What are ADCC antibodies? Work on influenza ADCC antibodies Greenberg et al, Hashimoto et al. First describes Fluspecific

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1 14/7/214 Acknowledgement antibodies against diverse influenza strains: Implications for universal vaccination Sinth Jegaskanda PhD Prof Stephen Kent University of Melbourne What are antibodies? Work on influenza antibodies Greenberg et al, Hashimoto et al. First describes Fluspecific nothing! 24: Jegerlehner et al. to extracellular portion of M2 protein may help control influenza infection in mice. 211: Corti et al. Reduced protection of mice when neutralizing antibody mutated to remove Fc receptor binding. Note: process can both kill virus infected cells AND result in liberation of cytokines/chemokines that induce an antiviral state (non-cytolytic mechanism) 214: DeLillio et al: Anti-stem broadly neutralizing antibodies effective in mice in part related to Fc receptor presence/ activity 212-4: Jegaskanda/Kent: induced by infection recognizes broad range of influenza strains in humans and macaques MicobelLibrary, Gary Kaiser, Biology Department, Community college of Baltimore County 1

2 % NK cells expressing CD7a % NK cells expressing CD7a 14/7/214 Outline Ability of influenza infection to induce cross reactive Macaques Humans Individuals pre and post 29 of different ages Pooled IVIG made pre and post 29 Ability of vaccines to induce cross reactive in macaques TIV Whole inactivated virus with adjuvant MVA vectored vaccine Do antibodies induced by influenza infection cross react with other strains? Seasonal 21 H1N1 (A/Kawasaki/173/21) sh1n1 Naive group 4 months later 29 H1N1pdm virus (A/California/4/29) ph1n1 ph1n1 Primed group Jason Weinfurter, Tom Friedrich U. Wisconsin. (212) PloS Path Seasonal H1N1 infection induced cross-reactive antibodies to 29-pandemic HA protein. Day pre infection. Day 14 post seasonal H1N1 Seasonal Pandemic A/New Caledonia/2/99 A/California/4/29 Why did older subjects have less mortality during the H1N1pdm pandemic?. rh236 r72 r227 r28 r rh236 r72 r227 r28 r rh236 r72 r227 r28 r379 2 rh236 r72 r227 r28 r379 Anamnestic response after subsequent H1N1pdm infection was faster than CTLs or Nabs Associated with protection from high levels of influenza virus recovery Jegaskanda et. al. (213) J. Virol. Chowell et. al. (29) NEJM 2

3 % NK cells expresing IFN-g % NK cells expresing IFN-g 14/7/214.. Older individuals (>45 years) had preexisting H1N1pdm-specific Seasonal 27 H1N1(A/Brisbane/59/27) HA Pre Post Pre Post Pre Post >45 Ages (years).. 29 H1N1pdm (A/California/7/29) HA p<.1 p<.1 p=.1 p=.8 Pre Post Pre Post Pre Post >45 Ages (years) Intravenous Ig (IVIG) Pooled polyvalent IgG extracted from plasma from over -4, donors Used to treat immunodeficiencies and autoimmune diseases Provides a snap-shot of herd immunity towards influenza viruses Could have therapeutic benefits against new pandemic influenza viruses if it contains cross-reactive antibodies Jegaskanda et. al. (213) JID IVIG manufactured pre-29 and post- 29 Pre 29 Pandemic Sample No. Date manufactured Post 29 Pandemic Sample No. Date manufactured IVIG made prior to 29 contains cross-reactive 29 H1N1pdm-specific mediating antibodies pre post pre post pre post pre post pre post Studied recombinant influenza HA proteins from either the 29 H1N1 pandemic (CA/9), seasonal H1N1 (SI/6, NC/99, BR/7) or HIV gp14 (negative control) Kindly provided by Steve Rockman at biocsl 3

4 14/7/214 IVIG can recognise a broad range of influenza subtypes How cross-reactive is IVIG to different influenza subtypes? H1 H2 H3 H4 H5 H7 gp14 Tested recombinant influenza HA proteins from either H1N1 (A/California/4/29), H2N2 (A/Japan/35/1957), H3N2(A/Brisbane//27), H4N6(A/Swine/Ontario/1911-1/1999), H5N1(A/Vietnam/123/24) and H7N7 (A/Chicken/Netherlands/1/23) or gp14 negative control Is broadly-reactive induced by current protein vaccine strategies? No induced by TIV in macaques Stimulated with H1N1pdm HA protein Pre-vaccine Post-vaccine Post-infection Gated on NK cells Jegaskanda et. al. (213) J Virol 4

5 14/7/214 No induced by TIV in macaques Post-vaccine (4 weeks post 2 nd dose) 4 weeks Post-H1N1 (SI/6) infection TIV-vaccinated Unvaccinated Summary antibodies that cross react to diverse influenza strains are induced by influenza infection in both macaques and humans May have played a role in reduced mortality of elderly during H1N1pdm pandemic Are present in IVIG, even prior to epidemics Modest or no induction of by current protein vaccines without adjuvant in macaques (and humans) Improved vaccine strategies needed to induce Will broadly reactive antibodies induced by a universal influenza vaccine offer partially protection from disease following infection with divergent strains? Much still to learn. A few key research priorities Difficult to measure responses in mice and ferrets Mice: NK cells more resistant to activation. Use of Fc-knock out mouse models current standard. Ferrets: few immunologic reagents Need for passive transfer experiments in larger animal models only antibodies without Nab function Need for incisive studies of human vaccinees Pre-existing from prior influenza infections can make interpretation difficult in adults Analyses of younger children or subjects pre-screened for needed Need protection studies in humans with vaccines inducing Better understanding of the best antigens to target by Surface proteins - HA, NA, M2e obvious targets. Further targeting of the most conserved elements of HA and NA. internal proteins also induce and might be useful NP, M1. Better understanding of role of other Fc-mediated functions phagocytosis etc University of Melbourne Sinthujan Jegaskanda Pat Reading Andrew Brooks Matt Parsons T. Amarasena S. Alcantara Wendy Winnall Marit Kramski Rob De Rose Emma Job Gamze Isitman Ivan Stratov Rest of Kent lab Acknowledgements University of Wisconsin Tom Friedrich Jason Weinfurter BioCSL Melbourne Steve Rockman Kirsten Vandenberg and Clinical investigators Clinical trial participants, blood donors and funders WHO influenza center Melbourne Karen Laurie Aeron Hurt Ian Barr Anne Kelso 5

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