Update on InfluenzaVirus Surveillance Findings in 2008/09

Season in Northern Hemisphere

Who Collaborating Center for Reference and Research on Influenza at Center for Influenza Virus Research, National Institute of Infectious Diseases,

Tokyo, Japan

Takato Odagiri, Ph.D.

Weekly report of virus isolation/detection in 2008-09 season

By NIID

By CDC

Pandemic A/H1N1 viruses

3584(32%)

2071(19%)

2475(22%)

2949(27%)

H1N1

H1N1pdm

H3N2

BAs of 11 Aug

248(11%)

201 (9%)

625(28%)

1186(52%)

H1N1

H1N1pdm

H3N2

B

As of week 30

A(H1N1) viruses September 2008-January 2009

H1N1, Sept 2008 – Jan 2009

H1N1 antigenic analysis

By NIID

H1N1 antigenic analysis

By CDC

Summary of antigenic analysis of A/H1N1 in four WHO CCs

A/Br/59/07-like low-reactors (≥ 8-f)

• CDC (Oct-) 179 [100%] 0 [0%]• NIID (Oct-) 72 [95%] 4 [5%]• NIMR (Oct-) 62 [87%] 9 [13%]• VIDRL (Oct-) 119 [97.5%] 3 [2.5%]• Total 432 [96%] 16 [4%]

March-July 2009

By CDC

By NIMR

Antigenic characterization of AH1N1 viruses isolated after March

By NIID

By CDC

By CDC

Number of oseltamivir resistance viruses isolated in Japan

（Oct 2008-May 2009）

By NIID

H1N1 viruses - SummaryOct 2008-Feb 2009• Influenza A(H1N1) viruses predominated in many Asian and North

American countries• The majority of viruses were antigenically closely related to

A/Brisbane/59/2007 by HI analysis• Phylogenetically the majority of H1N1 viruses grouped into the

A/Brisbane/59/2007 clade (2B) based on their HA gene. • Recent 2B viruses share H275Y indicative of resistance to

oseltamivir. They were antigenically similar to osel-sensitive viruses.• Recent viruses from China fall within the A/Hong Kong/2652/2006

clade (2C) and these viruses were similar antigenically to clade 2B viruses. Clade 2C viruses are sensitive to oseltamivir and resistant to amantadine/rimantadine

Mar - July 2009• The features of viruses were similar to those observed earlier in

2009.• Majority of recent A(H1N1) viruses were antigenically closely related

to A/Brisbane/59/2007 and formed clade 2B sharing H275Y.• In the few clade 2C viruses, dual resistance to oseltamivir and

amantadine/rimantadine were detected.

A(H3N2) viruses September 2008-January 2009

H3N2, Sept 2008 – Jan 2009

By NIID

Table 4B. Antigenic analyses of influenza A H3N2 viruses - Guinea Pig RBCs (05/02/2009)

Viruses Collection A/Wis A/Trieste A/Wis A/Bris A/Uru A/Fin A/JHB A/Eng A/BrisDate 67/05 25c/07 3/07 10/07 716/07 9/08 15/08 394/08 24/08

F18/08 F5/07 F15/07 F29/08 F26/08 F12/08 F22/08 F32/08 F3/09

REFERENCE VIRUSES

A/Wisconsin/67/2005 31/08/2005 SpfCk3E3 \3 1280 1280 640 1280 2560 640 640 80 1280A/Trieste/25c/2007 Jan-07 MDCKx \2 80 160 160 320 160 80 160 160 160A/Wisconsin/3/2007 21/01/2007 Ex \2 5120 5120 2560 5120 5120 2560 2560 640 5120A/Brisbane/10/2007 06/02/2007 E2 \3 1280 2560 640 1280 2560 1280 1280 320 2560A/Uruguay/716/2007 21/06/2007 spfck1, E3 \2 1280 2560 640 1280 2560 2560 1280 320 1280A/Finland/9/2008 07/01/2008 MDCK2 \1 160 160 320 320 320 320 320 320 320A/Johannesburg/15/2008 25/06/2008 MDCKx \2 160 320 320 320 320 320 320 320 640A/England/394/2008 22/09/2008 MDCK2 \7 160 320 320 320 640 160 320 640 320A/Brisbane/24/2008 23/06/2008 E5 \1 2560 2560 2560 5120 5120 2560 2560 320 5120

