Post on 30-Jul-2015
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