Rhiza Labs FluTracker Forum

Although much of the H5N1 transmission story has centered on the manufactured side show of terrorist recipes for a bioweapon, the evolution in ferrets raises serious concerns about the natural evolution of H5N1 in mammalian hosts. One change that has received little attention is the appearance of G225E in 4 of the 10 ferrets inncoulated with N158D/V318I (supplemental table 5)

http://www.nature.com/nature/journal/va ... 831-s1.pdf

It should be noted that the Kawaoak H5 was on an H1N1pdm09 background, and D225E was present in a subset transmitting at high levels in 2009, largeky in Europe.

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Above figure notes G225E in receptor biding domain (figure 1a).

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Sequence comparison of viruses from inoculated and contact animals identified mutations at positions 225 and 242 as well as a reversion at position 224 (Fig. 1a and Supplementary Table 5)

http://www.nature.com/nature/journal/va ... 10831.html

The above descibes H5 isolated from ferrets infected with N158D and V318I.

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G225E

EPI214881 A/chicken/Egypt/0920-NLQP/2009

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Top 100 matches (human emphasis)

EPI214881 A/chicken/Egypt/0920-NLQP/2009 (A/H5N1) segment 4 (HA) 3147.8 0.000000e+00 1704/1704 (100%)
EPI255398 A/Egypt/N09174/2009 (A/H5N1) segment 4 (HA) 3120.1 0.000000e+00 1699/1704 (99%)
EPI184804 A/Egypt/N00585/2009 (A/H5N1) segment 4 (HA) 3120.1 0.000000e+00 1699/1704 (99%)
EPI255400 A/Egypt/N09539/2009 (A/H5N1) segment 4 (HA) 3109.0 0.000000e+00 1697/1704 (99%)
EPI116470 A/Egypt/2621-NAMRU3/2007 (A/H5N1) segment 4 (HA) 3109.0 0.000000e+00 1697/1704 (99%)
EPI283813 A/duck/Egypt/1068s/2010 (A/H5N1) segment 4 (HA) 3103.5 0.000000e+00 1696/1704 (99%)
EPI283791 A/chicken/Egypt/1034/2010 (A/H5N1) segment 4 (HA) 3103.5 0.000000e+00 1694/1701 (99%)
EPI255371 A/Egypt/N02127/2010 (A/H5N1) segment 4 (HA) 3103.5 0.000000e+00 1696/1704 (99%)
EPI214880 A/chicken/Egypt/092-NLQP/2009 (A/H5N1) segment 4 (HA) 3103.5 0.000000e+00 1696/1704 (99%)
EPI182355 A/chicken/Egypt/0894-NLQP/2008 (A/H5N1) segment 4 (HA) 3103.5 0.000000e+00 1696/1704 (99%)
EPI255367 A/Egypt/N01982/2010 (A/H5N1) segment 4 (HA) 3099.8 0.000000e+00 1695/1704 (99%)
EPI184806 A/Egypt/N00605/2009 (A/H5N1) segment 4 (HA) 3098.0 0.000000e+00 1695/1704 (99%)
EPI214908 A/duck/Egypt/0990SM-NLQP/2009 (A/H5N1) segment 4 (HA) 3094.3 0.000000e+00 1693/1702 (99%)
EPI315697 A/chicken/Egypt/0918-NLQP/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1695/1705 (99%)
EPI287352 A/chicken/Egypt/1553-15/2010 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI283821 A/duck/Egypt/1063s/2010 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1692/1701 (99%)
EPI283815 A/duck/Egypt/106d/2010 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1692/1701 (99%)
EPI283782 A/chicken/Egypt/091317s/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI255394 A/Egypt/N08835/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI255392 A/Egypt/N07908/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI255388 A/Egypt/N05912/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI214906 A/duck/Egypt/0972-NLQP/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI214895 A/duck/Egypt/0918-NLQP/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1695/1705 (99%)
EPI184845 A/Egypt/N04823/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1694/1704 (99%)
EPI184835 A/Egypt/N04395/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1695/1705 (99%)
EPI184828 A/Egypt/N03439/2009 (A/H5N1) segment 4 (HA) 3092.4 0.000000e+00 1695/1705 (99%)
EPI214910 A/goose/Egypt/0929-NLQP/2009 (A/H5N1) segment 4 (HA) 3088.7 0.000000e+00 1692/1702 (99%)
EPI340165 A/duck/Egypt/10228SF/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI340157 A/turkey/Egypt/10453F/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI340139 A/chicken/Egypt/115/2011 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI310236 A/chicken/Egypt/F7/2009 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI283811 A/goose/Egypt/1057/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1691/1701 (99%)
EPI283792 A/chicken/Egypt/1035/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1691/1701 (99%)
EPI283789 A/chicken/Egypt/1029/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1694/1705 (99%)
EPI283784 A/duck/Egypt/103swf/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1691/1701 (99%)
EPI255377 A/Egypt/N03071/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI255375 A/Egypt/N02770/2010 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI214898 A/duck/Egypt/0926-NLQP/2009 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI184832 A/Egypt/N04316/2009 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI184812 A/Egypt/N02039/2009 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1694/1705 (99%)
EPI184810 A/Egypt/N01310/2009 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1694/1705 (99%)
EPI115007 A/Egypt/0636-NAMRU3/2007 (A/H5N1) segment 4 (HA) 3086.9 0.000000e+00 1693/1704 (99%)
EPI315963 A/duck/Egypt/0897-NLQP/2008 (A/H5N1) segment 4 (HA) 3083.2 0.000000e+00 1692/1704 (99%)
EPI184802 A/Egypt/N00001/2009 (A/H5N1) segment 4 (HA) 3083.2 0.000000e+00 1692/1704 (99%)
EPI340162 A/duck/Egypt/10255AG/2010 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1692/1704 (99%)
