EBOLA VIRUS

(1)

第 13 組：伊波拉病毒介紹

蔡昀哲

劉士銘

林新凱

陳彥君

何念青

孫義強

施懷勛

(2)

EBOLA VIRUS

^蔡昀哲

(3)

INTRODUCTION

Ebola is a rare but deadly infect

ion that causes bleeding inside an d outside the body.

Ebola strikes mainly in remote v

illages of Central and West Africa , but it has spread to some Africa n cities, too.

(4)

INTRODUCTION

EBOV is a select agent, World Health Organization Risk Group 4

Pathogen (requiring Biosafety Level 4-equivalent containment).

Biosafety level 1: canine hepatitis, non-pathogenic Escherichi

a coli

Biosafety level 2: HIV, MRSA, Salmonella

Biosafety level 3: Bacillus anthracis

, SARS coronavirus

(5)

INTRODUCTION

Biosafety level 4: Ebola virus, Marburg virus

(6)

STRUCTURE

Tubular in general, shepherd's crook or eyebolt, as a U or a 6, or co iled, circular, or branched

80 nm in diameter

800 nm in general, but may be up to 1000 nm long

In the center of the virion is formed by the helically wound viral

genomic RNA complexed with the proteins NP, VP35, VP30, and L.

(7)

STRUCTURE

Virally encoded glycoprotein (G

P) spikes 10 nm long and 10 nm apart are present on the outer viral envelope of the virion.

Between envelope and nucleocaps

id, in the so-called matrix spa ce, the viral proteins VP40 and VP24 are located.

(8)

STRUCTURE

(9)

GENOME

Each virion contains one molecule of linear, single-stranded,

negative-sense RNA, 18,959 to 18,961 nucleotides in length.

The 3′ terminus is not polyadenylated and the 5′ end is not

capped.

(10)

GENOME

The gene order is 3′ – leader – NP – VP35 – VP40 – GP/sGP –

VP30 – VP24 – L – trailer – 5′.

(11)

CLASSIFICATION - ZAIRE EBOLA VIRUS (ZEBOV)

highest case-fatality rate, up to 90% in some epidemics average case: 83% over 27 years

The symptoms resembled malaria.

Transmission has been attributed to reuse of unsterilized need

les and close personal contact.

(12)

CLASSIFICATION - SUDAN EBOL AVIRUS (SEBOV)

SEBOV emerged in 1976.

The most recent outbreak occurred in May, 2004.

The average fatality rates for SEBOV were 54%

(13)

CLASSIFICATION - RESTON EBOL AVIRUS (REBOV)

Discovered during an outbreak of simian hemorrhagic fever viru

s (SHFV) in crab-eating macaques from Hazleton Laboratories (n ow Covance) in 1989.

Despite its status as a Level 4 organism and its apparent path

‑

ogenicity in monkeys, REBOV did not cause disease in exposed h uman laboratory workers.

(14)

CLASSIFICATION - CÔTE D'IVOI RE EBOLAVIRUS (TAFV)

It was first discovered among chimpanzees from the Taï Forest

in Ivory Coast, Africa, in 1994.

Necropsies showed blood within the heart to be brown; no obvio

us marks were seen on the organs; and one necropsy showed lung s filled with blood.

One of the scientists performing the necropsies on the infect

ed chimpanzees contracted Ebola.

(15)

CLASSIFICATION: BUNDIBUGYO E

BOLAVIRUS (BDBV)

(16)

SIGNS & SYMPTOMS

Symptoms show up 2 to 21 days after infected.

Ebola damages immune system and organs.

Ebola causes platelets to fall, which can lead to severe bleed

ing.

(17)

SIGNS & SYMPTOMS

Early symptoms of Ebola look like flu, including:



Fever



Headache



Muscle aches



Sore throat



Weakness



Diarrhea

• As the disease gets worse:

• Bleeding inside and outside of the body

• Rash

• Trouble breathing

(18)

ENTRY

Niemann–Pick C1 (NPC1) appears to be essential for Ebola infection.

When cells from Niemann Pick Type C1 patients were exposed to Ebola viru

s in the laboratory, the cells survived and appeared immune to the virus , further indicating that Ebola relies on NPC1 to enter cells.

The same studies described similar results with Ebola's cousin in the fi

lovirus group, Marburg virus, showing that it too needs NPC1 to enter ce lls.

(19)

PATHOPHYSIOLOGY

Endothelial cells, mononuclear phagocytes, and hepatocytes are the main targets of infection.

Ebola replication overwhelms protein synthesis of infected cell s and host immune defenses.

The sGP forms a dimeric protein that interferes with the signal

ing of neutrophils, which allows the virus to evade the immune

system by inhibiting early steps of neutrophil activation.