TEST VIRUSES

A/Singapore/N474/2008 21/05/2008 Cx \1 1280 2560 1280 2560 5120 1280 2560 640 2560A/Singapore/N486/2008 21/05/2008 Cx \1 640 1280 320 1280 1280 640 640 320 640A/Singapore/N510/2008 04/06/2008 Cx \2 1280 2560 1280 2560 5120 2560 2560 640 1280A/Singapore/N593/2008 15/09/2008 Cx \1 1280 2560 640 1280 2560 1280 1280 640 2560A/Sweden/3/08 15/10/2008 MDCK2 \1 1280 1280 640 1280 1280 1280 1280 320 640A/England/694/2008 05/12/2008 SIAT P1 \1 1280 1280 640 1280 2560 640 1280 640 640A/England/687/2008 30/12/2008 MDCK P1 \2 640 1280 320 1280 2560 1280 1280 320 640A/Switzerland/1370948/2008 12/12/2008 MDCK2 \2 640 640 320 640 1280 320 320 160 320A/Seoul/4436/2008 05/12/2008 MDCK-P2 \1 640 640 320 640 1280 640 640 320 640A/Slovenia/60/2009 06/01/2009 MDCKx \1 640 640 320 640 1280 640 640 160 640A/Slovenia/51/2009 05/01/2009 MDCKx \1 160 80 160 160 320 160 160 160 160A/Baden-Württemburg/199/2008 15/12/2008 C4 \2 320 320 160 320 640 160 320 160 320A/Parma/17/09 12/01/2009 I MDCK \1 80 80 160 160 160 80 80 160 160A/Singapore/N565/2008 12/08/2008 Cx \2 1280 1280 320 1280 1280 640 1280 320 640A/Switzerland/1409256/2008 29/12/2008 cs \2 1280 1280 320 1280 1280 640 640 160 640A/Switzerland/1409281/2008 31/12/2008 cs \2 640 1280 320 640 1280 320 640 160 640A/Leon/5/2009 13/01/2009 MDCK1 \1 1280 2560 640 1280 2560 1280 2560 640 640A/Belgium/G211/2009 30/12/2008 MDCK1 \1 1280 2560 640 1280 2560 1280 1280 640 640A/Belgium/G228/2009 30/12/2008 MDCK1 \1 1280 1280 320 1280 1280 640 1280 320 640A/Belgium/G278/2009 08/01/2009 MDCK1 \1 1280 2560 640 1280 2560 1280 1280 640 640A/Belgium/G298/2009 12/01/2009 MDCK1 \1 1280 2560 640 1280 2560 1280 2560 640 640

1. < = <40 Vaccine strains

Passage History

Haemagglutination inhibition titre1

Post infection ferret sera

By NIMR

Table 5B. Antigenic analysis of influenza A H3N2 viruses (Neutralisation)

Viruses Collection A/Well A/Bris A/Uru A/Eng A/BrisDate 1/04 10/07 716/07 394/08 24/08

F13/04 F18/07 F3/07 F32/08 F3/09

REFERENCE VIRUSES

A/Wellington/1/2004 26/01/2004 E3 \1 1280 320 320 320 320A/Brisbane/10/2007 06/02/2007 E2 \3 320 2560 5120 320 2560A/Uruguay/716/2007 21/06/2007 spfck1, E3 \1 320 1280 2560 160 1280A/England/394/2008 22/09/2008 TC P2 \1 80 2560 2560 2560 1280A/Brisbane/24/08 23/06/2008 E5 160 1280 2560 320 2560

TEST VIRUSES

A/Paris/130/2008 20/10/2008 C2 \1 160 640 1280 320 1280A/Egypt/59/2008 16/11/2008 C1 \1 160 1280 1280 320 1280A/Toulouse/1317/2008 14/11/2008 P3MDCK \1 160 1280 2560 640 2560A/Norway/2308/2008 15/12/2008 MDCK1 \1 320 2560 2560 640 2560A/Denmark/191/2008 13/11/2008 MDCK3 \1 160 1280 2560 320 2560A/Sweden/6/2008 08/12/2008 MDCK2 \1 160 1280 1280 320 1280A/Paris/458/2008 1/12/2008 C1 \1 320 2560 5120 640 5120A/HongKong/3197/2008 26/11/2008 MDCK2 \1 160 1280 1280 320 1280A/England/669/2008 30/12/2008 MDCK P1 \1 80 1280 1280 320 1280A/Norway/1/2009 30/12/2008 MDCK1 \1 160 640 1280 320 1280

1 Based on 50% plaque reduction compared to serum negative controls2 RDE treatedAgglutinate guinea pig RBCs only

Vaccine strains

Passage History

Neutralisation titre1

Post-infection ferret sera2

By NIMR

A/Brisbane/10/07-like low-reactors (≥ 8-f)

• CDC (Oct-) 45 [90%] 5 [10%]

• NIID (Oct-) 32 [56%] 25 [44%]

• NIMR (Oct-) 160 [90%] 18 [10%]*

• VIDRL (Oct-) 145 [93%] 11 [ 7%]

• Total 382 [87%] 59 [13%]

H3 low reactors in HI assays in WHOCCs

*0% by virus neutralization

Vaccine virus

March-July 2009

By NIMR

By NIID

Vaccine virus

By Cambridge Univ.

