INFORMATION FOR THE VACCINE AND RELATED BIOLOGICAL PRODUCTS ADVISORY COMMITTEE CBER, FDA FEBRUARY 22, 2010 Nancy J. Cox, Ph. D Director WHO COLLABORATING CENTER FOR SURVEILLANCE, EPIDEMIOLOGY AND CONTROL OF INFLUENZA Influenza Division National Center for Immunization and Respiratory Diseases Coordinating Center for Infectious Diseases Centers for Disease Control and Prevention Atlanta, GA 30333

Specimens Received by CDC in 2009 & /08/2010

Specimens Received by CDC in 2009 & 2010 ( by Date Collected ) Feb Apr Jun Aug Oct Dec Jan Mar May Jul Sep Nov Jan Feb Specimens Received by CDC in 2009 & 2010 ( by Date Collected ) B H1 H3 H1pdm Misc.

Percentage of Global Influenza Surveillance Network Influenza Viruses by Types/Subtypes (From 1st September 2009 to 30th January 2010)

Influenza A (H1N1) 2009 Pandemic Viruses

Pandemic A(H1N1) 2009 viruses September 2009 to January 2010 Viruses from ~100 countries examined by WHO CCs –Antigenic analysis –Sequence Analysis –Antiviral resistance assessment by phenotypic and sequence analysis

Pandemic A(H1N1) 2009 viruses Vast majority of viruses antigenically homologous to the A/California/07/2009 vaccine virus A minority of viruses were observed with reduced titres against reference antiserum panels Examination of clinical samples from low reactor group indicated that isolation substrate affected the antigenicity of the virus isolates

Pandemic A(H1N1) viruses- HI analysis (2) 1.Viruses showing low reactivity in HI assays were associated with selection in a variant MDCK cell line. 2.HA AA substitutions in isolates NOT seen in clinical samples

Antigenic cartography illustrates very little change in antigenicity September 2009 to December 2009

Limited sequence diversity Oseltamivir resistance falling throughout the tree H1N1 pandemic viruses- Sequence analysis of HA gene

02/19/10 Vaccine Candidates #: Egg Isolate ◄: Adamantane H275Y Oseltamivir Resistant LR- low reactor to A/California/07/2009 (≥ 8 fold) F: Ferret Antiserum $: Serology Antigen H1 Numbering Evolutionary Relationships Among Pandemic Influenza A (H1N1) Hemagglutinin Genes, September 2009 October 2009 November 2009 December 2009 January 2010 North American Swine H A/Kansas/01/2010 ◄ Jan namru3 A/Egypt/N14645/2009 Dec FATAL A/Bangladesh/3009/2009 ◄ Dec # A/Ayutthaya/568/2009 ◄ Nov afioh A/Korea/3160/2009 Oct A/California/42/2009 ◄ Dec A/Sendai-H/41/2010 Jan A/Vermont/29/2009 ◄ Dec A/Nigeria/944/2009 ◄ Oct A/Puerto Rico/51/2009 ◄ Dec A/Hebei-Yuhua/Swl1441/2009 ◄ Dec A/Hebei-qiaodong/SWL1523/2009 Dec A/Guam/6687/2009 ◄ Oct A/Vietnam/2043/2009 Dec A/Philippines/2810/2009 ◄ Aug LR A/Arizona/20/2009 ◄ Dec A/Iowa/01/2010 ◄ Jan A/Tanzania/393/2009 ◄ Sep LR H1 PDM 2009/10 Consensus 2412 namru3 A/Egypt/N14644/2009 Dec FATAL A/Oita/1/2010 Jan A/Norway/3206/2009 ◄ Sep LR FATAL A/Uganda/0543/20009 ◄ Oct # A/Nordrhein-Westfalen/79/2009 Jul LR A/Lviv/N6/2009 Oct LR FATAL A/India/8489/2009 ◄ Oct A/Utah/42/2009 ◄ Jul FATAL F A/Florida/31/2009 ◄ Dec A/Brisbane/115/2009 Nov A/South Carolina/18/2009 ◄ Sep LR $ namru3 A/Egypt/N14648/2009 Dec FATAL A/Iraq/8529/2009 ◄ Sep # $ A/Iraq/8531/2009 ◄ Sep # A/Ontario/RV3226/2009 ◄ Oct $ A/Minnesota/38/2009 ◄ Dec afioh A/Alaska/3255/2009 Nov A/Bayan-ulgii/9912/2009 Dec A/New York/18/2009 ◄ Apr # F A/HunanChangsha/SWL4346/2009 Nov A/Hungary/36/2009 Jul # A/Korea/9741/2009 Sep # A/Utah/20/2009 ◄ Jul LR F A/Texas/05/2009 ◄ Apr F A/Bayern/69/2009 Jul LR NYMC X181 A/California/07/2009 # $ NYMC X181A A/California/07/2009 # NYMC X179A A/California/07/2009 # F IVR-153 A/California/7/2009 # F A/California/07/2009 ◄ Apr # F NIBRG-121 A/California/07/2009 # A/California/04/2009 ◄ Apr F S83P V321I K209T N129D S203T Q223R G155E Q293H N156D D127E S128L I295V S157L G155G/E N156K D222G D86G D222E E222G D222N P297S S128P D97N E374K G155E N156D

