘prion’, especially on the linkage of biological phenotypes of prion to biochemical properties of disease-specific prion protein. This includes the understanding of mechanism how prion strains are determined. 2) Elucidating molecular mechanisms of prion propagation and prion transmission between cells. 3) The establishment of treatment for prion diseases. We are now trying a combination of compounds that inhibit PrPSc formation and regenerative medicine using mesenchymal stem cell. 4) The elucidation of functions of cellular prion protein (PrPC). Our laboratory has also interested in infectious diseases that cause encephalopathy/encephalitis. We are now attempting to establish reverse genetics of West Nile virus and to establish a differential diagnosis of flavivirus infections. We have also started research on porcine progressive enteritis.

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Propagation and inhibition of PrPSc formation in vitro and in vivo Motohiro Horiuchi Professor Laboratory of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, JAPAN

The major component of the causative agent of prion diseases is thought to be a disease-specific isoform of prion protein (PrPSc). PrPSc is generated from cellular isoform of prion protein (PrPC) and the process in the conversion of PrPC to PrPSc is a central event in pathogenesis of prion diseases. Not only PrPC but also other host factors are believed to be involved in the PrPSc formation, i.e., prion replication. Finding of such factors will be valuable for understanding the molecular mechanism of prion propagation. In addition, understanding of the mechanism of prion propagation will provide new therapeutic targets. In the symposium, I will present two of our ongoing researches, one is an attempt to identify host factor(s) that will be involved in prion replication, and the other is the effect of anti-PrP antibodies on the propagation of prion and neurodegeneration in prion-infected mice Comparison between the circumstance(s) permitting prion replication and that not permitting would be one of the ways to identify the host factor(s) that will be involved in prion replication. To address this issue, we established prion-susceptible and prion-unsusceptible Neuro2a (N2a) mouse neuroblastoma cell subclones, and compared gene expression profiles among them using DNA microarray analysis. We first selected 44 genes that showed more than two-fold higher expression in susceptible N2a-5 than in unsusceptible N2a-1 subclone base on DNA microarray analysis. Quantitative RT-PCR analysis using TaqMan probe revealed that expression patterns of 38 genes up-regulated in N2a-5 were consistent with the results of DNA microarray analysis. Next, we attempted to investigate the effect of these genes on PrPSc formation by knocking down the targeted gene expression using small interfering RNA (siRNA) technology. So far now, we have found that 6 siRNA reduced the formation of PrPSc in ScN2a-5 to nearly half of that in ScN2a-5 cells transfected with control siRNA. These results suggested that these are possible candidates for the host factor(s) being involved in prion replication. Experiments are now under way to examine whether the expression of these genes confers prion-susceptibility to prion-unsusceptible N2a subclones. A couple of laboratories including us has shown that anti-PrP antibodies inhibit PrPSc formation in cell-free conversion reaction and in the cells persistently infected with prion, although the mechanism for the inhibition is still controversial. We examined whether intra-cranial infusion of anti-PrP antibodies prevents prion propagation and neurodegeneration. We administrated anti-PrP mAbs to mice infected with prion during 120 to 148 dpi by Alzet osmotic pump and brains were examined for the accumulation of PrPSc and histopathology. The results showed that intracranial infusion of anti-PrP mAb retarded the PrPSc accumulation. In addition, spongiform degenerations in hippocampus and thalamus of mice treated with anti-PrP mAbs were apparently milder than those treated with negative-control mAb. Although adverse effects by anti-PrP mAbs should be carefully investigated, these results suggested that mAb treatment is one of the candidates for therapeutics for prion diseases.

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Profile Nam-Shik Shin Assistant Professor College of Veterinary Medicine, Seoul National University San56-1 Shillim9-dong, Gwanak-gu, Seoul, 151-742, Korea

ACADEMIC DEGREES: B.S. Ph.D.