H3N2 viruses - Summary Oct 2008-Feb 2009• H3N2 viruses co-circulated with H1N1 and B viruses in many countries• Majority were antigenically similar to A/Brisbane/10/07 and

A/Uruguay/716/07 by HI tests with turkey RBCs or guinea pig RBCs.• The ‘low reactors’ were antigenically indistinguishable from vaccine

viruses by Neutralization tests. • HA and NA sequences fairly homogeneous and fell within the

A/Brisbane/10/07 clade • M2 sequences possessed S31N mutation, indicative of amantadine

resistance• Sensitive to oseltamivir and zanamivirMar - July 2009• Majority were antigenically similar to vaccine viruses.• An increasing number of H3N2 isolates have been shown to be

antigenically distinct from vaccine viruses and related to A/Hawaii/7/09, A/HK/1985/09 and A/Perth/16/09.

• The majority of HA sequences of those viruses formed a separate clade characterized by aa subsitutions E62K, N144K, K158N and N18K.

• They retained resistance to amantadine/rimantadine and were sensitive to oseltamivir and zanamivir.

B viruses September 2008-January 2009

B, Sept 2008 – Jan 2009

Numbers of 2 lineages of B viruses detected in GISN since week 40

2008

0

5

10

15

20

25

30

35

40

45

40 41 42 43 44 45 46 47 48 49 50 51 52 1 2 3 4

week #

B-Y B-V

B-Victoria lineage

B-Yamagata lineage

HI reactions of influenza type B viruses

By NIID

HI reactions of influenza type B virusesB/Victoria-lineage

By NIMR

HI reactions of influenza type B viruses B/Yamagata lineageTable 7. Antigenic analyses of influenza B viruses (Yamagata lineage)

Viruses Collection B/Fl3 B/Eg B/Fl B/Bris B/Barc2 B/Eng2 B/Vall2 B/Ban B/Indi

date 4/06 144/05 4/06 3/07 143/08 145/08 18/08 3333/07 1/08Sh 478 F3/07 F19/07 F24/07 F11/08 F9/08 F10/08 F21/08 CDC174/08

REFERENCE VIRUSES

B/Egypt/144/2005 01/05/2005 E2 \2 2560 160 320 640 10 40 20 160 1280B/Florida/4/2006 15/12/2006 E2 \1 5120 160 640 640 10 80 20 160 1280B/Brisbane/3/2007 03/09/2007 E2 \2 5120 160 640 640 10 80 20 160 1280B/Barcelona/143/2008 06/01/2008 MDCK1 \2 5120 80 160 320 320 160 320 320 640B/England/145/2008 Ex \2 320 40 < 40 10 160 10 20 640B/Valladolid/18/2008 18/02/2008 MDCK1 \2 2560 80 160 80 160 160 320 160 1280B/Bangladesh/3333/2007 19/08/2007 E4 \1 2560 80 160 160 20 40 20 160 1280B/Indiana/1/2008 10/03/2008 E2 \1 1280 40 80 160 20 40 20 320 640