Residues 155/156 Receptor binding site Model of the Texas/05/2009 H1 HA trimer

Gly155 Receptor binding site Asn156

H1N1 pandemic viruses- Mapping of common substitutions onto H1HA structure Residues 203 and 222 differing from the vaccine virus have little effect on antigenicity

H1N1 pandemic viruses- Sequence analysis of NA gene 1.Limited sequence diversity 2.Oseltamivir resistance grouping in the tree 3.Caused by the H275Y substitution

02/19/10 Vaccine Candidates #: Egg Isolate ◄: Adamantane H275Y Oseltamivir Resistant LR- low reactor to A/California/07/2009 (≥ 8 fold) F: Ferret Antiserum $: Serology Antigen H1 Numbering Evolutionary Relationships Among Pandemic Influenza A (H1N1) Neuraminidase Genes, September 2009 October 2009 November 2009 December 2009 January Eurasia Swine N1NA A/Iraq/8531/2009 ◄ Sep # A/Lviv/N6/2009 Oct FATAL $ A/Iraq/8529/2009 ◄ Sep # $ A/Florida/13/2009 ◄ Aug # A/Bangladesh/3009/2009 ◄ Dec # A/Vietnam/2043/2009 Dec A/Puerto Rico/51/2009 ◄ Dec A/Vermont/29/2009 ◄ Dec A/Bayan-ulgii/9912/2009 Dec A/Hebei-Yuhua/Swl1441/2009 ◄ Dec A/Utah/20/2009 ◄ Jul LR F A/Ayutthaya/568/2009 ◄ Nov A/Korea/9741/2009 ◄ Sep # A/South Carolina/18/2009 ◄ Sep LR $ A/Utah/42/2009 ◄ Jul FATAL F A/India/8489/2009 ◄ Oct A/Philippines/2810/2009 ◄ Aug LR F A/Arizona/20/2009 ◄ Dec A/Guam/6687/2009 ◄ Oct A/Kansas/01/2010 ◄ Jan A/Ontario/RV3226/2009 ◄ Oct $ A/New York/18/2009 ◄ Apr # F A/Minnesota/38/2009 ◄ Dec A/Nigeria/944/2009 ◄ Oct N1 PDM 2009/10 Consensus 1684 A/Iowa/01/2010 ◄ Jan A/Tanzania/393/2009 ◄ Sep LR A/Nordrhein-Westfalen/79/2009 Jul LR A/Brisbane/2013/2009 Jul A/Norway/3206/2009 ◄ Sep FATAL A/Uganda/0543/20009 ◄ Oct # A/Oita/1/2010 Jan IVR-153 A/California/7/2009 # F NYMC X181 A/California/07/2009 # $ NIBRG-121 A/California/07/2009 # NYMC X181A A/California/07/2009 # F NYMC X179A A/California/07/2009 # F A/California/07/2009 ◄ Apr # F A/California/04/2009 ◄ Apr F A/Texas/05/2009 ◄ Apr F A/Bayern/69/2009 Jul LR V106I N248D

Pandemic A(H1N1) viruses – antiviral surveillance Antiviral resistance assessment by phenotypic and sequence analysis Regular assessment of samples at WHO CCs for resistance to sialidase inhibitors. –Approx 7000 samples analyzed- ~100 oseltamivir resistant viruses - H275Y 0 zanamivir resistant viruses Pandemic viruses remained predominantly resistant to amantadine/rimantadine.