(20)

PATHOPHYSIOLOGY

The loss in vascular integrity is

furthered with synthesis of GP, wh ich reduces specific integrins res ponsible for cell adhesion to the inter-cellular structure, and dama ge to the liver, which leads to co agulopathy.

(21)

TRANSMISSION

Ebola is introduced int

o the human population through close contact w ith the blood, secretio ns, organs or other bod ily fluids of infected animals.

(22)

TRANSMISSION

Ebola then spreads in the community through human-to-human tra

nsmission, with infection resulting from direct contact with b lood, secretions, organs or other bodily fluids of infected pe ople, and indirect contact with environments contaminated with such fluids.

Men who have recovered from the disease can still transmit the

virus through their semen for up to 7 weeks after recovery fro m illness.

(23)

TRANSMISSION

Burial ceremonies in which

mourners have direct conta ct with the body of the de ceased person can also pla y a role in the transmissi on of Ebola.

(24)

VACCINE AND TREATMENT

No licensed vaccine for EVD is available. Several vaccines are

being tested, but none are available for clinical use.

Patients are frequently dehydrated and require oral rehydratio

n with solutions containing electrolytes or intravenous fluids .

No specific treatment is available. New drug therapies are bei

ng evaluated

(25)

劉士銘

HISTORY AND

CASES

(26)

HISTORY

Zaire ebolavirus Sudan ebolavirus

Taï Forest ebolavirus (Côte d'Ivoire)

(27)

HISTORY

Zaire ebolavirus



August 1976, in Yambuku village



Name of river

 First case: Mabalo Lokela

(28)

HISTORY

Zaire ebolavirus

 Death rate: 1976 -> 88%, 1977 -> 100%, 1994 -> 59%, 1995 -> 81%, 1996 ->

93%, 2001~02 -> 80%, 2003 -> 90%, 2007 -> 83%



Over 55 neighbor villages, hundreds of people died (1976)



41 cases, 31 of them died (1994)



Total 1100 infected, 793 died

(29)

HISTORY

Sudan ebolavirus

 1976, in Nzara, Sudan



The most recent outbreak: May, 2004 in Yambio County

 First case: worker exposed to a potential natural reservoir

(30)

HISTORY

Sudan ebolavirus

 Death rate: 1976 -> 54%, 1979 -> 68%, 2000~01 -> 53%



The lack of barrier nursing (bedside isolation) facilitated the spread of the disease

(31)

HISTORY

Taï Forest ebolavirus (Côte d'Ivoire)

 1994, in the Tai Forest in Côte d'Ivoire

 First case: Chimpanzees -> scientist



similar to those of dengue fever



2 weeks -> discharged from hospital, 6 weeks -> fully recovered

(32)

OUTBREAK LOCATION

R. Congo 1976, 1977, 2007, 2008,

2012 R. Congo

1976, 1977, 2007, 2008,

2012

Sudan 1976, 1979,

2004 Sudan

1976, 1979,

2004

Uganda 2000, 2007, 2011,

2012 Uganda

2000, 2007, 2011,

2012 Guinea

2014 Guinea

2014

Côte d'Ivoire

1994 Côte d'Ivoire

1994

(33)

CASES

Movie: outbreak( 危機總動員 )

(34)

CASES

Republic of Uganda, 2000



10/12, appeared in Gulu



10/13: 30 death -> 10/27: 191 death (government)



12/8, administrator of hospital died

 Total 1 doctor, 12 nurses, 2 hospital workers died

(35)

CASES

Republic of Uganda, 2000



10/16, WHO, CDC intervened to investigate the cause of the source of dese ase



10/17, the government isolated the infected area by assigning soldiers to prevent patient leaving the infected area

(36)

SUMMARY

Like HIV, but more fatal

High death rate cause limited spreading speed

Only outbreak in region country, not globally

Intermittent, not continuous

(37)

EBOLA VIRUS

林新凱

CIES SPE

(38)

(39)

(40)

Ebola Virus

Bat MonkeyDuikers Antelope Chimpanzee

Human

Human Human Human Human Human

?

(41)

(42)

A leading suspect is fruit bats.

The little collared fruit bat, Myonycteris torquata, has a rang e that stretches as far west as Guinea.

(43)

Although bats may have carried the virus west from Central Afri ca, they may not be infecting humans directly.

No clear case of bat-to-human transmission of Ebola has ever be en proven.

