Adolfo García-SastreIcahn School of Medicine at Mount Sinai, New YorkCRIP, Center for Research on Influenza Pathogenesis

INFLUENZAEPIDEMICS PANDEMICS

YoshiKawaoka

TakeshiNoda

H1N1

INFLUENZA VIRUSES

PAx

‘40 ‘50 ‘60 ‘70 ‘80 ‘00‘901918

H1N1

H2N2

H1N1

H3N2

A

B

EPIDEMIOLOGY OF HUMAN INFLUENZA VIRUSES

‘10

1957

1968

1977

pH1N1

2009

INFECTIONS IN HUMANS WITH AVIAN AND SWINE INFLUENZA A VIRUSES

H5N1 H9N2 H5N1 H5N1H7N7

‘40 ‘50 ‘60 ‘70 ‘80 ‘00‘901918

H1N1

H2N2

H1N1

H3N2

A

B

‘10

1957

1968

1977

pH1N1

2009

H7N9H3N2v

Evolution and spread of flu viruses

aquatic birds

poultry

pigs

humans

horsesfecal/oral

respiratory

dogs

cats

H1N11918

Alicia SolórzanoAdolfo García-Sastre

Patty AguilarChris Basler

Peter Palese

Mount Sinai

Terry TumpeyHui Zeng

Nancy CoxJacky Katz

CDC

David SwayneUSDA

Jeff TaubenbergerAFIP

U.S. Life ExpectancyBy age

70

60

50

40

30

1900 ‘30 ‘50 ‘70 ‘90

Lung tissue samples (1918)

1918 influenza AFIP lung block

Extract RNA, sequence, clone the virus

Gene sequencing

Gene reconstruction

Pathological specimen (circa 1918). . . . .

Reverse genetics

Signatures of virulence of the 1918 influenza virus

Phenotypic characterization in: Tissue culture Animal models

Influenza A/CDC/1918 virus

0

20

40

60

80

100

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

% s

urv

ival

Days after infection

Tx/91

1918 5:3 Tx/911918

1918 7:HA Tx/91

Intranasal inoculation of mice, 106 pfuViral titers in lungs, day 4

103 pfu

106 pfu108 pfu

105 pfu

1918 VIRUSWhat do we know now?

1. The 1918 virus is the only known human influenza virus lethal to mice, ferrets, macaques and embryonated eggs

2. The glycoprotein (HA and NA) and non-structural (NS1 and PB1-F2 genes) of the virus contribute to enhanced virulence

3. Viruses containing 1918 genes are sensitive to existing antivirals

4. H1N1 based vaccines are protective

Would a 1918-like HIN1 virus be today as lethal as in 1918?

H1N12009

Rafa MedinaBalaji ManicassamyEstanis Nistal-VillánAdolfo García-Sastre

Tshide TsibaneChris Basler

Silke StertzPeter Palese

Mount Sinai

Petra ZimmermannOsvaldo Zagordi

Silke StertzUniversity of Zurich

Hanni Uusi-KerttulaRafa Medina

Universidad Católica de Chile

Xiangjie SunTerry Tumpey

CDC

Sharon FreyBob Belshe

SLU

‘40 ‘50 ‘60 ‘70 ‘80 ‘00‘901918

H1N1

H2N2

H1N1

H3N2

A

B

EPIDEMIOLOGY OF HUMAN INFLUENZA VIRUSES

‘10

1957

1968

1977

pH1N1

2009

SWINE FLU

Human H3N2

Swine H1N1

Avian virus

Human pandemic H1N1

NO 1918 PB1-F2

1918 “Spanish” flu

Pigs Humans

Classical swine flu Modern human H1N1

Sa

Ca2

Ca1

Cb

Receptor binding site

Sb

Fusion peptide

Antigenic sites

Pandemic H1N1 infections in humans

• Infections are primarily seen in children and young adults

• Serology studies show the presence of neutralizing antibodies against 2009 H1N1 virus in people older than 65 yrs

Due to prior exposure to an H1 virus similar to 2009 H1N1?

Do antibodies to any specific H1N1 virus protect against SOIV 2009 H1N1?

Virus Type Year (lineage)

A/California/04/09 (6:2) H1N1 2009

1918-like or classical H1N1

1918 VLP H1N1 1918

A/swine/Iowa/30 H1N1 1930 (Classical)

A/Weiss/43 H1N1 1943 (1918-like)

A/New Jersey/8/76 H1N1 1976 (Classical?)