EPI283820 A/chicken/Egypt/1021L/2010 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1690/1701 (99%)
EPI283807 A/duck/Egypt/1020sd/2010 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1691/1702 (99%)
EPI283799 A/duck/Egypt/1011d/2010 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1690/1701 (99%)
EPI255369 A/Egypt/N02038/2010 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1692/1704 (99%)
EPI245999 A/goose/Egypt/0910sm-NLQP/2009 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1686/1695 (99%)
EPI184816 A/Egypt/N02563/2009 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1692/1704 (99%)
EPI182356 A/chicken/Egypt/0896-NLQP/2008 (A/H5N1) segment 4 (HA) 3081.3 0.000000e+00 1692/1704 (99%)
EPI255379 A/Egypt/N03072/2010 (A/H5N1) segment 4 (HA) 3077.6 0.000000e+00 1691/1704 (99%)
EPI184826 A/Egypt/N03438/2009 (A/H5N1) segment 4 (HA) 3077.6 0.000000e+00 1691/1704 (99%)
EPI340174 A/duck/Egypt/1046SF/2010 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1691/1704 (99%)
EPI340151 A/chicken/Egypt/10513S/2010 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1691/1704 (99%)
EPI310237 A/chicken/Egypt/F8/2009 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1691/1704 (99%)
EPI283786 A/environment/Egypt/105swf/2010 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1689/1701 (99%)
EPI255361 A/Egypt/N00270/2010 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1691/1704 (99%)
EPI245988 A/chicken/Egypt/0945-NLQP/2009 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1685/1695 (99%)
EPI237644 A/chicken/Egypt/0987-NLQP/2009 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1692/1705 (99%)
EPI184837 A/Egypt/N04396/2009 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1692/1705 (99%)
EPI184814 A/Egypt/N02407/2009 (A/H5N1) segment 4 (HA) 3075.8 0.000000e+00 1692/1705 (99%)
EPI255365 A/Egypt/N01644/2010 (A/H5N1) segment 4 (HA) 3072.1 0.000000e+00 1690/1704 (99%)
EPI348152 A/chicken/Egypt/11VIR4453-268/2010 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1690/1704 (99%)
EPI340149 A/duck/Egypt/10336SF/2010 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1690/1704 (99%)
EPI340138 A/duck/Egypt/1125S/2011 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1690/1704 (99%)
EPI340112 A/chicken/Egypt/1197AG/2011 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1690/1704 (99%)
EPI283803 A/chicken/Egypt/1012sf/2010 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1688/1701 (99%)
EPI283785 A/environment/Egypt/104swf/2010 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1689/1702 (99%)
EPI255373 A/Egypt/N02554/2010 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1690/1704 (99%)
EPI246005 A/goose/Egypt/09324sm-NLQP/2009 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1685/1696 (99%)
EPI245985 A/chicken/Egypt/0910-NLQP/2009 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1684/1695 (99%)
EPI125660 A/duck/Egypt/R5/2007 (A/H5N1) segment 4 (HA) 3070.3 0.000000e+00 1690/1704 (99%)
EPI348153 A/chicken/Egypt/11VIR4453-266/2010 (A/H5N1) segment 4 (HA) 3066.6 0.000000e+00 1689/1704 (99%)
EPI369180 A/duck/Egypt/1120SMF/2011 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340173 A/chicken/Egypt/1021AD/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340160 A/chicken/Egypt/10250SF/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340154 A/duck/Egypt/10118/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340153 A/duck/Egypt/10283SF/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340142 A/chicken/Egypt/119S/2011 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340137 A/duck/Egypt/1130AG/2011 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340135 A/chicken/Egypt/1113/2011 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI340104 A/chicken/Egypt/117AD/2011 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI338733 A/duck/Egypt/M2583B/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI338732 A/duck/Egypt/M2583C/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI338616 A/duck/Egypt/M2583D/2010 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI246003 A/duck/Egypt/09238sm-NLQP/2009 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1683/1695 (99%)
EPI125656 A/chicken/Egypt/R3/2007 (A/H5N1) segment 4 (HA) 3064.7 0.000000e+00 1689/1704 (99%)
EPI116452 A/Egypt/1394-NAMRU3/2007 (A/H5N1) segment 4 (HA) 3062.9 0.000000e+00 1688/1704 (99%)
EPI348160 A/chicken/Egypt/11VIR4453-267/2010 (A/H5N1) segment 4 (HA) 3061.0 0.000000e+00 1687/1702 (99%)
EPI348157 A/chicken/Egypt/11VIR4453-15/VRLCU/2010 (A/H5N1) segment 4 (HA) 3061.0 0.000000e+00 1687/1702 (99%)
EPI348156 A/chicken/Egypt11VIR4453-269/2010 (A/H5N1) segment 4 (HA) 3061.0 0.000000e+00 1687/1703 (99%)
EPI255382 A/Egypt/N04434/2010 (A/H5N1) segment 4 (HA) 3061.0 0.000000e+00 1688/1704 (99%)
EPI340152 A/duck/Egypt/10290SF/2010 (A/H5N1) segment 4 (HA) 3059.2 0.000000e+00 1688/1704 (99%)
EPI340147 A/chicken/Egypt/10132/2010 (A/H5N1) segment 4 (HA) 3059.2 0.000000e+00 1688/1704 (99%)
EPI340144 A/chicken/Egypt/10413SF/2010 (A/H5N1) segment 4 (HA) 3059.2 0.000000e+00 1688/1704 (99%)
EPI340140 A/chicken/Egypt/1115S/2011 (A/H5N1) segment 4 (HA) 3059.2 0.000000e+00 1688/1704 (99%)
EPI340133 A/goose/Egypt/1117SF/2011 (A/H5N1) segment 4 (HA) 3059.2 0.000000e+00 1688/1704 (99%)
EPI340132 A/duck/Egypt/1123SF/2011 (A/H5N1) segment 4 (HA) 3059.2 0.000000e+00 1688/1704 (99%)

