Research and Development Priorities for Emerging Infections and Biodefense

<richard.frothingham@duke.edu>

Influenza pandemic 1918-1919 20 million deaths

worldwide 549,000 deaths in

US Greatest pandemic

in world history?

HIV/ AIDS, 1981 to date

World Health Organization. 40 million persons living with HIV in 2006.

Marburg Hemorrhagic Fever

Outbreak in Northern Angola

Onset October 2004 Recognition March 2005 275 cases 255 deaths

SARS, 2003

Avian influenza, 2003 to 2007

World Health Organization. Confirmed human cases since 2003.

Anthrax attack, 2001

Anthrax spores delivered in US mail. 22 cases, 5 deaths.

Economic impact

European shares have extended sharp opening losses, after fears over the impact of the anthrax bacterium on consumer confidence sent US and Asian stock markets tumbling. . . . Observers blamed the drops on the growing unease surrounding the anthrax scares, which have raised fears of a "further chill" in US consumption.

BBC News, October 18, 2001

Economic impact WASHINGTON, March 15 - Health officials

believe that a mix-up of samples in a Defense Department contractor's laboratory was behind an anthrax scare Monday and Tuesday that rattled the stock market, set the White House on alert, shut three post offices in the Washington area and led to more than 800 people being offered antibiotics.

Scott Shane, New York Times, March 16, 2005

December, 2001: “Prices for gas masks and antibiotics grew by as much as 1000% in some areas, including New York and Washington.”

Biodefense.com?

Research priorities

NIAID Category A, B, and C Priority Pathogens

List found on NIAID web site

CDC bioterror list

CDC/ USDA Select Agent

List

NIAID priority pathogens

Category A Diseases/Agents High-priority agents include organisms that pose a risk to

national security because they:– can be easily disseminated or transmitted from person to person; – result in high mortality rates and have the potential for major public

health impact; – might cause public panic and social disruption; and

– require special action for public health preparedness. •Anthrax (Bacillus anthracis)•Botulism (Clostridium botulinum toxin)•Plague (Yersinia pestis)•Smallpox (variola major)•Tularemia (Francisella tularensis)•Viral hemorrhagic fevers (filoviruses [e.g., Ebola, Marburg] and

arenaviruses [e.g., Lassa, Machupo])

•Category B Diseases/Agents Second highest priority agents include those that

– are moderately easy to disseminate; – result in moderate morbidity rates and low mortality rates; and – require specific enhancements of CDC's diagnostic capacity and enhanced

disease surveillance. Brucellosis (Brucella species)•Epsilon toxin of Clostridium perfringens Food safety threats (e.g., Salmonella species, Escherichia coli O157:H7,

Shigella)•Glanders (Burkholderia mallei)•Melioidosis (Burkholderia pseudomallei)•Psittacosis (Chlamydia psittaci)

Q fever (Coxiella burnetii) Ricin toxin from Ricinus communis (castor beans) Staphylococcal enterotoxin B Typhus fever (Rickettsia prowazekii) Viral encephalitis (alphaviruses [e.g., Venezuelan equine encephalitis,

eastern equine encephalitis, western equine encephalitis]) Water safety threats (e.g., Vibrio cholerae, Cryptosporidium parvum)

•Category C Diseases/Agents Third highest priority agents include emerging pathogens that could be

engineered for mass dissemination in the future because of– availability;

– ease of production and dissemination; and

– potential for high morbidity and mortality rates and major health impact.

•Emerging infectious diseases such as Nipah virus and hantavirus

HSPD-18; Medical Countermeasures against Weapons of Mass Destruction, January, 2007

Biological Threats– Traditional agents—Natural (e.g., anthrax)– Enhanced agents—Traditional plus modification

or selection (e.g., XDR-TB, MDR-plague)– Emerging agents (e.g. SARS, avian influenza)– Advanced agents—Novel pathogen artificially

engineered in the laboratory (No example here)

Evolution in the NIAID biodefense program

Original Current

Drugs, diagnostics, and vaccines for Category A bioterror pathogens

Goals refined based on pathogenAlso high quality basic researchCategories A, B, and C

Biodefense and emerging infections

Emerging infections and biodefense

Pathogen-driven Addition: Antimicrobial researchAddition: Innate immunity

NIAID Strategic Plan for Biodefense Research

Broad spectrum activity Broad spectrum

technology Broad spectrum

platforms

Unique scientific paradigms

Potent toxins (botulinum toxin, anthrax) Low infectious doses (tularemia, TB, Q fever) High mortality rates (avian influenza, Ebola) Type three secretion system (plague) Persistence in host (TB) Persistence in environment (anthrax)

Mandate for RBLs Provide BSL3 containment to support work

with NIH priority pathogens, support RCE research programs, and support NIAID biodefense program

Be available and prepared to assist national, state, and local public health efforts in the event of a bioterrorism emergency

Existing 4-story Building

Entry/ Admin

BSL 2

BSL 3

Animal Housing/ Aerobiology

Loading Dock

GHRB timeline February 2003: Grant submission September 2003: Grant award May 2005: Groundbreaking November 17, 2006: Certificate of occupancy December 1, 2006: BSL2 labs open February 16, 2007: Ribbon-cutting

GHRB Ribbon Cutting Ceremony including Michael Kurilla, Brian Letourneau, Buck Lewis, Nancy Boyd, Bill Angus, Bill Bell, Bart Haynes, Mary Ann Black,

Victor Dzau, Rich Frothingham, Richard Broadhead, and Sandy Williams.