Pandemic A(H1N1) viruses – summary Pandemic A(H1N1) 2009 viruses pose a public health risk in the forthcoming northern hemisphere influenza season. Currently circulating pandemic A(H1N1) 2009 viruses remain similar to the WHO recommended vaccine virus Recommendation: –A/California/7/2009-like virus remains suitable for use in influenza virus vaccines for northern hemisphere

Influenza A (H1N1) Viruses

Seasonal A(H1N1) influenza viruses Very few seasonal H1N1 viruses detected From a small number of countries Antigenic, genetic and drug resistance assays were carried out Most seasonal H1N1viruses were antigenically similar to the previous A/Brisbane/59/2007 vaccine virus

Seasonal A(H1N1) viruses – sequence analysis Most recent viruses were of the Brisbane/59/2007 genetic clade, 2B Viruses retained resistance to oseltamivir associated with H275Y in NA Some viruses were resistant to oseltamivir and amantadine/rimantadine

Evolutionary Relationships Among Seasonal Influenza A (H1N1) Hemagglutinin (HA1) Genes, September 2009 October 2009 November 2009 December /19/10 LR- low reactor to A/Brisbane/59/2007 (≥ 8 fold) Current Northern Hemisphere Vaccine Strain #: Egg Isolate F: Ferret Antiserum $: Serology Antigen ◄: Adamantane Oseltamivir Resistant A/Hong Kong/33774/2009 Oct A/Hong Kong/33773/2009 Oct # A/Hong Kong/26552/2009 Sep nimr A/Hong Oct LR A/Hong Kong/33775/2009 Oct LR nimr A/Hong Oct LR aus A/Macau/2517/2009 Jul nimr Aug LR cnic A/SichuanDongqu/1827/09 Nov A/Texas/57/2009 Aug LR niid Aug LR A/Sichuan-Dongqu/1836/2009 Nov niid Aug LR A/Beijing-Chaoyang/11405/2009 Nov A/Hong Jul A/Fujian-Gulou/1581/2009 May LR # Feb LR # A/Fujian-Gulou/1896/09 Jul # $ A/Hunan-Changning/52/2009 Jul A/Hong Kong/13503/2009 ◄ Aug Jun nimr Oct LR nimr Sep LR Aug H1 HA 2010 Consensus 78 niid Sep aus Jun Oct F A/Texas/38/2009 Feb Oct A/Rhode Sep Aug Aug # Jul # A/Alberta/RV2859/2009 Sep Dec Feb LR # Aug # Sep # Sep # Oct A/South Dakota/06/2007 Jul # F Aug # F A/Brisbane/59/2007 # F aus A/Brisbane/59/2007 IVR148 # $ A/Vladivostok/45/2009 ◄ Feb A/Hong Kong/1870/2008 ◄ Jul F cnic A/JiangxiYuanzhou/11218/09 ◄ Nov A/Liaoning-Huanggu/1144/2009 Mar C 2B Dual Resistant Reassortants S36N I47K E68G K82R R188M E273G A189T T193K K273E I417V S450N N495S T82K Y94H K140E R145K D186D/N/I/V/S/G R208K T266N L415I N472D D35N K145R R188K E273K V131A S141N G185A A189T G185V H192R N183S G185S Q222R

