Novel Nanotechnology-Based Antiviral Agents: - The Silver Edge

22 Pages·2013·2.81 MB·English

UNCLASSIFIED
Novel Nanotechnology-Based
Antiviral Agents:
Silver nanoparticle neutralization of hemorrhagic
fever viruses
Janice Speshock, Ph.D.
Saber Hussain, Ph.D.
Applied Biotechnology Branch
711th Human Performance Wing
Air Force Research Laboratory
Cleared for public release 88ABW-2009-4491
UNCLASSIFIED
UNCLASSIFIED
Nanomaterials
Unique Properties DOD Applications Challenges
•
Toxicity
• •
Size (< 100nm) Biosensors
•
• Reproducibility
Optical (metal &
•
Antimicrobial
Semiconductors
•
Stability of
Agents
•
Magnetic (metal) coatings/functional
• groups
• Surface reactivity Munitions
•
• bioaffinity
Catalytic activity
•
Propellants
(high surface area)
•
Effects on protein
•
• Bioaffinity Coatings activity
•
Surface modification • Smart Suits • Effects on gene
expression
UNCLASSIFIED 2
UNCLASSIFIED
Silver Nanoparticles
POLYSACCHARIDE
UNCOATED
COATED
10 nm
12.78 ± 0.13 9.48 ± 4.286
Dr. Steve Oldenburg, NanoComposix Dr. Dan Goia, Clarkston University
25 nm
27.474 ± 9.062 25.98 ± 8.38
Dr. Karl Martin, Novacentrix Dr. Dan Goia, Clarkston University
UNCLASSIFIED 3
UNCLASSIFIED
Hemorrhagic Fever Viruses
Arenaviridae Filoviridae
• South american HFV, • Ebola and Marburg
Lassa Fever, LCMV
• Enveloped, RNA viruses
• Enveloped, RNA viruses
• No effective therapies
• No effective therapies
• Vaccine in Phase I trials
• Candid#1 vaccine
• Up to 90% fatality rate
• 5-35% fatality rate
UNCLASSIFIED 4
UNCLASSIFIED
Tacaribe Virus
• New World (Tacaribe) Complex
– Junin, Machupo, Guanarito, and
Sabia
• Tacaribe virus is a biochemically and
serologically close relative of the
CDC category A arenaviruses but
has a low pathogenic potential for
humans
• Experimentally:
– Cytopathic effect in vero cells
– lethal meningoencephalitis in mice
UNCLASSIFIED 5
UNCLASSIFIED
Arenavirus Experimental Setup
2 h.p.i 12 h.p.i 8 d.p.i
Confocal Microscopy TEM Harvest Progeny Virus
Cell surface expression Virus internalization TCID determination
50
0 hour
1 h.p.i
4 h.p.i 4 d.p.i
Confocal Microscopy qRT-PCR
Virus internalization
Vero cells
UNCLASSIFIED 6
UNCLASSIFIED
TCRV Progeny Virus Production
UNCLASSIFIED 7
UNCLASSIFIED
Cell Surface TCRV Expression
Negative Control 10 nm 50 µg/ml 25 nm 50 µg/ml
PositiveControl 10 nm 10 µg/ml 25 nm 10 µg/ml
UNCLASSIFIED 8
UNCLASSIFIED
Dynamic Light Scattering
UNCLASSIFIED 9
UNCLASSIFIED
TCRV Internalization into Vero Cells
Untreated TCRV TCRV + 10nm Ag TCRV + 25nm Ag TCRV + 25nm Ag
• Ag-NP-treated TCRV is internalized into infected Vero cells
• Ag-NPs and TCRV interact inside the cell lysosomes
UNCLASSIFIED 10