Control of avian influenza and preparedness for pandemic influenza

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Kida, DVM, PhD, MJA University Professor, Hokkaido University Head, Research Center for Zoonosis

1 Asia-Pacific WHO/OIE/FAO Joint Workshop on Surveillance, Prevention and Control of Zoonotic influenza Paro, Bhutan, 30 August 2016 Control of avian influenza and preparedness for pandemic influenza Hiroshi Kida, DVM, PhD, MJA University Professor, Hokkaido University Head, Research Center for Zoonosis Control Head, OIE Reference Laboratory for Animal Influenza Head, WHO Collaborating Centre for Zoonoses Control

2 Control of avian influenza and preparedness for pandemic influenza 1. How do highly pathogenic avian influenza viruses (HPAIVs) and human pandemic influenza virus strains emerge? 2. Why have the H5N1 HPAIV viruses persisted in poultry for 20 years? 3. Why are antigenic variants of different clades circulating in poultry birds? 4. Will the HPAIVs that have returned to migratory birds persist in nature? 5. Does influenza vaccine confer complete protective immunity? 6. How should HPAI be controlled? 7. Will H5N1 HPAIV cause pandemic influenza? 8. Then will H7N9 low pathogenic AIV that appeared in China cause pandemic? 9. Are the measures for the control of seasonal flu satisfactory? 10.How should be prepared for pandemic influenza? To answer to these questions, it is prerequisite to understand Ecology of influenza viruses in nature, birds and mammals; Origin of, perpetuation in nature and evolution of influenza viruses, and mechanisms of the emergence of HPAIVs and pandemic strains in humans.

3 How do highly pathogenic avian influenza viruses (HPAIVs) emerge? Human pandemic influenza Hong Kong flu Spanish flu Asian flu 2009 (H1N1) pdm flu H2N8 H3N8 H1N1 H2N2 H1N1(Russian type) H3N First Isolation of influenza virus from human 1925 In 16 countries Human cases of H5N1 HPAIV infection in Hong Kong in 1997 and 2003~present Highly pathogenic avian influenza viruses (H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N3, H7N4, H7N7) 2020 (year) 2014 (H5N8) (H5N1) 2007(H5N1) 2004 (H5N1) Outbreak of HPAI caused by H7N7 virus in Japan Highly pathogenic avian influenza Outbreaks of HPAI caused by H5 AI viruses in Japan

4 Host range, and HA and NA subtypes of influenza A virus ー Influenza is a typical zoonosis ー H1N1 H1N2 H3N2 H2N3, H3N1, H3N3, H3N8, H4N6, H5N1, H5N2, H9N2 H1-H12 N1-9 H1N1 H2N2 H3N2 (H2N8,H3N8) H5N1,H7N7, H9N2 H3N8,H5N1 H3N8 H7N7 H1-10 N1-9 H1-16 N1-9 H1-7, H9-16 N1-9 H3N3 H4N5 H7N7 H5N1 H1-7, H9-11 N1-4, N6-8 H1N3, H13N2, H13N9 H3N2 H5N1 H10N4 4

5 Intestinal replication of influenza virus in ducks Tissue Virus recovery F A (log EID 50 ) Brain - - Pharynx Larynx Trachea, Lungs - - Esophagus, Proventriculus - - Duodenum, Jejunum - - Ileum Cecum Colon Rectum Bursa Pancreas, Liver, Kidney, - - Genitals, Heart, Blood, - - Spleen, Thymus - - Feces 7.7 Ducks were infected with A/duck/Hokkaido/5/77 (H3N2) Kida et al (1980) Infect Immun

6 Ito T et al (1995) Arch Virol

Irkutsk(290) Kenkeme(32) Kharyyalah(146) 40 Ilands(1321) Yakutsk(232) H3N8 H13N6 Khabarovsk(23) Buotama(51) Elavga(66) Kobyaysky(820) H4N6 H4N9 H11N1 H11N6 H11N9 White Lake(1136) H3N8 Magadan(295)