Evolutionary Relationships Among Seasonal Influenza A (H1N1) Neuraminidase Genes, September 2009 October 2009 November 2009 December /19/10 LR- low reactor to A/Brisbane/59/2007 (≥ 8 fold) Current Northern Hemisphere Vaccine Strain #: Egg Isolate F: Ferret Antiserum $: Serology Antigen ◄: Adamantane Oseltamivir Resistant A/Hong Kong/33773/2009 Oct # A/Hong Kong/33775/2009 Oct LR nimr A/Hong Oct LR nimr A/Hong Oct LR A/Hong Kong/26552/2009 Sep A/Hong Kong/13503/2009 ◄ Aug A/Fujian-Gulou/1896/09 Jul # $ cnic A/SichuanDongqu/1827/09 Nov A/Hong Jul Feb LR # A/Texas/57/2009 Aug LR A/Fujian-Gulou/1581/2009 May LR # A/Beijing-Chaoyang/11405/2009 Nov A/Sichuan-Dongqu/1836/2009 Nov aus A/Macau/2517/2009 Jul nimr Aug LR Aug nimr Sep LR Jun aus Jun N1 NA 2010 Consensus 72 Aug # Jul # Oct F A/Texas/38/2009 Feb A/Liaoning-Huanggu/1144/2009 Mar Aug A/Rhode Sep Oct Aug # F Feb LR # Sep # Aug # A/Alberta/RV2859/2009 Sep Dec Sep # Oct A/South Dakota/06/2007 Jul # F A/Brisbane/59/2007 # F cnic A/JiangxiYuanzhou/11218/09 ◄ Nov A/Hong Kong/1870/2008 ◄ Jul F A/Vladivostok/45/2009 ◄ Feb C 2B V48I E214G R222Q V234M K329E D344N D382N D461G L466F H45N K78E G249K T287I S82P Y100H R130K M188I T264I H275Y G354D A250T T81N I48M T81K N28S V83M Dual Resistant Reassortants I267M L367I V393I T453I

Seasonal A(H1N1) viruses - Summary Seasonal A(H1N1) viruses showing little evidence of circulation Mostly likely pose a low risk in the forthcoming Northern Hemisphere influenza season WHO is not recommending seasonal A(H1N1) as a component for vaccines for the northern hemisphere

Influenza A (H3N2) Viruses

Seasonal A(H3N2) Viruses - Summary Influenza A(H3N2) viruses have continued to circulate globally, but at reduced levels compared with previous years H3N2 viruses likely pose a risk in the forthcoming Northern Hemisphere influenza season Antigenic, genetic and drug resistance assays were carried out H3N2 viruses analyzed are antigenically closely related to the vaccine virus recommended for the Southern Hemisphere 2010, A/Perth/16/2009

September 09 – January 10, Influenza A(H3N2) 17 February 2010 The boundaries and names shown and the designations used on this map do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement. Data source: WHO, GISN Map production: GIP/VMV World Health Organization © WHO All rights reserved. No activity or no data Sporadic activity Local activity Regional outbreaks Widespread outbreak

