ISIS Report 16/03/05
No Biosecurity without Biosafety
Biodefence Research Endangers the Public
Ensuring the safe use of genetic engineering is much more important
than preventing or defending the nation from bioterrorist attacks
Dr. Mae-Wan Ho
referenced version is posted on ISIS Members website.
Biosecurity & biosafety
Biosecurity originated from a small group of scientists who
met in 2001 to discuss how to keep diseases affecting crops and livestock from
crossing national boundaries. Then, came the anthrax attacks post September 11,
and the term came to be used for measures aimed at countering terrorist attacks
involving biological agents or toxins. Suddenly, thousands of US scientists are
caught in a web of new rules for keeping dangerous agents and substances, and
even scientific knowledge, out of reach of bioterrorists. Biosecurity should
come under the Biological Weapons Convention (BWC) to which the US is
signatory; but the US has rejected the Conventions remit to establish a
procedure to verify compliance with the Convention ("Bioweapons Convention
no progress in sight",
Biosafety refers to a set of measures aimed at regulating
and ensuring the safe use of genetic engineering and transnational movements of
genetically modified organisms. It falls within the scope of the Cartagena
Biosafety Protocol under the Convention of Biological Diversity. The US is not
a party to the Biosafety Protocol and has steadfastly refused to acknowledge
it. The US position is that genetic engineering biotechnology is inherently
safe, and only its misuse needs to be prevented.
It is clear that the BWC and Cartagena Biosafety Protocol overlap, and
are both needed for effective control of genetic engineering and biological
weapons. Of the two, biosafety is the more critical, although most of the
attention is focussed on biosecurity.
Biosecurity and biotechnology research
A report from the National Academy of Sciences, Biotechnology Research in an Age of Terrorism: Confronting
the Dual Use Dilemma, grew out of a meeting in January 2003 chaired by
Gerald Fink, Professor of Genetics at the Whitehead Institute for Biomedical
Research at Massachusetts Institute of Technology.
The Fink Report recommends a review process both at the research stage
and at the stage of publication. Its stated aim was to "safeguard the integrity
of science", and not "to censor research or research publications". It proposes
"a system of filters" to help determine whether a particular study should be
done, and if so, how the finding might best be published to avoid potential
misuse. It emphasizes "voluntary self-governance by the scientific community".
The editors of Cell, Science, Nature and PNAS have
already agreed to review and "filter" out "sensitive" papers or information.
The Bush administration has adopted the Fink Report, but its
implementation is mired in difficulty. The system relies on Institutional
Biosafety Committees that frequently fail to comply with federal rules and on a
National Science Advisory Board on Biosecurity announced a year ago, but has
yet no members.
Biodefence is bad for biosecurity and biosafety
The US has been caught in a dilemma of its own making because the
government has been pouring billions into biodefence research on
biological weapons agents. It sets up some 40 new and upgraded hot
zones across the country with hundreds of biosafety level 3 or 4 labs,
designed to research the most dangerous pathogens such as Ebola, plus at least
11 doing classified, secretive research, 6 major aerosol facilities and 3
open-air testing facilities. These labs and facilities include not just
established bio-weapons research institutions, but also former nuclear weapons
installations and top universities, like Boston, Harvard, John Hopkins,
California, Illinois, Texas and New York; many situated in the middle of
densely populated areas, or in some cases, deliberately sited in remote regions
where they can operate in secrecy (e.g., Dugway Proving Ground, Utah or Rocky
Mountain Labs, Montana).
Critics call attention to accidental releases of dangerous pathogens and
failures of biosafety containment that could spell disaster for residents
("Bio-defence mania grips United States", and "Biodefence contravenes
19). It is difficult to distinguish bio-defence research from bio-weapons
research. In order to make vaccines against deadly biological agents, the
deadly biological agents have to be created. The US programme, particularly in
the Departments of Defense and Homeland Security, is increasingly focussed on
"threat assessment" studies where researchers deliberately create the "threat",
i.e., the weapon, claiming they are learning to defend against it.
There is also the danger that researchers will be trained in precisely
the techniques needed for bio-terrorism. The post-September 11 anthrax attacks
have been traced to the federal bio-defence lab in Fort Detrick Maryland.
