Download THE BIOLOGICAL AND TOXIN WEAPONS CONVENTION

Dual-Use Examples
Lecture No. 15
1. Outline
• Contentious Research
– Slides 2 - 8
• Mousepox
– Slides 9 - 14
• Synthesis of Polio Virus
– Slides 15 – 18
• Influenza Virus
– Slide 19
• Virulence in Smallpox
– Slides 20-21
2. Contentious Research (i)
• Fink Chapter 1
–
–
–
–
The Life Sciences Today
The “Dual use” Dilemma
A Brief History of Biological Warfare*
U.S. Policy and the Creation of the Biological and
Toxin Weapons Convention
– *Annex: Biological Warfare in History
3. Contentious Research (ii)
• Fink Chapter 1 (continued)
– The New Threat
– Recent Examples of “Contentious Research”
in the Life Sciences
– The Response of the Life Science Community
to Previous Challenges
– Committee Charge and Process
4. Contentious Research (iii)
• The Life Sciences Today
– “The biological sciences have experienced enormous
growth over the last century…”
– “The ever-expanding research activity has resulted in
numerous new…products that are transforming
medicine…”
– “Biotechnology research is now a truly global
enterprise…”
– “In addition to the dispersed research enterprise,
publications and personnel are also widely spread…”
5. Contentious Research (iv)
• The Life Sciences Today (continued)
– “The rapid spread of scientific knowledge and
applications owes much to a research culture in which
knowledge and biological materials are shared among
scientists and people move freely between
universities, government agencies, and private
industry. Large numbers of foreign graduate students
and postdoctoral associates have been an essential
ingredient of the success of the biological research
enterprise. The scientific workforce is increasingly
international…”
6. Contentious Research (v)
• The Dual Use Dilemma
– “The regulation of dual use biotechnology research is
a highly contentious technical, political, and societal
issue. In the language of arms control and
disarmament, dual use refers to technologies
intended for civilian application that can also be used
for military purposes…”
– “…The key issue is whether the risks associated with
misuse can be reduced while still enabling critical
research to go forward.”
7. Contentious Research (vi)
• The New Threat
– “Every major technology - metallurgy,
explosives, internal combustion, aviation,
electronics, nuclear energy - has been
intensively exploited, not only for peaceful
purposes but also for hostile ones. Must this
also happen with biotechnology, certain to be
a dominant technology of the coming
century…?”
8. Contentious Research (vii)
• The New Threat (continued)
– “…During the century just begun, as our ability to
modify fundamental life processes continues its rapid
advance, we will be able not only to devise additional
ways to destroy life but…also…to manipulate it including the processes of cognition, development,
reproduction, and inheritance. A world in which these
capabilities are widely employed for hostile purposes
would be a world in which the very nature of conflict
has radically changed. Therein could lie
unprecedented opportunities for violence, coercion,
repression, or subjugation.”
9. Mousepox (i)
• “…Probably the most celebrated recent case
involving the dissemination of research with the
potential for bioterrorist uses was the report of
an unexpected effect of the bioengineering of a
strain of ectromelia virus (mousepox) that was
intended to help eradicate mice in Australia….
Some have felt that the publication of this paper
provides a blueprint or roadmap for terrorist to
engineer a more virulent strain of smallpox that
could overwhelm the human immune system in
even well-vaccinated individuals…”
10. Mousepox (ii)
• “The authors of the paper had originally set out
to make an infectious immunocontraceptive for
wild mice by incorporating a gene encoding an
antigen from fertilized mouse eggs into the
genome of ectromelia virus. Since the
expression of this egg antigen of the virus did
not result in infertility, the authors attempted to
increase the virulence of ectromelia with the
hope that this would increase the immune
response…”
11. Mousepox (iii)
• “They drew on previously published work…in
which it had been shown that incorporating the
gene for…IL-4 into the viral genome and thus
overexpressing it in vivo enhanced the virulence
of vaccinia virus in mice. The increased
virulence is probably due to suppression of the
antiviral immune response mediated through
competing cytokines like IL-2…which work by
stimulating immune effector cells to kill virusinfected cells and thus control the virus
infection.”
12. Mousepox (iv)
• “…They then demonstrated that this engineered
mousepox virus was much more virulent than
the parent virus and killed 60% of infected mice,
even if the mice were from a genetically resistant
strain. Even more unexpected was their
observation that mice that had been vaccinated
and were completely resistant to the parent
virus…were now killed by the IL-4 geneexpressing virus.”
