Emerging virus infections“in a changing world”

Animals as reservoirs of human viruses

Albert Osterhaus

Bruxelles June 2004

West Nile Yellow fever Dengue Avian influenza Lassa MeningitisJapanese encephalitis Ebola/Marburg Rift valley Polio Nipah SARS

Crimean-Congo HFVenez. Eq. encephalitis VHF HantaMonkeypox Enterovirus

Virus threats worldwide- 1996-2003, WHO outbreak reports -

Emerging virus infections in the last decennia- Part 1 -

Facilitated by changes in:

Social environment- Behaviour (taboos, mores, i.v. drug abuse)- Mobility (air travel, wars …)- Demography (population density, urbanisation…)- Public health measures (breakdown…)

Technology- Medical (transfusion, transplantation, vaccination)- Food production (recycling…)

Emerging virus infections in the last decennia- Part 2 -

Facilitated by changes in:

Ecology:- Animal contacts (wildlife, zoo, pets…)- Agriculture (deforestation…)- Fisheries (predator migration…)- Environmental pollution (immune suppression…)- Global warming (flora and fauna…)

Virus:- Mutation (host range, virulence…)- Recombination/reassortment

Bioterrorism?

Lentiviruses- Host range -

Influenza pandemics of the 20th century

Virus Subtype Name Est. # deaths

1918 H1N1 “Spanish Flu” > 40 million

1957 H2N2 “Asian Flu” > 1 million

1968 H3N2 “Hong Kong Flu” > 1 million

Bird markets- Asia -

Claas & Osterhaus, Nature Medicine 4: 1122-1123 (1998)

Human virusReassortant virus

Avian virus Avian virus

Influenza A virus- Man as a “mixing vessel” -

Highly pathogenic avian influenza (HPAI) seriously suspected clinically: February 28th 2003

Reports of conjunctivitis- By date of onset of symptoms -

Suspect A/H7 conjunctivitisInfluenza A-conjunctivitisConfirmed A/H7 conjunctivitis

Date of onset

June

12

Feb

28

0

5

10

15

20

25

# of

cas

es

Mar

7

Mar

14

Mar

21

Mar

28

Apr

4

Apr

11

Apr

18

Apr

25

May

2

May

9

May

16

May

23

May

30

Jun

6

Koopmans et al., Lancet 2004

H7N7 in The Netherlands- Fatal case chest X-ray upon admission -

X-ray fatal H7N7 infectionday of admission

Virus # substitutions inGene A/Netherlands/33/03 (conj.) A/Netherlands/219/03 (fatal case)

Segment Nucleotide Amino acid Nucleotide Amino acid1 (PB2) 2 0 8 5 (S79I,V297I,R355K,Q563R,E627K)2 (PB1) 0 0 2 03 (PA) 0 0 5 1(F666L)4 (HA) 0 0 4 3(I13S,A143T,K416R)5 (NP) 0 0 1 06 (NA) 0 0 5 4(N308S, A346V, T442A, P458S)7 (MA) 0 0 0 08 (NS) 1 1 (K126R)2 1 1(V137I)2

H7N7 in The Netherlands- Sequence analysis of full-length genomes -

1. Sequences were compared to those of A/Chicken/Netherlands/1/032. Both mutations in NS1

Fouchier et al., PNAS, 2004

?

?

H7N7 in The Netherlands- Chain of transmission -

LPAI

HPAI

Order Mononegavirales, family Paramyxoviridae

0.1

NiVHeV

TuV CDV RPVMV

hPIV3bPIV3

SeV

hPIV1

NDV

MuVSV5hPIV2

hMPVAPV

hRSV

bRSV

SV41

LPMV

Pneumovirinae

Paramyxovirinae Respirovirus

Rubulavirus

Morbillivirus

Metapneumovirus

Pneumovirus

DNA Maximum likelihood, Polymerase ORF

PDV

Avulavirus

Henipahvirus

- Pleiomorph

- Average size: 100-600 nm

- Envelope projections: 13-17 nm

- Paramyxovirus

Newly discovered paramyxovirus- Electron microscopy -

B.G. van den Hoogen et al.Nat. Med. 7:719-724, 2001

- (Young) children > 7-8 % of children with RTI?

- Immunocompromised individuals

- Elderly

- Normal individuals> 2-3 % of RTI in community surveillance studies

Human metapneumovirus- Risk groups -

Osterhaus and Fouchier, Lancet 2004

Severe acute respiratory syndrome (SARS)- A world-wide threat -

The same week: Hong Kong researchers announce fingings of Paramyxoviruses in SARS patients.Similar findings communicated from Canada through SARS etiology network

Novel coronavirus identified in SARS patients

SARS coronavirus phylogeny

P.Rota et al., Science, 2003

HCoV-NL: a fourth coronavirus of humans- phylogeny -

Full

geno

me

TM g

lyco

prot

ein

E

Rep

licas

e 1a

bM

atrix

Spik

eN

ucel

ocap

sid

Fouchier et al., PNAS 2004

SARS and the Koch’s postulates

Two viruses have been isolated from SARS patients: SCV and hMPV

Koch’s postulates modified for viruses by Rivers (1937):1. Isolation from diseased hosts

2. Cultivation in host cells

3. Proof of filterability

4. Production of comparable disease in host or related species

5. Reisolation of the virus

6. Detection of specific immune response

Cynomolgus monkeys (Macaca fascicularis)

Simultaneous SCV-hMPVinfection experiments:

•SCV alone•hMPV alone•SCV followed by hMPV

Fouchier et al.,Nature 2003

SARS coronavirus (SCV) in macaques

Histological lesions in lungs from cynomolgus macaques infected with SCV

Kuiken et al. The Lancet 2003

Time afterinoculation

(days)

02468

10121416

Throat

-++------

Nose

-++------

Faeces

N.T.N.T.