7 Irkutsk(290) Kenkeme(32) Kharyyalah(146) 40 Ilands(1321) Yakutsk(232) H3N8 H13N6 Khabarovsk(23) Buotama(51) Elavga(66) Kobyaysky(820) H4N6 H4N9 H11N1 H11N6 H11N9 White Lake(1136) H3N8 Magadan(295) Ptropavlovsk- Kamchatsky(58) Lake Kanicheva(95) Malyshevo(90) Taiwan H1N1 H4N5 H4N6 H7N7 Wakkanai (958) H1N1 H3N8 H5N3 H5N4 H6N2 H6N7 H8N1 H8N3 H9N2 H11N Okazaki et al (2000) Arch Virol

8 Duck influenza Each of the known subtypes (H1-16, N1-9) of influenza A virus has been isolated from ducks. In ducks, viruses replicate in the colon, being shed with feces in a week, and non-pathogenic. Water-borne fecal-oral transmission Viruses are preserved in frozen water of the lakes, where ducks nest in summer, in winter such as in Siberia, Alaska and Canada. Ducks carry and provide viruses during migration and over-wintering. Influenza viruses circulating in ducks are highly stable antigenically and genetically. Migratory duck is the natural host of influenza A viruses. Kida et al (1980) Infect Immun; (1987) Virology; Ito et al (1995) Arch Virol

9 Acquisition of pathogenicity of avian influenza viruses in chickens APAIV > 6 Months LPAIV Live bird markets LBM & Farms HPAIV (H5 or H7)

10 Amino acid sequences at the cleavage site of influenza A virus HAs Subtype Strain A A sequence H1 H2 H3 H4 H5 H5 H6 H7 H7 H8 H9 H10 H11 H12 H13 H14 H15 Dk/Alberta/35/76(H1N1) b Mal/MT/Y61(H2N2) b Dk/Menphis/928/74(H3N8) b Dk/Czechoslovakia/56(H4N6) b Ck/Scotland/59(H5N1) b Ty/MN/3/92(H5N2) a Shw/Australia/1/72(H6N5) b FPV/Rostock/34(H7N1) b Mal/Alberta/195/89(H7N3) a Ty/Ontario/6118/68(H8N4) b Ty/Wisconsin/66(H9N2) b Ck/Germany/N/49(H10N7) b Dk/England/56(H11N6) b Dk/Alberta/60/76(H12N5) b Gl/Maryland/704/77(H13N6) b Mal/Gurjev/263/82(H14N5) b Shw/Australia/2576/79(H15N9) b IQSR GLF IESR GLF KQTR GLF KASR GLF RKKR GLF RETR GLF IETR GLF KKRKKR GLF KKTR GLF VEPR GLF RSSR GLF VQGR GLF IASR GLF VQDR GLF ISNR GLF KQAK GLF IRTR GLF a Senne et al (1996), b Kovacova et al (2002)

Extensive virus multiplication induces cytokine storm causing break down and increase of

11 Pathological findings of abnormal coagulation in the brain of the chickens infected with HPAIV Ty/Italy/99 Vessel Vessel line Red blood cells thrombus 50um hemorrhage 100um Extensive virus multiplication induces cytokine storm causing break down and increase of permeability of capillary, leading multiple organ failure. Kuribayashi et al (2013) PLoS ONE

12 Return of the HPAIV from domestic poultry to migratory water birds APAIV > 6 months LPAIV Live bird markets LBM & Farms HPAIV (H5 or H7)

swan/mongolia/2/09 (H5N1) A/whooper swan/mongolia/9/09 (H5N1) A/bar-headed goose/mongolia/x53/09 (H5N1) A/rubby

13 HPAI viruses isolated from wild birds in Mongolia A/whooper swan/mongolia/3/05 (H5N1) A/bar-headed goose/mongolia/1/05 (H5N1) A/common goldeneye/mongolia/12/06 (H5N1) Ugii Lake A/whooper swan/mongolia/2/06 (H5N1) Qinghai Lake A/whooper swan/mongolia/2/09 (H5N1) A/whooper swan/mongolia/9/09 (H5N1) A/bar-headed goose/mongolia/x53/09 (H5N1) A/rubby sholduck/mongolia/x42/2009 (H5N1) A/common goldeneye/mongolia/x60/09 (H5N1) A/whooper swan/mongolia/1/10 (H5N1) A/whooper swan/mongolia/7/10 (H5N1)