1974 1998

Department of Veterinary Medicine, University of Seoul, College of Veterinary Medicine, Seoul National University

PROFESSIONAL APPOINTMENTS: 2002.09-present Assistant Professor, College of Veterinary Medicine, Seoul National University

RESEARCH INTERESTS: - Zoo and Wildlife Medicine - Zoo Management - Human-Animal Bond - Small Animal Behavior - Reproduction and Reintroduction for Endangered Wild Animals

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Clostridium Infections in Captive Wild Animals at the Seoul Grand Park Zoo in Korea Nam-Shik Shin Assistant Professor Department of Wild Animal Diseases, College of Veterinary Medicine and Zoonotic Disease Insitute (ZooDI, KRF Priority Research Institute), Seoul National University, San 56-1, Sillim-dong, Gwanak-Gu, Seoul 151-742, KOREA

Under Seoul Metropolitan Government management, Seoul Grand Park Zoo keeping over 350 species, around 3,200 animals is the largest zoo in Korea. There are 17 veterinarians working for breeding and health management of captive wild animals. This report is on the case of Clostridium infections caused death in birds and hippopotamus in Seoul Grand Park Zoo. In June of last year and May of this year, Clostridium infections occured at the large aviary exhibited about 20 species, over 200 waterfowls such as cranes, swans and wild geese. Last year, 2 mute swans and a white goose were dead showing depression, anorexia, lolling head out, and paralysis of feathers and legs. Gross findings appeared petechial lesions in myocardium, congestion and hemorrhage in small and large intestine membranes, also lots of bloody mucosal fluid in intestines. C. botulinum was detected in small intestine and the colon by microbial culture and C. botulinum type C was identified by PCR. This year, 4 hopper swans, 5 mute swans, 4 canadian geese, 7 mallards, 2 white naped cranes, 2 spoonbills and a pintail were dead with presented signs of depression, anorexia, congestion of eyeballs, lolling head out and paralysis of feathers and legs which was similar signs occurred in mute swans and a white goose last year. Gross findings showed swelling of liver and spleen, congestive hemorrhage of the duodenum and small intestine membranes. C. perfringens was detected in microbial culture of organs showed clinical signs. But, Newcastle (ND), Infectious Bronchitis (IB) and Avian Influenza (AI) were not detected. We assumed that feed for chicken and feces of about 200 General herons living outside of aviary loop contaminated water of inside aviary and produced Clostridium infection. A 2-day-old hippo that could not suckle colostrum died of C. perfringens type A infection. Clinical features of the hippopotamus have showed lethargy and anorexia before death. Gross post-mortem findings of hippopotamus were hemorrhagic enteritis of intestine. Ruminant stomach and hoof showed hemorrhagic lesions and the lumen of the small intestine was filled with mucoid and hemorrhagic fluid. Also, Intestine and stomach of hippopotamus were distended with gas and hemorrhagic fluid. C. perfringens was isolated in culture of small intestine and the presence of C. perfringens type A was confirmed by PCR. It is assumed that the hippopotaums in decreased immunity have drunken water infected with C. perfringens, which rapidly proliferated and caused death. Clostridium infections are known to be one of the life-threatening diseases in zoo animals. Most Clostridium spp. are ubiquitous bacteria present in nature. In order to control the microbial hazards, it is more important to sustain immune functions as maintaining hygienic environment and keeping healthy population size that reduce predisposing factors such as stress rather than eliminating the lethal pathogens. 11

Profile Ken Katakura Professor Laboratory of Parasitology Department of Disease Control Graduate School of Veterinary Medicine Hokkaido University, Japan

ACADEMIC DEGREES: B.S. 1977 M.V.M. 1979 Ph.D. 1986

Hokkaido University, Sapporo (Veterinary Medicine) Hokkaido University, Sapporo (Veterinary Parasitology) Jikei University School of Medicine, Tokyo (Medical Parasitology)

PROFESSIONAL APPOINTMENTS: 1979.4-1987.6 1987.7-1989.12 1990.1-1992.9 1992.10-1998.9 1998.10-2004.5 2004. 6-present

Research Fellow, Jikei University School of Medicine, Tokyo Research Associate, Chicago Medical School, North Chicago, USA Research Fellow, Jikei University School of Medicine, Tokyo Assistant Professor, Jikei University School of Medicine, Tokyo Associate Professor, Graduate School of Medicine, Gunma University, Maebashi Professor, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo

RESEARCH INTERESTS: Host-parasite interactions Parasitism and evolution of organelle Drug resistance and ABC transporters Immunity against parasitic infections and escape mechanisms by parasites Development of new diagnostic tools of parasitic diseases Molecular epidemiology of parasitic diseases