Human H1N1(1977-2007)

A/USSR/92/77 H1N1 1977

A/Houston/20593/84 H1N1 1984

A/Texas/36/91 H1N1 1991

A/Brisbane/59/07 H1N1 2007

Control (H3N2)

A/NT/60/68 H3N2 1968

A/Brisbane/10/07 H3N2 2007

Vaccination and challenge experiment

Vaccination15mg

-28

Boost15mg

Challenge (50 LD50)

-14

Body weight & Survival

0Day

- 5 week old female C57B/6 mice- Mice were vaccinated with 11 different inactivated viruses- Challenged with Neth/09 strain

14

Inactivated vaccines based in classical swine viruses (1930-2009) and in human H1N1 viruses (1918-1943) protect against lethal infection with the new H1N1

New H1N1

Seasonal H1N1

1918

1918New H1N1

Seasonal H1N1

Sa

Conclusions

-People carrying antibodies against H1N1 viruses that circulated 1918-1950 and A/NJ/76 (vaccination) are likely to be protected

- Pigs act as reservoirs for strains that become antigenically “frozen” H3 and H1 viruses have also established lineages in pigs in 1997-1998 and 2003-2005. Swine H3N2v jumps frequently to humans.

Sample AgeCal/09 1918 Vaccinated with

Pre PrePost Post 2009 H1N1 Seasonal TIV

123456789

101112131415

192133343442434955606767825174

<10<10<10<10<1010 <10<1010

<101010 <10<10<10

801280

802560

80320 32040

12801280640640 2560<1010

10<1020

<10<10<10 20

<10<1010

<1080 <10<1010

201280

402560

80320 64020

128012801280640 640<1040

YesYesYesYesYesYesYesYesYesYesYesYesYesNoNo

YesYesNoYesYesYesYesYesYesYesYesYesNoYesYes

New H1N1 vaccines induce 1918 HI antibodies in humans

In collaboration with Bob Belshe

Passive administration of sera from new H1N1 vaccinated humans

In collaboration with Bob Belshe

Passive administration of sera from new H1N1 vaccinated humans

In collaboration with Bob Belshe

Vaccination (or exposure) to new H1N1 virus generates protective immunity against 1918 virus

AVIAN FLU

HIGHLY PATHOGENICAVIAN INFLUENZA VIRUSES

LOW PATHOGENIC H5N1 VIRUSES

H5N1 ORIGIN

LOW PATHOGENIC H5N1 VIRUSES

HIGLY PATHOGENIC H5N1 VIRUSES

Mutation

H7N9

India

Nepal

Russia

Mongolia

Bangladesh

Bhutan

Burma

Thailand

Laos

0

0

500 Kilometers

500 Miles

Vietnam

Philippines

SouthKorea

NorthKorea

Pakistan

INNER MONGOLIA

HENAN

ANHUI

ZHEJIANG

SHANGHAI

JIANGSU

FUJIAN

JIANGXI

HUBEI

SHANXI SHANDONG

HEBEI

BEIJING

TIANJIN

LIAONING

Taiwan

Location of H7N9 Influenza in China (Feb-Oct 2013)

SHAANXI

137 total cases/45deaths

Province/City

Number of Cases

Anhui 4

Beijing 2

Fujian 5

Guangdong 1

Guangxi 0

Hebei 1

Henan 4

Hunan 2

Jiangsu 28

Jiangxi 6

Jilin 0

Shandong 2

Shanghai 33

Zhejiang 48

GUANGDONG

HUNAN

GUANGXI

CHONGQING

GUIZHOU

NINGXIA

GANSU

SICHUAN

YUNNAN

JILIN

HEILONGJIANG

QINGHAI

XIZANG

XINJIANG

HAINAN

Source WHOAdapted from

Taiwan 1

India

Nepal

Russia

Mongolia

Bangladesh

Bhutan

Burma

Thailand

Laos

0

0

500 Kilometers

500 Miles

Vietnam

Philippines

SouthKorea

NorthKorea

Pakistan

INNER MONGOLIA

HENAN

ANHUI

ZHEJIANG

SHANGHAI

JIANGSU

FUJIAN

JIANGXI

HUBEI

SHANXI SHANDONG

HEBEI

BEIJING

TIANJIN

LIAONING

Taiwan

Location of H7N9 Influenza in China (8/4/14)

SHAANXI

452 total cases/124deathsProvince/City

Number of Cases

Anhui 18

Beijing 5

Fujian 22

Guangdong 118

Guangxi 3

Hebei 1

Henan 4

Hunan 24

Jiangsu 61

Jiangxi 8

Jilin 2

Shandong 4

Shanghai 42

Zhejiang 140

GUANGDONG

HUNAN

GUANGXI

CHONGQING

GUIZHOU

NINGXIA

GANSU

SICHUAN

YUNNAN

JILIN

HEILONGJIANG

QINGHAI

XIZANG

XINJIANG

HAINAN

Source WHO, flutrackers, newsAdapted from

Taiwan 1

Cases Deceased

Cases Alive

Source: WHO, Flutrackers, news reports

There have now been 9 human cases from Guangdong Province over the past 7 days.

