1. MAJ Jittawadee R. Murphy, PhD
Entomologists’ Role in the Clinical Trials of Investigational Malaria Vaccines and Drugs
MAJ Jittawadee R. Murphy, PhD
Department of Mosquito Biology
Division of Entomology
Walter Reed Army Institute of Research
Silver Spring, MD
DoD Pest Management Workshop, 9 FEB 10

2. Outline Human malaria challenge
Production of malaria infected mosquitoes
Preparation for the challenge
Challenge day
Challenge using imported infected mosquitoes
Summary
DoD Pest Management Workshop, 9 FEB 10

3. Human malaria challenge
Malaria Life Cycle
Background
R&D of anti-malarial vaccines and drugs
Evaluation of effectiveness
mouse, non-human primate model  human challenge
Clinical Trial Center (CTC) and Entomology
Scheduling is critical
CTC- recruit volunteers, Human Use Protocol, administer vaccine or drug, close contact with Ento for challenge
DoD Pest Management Workshop, 9 FEB 10

4. Plasmodium falciparum WT
Production of malaria infected mosquitoes
Axenic cultures of malaria parasites from asexual to sexual stages i. e. rings to gametocytes to infect mosq.
2. Mosquito colonization- Anopheles stephensi
Plasmodium falciparum WT
NF weeks
3D wks
DoD Pest Management Workshop, 9 FEB 10

5. Prod. of mal. infected mosquitoes (2 of 3)
3. Membrane feeding of mosquitoes with human blood/serum mixed with gametocyte cultures
4. Incubating infected mosquitoes (26°C, 80%RH)
DoD Pest Management Workshop, 9 FEB 10

6. Prod. of mal. infected mosquitoes (3 of 3)
Dissection of mosquitoes:
midguts for oocysts at 7 days post-feed
salivary glands for sporozoites at 14 days post-feed
Gold Standard sporozoite ratings: 0 -4
DoD Pest Management Workshop, 9 FEB 10

7. Preparation for the challenge
Select the highest infectious batches of mosquitoes- starve overnight
Preparation of challenge room
Volunteers’ seats
Timers, surgical towels
Volunteers’ sign-in sheets
Physician and crash cart space
Preparation of dissection room
Dissection scopes and tools
Dissection result forms
DoD Pest Management Workshop, 9 FEB 10

8. Challenge Day Objective – 5 infective bites/volunteer
Aspirate 5 mosquitoes / carton (pint-size)
Volunteers arrive, sign-in
Moderator, physician, dissectors
DoD Pest Management Workshop, 9 FEB 10

9. Challenge Day (2 of 2) Expose volunteer’s arm to bites for 5 minutes
Dissect for sporozoite ratings
Ratings 2-4 (11- >1,000 spz.) = infective bites
DoD Pest Management Workshop, 9 FEB 10

10. Challenge using imported infected mosquitoes
Plasmodium vivax – infected mosquitoes imported from Thailand
AFRIMS (Armed Forces Research Institute of Med.Sciences)
Hand carry infected mosquitoes
Permits: CDC, USDA APHIS, DOT
Airport & Airlines notified/approved in advance:
X-ray exemption at check-in, lay over
2. Courier arrival
DoD Pest Management Workshop, 9 FEB 10

11. Challenge using imported infected mosquitoes (2 of 2)
3. Mosquitoes: ideally at oocyst stage
Unpack- cartons in IATA-approved EXAKT packs,
in pelican case
Dissect, rate sporozoite infection
Select the highest infectious mosquitoes for challenge
Perform malaria human challenge

12. Performing Challenges Off Site
Challenges at: Oxford Univ., England,
U. of MD, Baltimore, Kenya?
Produce malaria-infected mosquitoes
Pack in 4-layered IATA-approved containers
Hand carry at sporozoite stage in pelican case
Permits- DOT, TSA, Airlines, X-ray exemption
DoD Pest Management Workshop, 9 FEB 10

13. Summary Entomologists play an important role from beginning to end
Challenging job for entomologists
Pressure for successful challenges
The opinions or assertions contained herein are the private views of the
author, and are not to be construed as official, or as reflecting true views
of the Department of the Army or the Department of Defense.
DoD Pest Management Workshop, 9 FEB 10

14. Acknowledgements WRAIR Mosquito Biology Dept.
Megan Dowler, Tatyana Savranskaya, Brooke Bozick, Michael Porter, Kyle Loughlin, Dr. Jackie Williams, and LTC Michael O’Neil
WRAIR Clinical Trial Center
USMMVP (US Military Malaria Vaccine Program)
AFRIMS Entomology Dept.
NMRC: CDR David Fryauff
DoD Pest Management Workshop, 9 FEB 10

15. Questions?
DoD Pest Management Workshop, 9 FEB 10