(44)

(45)

Ebola Retrovirus

(46)

(47)

(48)

1989 Reston ebolavirus

crab-eating macaques

(49)

(50)

1994 Côte d'Ivoire ebolavirus

(51)

(52)

2009 Reston ebolavirus

Weingartl HM, Nfon C, Kobinger G (2013) Review of Ebola virus inf

ections in domestic animals. Dev Biol (Basel). 2013;135:211–218

(53)

IMMUNE PARAMETERS CORRELATE WITH PRO

TECTION

Speaker: 陳彥君

(54)

INTRODUCTION

Zaire ebolavirus (ZEBOV)

Immunoglobulin G (IgG)

ZEBOV glycoprotein (ZGP)

(55)

IMMUNOGLOBULIN G (IGG)

http://en.wikipedia.org/wiki/Immunoglob ulin_G

(56)

RESULTS

knockout mice guinea pigs

Humoral immune responses in NHPs Cellular immune responses in NHPs

Innate immune response in NHPs

(57)

RESULTS

Innate immune response in NHPs

(58)

KNOCKOUT MICE

Immune Parameters Correlate with Protection, Gary Wong et al., Science

(59)

KNOCKOUT MICE

(60)

RAG-1 & IFN-GAMMA

Rag-1 (Recombination-activating gene) IFN-gamma (Interferon)

http://en.wikipedia.org/wiki/Interferon_ga mma

(61)

CD8+ T cell

CD4+ T cell

B cell

(62)

RESULTS

Innate immune response in NHPs

(63)

GUINEA PIGS

(64)

GUINEA PIGS

(65)

GUINEA PIGS

(66)

RESULTS

Innate immune response in NHPs

(67)

(68)

(69)

(70)

DISCUSSION

A correlate of protection induced by vaccination is not necess

arily the same as the mechanism that eliminates infection.

Failure to generate ZGP-specific B cell–mediated immunity is

an indicator of mortality from ZEBOV challenge in mice.

(71)

MABS AND AD-VECTORED IFN-A THERAPY RESCUE EBOLA-INFECTED NONHU MAN PRIMATES WHEN AD MINISTERED AFTER THE DETECTION OF VIREMIA

AND SYMPTOMS

何念青

(72)

PREVIOUS WORK

ZMAb is a promising treatment against Ebola virus (EBOV) disea

se that has been shown to protect 50% (two of four) of nonhuma n primates (NHPs) when administered 2 days post-infection (dp i).

Ad-IFN has been developed as a broad spectrum antiviral and sh

own to enhance the EBOV-specific adaptive immune response as w ell as inhibit viral replication.

(73)

PREVIOUS WORK

Combination therapy with ZMAb and Ad-IFN was evaluated previou

sly in guinea pigs, where initiation of treatment at 3 dpi pro vided 100% survival. Early administration of Ad-IFN within 1 d pi permitted later mAb use up to 7 dpi with full survival, con stituting a significant improvement over either treatment alon e

(74)

PURPOSE

To develop and optimize a combination therapy, ZMAb with Ad-IF

N, that would be efficacious in NHPs after positive identifica tion of EBOV infection.

(75)

MATERIALS AND METHODS

Zaire ebolavirus

ZMAb, antibody therapy cocktails

Ad-IFN, a replication-deficient recombinant adenovirus serotyp

e 5 that drives the expression of human IFN-a

cynomolgus macaques, rhesus macaques

(76)

RESULTS

Exp. 1 : cynomolgus macaques

(77)

RESULTS

Exp. 2 : cynomolgus macaques

(78)

RESULTS

Exp. 3 : rhesus macaques

(79)

FUTURE WORK

Focus on a few selected optimal treatments with increase numbe

rs of NHPs

per group

(80)

THERAPEUTIC OF I NTERVENTION OF E BOLA

孫義強

(81)

THERAPEUTIC OF INTERVENTION OF EBOLA

Ebola cures with MB-003 combined of three Monoclonal Antibody.

Survived rate from 10% to 43% in NHPs

(82)

MB-003

EBOV glycoprotein epitopes 13C6

13F6

6D8

(83)

EXPERIMENT DESIGN

Phosphate-buffered saline (PBS) control The irrelevant mAb-treated control

MB-003

(84)

(85)

CLINICAL RESULT FROM DAYS 0

TO 28

(86)

SURVIVAL RATE

(87)

CLINICAL ANALYSIS

AND OBSERVATIONS

(88)

FDA-APPROVED SE LECTIVE ESTROGE N RECEPTOR MODU LATORS INHIBIT

EBOLA VIRUS INF ECTION

施懷勛

(89)

SERMS

Johansen et al

have found that FDA- proved selective estrogen receptor modulators (SERMs) could potentiall y be repurposed to treat Ebola viru s infection.

(90)

CLOMIPHENE & TOREMIFENE

(91)

VITRO TESTING

(92)

MICE TESTING

(93)

FUTURE

The approved drug status of clomiphene and toremifene may allo

w them to be rapidly developed for use with filovirus disease.

The oral availability for resource-constrained geographical re

gions