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Leishmaniasis Ken Katakura Professor Laboratory of Parasitology, Department of Disease Control Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan

Leishmaniasis is a protozoan disease caused by the genus Leishmania species. Leishmaniasis is a vector-borne disease that is transmitted by the bite of some species of phlebotomine sandflies. Leishmaniasis is a zoonosis as the parasites can infect numerous mammals, including humans, carnivores and rodents. The parasites are obligatory intracellular and multiply within the phagolysosomes of infected macrophages in the hosts. The disease commonly manifests as either a skin form (cutaneous leishmaniasis, CL) or an internal organ form (visceral leishmaniasis, VL). The dog is the main reservoir of visceral leishmaniasis. Leishmaniasis mostly affects poor and marginalized populations and is found in approximately 90 countries in the tropics and subtropics around the world. There are 1.0-1.5 million cases of CL and 500,000 cases of VL each year. In several regions, there is a clear and worrying increase in the number of cases. In Ecuador, we have conducted epidemiological studies on leishmaniasis for more than 25 years. We found that the main causative agent of CL in the lowland Pacific regions is L. (V.) panamensis, whereas L. (L.) mexicana is the main species of CL in the highland Andean regions. Sandfly species are different with these regions. In Pakistan, we found that the main causative agent of CL in the western mountainous areas is L. (L.) tropica, while recent outbreaks of CL in southern dry regions along with the Indus River are due to L. (L.) major infections. Diagnosis of CL is generally based on clinical criteria alone in rural endemic areas, where laboratory facilities are limited. Development of a simple and rapid diagnostic tool is required for differential diagnosis of CL from other skin diseases. To find out diagnostic antigens of CL, Western blot analysis was performed on sera from CL patients in Ecuador and Pakistan. Antigens of approximately 120, 95, 85 and 75 kDa were frequently recognized by sera from patients, who are living in endemic areas of L. (V.) Panamensis in Ecuador. These antigens were analysed by Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (TOF-MS). Production of these recombinant proteins using the wheat germ cell-free system in underway. Understanding of drug resistance mechanisms in Leishmania is important for more rational use of drugs and drug combinations to minimize or circumvent resistance. Studies on multidrug resistance in mammalian cancer cells lead the discovery of the ATP-binding cassette (ABC) proteins, including MDR and MRP subfamily, which are conserved among organisms from bacteria to human. In L. (L.) amazonensis, we have isolated two different ATP protein genes of MDR subfamily, encoding LaMDR1 and LaMDR2 protein, respectively. We found that LaMDR1 is a homologue of mammalian MDR1 and can confer resistance to some anticancer drugs, and that LaMDR2 can mediate 5-fluorouracil resistance. These ABC proteins appear to be localized in the multivesicular tubules of lysosome and mediate drug resistance via vesicular transportation and exocytotic pathway of the parasite. 13

Profile Hyuk-Joon Kwon Assistant Research Professor, DVM, PhD Laboratory of Avian Diseases, Zoonotic Disease Institute (ZooDI) College of Veterinary Medicine, Seoul National University San56-1 Shillim9-dong, Gwanak-gu, Seoul, Korea

ACADEMIC DEGREES: B.S. M.S. Ph.D.

1993 1995 2000

Seoul National University, College of Veterinary Medicine Seoul National University, College of Veterinary Medicine (Microbiology) Seoul National University, College of Veterinary Medicine (Microbiology)

PROFESSIONAL APPOINTMENTS: 2004.12-present

Assistant Professor for Research, ZooDI, Seoul National University

RESEARCH INTERESTS: Molecular epizootiology and reverse genetics of Avian Influenza virus and Newcastle disease virus Bacteriophage applications for prevention and therapy of avian diseases Development of anti-viral drugs from plant extracts

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Low Pathogenic Avian Influenza by H9N2 subtype virus in Korea Hyuk-Joon Kwon, Sun-Hee Cho, Sun-Joong Kim Assistant Research Professor Laboratory of Influenzavirus, College of Veterinary Medicine, Zoonotic Disease Insitute (ZooDI, KRF Priority Research Institute), and BK 21 for Veterinary Science, Seoul National University, San 56-1, Sillim-dong, Gwanak-Gu, Seoul 151-742, KOREA