Antigenic cartography of A(H3N2) A/Brisbane/10/07 A/Perth/16/09

02/19/10 Current Northern Hemisphere Vaccine Strain #: Egg Isolate ◄: Adamantane Resistant LR- low reactor to A/Perth/16/2009 (≥ 8 fold) F: Ferret Antiserum $: Serology Antigen Evolutionary Relationships Among Influenza A (H3N2) Hemagglutinin Genes, September 2009 October 2009 November 2009 December 2009 January 2010 A/Guangdong-Luohu/1257/2009 ◄ Jun # A/Guangdong-Luohu/1256/2009 X185 # A/Guangdong-Luohu/1256/2009 X185XP # A/Guangdong-Luohu/1256/2009 ◄ Jun # A/Hunan-Xiangxiang/442/2009 ◄ Oct A/Hong Kong/26560/2009 ◄ Sep # F $ A/Beijing-Huairou/11088/2009 ◄ Nov A/Hong Kong/34341/2009 ◄ Nov A/Tianjin-Hexi/1471/2009 ◄ Nov cnic A/TianjinHexi/1530/2009 ◄ Dec niid A/Taiwan/948/2009 Nov A/Oregon/36/2009 ◄ Oct cnic A/HebeiYuhua/1435/2009 ◄ Dec A/Hawaii/07/2009 ◄ Mar F A/Hawaii/07/2009 RG-46V3 # F A/Philippines/2191/2009 ◄ May # F A/Wisconsin/15/2009 ◄ Jun # F A/Wisconsin/15/2009 X183 # F $ H3HA 2009 Consensus 264 A/Bangkok/561/2009 ◄ Nov A/Cayman Islands/9266/2009 ◄ Oct niid A/Laos/I258/2009 Oct A/Perth/16/2009 NIB64 # F A/Perth/16/2009 ◄ # F namru3 A/Egypt/N11396/2009 Oct namru3 A/Egypt/N11830/2009 Nov A/Surat Thani/559/2009 ◄ Nov A/Victoria/210/2009 ◄ Jun # F A/Victoria/210/2009 X-187 # usafsam A/Kuwait/4562/2009 Dec usafsam A/Afghanistan/2950/2009 Oct niid A/Shizuoka/736/2009 May $ Canada A/Toronto/H42220/2009 Dec A/New York/01/2010 ◄ Jan A/India/2710/2009 ◄ Sep LR A/BritishColumbia/RV3075/2009 ◄ Nov A/Hong Kong/4547/2009 ◄ Jul # F A/Florida/26/2009 ◄ Nov A/Victoria/208/2009 ◄ Jun # F $ A/Rhode Island/01/2010 Jan A/Ohio/17/2009 ◄ Oct LR namru3 A/Egypt/N12479/2009 Dec A/Nigeria/946/2009 ◄ Nov A/Taiwan/713/2009 ◄ May F A/Vietnam/409/2009 ◄ Jul LR niid A/Yamaguchi/30/2008 Dec A/Brisbane/10/2007 IVR147 # F A/Brisbane/10/2007 ◄ # F A/Uruguay/716/2007 ◄ # F A/Uruguay/716/2007 X175C # F $ G50E K140I K173Q I361R T212A K158N N189K E62K N144K V213A A138S L157S L194P Vic/208 Perth/16 Bri/10 N312S R261Q I260M H183L I214S D487A L194P

02/19/10 Current Northern Hemisphere Vaccine Strain #: Egg Isolate ◄: Adamantane Resistant LR- low reactor to A/Perth/16/2009 (≥ 8 fold) F: Ferret Antiserum $: Serology Antigen Evolutionary Relationships Among Influenza A (H3N2) Neuraminidase Genes, September 2009 October 2009 November 2009 December 2009 January 2010 A/Guangdong-Luohu/1256/2009 X185XP # A/Guangdong-Luohu/1256/2009 X-185 # A/Guangdong-Luohu/1257/2009 ◄ Jun # A/Guangdong-Luohu/1256/2009 ◄ Jun # A/Hong Kong/26560/2009 ◄ Sep # F $ cnic A/HebeiYuhua/1435/2009 ◄ Dec A/Hunan-Xiangxiang/442/2009 ◄ Oct A/Tianjin-Hexi/1471/2009 ◄ Nov A/Hong Kong/34341/2009 ◄ Nov cnic A/TianjinHexi/1530/2009 ◄ Dec A/Oregon/36/2009 ◄ Oct A/Hawaii/07/2009 ◄ Mar F A/Hawaii/07/2009 RG-46V3 # F A/Surat Thani/559/2009 ◄ Nov A/Cayman Islands/9266/2009 ◄ Oct A/Bangkok/561/2009 ◄ Nov N2NA 2009/10 Consensus 180 A/Philippines/2191/2009 ◄ May # A/Wisconsin/15/2009 ◄ Jun # F A/Wisconsin/15/2009 X183 # F $ A/Victoria/210/2009 ◄ Jun # F A/Victoria/210/2009 X-187 # A/Perth/16/2009 NIB64 # F A/Perth/16/2009 ◄ # A/Florida/26/2009 ◄ Nov A/Nigeria/946/2009 ◄ Nov Canada A/Toronto/H42220/2009 Dec A/Hawaii/31/2009 ◄ Sep A/New York/01/2010 ◄ Jan A/India/2710/2009 ◄ Sep LR A/Ohio/17/2009 ◄ Oct LR A/BritishColumbia/RV3075/2009 ◄ Nov A/Hong Kong/4547/2009 ◄ Jul # F niid A/Shizuoka/736/2009 May $ A/Taiwan/713/2009 ◄ May F A/Victoria/208/2009 ◄ Jun # F $ A/Vietnam/409/2009 ◄ Jul niid A/Yamaguchi/30/2008 Dec A/Brisbane/10/2007 IVR147 # F A/Brisbane/10/2007 ◄ # F A/Uruguay/716/2007 ◄ # F A/Uruguay/716/2007 X175C # F $ Perth/16 Bris/10 E258K I26T Y40C I312T D147N I215V S367N K369T I464L R60K I469V T148T/I/L D151/D/N/V/G/S/A Vic/208