Under the newly formed Department of Homeland Security (DHS), much
defence-related research and development will be exempt from the Freedom of
Information Act, and hence there will be little or no mandatory public
Biodefence is bad for science
Nevertheless, the Sunshine Project (www.sunshine-project.org), a
bioweapons watchdog, has developed and released a new webtool (CRISPER)
to search and organize research grant data from the National Institutes of
Health (NIH). It revealed that the decision of NIH National Institute for
Allergy and Infectious Diseases (NIAID) to give priority to research of high
biodenfence but low public health significance resulted in 1500% increase in
the number of grants awarded for biological weapons agents, while the number of
grants for model microorganisms and non-bioweapons pathogens decreased by 41%
and 27% respectively.
More than 750 prominent scientists signed an open letter, published in
Science, to the director of NIH to express their concern. Richard
Ebright of Rutgers University, who initiated the letter, points out that
prioritising research on poorly known agents could backfire, not least because
of the need for strict containment and new experimental tools. The biodefence
money would be better spent researching related, but less pathogenic organisms.
He also believes that increasing the number of labs and people working on
bioterror agents would increase the risk of an accidental release or deliberate
NIAIDs biodefence budget shot up from $42m in 2001 to $1.5bn in
2004, with $1.6 projected for 2005. The government wide total spending on
biodefence before 2001 was less than $1 billion/year; between 2001 and 2005,
the total spending comes to $22 billion, and the projected total spending
between 2001 and 2006 is $30 billion.
There is no effective biodefence
One should not underestimate the increased possibilities of creating
powerful biological weapons in the post-genomics era ("GM & bioweapons in
the post-genomics era", SiS15) (see Box
1). But thats precisely also why there can be no effective biodefence
("Biodefence in tatter", SiS 15) (see Box
2). The agents are unknown and unpredictable. They can target the immune system
directly to undermine the bodys defence. Vaccines will be ineffective, or
worse than useless as the smallpox vaccine may prove to be; and there is no way
to adequately test for efficacy or safety. Theoretical studies indicate that
partially effective vaccines may increase the virulence of pathogens. There is
no known defence against agents that target buildings or structures.
Post genomics possibilities for bioweapons
- Stealth viruses targeting specific populations
- Designer diseases
- Agents targeting the immune system
- Non-lethal agents targeting agriculture
- Non-lethal agents targeting buildings and structure
- Interfering RNAs that turn genes off
- Completely novel disease agents made in the lab
Futility of biodefence
- Agents unknown
- Immune system targeted
- Vaccines ineffective or worse than useless and there is no way
to test for efficacy or safety
- Partially effective vaccines may increase virulence of
- No known defence against agents that target buildings or
The dangers of biodefence were highlighted in January 2005 when three
lab workers in Boston University were reported to have fallen ill from being
infected with tularaemia. The infections happened between May and September
2004, but did not become public knowledge until a week after Bostons
Zoning Commission approved the construction of a biosafety level 4 laboratory
in the University.
The workers had handled a live strain of the tularemia bacterium instead
of the non-infectious one typically used. They were trying to find a vaccine
for rabbit fever.
Before that, there were three lab accidents involving the SARS virus.
The Washington Post (29 May 2004) commented: "Scientists still do not
fully understand exactly where or how SARS emerged 18 months ago. But it is now
clear that the most threatening source of the deadly virus today may be places
they know intimately their own laboratories."
The three lab outbreaks of the disease since September 2003 - in
Singapore, Taiwan, followed by the 9 cases linked to Chinas National
Institute of Virology - have wider implications. They "highlighted the unique
hazards to public health that arise from accidental releases of germs that no
longer exist or barely exist in the wild."
The article described the notorious release of smallpox in Birmingham,
England, in August 1978, 10 months after the last infection occurred in
Henry S. Bedson, head of the microbiology department at a medical
school, was rushing to finish his experiments before the deadline to turn in or
destroy his stocks of smallpox, as his lab had been judged unsatisfactory by
the World Health Organisation inspectors. The smallpox virus apparently became
airborne, and transported up one floor through air ducts to a photographic
studio and darkroom to infect a 40-year-old photographer who died, even though
she had been vaccinated 12 years earlier. But not before she transmitted the
virus to her mother, who also became ill but survived. Her father did not
become infected but died from a heart attack. Bedson slashed his throat,
leaving a note that said, "I am sorry to have misplaced the trust which so many
of my friends have placed in me and my work."