The first outbreak of low pathogenic avian influenza (LPAI), H9N2 virus subtype, in 1996 prompted an eradication response, but LPAI returned to Korea in 1999. The relationship between the first and the recurrent viruses is unclear. To determine the molecular epizootiology of recurrent LPAI, we performed phylogenetic analysis with partial nucleotide sequences (HA, NA, NP and PB2) of eight chicken H9N2 viruses. The recurrent H9N2 viruses showed higher nucleotide similarity in HA and NA genes to the 1996 Korean isolates than other Eurasian viruses, and formed a distinct cluster with the early Korean isolates and some isolates from migratory and domestic ducks in Hokkaido, Japan, and China. Phylogenetic analysis with internal genes showed that some Korean isolates formed a cluster with other subtypes, such as H5N1, H6N1, and H6N2 in China and Taiwan. Relatively high nucleotide variation between recurrent isolates indicated frequent viral infection and replication. Therefore, appropriate vaccine programs should be considered to minimize economic losses in poultry industry and to slow down the uncontrolled rapid evolution of H9N2 viruses in Korea, urgently. In development of effective vaccine for LPAI the major obstacle was poor productivity of vaccine virus in embryonated chicken eggs (ECE). Recently we isolated highly productive virus strain, KBNP-0028 by a specialized ECE passage protocol and characterized antigenicity and molecular properties. KBNP-0028 was similar to recent LPAI viruses in antigenicity and thought to be safe based on the amino acid (aa) sequences related to virulence such as cleavage site amino acid sequence of HA, aspartic acid at 92nd aa of NS1, and glutamic acid at 627th aa of PB2. In conclusion the epizootiological data and the highly productive vaccine strain (KBNP-0028) in the present study may be useful in improving productivity of poultry industry.

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Profile Jae-Ho Shin, DVM, PhD COE researcher Laboratory of Comparative Pathology Graduate School of Veterinary Medicine Hokkaido University Kita 18 Nishi 9, Kitaku, Sapporo, 060-0818, Japan

ACADEMIC DEGREES: B.S. M.S.

1987 1989

Ph.D.

1995

Seoul National University, College of Veterinary Medicine (Veterinary Medicine) Seoul National University, Graduate School of Veterinary Medicine (Veterinary Pathology) Seoul National University, Graduate School of Veterinary Medicine (Veterinary Pathology)

PROFESSIONAL APPOINTMENTS: 2004.3-present 1990.9-present. 1996.8-1999.10

Laboratory of Comparative Pathology, Graduate School of Veterinary Medicine, Hokkaido University National Institute of Toxicological Research, Korea Food and Drug Administration Department of Anatomy and Embryology, Graduate School of Medicine, Kyoto University

RESEARCH INTERESTS: 1. Mechanisms of transneural spread of neurotropic viruses 2. Application of intracerebral immunization in animal models for protection or treatment of the infected mice

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Efficacy of intracerebral immunization against neurotropic viruses in mice Jae-Ho Shin COE researcher Laboratory of Comparative Pathology, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18 Nishi 9, Kitaku, Sapporo, 060-0818, Japan

To evaluate the efficacy of intracerebral (IC) immunization, mice were immunized with by subcutaneous (SC), intramuscular (IM) or IC route, and then infected with two neurotropic viruses, pseudorabies virus (PRV) and rabies virus (RBV) into the hindleg of the immunized or non-immunized mice. At first, ICR mice were immunized with formalininactivated PRV by either SC or IC route, and then infected with 106 plaque-forming units of PRV (Yamagata S-81 strain). The antibody titers in serum were elevated and boosted by additional immunization via both the SC and IC routes, but were higher after IC immunization. Intracerebrally immunized mice were completely protected from mortality and neurological signs, whereas all the non-immunized and 80 % of the subcutaneously immunized mice died after developing neurological signs. Second, mice were immunized with inactivated RBV vaccine (TC origin) by SC, IM or IC route, and then infected with 109 fluorescent-focus units of RBV (CVS strain). Intracerebrally immunized mice were completely protected from mortality and neurological signs as similar as the PRV challenge study, whereas all the nonimmunized, 70 % or 30 % of the subcutaneously or intramuscularly immunized mice, respectively, died after developing neurological signs. In mouse models, IC immunization is more effective at inducing a protective immune response against the transneural spread like PRV and RBV than immunization by SC or IM route.

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