Seasonal A(H3N2) viruses - Summary Influenza A(H3N2) activity relatively low worldwide A(H3N2) viruses circulating since September 2009 were antigenically closely related to A/Perth/16/2009 Hemagglutinin genes of recent A(H3N2) viruses fell into 2 distinct genetic clades represented by the A/Perth/16/2009 and A/Victoria/208/2009 viruses A/Perth/16/2009 and A/Victoria/208/2009 clade viruses were antigenically similar All A(H3N2) viruses were resistant to amantadine and rimantadine, but sensitive to neuraminidase inhibitors

Influenza B Viruses

Influenza B Viruses - Summary Influenza B viruses of both the B/Victoria and B/Yamagata lineages continued to circulate at low levels globally; resurgence of influenza B activity reported by China with B/Victoria lineage viruses predominating Influenza B viruses likely pose a continuing risk in the forthcoming Northern Hemisphere influenza season Antigenic, genetic and drug resistance assays have been carried out WHO recommendation is for the B/Victoria lineage only and to retain B/Brisbane/60/2008

Number of B viruses detected in GISN 13,247 positives detected 31,519 positives detected 25,880 positives detected 4,927 positives detected

September 2009 – January 2010, Influenza B 17 February 2010 The boundaries and names shown and the designations used on this map do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement. Data source: WHO, GISN Map production: GIP/VMV World Health Organization © WHO All rights reserved. No activity or no data Sporadic activity Local activity Regional outbreaks Widespread outbreak

B activity in China Since week 48, the percentage of influenza B virus began to increase and became the predominant virus in week 2. In week 4, the percentage of influenza B virus increased to 75.1%.

Number of 2 lineages of B viruses detected in GISN 2009 and 2010 (until 30th January 2010)

B Victoria antigenic cartography ML/2506/04 BR/60/08

Evolutionary Relationships Among Influenza B Victoria Lineage Hemagglutinin (HA1) Genes, /10/10 LR to B/Brisbane/60/2008 (≥ 8 fold) # Egg Isolate F: Ferret Antiserum $: Serology Antigen ____ 165N September 2009 October 2009 November 2009 December 2009 January 2010 B/BritishColumbia/RV1072/2009 Mar LR B/Hawaii/02/2009 Mar LR B/Washington/01/2009 Feb LR # B/Cambodia/6535/2009 Jul # nimr B/Norway/1850/2009 Jun B/Nevada/05/2009 Sep B/Brisbane/33/2008 Jul # F B/Massachusetts/03/2009 Oct B/Brisbane/60/2008 Aug # F $ B/Pennsylvania/10/2009 Oct # usafsam B/California/2986/2009 Oct B/Haiti/583/2009 Nov B/Nigeria/0958/2009 Nov aus B/Chantahburi/329/2009 Jul B/Incheon/4293/2009 Nov niid B/Niigata/755/2009 Jun usafsam B/Japan/3280/2009 Nov B/Uganda/0514/2009 Oct B/Bangladesh/5945/2009 Sep # $ B/Bangladesh/7055/2009 Oct nimr B/Ghana/FS-1037/2009 Jul B/Guatemala/1317/2009 Jun # BHA VIC 2009 Consensus 61 nimr B/Madagascar/4254/2009 Jun B/Ontario/RV2893/2009 Oct cnic B/AnhuiBaohe/136/2010 Jan B/Shanxi-Zuoquan/311/2009 Oct B/Shanghai-Pudongxin/1465/2009 Dec cnic B/ShanghaiSongjiang/1349/2009 Dec cnic B/HebeiYuhua/14/2010 Jan cnic B/JiangsuBeitang/1893/2009 Dec B/Bangladesh/4008/2008 Jun # F aus B/Victoria/304/2006 # B/Dominican Republic/1078/2009 May B/Minnesota/06/2009 May B/Hong Kong/537/2009 Oct LR B/Hong Kong/230/2009 Jul LR $ B/Hubei-Jiangan/1183/2009 Jun LR B/Guangxi-Yizhou/312/2009 Apr LR niid B/Laos/1232/2009 Jul LR $ niid B/Laos/1231/2009 Jul B/Taiwan/55/2009 Nov LR aus B/Singapore/22/2009 Jun aus B/Nong Khai/332/2009 Jul LR B/Vietnam/244/2009 Jun LR B/Vietnam/532/2009 Aug LR N75K N165K S172P L58P V146I K203T N197D K257R T37I N165K