62 Countries where H5N1 HPAIV infections were reported in wild birds, poultry, and both Japan, Republic of Korea, China, Mongolia, Myanmar, Lao PDR, Thailand, Cambodia, Viet Nam, Malaysia, Indonesia,

14 62 Countries where H5N1 HPAIV infections were reported in wild birds, poultry, and both Japan, Republic of Korea, China, Mongolia, Myanmar, Lao PDR, Thailand, Cambodia, Viet Nam, Malaysia, Indonesia, Bangladesh, India, Pakistan; Afghanistan, Iran, Azerbaijan, Georgia, Iraq, Kuwait, Saudi Arabia, Turkey, Israel; Russian Federation, Kazakhstan, Ukraine, Romania, Bulgaria, Albania, Serbia, Hungary, Slovakia, Czech Republic, Croatia, Poland, Slovenia, Bosnia & Herzegovina; Greece, Switzerland, Austria, France, Italy, Germany, Netherlands, Denmark, Sweden, Spain, England, Ireland; Djibouti, Gaza Strip, Egypt, Sudan, Nigeria, Niger, Cameroon, Burkina Faso, Cote d Ivoire

03 04 05 06 07 08 09 10 11 12 13 14 15 03 04 05 06 07 08 09 10 11 12 13 14 15 Confirmed human cases of H5N1 HPAIV

Turkey Djibouti Nigeria Myanmar Pakistan Bangladesh Canada 31 64 167 116 37 2 17 2 5 4 0 1 0 1 1 1 / / / / / / / / / / /

15 Confirmed human cases of H5N1 HPAIV infection Country Deaths/Cases China Egypt China Vietnam Indonesia Egypt Cambodia Lao PDR Thailand Iraq Azerbaijan Turkey Djibouti Nigeria Myanmar Pakistan Bangladesh Canada / / / / / / / / / / / / / / / / Viet Nam Indonesia Thailand Cambodia Total 450 / 854 As of 19 July 2016 WHO (2016)

16 Avian influenza vaccines Vietnam: H5N2 and H5N1 (Adjuvant inactivated vaccines) China: H5N1 and recombinant NDV ( Reverse genetics inactivated vaccines) Indonesia: H5N1, H5N2, H5N9 and recombinant H5N1 (inactivated vaccines) Egypt: since 2006 Thailand: Officially prohibited vaccination in 2006 As a stockpile, Singapore: H5N2 ( Inactivated, adjuvant vaccine) Japan: H5N1 and H7N7 (Inactivated oil-adjuvant vaccines) Pakistan: H5N1, H5N2, H5N9, and H5N3 (Inactivated alum hydroxide- and oiladjuvant vaccines)

17 Vaccine for avian Influenza may prevent manifestation of disease signs and decrease the amount of virus shed, but does not confer protective immunity from infection. Stamping-out policy is recommended for the control of avian influenza. Vaccination was not primarily recommended but later approved as one of the options applied only under DIVA based strategy. Country where vaccine is used is not designated as HPAI-free. leads silent spread of virus.

18 26TH CONFERENCE OF THE OIE REGIONAL COMMISSION FOR ASIA, THE FAR EAST AND OCEANIA Shanghai, People s Republic of China, November 2009 RECOMMENDATION FOR THE CONTROL OF AVIAN INFLUENZA It is considered that; Highly pathogenic avian influenza H5N1 virus strains have persisted in domestic poultry for 14 years and antigenic variants have been selected mainly due to the misuse of vaccine. HPAI has been put under control in several countries. Stamping out policy has been the most effective measures for the control HPAI. Vaccine is used in 4 countries where HPAI has not been controlled. Vaccine is used instead of stamping out in 2 countries and in the other 2 countries, basically in addition to stamping out. Sentinel bids are put in the vaccinated poultry population in Viet Nam and not in the other 3 countries where vaccine is used. Compensation for livestock owners is done in most countries in case of stamping out. It is recommended that; Since stamping out is the best and ultimate measure for the control of HPAI, vaccine should be used in addition to, not instead of stamping out when it is used. The OIE should continue and develop standards on animal influenza surveillance, prevention and control. Surveillance of swine flu is crucial in the countries where avian flu has not been controlled.