Confirmed human cases of H5N1(2003-2013)

Country Infections Deaths

H7N9 virus transmission

Source: CDC ILI and Vaccine Distribution Data

Pandemic H1N1 cases and vaccinations in US Sept 2009 – May 2010

0

1

2

3

4

5

6

7

8

9

0

20,000,000

40,000,000

60,000,000

80,000,000

100,000,000

120,000,000

140,000,000

ILI

Shipped Vaccine

% o

f V

isit

s fo

r IL

I

Nu

mb

er o

f H

1N1

Vac

cin

e S

hip

ped

Universal flu vaccines?

Neutralization of influenza viruses

Nat Struct Mol Biol. 2009 Mar;16(3):233-4.

HA1

HA2

Neutralizing antibodies

Hemagglutinin subtypes

UNIVERSAL FLU VACCINES?

Repeated vaccination with influenza virus chimeric HA vaccines induce protective antibodies against multiple subtypes of influenza virus.

Irina Margine Randy AlbrechtFlorian KrammerRong Hai Patrick WilsonGene Tan S.A. Andrews

Peter Palese Jon Runstadler

cH4/3 DNA cH5/3 proteinboost

H3 proteinboost

Shanghai (H7N9)challenge

Control groups: cH4/3 DNA + BSA + BSAnaïve (neg. contr.)matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Induction of protective levels of stalk-reactive antibodies using chimeric HA constructs in

mice

cH4/3 DNA cH5/3 proteinboost

H3 proteinboost

Shanghai (H7N9)challenge

Control groups: cH4/3 DNA + BSA + BSAnaïve (neg. contr.)matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Induction of protective levels of stalk-reactive antibodies using chimeric HA constructs in mice

YY

YYY

cH4/3 DNA cH5/3 proteinboost

H3 proteinboost

Shanghai (H7N9)challenge

Control groups: cH4/3 DNA + BSA + BSAnaïve (neg. contr.)matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Induction of protective levels of stalk-reactive antibodies using chimeric HA constructs in mice

YY

YYYY

YY

Y

YYY

cH4/3 DNA cH5/3 proteinboost

H3 proteinboost

Shanghai (H7N9)challenge

Control groups: cH4/3 DNA + BSA + BSAnaïve (neg. contr.)matched vaccine (pos. contr.)

4 weeks3 weeks3 weeks

Induction of protective levels of stalk-reactive antibodies using chimeric HA constructs in mice

YY

YYYY

YY

Y

YYY

YY Y

YY

YYY

YY

YYY

cHA vaccine protects against challenge with novel H7N9 virus

cHA vaccine protects against challenge with H10 and H3 viruses

cH4/3 DNA + cH5/3 protein + H3 protein cH4/3 DNA + cH5/3 protein + cH7/3 protein

Titers in mouse lungs, day 3 postinfection

Abs mediate protection

H3N2

Targeting group 1 HA viruses

cH9/1 DNA cH6/1 protein cH5/1 protein

Control groups: cH9/1 DNA + BSA + BSAmatched vaccine (pos. contr.)

YY

YY

YY

YY Y

YY

YYY

YYYY

YY

YYYYYY

Induction of protective levels of stalk-reactive antibodies using chimeric HA constructs in

mice

PR8 H1N1FM1 H1N1pH1N1H5N1H6N1challenge

Vaccination with cHA constructs protects from pH1N1

(A/Netherlands/602/09) challenge

positive control (matched inactivated)cH9/1 DNA + cH6/1 protein + cH5/1 proteincH9/1 DNA + BSA +BSA

Similar results for A/PR/8/34 H1N1 and A/FM/1/47 challenges

positive control (matched inactivated)cH9/1 DNA + H1 protein/cH6/1 protein + cH5/1 protein/H1 proteincH9/1 DNA + BSA +BSA

cHA constructs protect mice from heterosubtypic challenge

H5N1 challenge H6N1 challenge

cH5/1 (H5 challenge) or cH6/1 (H6 challenge) protein was replaced by full length H1 protein to exclude head-based protection

ELISA reactivity to Cal09 (pH1N1) protein

Protection is antibody mediated

cH9/1 + cH6/1 + cH5/1cH9/1 + BSA +BSAnaïve serum

NaïvePositive controlvector +BSA+BSAcH9/1 + cH6/1 + cH5/1

Passive transfer of serum protects from viral challenge

Days post challenge

cHA constructs protect ferrets from pH1N1 challenge

***

***ns

FLU, WHAT IS NEEDED?

SURVEILLANCE, ERRADICATION

NEW BROAD SPECTRUM ANTIVIRALS

BROADLY NEUTRALIZING VACCINES

TRAINING OF FUTURE INVESTIGATORS