ISIS submitted written questions to the upcoming ‘Open Meeting' of the
Advisory Committee for Novel Food Products (ACNFP) in which horizontal
gene transfer is to be discussed. Dr. Mae-Wan Ho, Director of ISIS, who
has written substantially on the subject, and made frequent
representations to the government, asked to be given time to present her
case at the meeting, but her request was refused. She has accordingly
submitted a written version of the case she would have presented
(reproduced below), and asked for a "fulsome" response to it from the
ACNFP at the ‘Open Meeting'.

Please circulate this as widely as possible as part of our contribution
to the ‘National Debate' on GM.



Recent Evidence Confirms Risks of Horizontal Gene Transfer

ISIS Contribution to ACNFP/Food Standards Agency Open Meeting 13 November
2002

Dr. Mae-Wan Ho, Institute of Science in Society
PO Box 32097
London NW1 0XR

Tel: 202-7272-5636

Horizontal gene transfer is one of the most serious, if not the most
serious hazard of transgenic technology. I have been drawing our
regulators' attention to it at least since 1996 [1], when there was
already sufficient evidence to suggest that transgenic DNA in GM crops
and products can spread by being taken up directly by viruses and
bacteria as well as plant and animals cells.

The oft-repeated refrain that "transgenic DNA is just like ordinary DNA"
is false. Transgenic DNA is in many respects optimised for horizontal
gene transfer. It is designed to cross species barriers and to jump into
genomes, and it has homologies to the DNA of many species and their
genetic parasites (plasmids, transposons and viruses), thereby enhancing
recombination with all of them [2]. Transgenic constructs contain new
combinations of genes that have never existed, and they also amplify
gene products that have never been part of our food chain [3].

The health risks of horizontal gene transfer include:

1.Antibiotic resistance genes spreading to pathogenic bacteria.
2.Disease-associated genes spreading and recombining to create new
viruses and bacteria that cause diseases.
3.Transgenic DNA inserting into human cells, triggering cancer.

The risk of cancer is highlighted by the recent report that gene therapy
- genetic modification of human cells - claimed its first cancer victim
[4]. The procedure, in which bone marrow cells are genetically modified
outside the body and re-implanted, was previously thought to avoid
creating infectious viruses and causing cancer, both recognized major
hazards of gene therapy.

The transgenic constructs used in genetic modification are basically the
same whether it is of human cells or of other animals and plants. An
aggressive promoter from a virus is often used to boost the expression
of the transgene, in animal and human cells, from the cytomegalovirus
that infects mammalian cells, and in plants, the 35S promoter from the
cauliflower mosaic virus (CaMV) that infects Cruciferae plants.

Unfortunately, although the CaMV virus is specific for plants, its 35S
promoter is active in species across the living world, human cells
included, as we discovered in the scientific literature dating back to
1989. Plant geneticists who have incorporated the promoter into
practically all GM crops now grown commercially are apparently unaware
of this crucial information [5].

In 1999, another problem with the CaMV 35S promoter was identified: it
has a ‘recombination hotspot' where it tends to break and join up with
other DNA [6]. Since then, we have continued to warn our regulators that
the CaMV 35S promoter will be extra prone to spread by horizontal gene
transfer and recombination [7-9]. The recent controversy over the
transgenic contamination of the Mexican landraces [10] hinges on
observations suggesting that the transgenic DNA with the CaMV 35S
promoter is "fragmenting and promiscuously scattering throughout the
genome" of the landraces, observations that would be consistent with our
expectations [11].

Similarly, I was not surprised by the research results released earlier
this year by the Food Standards Agency [12], indicating that transgenic
DNA from GM soya flour, eaten in a single hamburger and milk shake meal,
was found transferred to the bacteria in the gut contents from the
colostomy bags of human volunteers.

What I found unacceptable was the way the Agency dismissed the findings
and downplayed the risks. The comments, "it is extremely unlikely that
genes from genetically modified (GM) food can end up in bacteria in the
gut of people who eat them", and "the findings had been assessed by
several Government experts who had ruled that humans were not at risk",
are seriously misleading.

First, the experimental design stacked the odds heavily against finding
a positive result. For example, the probe for transgenic DNA covered
only a tiny fraction of the entire construct. So, only a correspondingly
tiny fraction of the actual transfers would ever be detected, especially
given the well-known tendency of transgenic constructs to fragment and rearrange.

Second, the scope of the investigation was intentionally restricted.
There was no attempt to check for transgenic DNA in the blood and blood
cells, even though scientific reports dating back to the early 1990s had
already indicated transgenic DNA could pass through the intestine and
the placenta, and become incorporated into the blood cells, liver and
spleen cells and cells of the foetus and newborn [13].

