Patricia Fitzgerald-Bocarsly

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1 FLU Patricia Fitzgerald-Bocarsly October 23, 2008

2 Orthomyxoviruses Orthomyxo virus (ortho = true or correct ) Negative-sense RNA virus (complementary to mrna) Five different genera Influenza A, B, C Thogotovirus - Tick-borne I i (i f ti l i i ) Isavirus (infectious salmon anemia virus) Segmented RNA allowing for reassortment, but only within genera Enveloped

3 Influenza Virus Influenza A, B and C are human pathogens but humans are not the natural host Named according to their genus (type), species isolated from (except human), location of isolate, number of isolate, year, and (for influenza A, the hemagglutinin(h1-16) and neuraminidase (N1-9) type)

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8 Influenza Viruses Usual host(s) Transmission Disease Distribution Influenza A Humans, Airborne Respiratory Worldwide birds, disease swine Influenza B Humans Airborne Respiratory disease Worldwide Influenza C Humans Airborne Respiratory disease Worldwide

9 Molecular Properties Influenza A, B: 8 gene segments, 14 kb Encodes 10 proteins Influenza C: 7 gene segments Encodes 9 proteins Receptor (all): sialic acid (but C uses a different form)

10 Entry Fuses in endocytic compartments Dependent on low ph Uncoating in endosomes Fusion requires structural change in the HA following cleavage of HA0 to HA1 and HA2; HA2 then allows fusion

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13 Replication Nucleus - unusual for RNA viruses 8 viral RNA fragments exist as complex with four proteins that all have NLS: viral ribonucleoprotein complexes (vrnp) RNA NP - nucleocapsid protein coats RNS PB1, PB2, PA: involved in cap recognition, RNA synthesis

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16 Assembly RNA exported Formation of virions Controversy over how RNA segments segregate: Random packaging of 10 or more segments Specific packaging g of 8 segments Bud from cytoplasmic membrane

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18 Heterogeneity of Flu Virion Forms

19 Neuraminadase Cleaves sialic acid residues Highly variable Function Prevents virus sticking back onto cells Prevents cell clumping

20 Major Immune Responses Innate immunity: NK, IFN-alpha, etc. Neutralizing ing antibodies against HA Great deal of variability in HA (also NA) CTL

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25 Cytotoxic T Cells in Viral Infection

26 Influenza NS1 Inhibits IFN induction Downregulates IRF-3 3, IRF-7 7, NF-kB Inhibits activation of PKR Flu with NS1 deleted d very sensitive to IFN

27 Epidemiology Influenza A is the most frequent infection of humans 10-20% world s population infected/year 250, ,000 deaths 20,000-30,000 deaths in the US Major reservoir is birds In birds, largely asymptomatic Not much pressure to mutate Human farming practices (pigs and fowl) lead to coinfection and reassortment of RNA

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30 Flu Vaccines Whole, inactivated - eggs or tissue culture Live, cold-adapted - FluMist intranasal Passaged to be heat sensitive Grows in upper airway Mimics natural infection - better CTL and antibody? 3 subtypes chosen in Spring: 2 A, 1B Last year missed This year, three different types in vaccine Future vaccine target conserved CTL epitope?

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32 Why do I need a flu vaccine every year? Antigenic shift and antigenic drift: virus escapes immune response Short incubation time (2 days) No time to activate memory cells No time to boost antibody levels Existing antibody might not be protective anyway

33 1918 Flu Pandemic Pandemic flu arises 3-4 times/century with influenza A (not B) Unusually high infection (30%) and high deaths ( million) Killed young people at high levels

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35 Reconstructed 1918 Virus Tissue samples from Armed Forces Institute of Pathology, London, and one frozen individual buried in permafrost in Alaska H1N1, virtually identical in all of the samples Most H1N1 non-pathogenic in mice, but 1918 more pathogenic In BSL-4 conditions in monkeys, see high replication rates and extensive spread in the lungs Altered innate imunity Inflammatory cytokines increased - cytokine storm Explains high mortality in young adults?

36 Bird Flu 18 people in Hong Kong infected and 6 died in 1997 Avian influenza H5N1 Destruction of 1.6 million domestic birds Reappeared in 2006 and has spread throughout h t Asia into Africa and Europe 50% mortality

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38 ? How is bird flu transmitted to people? At the molecular level, what would need to change to allow the virus to pass directly from human-to-human??

39 A smart virus does not wipe out its host species. What molecular properties of the H5N1 bird flu make it particularly pathogenic in birds? (Or, what is known about determinants for pathogenesis in flu?)

40 Is it possible to develop a protective vaccine against bird flu? What has already been done and what design would you propose for a future vaccine?

41 What are the antiviral drugs that are used in the US for flu infection? Describe their mechanisms of action and report whether they will be active against bird flu,and why. Do not discuss vaccines in your presentation.

42 Does Bird Flu constitute a viable terroristic threat? Why? How does the US classify potential threats?

43 The range of bird flu has been expanding; with the migration of birds over thousands of miles, what can/should the US be doing to protect Americans and American agriculture from bird flu? [Include the basis of molecular and immunological monitoring in your presentation.]

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