13. Mousepox (v)
• “Some have felt that the publication of this paper
provides a blueprint or roadmap for terrorists to
engineer a more virulent strain of smallpox that
could overwhelm the human immune system in
even well-vaccinated individuals…. It has been
suggested that either the paper should not have
been published, or at the very least the
‘materials and methods’ section…should have
been altered or omitted entirely from the
published article…”
14. Mousepox (vi)
• Reasons for publication?
– “…First, knowledge of these experiments allows the
scientific community to explore how to overcome such
engineered viruses…”
– “…Second, it suggests that we should be prepared to
treat infections caused by such an engineered virus
with antibodies that inactivate the relevant cytokine,
with gamma interferon that would counter the effect of
IL-4, or with both…”
15. Synthesis of Polio Virus (i)
• “Wimmer and colleagues reported that they had
reconstructed poliovirus from chemically
synthesized oligonucleotides that were linked
together and then transfected into cells. The
report attracted considerable attention in the
news media and concern in some segments of
the public….This…raised public concern about
bioterrorism because it suggested that the
Wimmer experiment provided a recipe for
terrorists to manufacture the virus…”
16. Synthesis of Polio Virus (ii)
• “Many scientists concluded that the Wimmer
experiment was neither a novel discovery nor a
potential threat. The general principle that one
could make live poliovirus from a DNA template
was already known in 1981, when Baltimore and
colleagues reported that a DNA copy of the
positive strand RNA genome of poliovirus could
be taken up into living cells under appropriate
conditions and result in the generation of
encapsulated, infectious virus…”
17. Synthesis of Polio Virus (iii)
• “We have improved upon the methodology and
dramatically shortened the time required for the
accurate assembly of a 5- to 6-kb segments of
DNA from synthetic oligonucleotides. As a test of
this methodology, we have established
conditions for the rapid (14-day) assembly of the
complete infectious genome of the
bacteriophage  X174 (5,386 bp) from a single
pool of chemically synthesized
oligonucleotides…”
18. Influenza Virus (iv)
• “Influenza A virus has been responsible for
widespread human epidemics because it readily
transmits form humans to humans by aerosol.
Recent events have highlighted the potential of
influenza A virus as a bioterrorist weapon: the
high virulence of influenza A virus that infected
people in Hong Kong in 1997: and the
development of laboratory methods to generate
influenza A viruses by transfection of DNAs
without a helper virus…”
19. Virulence in Smallpox (i)
• “Variola major virus causes smallpox, which has
a 30 - 40% mortality rate,whereas vaccinia virus,
which is used to vaccinate humans against
smallpox, causes no disease in
immunocompetent humans….Both viruses have
an inhibitor of immune response enzymes vaccinia virus complement control protein (VCP)
and smallpox inhibitor of complement enzymes
(SPICE). The authors focused on a comparison
of the genes encoding this inhibitor…”
20. Virulence in Smallpox (ii)
• “…As live variola is not available for study, they
used standard techniques to synthesize the
SPICE gene. They found that variola spice has a
greater degree of specificity for human
complement and is nearly a hundredfold more
active than VCP in inactivating this component
of the human immune system (human
complement component C3b)…”
Sample Questions
1. Do you agree with Meselson’s view that there is a grave
danger that the modern life sciences will be used in a
major way for hostile purposes? If you agree how do you
think this might be prevented? If you disagree set out
your reasons and discuss one of them in detail.
2. Outline the mousepox experiment. Should this work have
been reported in the scientific literature? Give the
reasons for your consideration.
3. “Wimmer’s synthesis of polio virus was not on example
of dual-use research of concern”. Discuss.
4. Could influenza virus be used as a serious
bioterrorism/biowarfare agent?
References
(Slide 2)
National Research Council (2004) Biotechnology Research in an Age of
Terrorism. Washington: National Academies Press. Available from
http://books.nap.edu/openbook.php?record_id=10827&page=15
(Slide 4)
National Research Council (2004) Biotechnology Research in an Age of
Terrorism. Washington: National Academies Press. Available from
http://books.nap.edu/openbook.php?record_id=10827&page=16
(Slide 5 and 6)
National Research Council (2004) Biotechnology Research in an Age of
Terrorism. Washington: National Academies Press. Available from
http://books.nap.edu/openbook.php?record_id=10827&page=18
(Slide 7)
Meselson, M (2000) Averting the Hostile Exploitation of Biotechnology,
The CBW Conventions Bulletin, June 48, 16-19. Available from
http://www.sussex.ac.uk/Units/spru/hsp/pdfbulletin.html
(Slide 8)
National Research Council (2004) Biotechnology Research in an Age
of Terrorism. Washington: National Academies Press. Available
from
http://books.nap.edu/openbook.php?record_id=10827&page=23
(Slide 9)
National Research Council (2004) Biotechnology Research in an Age
of Terrorism. Washington: National Academies Press. Available
from
http://books.nap.edu/openbook.php?record_id=10827&page=25
Jackson, R. J., Ramsay, A. J., Christensen, C. D., Beaton, S., Hall, D.