Evolutionary Relationships Among Influenza B Victoria Lineage Neuraminidase Genes, /10/10 LR to B/Brisbane/60/2008 (≥ 8 fold) # Egg Isolate F: Ferret Antiserum $: Serology Antigen September 2009 October 2009 November 2009 December 2009 January 2010 B/Brisbane/33/2008 Jul # F B/British Columbia/RV1072/2009 Mar LR B/Nevada/05/2009 Sep B/Washington/01/2009 Feb LR # B/Hawaii/02/2009 Mar LR aus B/Chantahburi/329/2009 Jul B/Massachusetts/03/2009 Oct B/Incheon/4293/2009 Nov niid B/Niigata/755/2009 Jun B/Nigeria/0958/2009 Nov B/Cambodia/6535/2009 Jul # B/Brisbane/60/2008 Aug # F $ B/Pennsylvania/10/2009 Oct # nimr B/Norway/1850/2009 Jun B/Guatemala/1317/2009 Jun # BNA VIC 2009 Consensus 50 nimr B/Madagascar/4254/2009 Jun nimr B/Ghana/FS-1037/2009 Jul B/Ontario/RV2893/2009 Oct B/Uganda/0514/2009 Oct B/Bangladesh/5945/2009 Sep # $ B/Bangladesh/7055/2009 Oct B/Shanghai-Pudongxin/1465/2009 Dec B/Shanxi-Zuoquan/311/2009 Oct cnic B/AnhuiBaohe/136/2010 Jan cnic B/HebeiYuhua/14/2010 Jan cnic B/JiangsuBeitang/1893/2009 Dec cnic B/ShanghaiSongjiang/1349/2009 Dec B/Bangladesh/4008/2008 Jun # F niid B/Victoria/304/2006 # B/Ohio/1/2005 # F B/Dominican Republic/1078/2009 May B/Haiti/583/2009 Nov B/Minnesota/06/2009 May aus B/Nong Khai/332/2009 Jul LR B/Vietnam/532/2009 Aug LR B/Vietnam/244/2009 Jun LR B/Taiwan/55/2009 Nov LR B/Guangxi-Yizhou/312/2009 Apr LR B/Hubei-Jiangan/1183/2009 Jun LR B/Hong Kong/230/2009 Jul LR $ B/Hong Kong/537/2009 Oct LR S41P N125K D320E D463N P42S V271I D342G M375K K404E T8M S15L T8M Q61H A389T N220K I204V A358E D329N D463N A465T P51S A465T L73F K343E N199D