GHRB Timeline February to August, 2007: “Three-week”

commissioning August 19, 2007: BSL3 vivarium IACUC approval August 28 to Sept 11, 2007: 14-day countdown to

BSL3 certification visit October 14, 2007: BSL3 certification by Global

Biohazard Technologies (GBT) October 22 -26, 2007: BSL3 training by SERCEB

core from Emory (Sean Kaufman and Lee Alderman) January 28, 2008: CDC Select Agent amendment June 3-4, 2008: CDC Select Agent inspection

Take nothing for granted

– Check your ducts– Check your commissioning documents– Validate the ductwork– Validate the cage racks– Validate the filters– Confirm the sealed penetrations– Check your PPE

Note found among Tyvek suits

The thread is detached from the seam in the lower part of the trousers. If this happens again, we will return the job to you and punish you severely.

Existing cores located in GHRB SERCEB protein production core

– (Larry Liao, PI) SERCEB monoclonal antibody core

– (Larry Liao, PI) SERCEB viral vector core

– (Liz Ramsburg, PI) Duke immune reconstitution core

– (Greg Sempowski, PI) Duke BSL3 flow cytometry core

– (John Whitesides, PI) Duke Select Agent Program

– (Rich Frothingham, PI)

Cage Racks

New cores

SERCEB Aerobiology and Animal Models Core

SERCEB In vivo imaging core

Major equipment provided through SERCEB grant awards. Ongoing core support proposed in re-competition.

SERCEB Aerobiology and Animal Models Core

Madison

Shower

Class II BSC Class III BSC

Animal Holding Room

Autoclave

Animal Holding Room

Anteroom

Cage Rack

Madison chamber attached to a Class III glove box

Pass-through to a Class II BSC in an animal holding room

Two suites in the RBL vivarium.

Duke ID Aerobiology core practices

Detailed Standard Operating Procedures (SOPs) Preparation and confirmation of the inoculum Real-time recording of exposure parameters Characterization of the aerosol particle distribution Aerosol sampling to determine concentration of

viable microbes (estimate dose inhaled) Sentinel animal necropsy to define dose delivered

SERCEB BSL3 animal models core

The BSL3 Animal Models Core will provide animal challenge models using multiple pathogens, animal species, and routes of infection.

Animal challenge models will be established using GLP-like conditions to include detailed SOPs, inoculum confirmation, & sentinel animal necropsy.

Mice, rats, hamsters, gerbils, ferrets, guinea pigs, rabbits, others (up to 8 species at once)

70

75

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100

105

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

Days after challenge

Avg

. % O

rigin

al W

eigh

t

Compound X

Control

Weight, last value carried forward.

Immunomodulator (compound X) protects against lethal plague infection

Survival

0

20

40

60

80

100

120

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

Days after infection

% S

urvi

val

Compound X

Control

Survival, dose administered two days prior to infection.

Greg Hopkins, Eva Click

Intranasal challenge with Yersinia pestis

Protection by immune serum– Survival– Weight (surv only)

ED50– Mortality 1.3 μl– Weight 2.8 μl

Use bead technology to measure 23 cytokine levels using 16 μl serum

Vaccinated mice have minimal cytokine response to infection

Serum cytokine levels after plague challenge

Eva Click, Greg Hopkins , Jeff Hale, Greg Sempowski

Activation of CD8 T cells in spleen 3 days after intranasal plague challenge.– Increased expression of CD69, but not CD25– Whitesides flow cytometry core– Responding cell functional analysis, cloning, etc.– Bacterial sorting capacity

Greg Hopkins, John Whitesides

Models for interaction with RBL

Researcher sends material to RBL for testing in an animal model– Experimental design, IACUC approval,

administration, challenge, endpoints, analysis, Researcher comes to RBL at Duke, works

collaboratively with RBL core Researcher comes to RBL at Duke and works

independently

Grants (partial list) Duke Center for Translational Research

(Frothingham, P30 AI 51445) Regional Biocontainment Laboratory Construction

Grant (Williams, UC6 AI 58607) Southeast Regional Center for Emerging Infections

and Biodefense (Sparling, U54 AI 57157) Alternative endpoints for plague challenge models

(Frothingham, R21 AI 59689)

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Email: <richard.frothingham@duke.edu>