19 Cases of infection with H5 and H7 viruses in Japan 1925 Chicken H7N7 HPAIV 2004 Chicken H5N1 (clade 2.5) HPAIV 2005 Chicken H5N2 LPAIV 2007 Chicken H5N1 (clade 2.2) HPAIV 2008 Swan H5N1 (clade 2.3.2) HPAIV 2009 Quail H7N6 LPAIV Chicken H5N1 (clade ) HPAIV Duck/swan H5N1 (clade ) HPAIV 2014 April Chicken H5N8 (clade ) HPAIV 2014 Nov present Duck/ Crane Chicken H5N8 (clade ) HPAIV

20 Surveillance of avian influenza in autumn (1991~2009) Mongolia (324) H1N1 (5) H2N2 (1) H2N3 (1) H3N2 (2) H3N6 (34) H3N8 (149) H4N2 (2) H4N3 (1) H4N6 (72) H4N7 (1) H4N8 (4) H5N2 (1) H5N3 (4) H7N1 (2) H7N6 (1) H7N7 (13) H7N9 (3) H8N4 (6) H9N2 (1) H10N3 (13) H10N5 (2) H10N7 (5) H12N5 (1) Australia (6) H2N5 (6) China (2) H3N8 (1) H4N6 (1) Russia (56) H3N8 (17) H4N6 (25) H4N9 (2) H10N7 (1) H11N1 (1) H11N6 (1) H11N9 (8) H13N6 (1) USA (111) H2N3 (1) H3N8 (39) H4N6 (57) H7N3 (1) H7N7 (1) H8N2 (1) H10N7 (11) Hokkaido (296) H1N1 (12) H2N2 (2) H2N3 (4) H2N5 (1) H3N2 (2) H3N6 (6) H3N8 (37) H4N2 (8) H4N5 (1) H4N6 (33) H4N9 (3) H5N2 (1) H5N3 (11) H6N1 (17) H6N2 (32) H6N5 (2) H6N8 (7) H6N9 (1) H7N1 (18) H7N7 (14) H8N4 (8) H9N2 (7) H9N4 (1) H9N5 (1) H9N9 (1) H10N2 (1) H10N4 (12) H10N5 (7) H10N6 (1) H10N7 (12) H10N8 (1) H10N9 (2) H11N6 (1) H11N9 (21) H12N2 (2) H12N5 (4) H13N6 (2) H5N1(1) 795 isolates from 22,744 samples

21 Surveillance of avian influenza in autumn 2010 Mongolia (36 isolates) H1N1 (1) H3N3 (1) H3N6 (7) H3N8 (14) H4N6 (8) H7N9 (1) H10N8 (4) Hokkaido (15 isolates) H3N8 (3) H5N2 (1) H6N2 (2) H7N7 (9) H5N1 (2) Hong Kong (3 isolates) H3N2 (1) H5N1 (2) Vietnam (1 isolate) H9N6 (1) Laos (none) Number of samples : 4,515 Influenza virus isolates : 55 As of 25 October 2010

22 Outbreaks of HPAI caused by H5N1 viruses in Japan in winter C 青森北海道 A A 福島 B 富山栃木島根鳥取京都兵庫愛知千葉 C 山口大分長崎鹿児島宮崎高知徳島三重奈良和歌山 Wild birds (63) Chicken farms (24) Okamatsu (2011)

23 Outbreak of HPAI caused by H5 influenza viruses of clade in Japan ( ) Wild bird (14 cases) Domestic poultry (5 cases) Yamaguchi Shimane Tottori Gifu Chiba Saga Okayama Miyazaki Kagoshima Domestic Oct Nov Dec Jan Feb Mar Apr Wild bird