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LOCUS GU002700 1704 bp cRNA linear VRL 04-JUN-2010
DEFINITION Influenza A virus (A/chicken/Egypt/0920-NLQP/2009(H5N1)) segment 4
hemagglutinin (HA) gene, complete cds.
ACCESSION GU002700
VERSION GU002700.1 GI:260104188
KEYWORDS .
SOURCE Influenza A virus (A/chicken/Egypt/0920-NLQP/2009(H5N1))
ORGANISM Influenza A virus (A/chicken/Egypt/0920-NLQP/2009(H5N1))
Viruses; ssRNA negative-strand viruses; Orthomyxoviridae;
Influenzavirus A.
REFERENCE 1 (bases 1 to 1704)
AUTHORS Arafa,A.-S.M., Sleim,A.A., Hassan,M.K. and Aly,M.M.
TITLE Phylogenetic analysis of 2009 H5N1 Egyptian viruses
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 1704)
AUTHORS Arafa,A.-S.M., Sleim,A.A., Hassan,M.K. and Aly,M.M.
TITLE Direct Submission
JOURNAL Submitted (25-SEP-2009) National Laboratory for Veterinary Quality
Control on Poultry Production, Animal Health Research Institute,
Nadi Elsaid Street, Dokki, Giza 12618, Egypt
FEATURES Location/Qualifiers
source 1..1704
/organism="Influenza A virus
(A/chicken/Egypt/0920-NLQP/2009(H5N1))"
/mol_type="viral cRNA"
/strain="A/chicken/Egypt/0920-NLQP/2009"
/serotype="H5N1"
/host="chicken"
/db_xref="taxon:678668"
/segment="4"
/country="Egypt: Monofia"
/collection_date="Feb-2009"
gene 1..1704
/gene="HA"
CDS 1..1704
/gene="HA"
/codon_start=1
/product="hemagglutinin"
/protein_id="ACX31993.1"
/db_xref="GI:260104189"
/translation="MEKIVLLLAIVSIVKSDQICIGYHANNSTEQVDTIMEKNVTVTH
AQDILEKTHNGKLCNLDGVKPLILRDCSVAGWLLGNPMCDEFLNVPEWSYIVEKINPA
NDLCYPGNFNDYEELKHLLSRINHFEKIQIIPKNSWSDHEASGVSSACPYQGRSSFFR
NVVWLTKKNNAYPTIKRSYNNTNQEDLLVLWGIHHPNDAAEQTRLYQNPTTYISVGTS
TLNQRLVPKIATRSKVNEQSGRMEFFWTILKSNDAINFESNGNFIAPENAYKIVKKGD
STIMKSELEYGNCNTKCQTPIGAINSSMPFHNIHPLTIGECPKYVKSNRLVLATGLRN
SPQGERRRKKRGLFGAIAGFIEGGWQGMVDGWYGYHHSNEQGSGYAADKESTQKAIDG
VTNKVNSIIDKMNTQFEAVGREFNNLERRIENLNKKMEDGFLDVWTYNAELLVLMENE
RTLDFHDSNVKNLYDKVRLQLRDNAKELGNGCFEFYHRCDNECMESVRNGTYDYPQYS
EEARLKREEISGVKLESIGTYQILSIYSTVASSLALAIMVAGLFLWMCSNGSLQCRIC
I"
ORIGIN
1 atggagaaaa tagtgcttct tcttgcaata gtcagtattg ttaaaagtga tcagatttgc
61 attggttacc atgcaaacaa ctcgacagag caggttgaca caataatgga aaagaacgtc
121 actgttacac acgcccaaga catactggaa aagacacaca acgggaaact ctgcaatcta
181 gatggagtga agcctctaat tttaagagat tgtagtgtag ctggatggct cctcgggaac
241 ccaatgtgcg acgaattcct caatgtgccg gaatggtctt acatagtgga gaagatcaat
301 ccagccaatg acctctgtta tccagggaat ttcaacgact atgaagaact gaaacaccta
361 ttgagcagaa taaaccattt tgagaaaatt cagatcattc ccaaaaattc ttggtcagat
421 catgaagcct caggagtgag ctcagcatgt ccataccagg gaagatcctc cttttttaga