F., and Ramshoaw, I. A. (2001) Expression of Mouse Interleukin-4
by a Recombinant Ectromelia Virus Suppresses Cytolytic
Lymphocyte Responses and Overcomes Genetic Resistance to
Mousepox. Journal of Virology, 75(3), 1205–1210. Available from
http://www.ncbi.nlm.nih.gov/pubmed/11152493
National Research Council (2004) Biotechnology Research in an Age
of Terrorism. Washington: National Academies Press. Available
from
http://books.nap.edu/openbook.php?record_id=10827&page=25
(Slide 12 and 13)
National Research Council (2004) Biotechnology Research in an Age of
Terrorism. Washington: National Academies Press. Available from
http://books.nap.edu/openbook.php?record_id=10827&page=26
(Slide 14)
National Research Council (2004) Biotechnology Research in an Age of
Terrorism. Washington: National Academies Press. Available from
http://books.nap.edu/openbook.php?record_id=10827&page=27
Bembridge, G. P., Lopez, J. A., Cook, R., Melero, J. A., and Taylor, G..
(1998) Recombinant Vaccinia Virus Coexpressing the F Protein of
Respiratory Syncytial Virus (RSV) and Interleukin-4 (IL-4) Does Not
Inhibit the Development of RSVSpecific Memory Cytotoxic T
Lymphocytes, whereas Priming Is Diminished in the Presence of
High Levels of IL-2 or Gamma Interferon, Journal of Virology
72(5):4080-7. Available from
http://www.ncbi.nlm.nih.gov/pubmed/9557697
Parker, S., Touchette, E., Oberle, C., Almond, M., Robertson, A., Trost, L.
C., Lampert, B., Painter, G., and Buller, R. M.(2008) Efficacy of
Therapeutic Intervention with an Oral Etherlipid Analogue of
Cidofovir (CMX001) in a Lethal Mousepox Model. Antiviral Research
77(1):39-49. Available from
http://www.ncbi.nlm.nih.gov/pubmed/17904231
(Slide 15)
National Research Council (2004) Biotechnology Research in an
Age of Terrorism. Washington: National Academies Press.
Available from
http://books.nap.edu/openbook.php?record_id=10827&page=27
Wimmer, E. (2007) The Test-Tube Synthesis of a Chemical Called
Poliovirus, EMBO Reports Special Issue 7, 3-9. Available from
http://www.nature.com/embor/journal/v7/n1s/full/7400728.html
Cello, J., Paul, A. V., and Wimmer, E. (2002) Chemical Synthesis of
Poliovirus cDNA: Generation of Infectious Virus in the Absence
of Natural Template, Science 297(5583), 1016 – 1018. Available
from http://www.sciencemag.org/cgi/content/abstract/1072266
(Slide 16)
National Research Council (2004) Biotechnology Research in an
Age of Terrorism. Washington: National Academies Press.
Available from
http://books.nap.edu/openbook.php?record_id=10827&page=28
(Slide 17)
Smith, H. O., Hutchison, C. A., Pfannkoch, C., and Venter, J. C. (2003)
Generating a Synthetic Genome by Whole Genome Assembly:
_X174 Bacteriophage from Synthetic Oligonucleotides, PNAS,
100(26), 15440–15445. Available from http://www.pnas.org/
Tian, J., Gong, H., Sheng, N., Zhou, X., Gulari, E., Gao, X., and Church,
G.. (2004) Accurate multiplex gene synthesis from programmable
DNA microchips, Nature, 432(23/30). Available from
http://www.nature.com/nature/journal/v432/n7020/full/nature03151.
html
(Slide 18)
Krug, M. R. (2003) The Potential Use of Influenza Virus as an Agent for
Bioterrorism, Antiviral Research 57, 147-150 Available from
http://www.elsevier.com/wps/find/journaldescription.cws_home/52
1852/description#description
(Slide 19)
Rosengard, A. M., Liu, Y., Nie, Z., and Jimenez, R. (2002) Variola virus
immune evasion design: Expression of a highly efficient inhibitor
of human complement, PNAS, 99(13), 8808–8813 Available from
http://www.pnas.org/