24 Antigenic characterization of H5 influenza viruses HI titer of the antisera to Clade Viruses Mal/Hok/ 24/09 Tn/SA/ 61 Pf/HK/ 810/09 Ck/Kum/ 1-7/14 - Duck/Hokkaido/101/10 (H5N2) 1,280 2, ,280 - Duck/Mongolia/194/11 (H5N3) 1,280 2, ,280 - Mallard/Hokkaido/24/09 (H5N1) 1,280 2, ,560 - Tern/South Africa/61 (H5N3) 2,560 10, Chicken/Hong Kong/156/97 (H5N1) 640 1, Peregrine falcon/hong Kong/810/09 (H5N1) <20 <20 2, Chicken/Kumamoto/1-7/14 (H5N8) Duck/Vietnam/1151/14 (H5N6)

26 Human cases of avian influenza A(H7N9) virus infection as of 23 February 2015, WHO

27 How do human pandemic influenza virus strains emerge? Human pandemic influenza Hong Kong flu Spanish flu Asian flu 2009 (H1N1) pdm flu H2N8 H3N8 H1N1 H2N2 H1N1(Russian type) H3N First Isolation of influenza virus from human 1925 Human cases of H5N1 HPAIV infection in Hong Kong in 1997 and 2003~present Highly pathogenic avian influenza viruses (H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N3, H7N4, H7N7) In 15 countries 2020 (year) 2014 (H5N8) (H5N1) 2007(H5N1) 2004 (H5N1) Outbreak of HPAI caused by H7N7 virus in Japan Outbreaks of HPAI caused by H5 AI viruses in Japan Highly pathogenic avian influenza

28 Type A influenza virus particle Hemagglutinin (HA, H1-16 subtypes) A/duck/Hokkaido/5/77(H3N2) Neuraminidase (NA, N1-9 subtypes) Envelope

29 Swine Epithelia l Cell Genetic reassortment A/duck/S China/x/67 (H3NX) A/S China/x/67 (H2N2) A/Hong Kong/1/68 (H3N2) Human Kida (1987, 1988, 1994 and 2008) Virology, J Gen Virol and Global Environmental Research; Yasuda (1991) J Gen Virol

1918 Spanish virus (H1N1) 1957 Asian virus (H2N2) and even 1918 Spanish flu virus (H1N1) must have

30 Rout of transmission of the genes of pandemic strains Migratory duck A/duck/ xx (H3N?) Domestic duck Pond Virus shedding Genetic reassortment A/Asian/67 (H2N2) 1968 HK A/HongKong/68 (H3N2) 1918 Spanish virus (H1N1) 1957 Asian virus (H2N2) and even 1918 Spanish flu virus (H1N1) must have appeared similarly. Kida (1987, 1988, 1994, and 2008) Virology; J Gen Virol and Global Environmental Research; Yasuda (1991) J Gen Virol

Library of pandemic flu vaccine strain candidates Influenza viruses of 75 combinations of the HA and NA subtypes have been isolated from fecal samples of ducks (black).

. Test vaccines prepared from H1N1, H5N1, H6N2, H7N7, H7N9 and H9N2 viruses in the library conferred sufficient immune response to protect chickens, mice, and macaques from the challenge with

31 Library of pandemic flu vaccine strain candidates Influenza viruses of 75 combinations of the HA and NA subtypes have been isolated from fecal samples of ducks (black). 69 other combinations have been generated by genetic reassertment procedure in the lab (red).. Test vaccines prepared from H1N1, H5N1, H6N2, H7N7, H7N9 and H9N2 viruses in the library conferred sufficient immune response to protect chickens, mice, and macaques from the challenge with isolates from poultry birds and humans. Thus we have vaccine strains for pandemic influenza. Thus, 2,900 avian influenza viruses of 144 combinations of HA and NA subtypes have been stocked as vaccine strain candidates. Their pathogenicity, antigenicity, genetic information and yield in chicken embryo have been analyzed, databased, and opened for Web site (

