Just a few viruses fell into subgroup 3B and group 6 (Fig. 4, Fig. S4). Some isolates from North America, Europe and Asia belonged to groups 5 and 6, which have signature AA substitutions D53N, Y94H, I230V and E280A in HA1, with group 6 isolates carrying an additional AA substitution S199A. Viruses with low HI titres to post-infection ferret antisera raised against cell-propagated A/Victoria/361/2011 viruses were scattered throughout the HA tree and did not form monophyletic groups or share common AA substitutions. Genetic analysis CCI-779 clinical trial of the HA sequences of several egg-propagated A/Victoria/361/2011-like

viruses were compared in order to see if low HI titres might be associated with amino acid substitutions linked to adaptation to growth in eggs. A number of such substitutions were noted in the HA of the initial egg-propagated A/Victoria/361/2011 wild-type virus, including a H156R substitution. A

subsequent R156Q change was acquired in the high growth reassortants IVR-165 and X221, although H156R was retained in the reassortant NIB-79. Changes in amino acid sequence in this area of the HA of A(H1N1)pdm09 viruses have been shown to alter their antigenic properties and, based on the ferret and human serology obtained for the egg-propagated A/Victoria/361/2011 virus, such substitutions may also have altered the antigenicity of this virus. Some egg-propagated viruses genetically similar to A/Victoria/361/2011, ABT-737 order such as A/Texas/50/2012, did not have similar adaptive substitutions in the 153–157 HA region. Egg-propagated A/Hawaii/22/2012 and the high growth reassortants made from this virus, X225 and X225A, had the substitution L157S and in HI assays antisera raised against A/Hawaii/22/2012 recognised the majority of test viruses with titres reduced more

than 4-fold compared to the homologous virus (Table 3). Vaccines containing influenza A/Victoria/361/2011 (H3N2)-like antigen stimulated anti-HA antibodies in all age groups that had reduced geometric mean HI titres to the majority of cell-propagated A(H3N2) viruses compared to the egg-propagated vaccine virus or other egg-propagated recent viruses (Fig. S5). The average reductions in HI GMT against cell-propagated A(H3N2) viruses compared to the egg-propagated vaccine virus were 66% not for adults, 68% for the elderly and 64% for children. Based on surveillance data available in February 2013, it was concluded that the A(H3N2) component of the 2012–2013 Northern Hemisphere influenza vaccine should remain as a A/Victoria/361/2011-like virus. However it was stipulated that the like virus should be antigenically like the cell-propagated prototype virus. For this reason a new vaccine virus A/Texas/50/2012 and its reassortants, X-223 and X223A, were recommended for use in vaccines that are based on egg propagation. From September 2012 to February 2013, 832 influenza B viruses were analysed by WHO CCs.