TEST VIRUSES

B/Mendoza/R530/2008 Aug-08 M2 /1 5120 160 160 320 160 160 160 320 NDB/Ghana/MH-31/2008 19/08/2008 MDCKx /1 2560 160 160 320 160 160 320 320 NDB/Dakar/06/2008 19/08/2008 MDCK2 /1 5120 160 640 640 320 640 320 640 NDB/Ghana/MH-36/2008 29/08/2008 MDCKx /1 2560 160 160 320 160 160 320 320 NDB/Ghana/NG-PK-01/2008 09/09/2008 MDCKx /1 5120 160 160 160 320 320 320 320 NDB/Hong Kong/1355/2008 09/09/2008 MDCK2 \2 2560 80 160 160 160 160 160 320 NDB/Ghana/MH-41/2008 12/09/2008 MDCKx /1 2560 80 40 40 80 160 160 160 NDB/Hong Kong/1372/2008 25/09/2008 MDCK2 \1 5120 160 160 160 160 320 160 160 640B/Ghana/FS301/2008 09/10/2008 MDCK2 \1 5120 80 160 320 320 160 320 320 640B/Hong Kong/1374/2008 17/10/2008 MDCK2 \1 2560 80 160 320 160 160 160 160 640B/Ghana/FS347/2008 31/10/2008 MDCK2 \1 2560 80 80 80 160 160 320 160 1280B/Ghana/FS351/2008 31/10/2008 MDCK2 \1 5120 80 160 80 320 160 320 320 640B/Paris/236/2008 03/11/2008 C2 \1 640 40 < < 10 80 40 40 NDB/Ghana/FS355/2008 04/11/2008 MDCK2 \2 640 < < < 80 80 160 80 NDB/Ghana/FS383/2008 17/11/2008 MDCK2 \1 5120 160 320 640 640 320 640 640 1280B/Ghana/FS385/2008 18/11/2008 MDCK2 \1 5120 160 160 160 320 320 320 320 1280B/Ghana/FS428/2008 09/12/2008 MDCK2 \1 5120 80 80 80 160 160 320 160 640B/Ghana/FS433/2008 11/12/2008 MDCK2 \3 2560 40 80 80 160 160 160 160 NDB/Ghana/FS435/2008 12/12/2008 MDCK2 \3 2560 40 80 80 160 80 160 160 NDB/Ghana/FS388/2008 18/11/2008 MDCK2 \2 2560 80 80 80 160 160 320 160 NDB/Switzerland/1334249/2008 04/12/2008 MDCK1 \1 2560 80 160 320 320 160 320 320 NDB/Egypt/133/2009 14/01/2009 Ex \1 160 < < < < 20 < 10 ND

1. < = <40; 2. < = <10; 3. hyperimmune sheep serum

ND = not done

Vaccine strains

Haemagglutination inhibition titre1

Passage

History

Post infection ferret sera

By NIMR

Victoria Yamagata• CDC (Oct-) 71 [60%] 48 [40%]

low reactors (77%) (23%)• NIID (Oct-) 18 [40%] 27[60%]

Low reactors (94%) (67%)• NIMR 17 [44%] 22[56%]

Low reactors (76%) (14%)VIDRL (Oct-) 340 [67%] 167 [33%]

Low reactors (94%) (62%)

Total 446 [63%] 264 [37%]Low reactors (91%) (51%)

B low reactors in HI assays in WHOCCs

10 nucleotides

B/Brisbane/60/2008-like

Evolutionary Relationships Among Influenza B (Victoria Lineage) HA Genes

B/Malaysia/2506/2004

K165N

Chinese clade

By Melbourne C.

Evolutionary Relationships Among Influenza B (Yamagata Lineage) HA Genes

10 nucleotides By Melbourne C.

March-July 2009

By NIMR

HI reactions of influenza type B virusesB/Victoria-lineage

By CDC

By Cambridge Univ.

B/Victoria-lineage HA

By NIID

Vaccine virus

Chinese viruses

By CDC

B/Yamagata-lineage HA

Vaccine virus

Summary for influenza B virusesOct 2008-Feb 2009

• Influenza B viruses circulated in many countries however regional outbreaks were limited.

• Viruses of both B/Victoria/2/87 and B/Yamagata/16/88 lineages continued to co-circulate in many countries

• B/Victoria-lineage viruses predominated in most countries and were mostly antigenically closely related to B/Brisbane/60/2008.

• Viruses mainly from China fell into a separate phylogenetic clade

• B/Yamagata-lineage viruses were antigenically closely related to B/Florida/4/2006 & B/Brisbane/3/2007 vaccine viruses.

• Three phylogenetic clades were apparent but were antigenically indistinguishable.

Mar - July 2009

• B/Victoria-lineage viruses predominated and were mostly antigenically closely related to vaccine strain B/Brisbane/60/2008.

• Many viruses isolated in China were closely related to B/Fujian Gulou/1272/08 and B/Hubei-Songzi/51/08.

• B/Yamagata-lineage viruses were antigenically closely related to B/Florida/4/2006 and mostly fell into their HA sequences of the B/Bagladesh/3333/07 clade.

WHO Collaborating Centers

(US-CDC, NIMR, Melbourne)

Cambridge University

NICs

(China-CDC, Korea-CDC, NHL-Myanmar,

NCLE-Lao PDR, NIH-Mongolia and other NICs of WHO GISN)

Taiwan-CDC

Local PHLs in Japan

Acknowledgement