32 Prediction of amino acid substitutions on the hemagglutinin molecule of influenza A viruses H3N2 The prediction of antigenic change of influenza A virus has been one of the major public health goals. HA amino acid sequences of 6,806 H3N2 human viruses were analyzed by multidimensional scaling. A mathematical formula on the relative distances were inferred. Retrospective prediction tests were conducted and showed that 70% of actual amino acid substitutions are correctly predicted. Result of prediction for the amino acid substitution on season. QKLPGNDNSTATLCLGHHAVPNGTIVKTITNDRIEVTNATE LVQNSSIGEICDSPHQILDGENCTLIDALLGDPQCDGFQNK KWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNNE SFNWTGVTQNGTSSACIRRSNSSFFSRLNWLTHLNFKYPAL NVTMPNNEQFDKLYIWGVHHPGTDKDQIFLYAQSSGRITVS TKRSQQAVIPNIGSRPRIRNIPSRISIYWTIVKPGDILLIN STGNLIAPRGYFKIRSGKSSIMRSDAPIGKCKSECITPNGS IPNDKPFQNVNRITYGACPRYVKQSTLKLATGMRNVPEKQT A three-dimensional map of 6,806 H3N2 human influenza viruses Amino acids substitutions occurred in the are shown in bold face. Positions with blue amino acids represent correctly predicted amino acid substitutions. Ito, K., Igarashi, M., Miyazaki, Y., Murakami, T., Iida, S., Kida, H., & Takada, A. (2011). Gnarled-Trunk Evolutionary Model of Influenza A Virus Hemagglutinin. PLoS ONE, 6(10), e25953

33 How to prepare for pandemic influenza should be based upon how to control seasonal influenza Drastic improvement of seasonal flu vaccine and biological standard Short term goal: 1. Improvement of the immunological potency of seasonal flu vaccines; Clinical evaluation of inactivated vaccine of purified whole virus particles Drastic revision of biological standard 2. Preemptive preparation of vaccine by forestall of antigenic variants on the basis of global surveillance and bioinformatics. Good bye backward selection of vaccine strains. Long term goal: 1. Development of effective and safe mucosal vaccines 2. Development of effective and safe adjuvants Points: 1. Influenza is a typical acute infection. 2. Influenza is not eradicable, every year seasonal influenza must come.

Current vaccines for influenza most widely used 1. Whole virus vaccines: purified inactivated viruses 2. Split virus vaccines: purified inactivated virus particles disrupted by detergent 3.

34 Current vaccines for influenza most widely used 1. Whole virus vaccines: purified inactivated viruses 2. Split virus vaccines: purified inactivated virus particles disrupted by detergent 3. Subunit or surface antigen vaccines: the hemagglutinin (HA) and neuraminidase (NA) further enriched from the other components in core 4. Live attenuated virus vaccines: live virus attenuated by reassortment of HA and NA onto cold-adapted master strain Most produced in embryonated eggs, some now in mammalian cell culture

35 Antigen presentation of optional peptides ー Cassette theory ー Hemagglutinin HA (conserved epitope) I-Ab binding component Hybrid peptide 46F/HA /54A P43-58 analog peptide 46F50E54A Ogasawara, K et al. PNAS 89, 8995 (1992) Naruse, H et al. PNAS 91, 9588, (1994)

Recent focus of attention has been conserved regions in the HA stem for the development of an universal vaccine Impagriazzo, Goudsmit, Wilson et al (2015) A stable trimeric influenza hemagglutinin

36 Recent focus of attention has been conserved regions in the HA stem for the development of an universal vaccine Impagriazzo, Goudsmit, Wilson et al (2015) A stable trimeric influenza hemagglutinin stem as a broadly protective immunogen. Science 349, Yassine, Okuno, Nabel, Graham et al (2015) Hemagglutinin-stem nanoparticles generate heterosubtypic influenza protection. Nat Med 21, On the basis of the concept of the mechanism of the antigen presentation, molecular structure of vaccine components is not important. Whole inactivated influenza virus particles are the best promising vaccine.