The Council for Responsible Genetics listed numerous breaches of
bio-containment of disease agents in the US: involving accidental and
deliberate environmental releases, failures of containment, loss of samples and
13 cases of exposures and infections of personnel between 1994 and 2004. The
agents included AIDS, Ebola virus, West Nile virus, glanders, plague, anthrax
Genetic engineering could be worse than bioweapons
I have stressed that the control of bioweapons and genetic engineering
must go together (SiS 13/14), and
that the hazards from genetic engineering could be worse than bioweapons: The
basic tools and materials for making bioweapons are the same as those used in
legitimate genetic engineering applications. But while bio-weapons
are made under strictly contained conditions, many dangerous experiments are
being done without adequate safety precautions, and hazardous gene products are
released into the environment as if they were safe.
More and more scientists are voicing concerns over genetically modified
crops (The Case
for a GM-free Sustainable World and "ISP letter to FDA" ): the
potential toxicity and allergenicity of transgenic products and their negative
impacts on biodiversity, as well as the horizontal transfer of transgenic DNA
and antibiotic resistance marker genes. The more serious and insidious hazards,
however, are associated with contained uses of genetic engineering
The Fink Report dismissed the risks of genetic engineering: "The initial
fears about the inadvertent creation of virulent microbes by gene splicing
techniques have abated because of overwhelming scientific evidence to the
contrary. There have been no reported cases of disease caused by recombinant
microorganisms despite the widespread use of gene splicing techniques in
academic laboratories and in the production of pharmaceuticals."
I co-authored a paper with six scientists entitled, "Gene technology and
gene ecology of infectious diseases", published in 1998, summarising all the
evidence suggesting that genetic engineering may have contributed to the
resurgence of infectious diseases since commercial scale genetic engineering
began, calling for an independent public enquiry.
I sent a preprint to the World Health Organisation and the UK Health and
Safety Executive. Eventually, the HSE wrote and said it would commission its
own review, and nothing more was heard. But the question we raised in the paper
remains as alive as ever ("SARS virus genetically engineers?"
The Fink Committeee identified seven kinds of "experiments of concern"
as those having "a greater likelihood for potential misuse", and hence would be
subject to further examination (Box 3).
Experiments of concern from Fink Report
- Demonstrate how to render a vaccine ineffective
- Confer resistance to therapeutically useful antibiotics or
- Enhance the virulence of a pathogen or render a nonpathogen
- Increase transmissibility of a pathogen
- Alter the host range of a pathogen
- Enable the evasion of diagnostic/detection modalities
- Enable the weaponization of a biological agent or toxin
Practically all seven classes of experiments are being done in genetic
engineering, in the course of genetic modification of bacteria, plants and
animals or human beings in gene therapy.
Antibiotic resistance marker genes have been released into the
environment with GM crops in field trials or for commercial growing. Bacteria
and viruses are routinely mutated and recombined in the laboratory, which could
change their host range, increase their virulence or transmissibility or render
existing vaccines ineffective. In fact, simply strip off
the coat from a virus, and the naked genome would be taken up by
non-host cells to generate infectious viruses. Researchers have been culturing
human viruses in animal cells for decades, which would certainly alter their
host range. They also culture viruses in mixtures of cells from different
species deliberately to obtain mutants that infect both species, the latest
involving the SARS virus. Similarly, gene therapists have been wrapping gene
transfer vectors in liposomes and other material to escape immune detection,
which is a major problem with gene therapy vectors. These vectors, though
disarmed, have the potential to regenerate live viruses or to cause insertion
mutagenesis. A third case of leukemia has surfaced among the children given
gene therapy for X-linked SCID in Paris. Finally, almost any bacterium can be
weaponised by transferring into it whole sets of virulence genes in
mobile pathogenicity islands or phage coding for toxins; in
experiments that are routinely done in genetic engineering.
Specific examples of dangerous experiments in genetic engineering
A research team in the State University of New York at Stony Brook made
the poliovirus by joining up chemically synthesized short DNA fragments into a
complete sequence that was then transcribed into the RNA viruses in a cell-free
system containing all the necessary enzymes and cell parts. The synthetic
viruses were capable of infecting cells.
The researchers demonstrated that one could synthesize any virus from
chemical reagents that can be purchased in the open market, using the public
database for the genome sequence. The experiment is not exactly new. David
Baltimore and colleague had shown in 1981 that a DNA copy of the RNA genome of
poliovirus could be taken up into living cells to generate infectious
Also in 2002, researchers in the University of Pennsylvania,
Philadelphia, showed that a gene in the smallpox variola virus is more than 100
times more potent than the version in vaccinia virus (used in vaccines against
smallpox) in inhibiting the human complement enzymes protecting the body
against viral attacks. And that could be why the smallpox virus is so much more
virulent than the vaccinia virus.