Evolutionary Relationships Among Influenza B Yamagata Lineage Hemagglutinin (HA1) Genes, /16/10 LR- Low Reactor to B/Florida/4/2006 (≥ 8 fold) Current Northern Hemisphere Vaccine Strain # Egg Isolate F: Ferret Antiserum $: Serology Antigen September 2009 October 2009 November 2009 December 2009 B/Hawaii/03/2009 Mar B/Florida/04/2006 # F B/Minnesota/08/2009 Aug B/Vietnam/256/2009 Jun B/Brisbane/03/2007 # F aus B/Johannesburg/450/2009 Jun B/Bangladesh/0802/2009 Oct B/Bangladesh/2249/2009 Dec B/Bangladesh/3750/2009 Oct namru3 B/Egypt/N10131/2009 Aug namru3 B/Egypt/N08946/2009 May nimr B/Tehran/8/2009 May namru3 B/Egypt/N11012/2009 Sep namru3 B/Egypt/N12566/2009 Nov B/Haiti/40/2009 Jun # BHA YAM 2009 Consensus 23 B/North Carolina/01/2009 Feb LR B/Washington/07/2009 Apr # B/Costa Rica/5141/2009 Jun # B/Texas/11/2009 Mar LR B/Mexico/2764/2009 Mar B/Georgia/02/2009 Apr B/Minnesota/05/2009 Apr B/Hong Kong/116/2009 Mar LR # niid B/Taiwan/40/2009 May cnic B/Hubei-wujiagang/158/2009 # LR B/Hong Kong/248/2009 Jul niid B/Sakai/68/2009 Nov cnic B/ShanghaiFengxian/1260/09 Dec LR niid B/Fujian-gulou/11410/2009 Sep niid B/Fujian-gulou/11386/2009 Sep B/Hong Kong/533/2009 Oct # B/Guangdong-Chikan/1482/2009 Oct niid B/Shanghai-fengxian/1299/2009 Dec B/Hong Kong/121/2009 Apr B/Guatemala/09/2009 Jul B/Egypt/5161/2009 May LR B/Bangladesh/3333/2007 # $ aus B/Perth/31/2009 May B/Shanghai-Pudongxin/1321/2009 Oct niid B/Shanghai-pudongxin/1335/2009 Oct G230S P108A V252M R48K S150I N166Y G230D M252V N203S A146S K254R N218D M252K 1 2 3

02/10/10 LR- Low Reactor to B/Florida/4/2006 (≥ 8 fold) Current Northern Hemisphere Vaccine Strain # Egg Isolate F: Ferret Antiserum $: Serology Antigen September 2009 October 2009 November 2009 December 2009 Evolutionary Relationships Among Influenza B Yamagata Lineage Neuraminidase Genes, B/Minnesota/08/2009 Aug B/Florida/04/2006 # F B/Hawaii/03/2009 Mar B/Vietnam/256/2009 Jun B/Brisbane/03/2007 # F B/Costa Rica/5141/2009 Jun # B/Washington/07/2009 Apr # B/North Carolina/01/2009 Feb LR B/Mexico/2764/2009 Mar B/Texas/11/2009 Mar LR nimr B/Tehran/8/2009 May B/Bangladesh/3750/2009 Oct B/Bangladesh/0802/2009 Oct B/Bangladesh/2249/2009 Dec B/Guatemala/09/2009 Jul B/Guangdong-Chikan/1482/2009 Oct B/Hong Kong/533/2009 Oct # BNA YAM 2009 Consensus 9 B/Haiti/40/2009 Jun # B/Hong Kong/248/2009 Jul cnic B/ShanghaiFengxian/1260/09 Dec LR B/Hong Kong/116/2009 Mar LR # niid B/Taiwan/40/20009 May B/Egypt/5161/2009 May LR B/Georgia/02/2009 Apr B/Minnesota/05/2009 Apr B/Hong Kong/121/2009 Apr B/Bangladesh/3333/2007 # $ B/Shanghai-Pudongxin/1321/2009 Oct I45T Q42R A68T T125K K186R D340N D463N A465T I248V L73P I45M 1 2 3

Summary of influenza B viruses Influenza B viruses of both the B/Victoria/2/87 – and B/Yamagata/16/88 - lineages viruses circulated in many countries: B/Victoria/2/87 lineage viruses continued to predominant The majority of recent B/Victoria/2/87- lineage viruses were antigenically closely related to the vaccine virus, B/Brisbane/60/2008 The majority of B/Yamagata16/88 - lineage viruses were closely related to the previous vaccine virus B/Florida/4/2006 Recent B/Yamagata16/88 lineage viruses from China were closely related to B/Hubei-Wujiagang/158/2009 Influenza B viruses remain sensitive to neuraminidase inhibitors