-------

IF

-N.T.N.T.N.T.N.T.

+N.T.N.T.

+

Throat

-++++----

Nose

-+++++---

Faeces

N.T.N.T.

+------

IF

-N.T.N.T.N.T.N.T.

+N.T.N.T.

+

Macaque # 3 Macaque # 4

SARS coronavirus in macaques

Exclusion of hMPV as the primary cause of SARS by macaque infection experiment

Koch’s postulates modified for viruses by Rivers:1.Isolation from diseased hosts2.Cultivation in host cells3.Proof of filterability4.No production of comparable disease in related species5.Reisolation of the virus6.Detection of specific immune response

SCV infected macaques subsequently inoculated with hMPV did not show disease exacerbation

Search for natural reservoir of SARS-CoV(Guan et al. 2003 Science)

111Raccoon dog

102Chinese ferret badger

366Himalayan palm civet

AntibodyPCR/

isolation

NSpecies

Hog badgers (3), beavers (3), domestic cats (3), Chinese hares (4), and Chinese muntjacs (2) all

tested negative.

Excretion of SCV in domestic cats and ferrets

SCV inoculated ferretsB

0 2 4 6 8 10 14 16 21 28

SCV exposed ferretsD

101

102

103

104

105

106

SCV inoculated catsA

N.D.

SCV

TCID

50/m

l

0 2 4 6 8 10 14 16 21 28101

102

103

104

105

106

SCV exposed catsC

N.D.

SCV

TCID

50/m

l

time (days)Martina et al. Nature, 2003.

Treatment of SARS-CoV infected macaqueswith pegylated interferon-α (Haagmans et al. 2004, Nature Medicine)

3 groups of cynomolgus macaquesControl (n = 4)Prophylactic (n = 4)Post-exposure (n = 4)

Treated with 3 mg/kg IFN, at-3, -1, +1, +3 dpi (prophylactic)+1, +3 (post-exposure)

Same infection protocol

Euthanasia at 4 dpi

Examination by:Pathology (gross, histology)ImmunohistochemistryRT-PCRVirus isolation

SARS-CoV excretion from pharynx

0

5 0

1 0 0

1 5 0

2 0 0

2 5 0

3 0 0

3 5 0

0 2 4 0 2 4 0 2 4 C o n tro l g ro u p

P ro p h yla c tic g ro u p

P o s t e xp o s u re g ro u p

D a y :* **

SC

V e

q/m

l

Haagmans et al., Nature Med. 2004

1 0 3

1 0 4

1 0 5

1 0 6

1 0 7

1 0 8

a

SCV

TCID

50/m

l

0

2 0

4 0

6 0

8 0

1 0 0b

SCV

infe

cted

cel

ls

(% S

CV

+m

icro

scop

ic fi

elds

)

0

1 0

2 0

3 0

4 0c

His

topa

thol

ogy

scor

e

C o n t r o l g r o u p

P o s t e x p o s u r e g r o u p

P r o p h y l a c t i c g r o u p

*

*

*

*

*

*

SARS-CoV replication, Ag-expression and histopathological lesions in lungs

Haagmans et al., Nature Med. 2004

A complex mix of socialsocial, technologicaltechnological, ecologicalecological and viralviral changes

Predisposes for the emergenceemergence of virus infections

Originating from animal reservoirsanimal reservoirs

Emerging virus infections- Conclusions 1 -

Extensive diagnosticdiagnostic and surveillancesurveillance networks

As well as novel vaccinevaccine- and antiviralantiviral development strategies

Should provide the safeguards to limit their impact

Emerging virus infections- Conclusions 2 -

Acknowledgements

Dept. Virology,Erasmus MC

Theo BestebroerBé NiemeyerGeorgina AronRobert Dias-d’UlloisGeert van AmerongenGerard van DoornumMartin SchuttenBert NiestersBart HaagmansByron MartinaGuus RimmelzwaanThijs KuikenRon FouchierAb Osterhaus

WHO & Members of the WHO SARS aetiology team

Centers for Disease Control & Prevention, Atlanta, USACentral Public Health Laboratory, London, UK

Public Health Laboratory Centre, Hongkong, SAR ChinaPrince of Wales Hospital, SAR China

Queen Mary Hospital, SAR ChinaSingapore General Hospital, Singapore

Federal Laboratories for Health Canada, Winnipeg, CanadaHealth Canada, Ottawa, Canada

Bernhard-Nocht Institute, Hamburg, GermanyInstitut Pasteur, Paris, France

National Institute of Infectious Disease, Tokyo, Japan

Emergingvirus infections

Department of Virology