The gene from the variola virus, therefore, has the potential to
increase the transmissibility of other viruses; but, as claimed, disabling it
may be "therapeutically useful if smallpox re-emerges".
Should scientists do this kind of experiments and report the results
openly? One could argue that open knowledge is always a good thing, because it
alerts people to the possibilities and encourages them to be vigilant and to
find means of overcoming the dangers. Whether they should do the experiment in
the first place is questionable.
The US is currently proposing to express variola genes in related pox
viruses, and to insert a reporter gene (expressing green fluorescent protein)
in variola itself. The World Health Assembly is to consider these controversial
proposals in May 2005.
- There are intentional creations of dangerous agents for supposedly
benign purposes that turned out not to be so benign. For example, numerous
vaccines against HIV/AIDS based on the envelope glycoprotein gp120 of the HIV
are proving worse than useless ("AIDS vaccines worse than useless?"
SiS 19). A
fierce row broke out between leading AIDS researchers in January 2004 over the
continued Phase III trial in Thailand of a vaccine made with a live-replicating
canarypox vector with a boost of gp120.
AIDS researchers have pointed out that the gp120 protein is strongly
immunogenic, but the antibodies fail to protect against the virus. Instead, it
ends up over-stimulating the immune system, leaving it less able to cope with
new infections. Moreover, the part of the gp120 molecule that plays the
dominant role in provoking an immune response is the V3 loop. The V3 loop and
flanking regions are similar in base sequence and structure to the
antigen-binding region of the human immunoglobulin (Ig). It has been suggested
since the early 1990s that this immunoglobulin-like domain in gp120 may
interfere with the immune regulatory network. The V3 loop and its flanking
regions are, moreover, located between recombination signals similar to those
found in human immunoglobulins, and to the Chi recombination hotpots
found in many viruses and bacteria. Consequently, the immunologically dominant
region of gp120 may be involved in recombining with human immunoglobulin genes
resulting in autoimmune responses, and may also recombine with co-infecting
viruses and bacteria to generate new pathogens. Evidence of recombination
between gp120 and bacterial DNA has subsequently been found in the sera of AIDS
The gp120 gene, spliced into numerous bacterial and viral vectors with
Chi or Chi -like sequences as vaccines, therefore multiply the
odds for recombination to generate new pathogens. The gp120 gene has been
incorporated into GM maize as a cheap edible vaccine against HIV; while the key
bioweapons institute in Russia has created an artificial protein consisting of
the env and gag antigens from HIV fused with the hepatitis B virus protein
HbsAg as a HIV/hepatitis B combined vaccine. This is tantamount to releasing
slow bioweapons on the populations. The new combinations of dangerous genes
will have plenty of opportunity for generating new pathogens through further
gene trafficking with bacteria and viruses in the environment, of which less
than 1% can be cultured and identified.
Also in connection with AIDS, researchers have created a hybrid simian
and human immunodeficiency virus (SIV +HIV = SHIV) for testing
vaccines, which kills victim primates in weeks.
Some of us have been warning that genetic engineering is inherently
dangerous because it greatly multiplies the scope and frequency of horizontal
gene transfer and recombination, the major route for creating new viruses and
bacteria that cause disease epidemics. This was brought home when researchers
in Australia accidentally transformed a harmless mousepox virus
into a lethal pathogen that killed all the mice, even those that were supposed
to be resistant to the virus.
Headlines in the New Scientist editorial: "The Genie is out,
Biotech has just sprung a nasty surprise. Next time, it could be catastrophic."
The lead article continued in the same vein: "Disaster in the making. An
engineered mouse virus leaves us one step away from the ultimate
The researchers added a gene coding for the immune signalling molecule
interleukin-4 to the virus, which they thought would boost antibody production;
instead, it suppressed both primary antiviral immune responses as well as
adaptive immune responses. The same gene, spliced into a vaccinia virus
previously, delayed the clearance of virus from the animals; so it may well
have the same immune suppressive effects for all viruses.
More surprisingly, a paper published in December 2003 described how
disrupting a set of virulence genes in Mycobacterium tuberculosis, the
tuberculosis bacterium, resulted in a hypervirulent mutant strain that killed
all the mice by 41 weeks, while all the control mice exposed to the wild-type
These two cases underscore the complexity of disease generation by
pathogens. They also leave us in no doubt that apparently innocent experiments
could end in nasty surprises.