37 All Japan Program of Excellence for the Development and Practical use of Seasonal Influenza Vaccines of Global Standard in Collaboration together with Industries, Universities, Academic Societies and Government Preclinical and clinical studies on seasonal influenza vaccines prepared from the inactivated whole virus particles 1) Each of 5 influenza vaccine producers in Japan participates in the program. 2) Comparison of immunogenicity and safety between whole virus particle vaccine and split vaccines. Viruses, cultured in the cells or embryonated chicken eggs and purified are subjected to prepare whole particle vaccines and ether-disrupted split vaccines by each of the vaccine producers and Hokkaido University. Vaccines prepared are subjected to immunological potency test and safety tests using in mice, ferrets, and monkeys for the preclinical tests, and humans for clinical tests. 3) Evaluation of the results of the tests. 4) Drastic revision of biological standard for vaccines for human use. 5) Each member independently studies on the route of inoculation of the vaccines and adjuvants. 6) Participants to this program: Industries; Kitasato-DaiichiSankyo, Takeda Chemical Ind, Biken, Kaketsuken, Denka Seiken Universities; Hokkaido Univ, Univ Melbourne Government; NIID, Ministry of Health and Labor, Ministry of Education, Sports, Science and Technology 7) Budget: MHLW, MEXT, JSPS, JST AMED etc

38 Test vaccines Producer A B C D E Strains A/California/07/09 (H1N1) A/California/07/09 (H1N1) A/California/07/09 (H1N1) A/California/07/09 (H1N1) A/California/07/09 (H1N1) A/Switzerland/ (H3N2) A/Switzerland/ (H3N2) A/Switzerland/ (H3N2) B/Texas/2/13 B/Texas/2/2013 B/Phuket/3073/13 B/Phuket/3073/13 Whole virus particle vaccine Inactivated with formalin or β-propiolactone Pyrogenic & Low-pyrogenic WPV Ether-split vaccine Monovalent Quadrivalent Monovalent Monovalent Monovalent Immune response in mice and monkeys Cytokine Innate immune cell response CD4 and CD8 T cell response Antibody Immunoglobulin Challenge test Potency and safety Potency in mice pyrogenic response in rabbits leukocyte number decrease other safety tests

39 Cells in draining LN (x10 6 ) Cytokines level in serum (pg/ml) SV 1000 WPV PBS CXCL10 MCP-1 IL-6 TNF-α Hours after vaccination (Vaccine C) 1.2 pdc 1.2 Macrophage Body weight loss (%) (PFU) E+06 Virus titer in the lungs Hours after vaccination (Vaccine C) Days after challenge 1.0E E E E E E+00 5 days after challenge (Vaccine A)

40 How to control avian influenza and how to prepare for pandemic influenza 1. How do HPAIVs and human pandemic strains emerge? Live bird markets, quails and chickens; genetic reassortants generated in pigs between human seasonal flu and avian viruses 2. Why have the H5N1 HPAIVs persisted in poultry for 20 years? 3. Why are antigenic variants of different clades circulating in poultry birds? Because of Misuse of Vaccine 4. Will the HPAIVs returned to migratory birds persist in nature? Contamination of HPAIVs in the nesting lakes of migratory ducks may have occurred. Prompt eradication of the H5N1 HPAIVs from poultry in Asia is urgently needed. 5. How should HPAI be controlled? For the containment and eradication of avian influenza viruses in the poultry flocks infected, enhanced surveillance, early detection, culling the flock, movement restriction, and control measures without misuse of vaccine should be done. Vaccine should be carefully used in addition to, not instead of stamping out. 6. Does AI vaccine confer complete protective immunity? No, it inhibit only manifestation of disease signs. 7. Will H5N1 HPAIV cause pandemic influenza? Unlikely 8. Then will H7N9 LPAIV that appeared in China cause pandemic? Unlikely 9. Are the measures for the control of seasonal flu satisfactory? No How to control pandemic flu should be based on the measures for the control of seasonal flu. Especially seasonal flu vaccines should be drastically improved since current split vaccines prepared by ether- or detergent-disruption are not immunogenic enough.

Title. Author(s)Kida, Hiroshi. Issue Date Doc URL. Type. File Information. example. Prevention of Infectious Diseases. 2 November 2009.

Title. Author(s)Kida, Hiroshi. Issue Date Doc URL. Type. File Information. example. Prevention of Infectious Diseases. 2 November 2009. Title Global Surveillance is the Key for the Control of Zo example Author(s)Kida, Hiroshi Sustainability Weeks 2009 Opening Symposium "Interna CitationRecommendations for Tackling the 5 Challenges of Glo