481 aatgtggtat ggcttaccaa aaagaacaat gcatacccaa caataaagag aagttacaat
541 aataccaacc aagaagatct tttggtacta tgggggattc accatccaaa tgatgcggca
601 gagcagacaa ggctttatca aaacccaact acctatattt ccgttgggac atcaacacta
661 aaccagagat tggtaccaaa aatagctact agatctaagg taaacgagca aagtggaagg
721 atggagttct tttggacaat tttaaaatcg aatgatgcaa taaactttga gagtaatgga
781 aatttcattg ctccagaaaa tgcatacaaa attgtcaaga aaggggactc aacaattatg
841 aaaagtgagt tggaatatgg taactgcaac accaagtgtc agactccaat aggggcgata
901 aactccagta tgccattcca caacatccac cctctcacca tcggggaatg ccccaaatat
961 gtgaaatcaa acagattagt ccttgctact gggctcagaa atagccctca aggagagaga
1021 agaagaaaaa agagaggact atttggagct atagcaggtt ttatagaggg aggatggcag
1081 ggaatggtag atggttggta tgggtaccac catagcaacg agcaggggag tgggtacgct
1141 gcagacaaag aatccactca aaaggcaata gatggagtca ccaataaggt caactcgatc
1201 attgacaaaa tgaacactca gtttgaggct gttggaaggg aatttaataa cttagaaagg
1261 agaatagaaa atttaaacaa gaagatggaa gacggattcc tagatgtatg gacttataat
1321 gctgaacttc tggttctcat ggaaaatgag agaactctag actttcatga ctcaaatgtc
1381 aagaaccttt acgacaaggt ccgactacag cttagggata atgcaaagga gcttggtaac
1441 ggttgtttcg agttctatca cagatgtgat aatgaatgta tggaaagtgt aagaaacgga
1501 acgtatgact acccgcagta ttcagaagaa gcaagattaa aaagagagga aataagtgga
1561 gtaaaattgg agtcaatagg aacttaccaa atactgtcaa tttattcaac agtggcgagc
1621 tccctagcac tggcaatcat ggtggctggt ctatttttat ggatgtgctc caatggatcg
1681 ctacaatgca gaatttgcat ttaa

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G225E also noted previously in Kawaoka H5

http://jvi.asm.org/content/84/13/6570.full

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Glycosylation at 158N of the Hemagglutinin Protein and Receptor Binding Specificity Synergistically Affect the Antigenicity and Immunogenicity of a Live Attenuated H5N1 A/Vietnam/1203/2004 Vaccine Virus in Ferrets ▿
Weijia Wang1, Bin Lu1,†, Helen Zhou1, Amorsolo L. Suguitan Jr.1, Xing Cheng1, Kanta Subbarao2, George Kemble1 and Hong Jin1,*

As shown in Table 1, the most frequently isolated large-plaque isolates contained sequence changes at or near the HA receptor binding site; no mutations in the NA were detected in any of these virus isolates. Some of the changes occurred as early as passage 1. Several substitution mutations, E190D, T219N, K222E, G225E, and S227N (H3 numbering) (27, 34), were detected in H5 HA at a much higher frequency.

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Thread on CEIRS meeting in New York

viewtopic.php?f=5&t=8160

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