There have been previous cases that went unnoticed. A paper published in
2001 described how a method established for recovering infectious Ebola virus
by reverse transcription enabled the researchers to produce a mutant
considerably more toxic to the host cells than the wild-type.
Finally, Willem Stemmer of Maxygen Inc. based in California invented a
DNA shuffling technique in which genes, viral and bacterial genomes, whatever,
could be chopped up into fragments, then made to join up at random into
millions of recombinants, out of which superior performing enzymes,
metabolically more efficient genomes or more infectious and virulent pathogens
could all be selected.
In one experiment, 6 mouse leukemia viruses (MLVs) were recombined in a
single round giving 5 million replicating recombinant viruses in a matter of
hours. Among them were completely new viruses that infected Chinese Hamster
Ovary cells, which none of the original MLV was capable of. Some recombinants
were 30 to 100 times more stable than the parental strains. There is no way to
characterize all of the 5 million recombinants and they may well include new
Researchers have given up trying to cope with the massive complexity of
the genome and gene function ("Life after the Central Dogma" series,
Instead of rational design, they have resorted to random genome shuffling to
improve industrial microbes, and the advantage is that the resultant microbes
arent even classified as genetically engineered, and therefore not
subject to usual regulation and can even be used in the food industry, we are
told. They even suggest using environmental libraries, containing the DNA of
the 99% bacteria unknown to science, in genome shuffling; but are at least
aware of the dangers involved: "New drug resistances" and emergence of
"accelerated or new pathogenic mechanisms or diseases".
It has been known for some time that bacterial and viral DNA can cause
immune reactions, because thats part of our innate immune response that
protects us from germs; and this has become a major obstacle to gene therapy
. Any fragment of double-stranded DNA or RNA down to 25bp is immunogenic.
There is now new evidence that certain sequences of single stranded RNA also
elicit specific immune reactions.
Biotechnological processes and genetic engineering labs are creating an
increasing variety of naked/free nucleic acids that are currently released
unregulated into the environment, where they can elicit immune reactions, as
well as undergo uncontrolled horizontal gene transfer and recombination to
generate new pathogens or trigger cancer, should some of them jump into the
genome of our cells.
Greater hazards of genetic engineering
Compare the list of "Dangerous experiments constructs and deliberate
releases in genetic engineering" (Box 4) with the "Experiments of concern" in
the Fink Report (Box 3). The list for genetic engineering is more extensive and
more insidious; as the deadly biological agents generated cannot be predicted,
nor the millions of cancers that may result from insertion of constructs with
strong viral promoters.
Dangerous experiments, constructs and deliberate releases in
- Creating lethal pathogens by accident, e.g., mousepox virus,
disarmed tuberculosis bacterium
- Making deadly viruses more lethal, e.g., ebola
- Making hybrid SIV-HIV virus (for testing vaccines) that kills
monkeys in weeks
- Releasing AIDS vaccines that are effectively slow
- Releasing gene therapy vectors that cause leukaemia
- Releasing antibiotic resistance genes and potentially toxic or
allergenic transgene products
- Manipulating genes associated with cancer and immune
suppression on a routine basis
- Creating cross-species viruses in cell cultures
- Generating millions of recombinant agents in hours by genome
- Creating huge varieties of vectors, DNA vaccines and endless
species of rDNAs and rRNAs that can generate new pathogens, cause immune
reactions, insertion mutations and cancers
Genetic engineering hazards preventable
The good news is that practically all the hazards of genetic engineering
can be prevented: by tightening the regulation of contained use as has been
done for deliberate release. I wrote a report with three colleagues in 2001,
the Regulatory Net: Naked and Free Nucleic
Acids . The report was reprinted 2002 and 2004, because our message has not
yet got through to the regulators.
Current European regulation allows users to release directly into the
environment certain live transgenic microorganisms considered nonpathogenic or
otherwise safe in liquid waste, although there is no agreement across European
countries as to which bacteria are pathogens.
Meanwhile, all killed microorganisms and cells containing transgenic DNA
are disposed of as solid waste, and are either recycled as food, feed and
fertilizer, or disposed of in landfill.
There is an urgent need for validated procedures to ensure that all
kinds of transgenic nucleic acids from contained use are thoroughly degraded
before transgenic wastes are released into the environment.
Meanwhile, there should be no environmental releases of GM crops unless
and until they can be proven safe beyond reasonable doubt.