The observation in the FSA report [12] that no transgenic DNA was found
in the faeces of the ‘healthy volunteers', far from being reassuring,
raises the worrying possibility that the transgenic DNA has all been
taken up into the intestinal cells and/or passed into the bloodstream.

Third, no attempt was made to address the limitations of the detection
method and the scope of the investigation, which grossly underestimated
the extent and frequency of horizontal gene transfer, and hence failed
completely in assessing the real risks. On the contrary, false
assurances were made that "humans were not at risk".

Another research project on horizontal gene transfer commissioned by the
Ministry of Agriculture, Fisheries and Food (MAFF), the predecessor to
the Food Standards Agency, concerns Agrobacterium tumefaciens, the soil
bacterium that causes crown gall disease, which has been developed as a
major gene transfer vector for making transgenic plants. Foreign genes
are typically spliced into T-DNA - part of a plasmid called Ti
(tumour-inducing) – that's integrated into plant genome.

It turns out that Agrobacterium injects T-DNA into plant cells in a
process that strongly resembles conjugation, ie, mating between
bacterial cells; and all the necessary signals and genes involved are
interchangeable with those for conjugation [14].

That means transgenic plants created by T-DNA vector system have a ready
route for horizontal gene escape, via Agrobacterium, helped by the
ordinary conjugative mechanisms of many other bacteria that cause
diseases [15].

A report submitted to MAFF in 1997 had indeed raised the possibility
that Agrobacterium tumefaciens could be a vector for gene escape [16,
17].

The researchers found that it was extremely difficult to get rid of the
Agrobacterium used in the vector system after transformation.

High rates of gene transfer are known to be associated with the plant
root system and the germinating seed [18]. There, Agrobacterium could
multiply and transfer transgenic DNA to other bacteria, as well as to
the next crop plant.

Agrobacterium was also found to transfer genes into several types of
human cells [19], and in a manner similar to that which it uses to
transform plant cells.

We have submitted two relevant ISIS reports together with some specific
questions to the ACNFP for consideration at the November 13 Open Meeting [20].

All the risks of horizontal gene transfer described above are real, and
far outweigh any potential benefits that GM crops can offer. There is no
case for allowing any commercial release of GM crops and food products.

The following experiments and tests should be done to address the risks
of horizontal gene transfer.

1.Feeding experiments similar to those carried out by Dr. Arpad
Pusztai's team should be done, using well-characterized transgenic soya
and/or maize meal feed, with full, adequate, monitoring for transgenic
DNA in the faeces, blood and blood cells, and post-mortem histological
examinations that include tracking transfer of transgenic DNA into the
genome of cells. As an added control, nontransgenic DNA from the same GM
feed sample should also be monitored.
2.Feeding trials on human volunteers should be carried out using
well-characterized transgenic soya and/or maize meal feed, with full,
adequate monitoring for transgenic DNA in the faeces, blood and blood
cells. Also as an added control, nontransgenic DNA from the same GM feed
sample should also be monitored.
3.The stability of transgenic plants in successive generations should be
systematically investigated, especially for those containing CaMV 35S
promoter, using adequate quantitative molecular techniques.
4.Full molecular characterisation of all transgenic lines must be
carried out to establish uniformity and genetic stability of the
insert(s).
5.All transgenic plants created by the Agrobacterium T-DNA vector system
should be tested for the persistence of the bacteria and vectors. The
soil in which they have been grown should also be monitored for gene
escape to soil bacteria. And the potential for horizontal gene transfer
to the next crop via the germinating seed and root system should be
carefully monitored.

References and Notes

1.Correspondences between myself and Ministry of Agriculture, Fisheries
and Food, and Health and Safety Executive, going back to 1996.
2.See Ho MW. Horizontal Gene Transfer. The Hidden Hazards of Genetic
Engineering, TWN Biotechnology Series, Third World Network, 2001
(available fom the ISIS online store); also Ho MW. Horizontal gene
transfer and genetic engineering, SCOPES website, AAAS, 2000.
3.Ho MW. Briefing to the Rt. Hon. Michael Meacher, Minister for the
Environment on the Special Safety Concerns of Transgenic Agriculture and
Related Issues. April 1999 ; published in Seminario Internacional sobtre
Direcito da Biodiversidade, Revista cej: Centro de estudos Judiciarios
do Conselho da Justica Federal, Brasil, pp.120-6, 1999.
4.Science, News of the Week, 4 October 2002; see also "Predicted hazard
of gene therapy a reality" by Mae-Wan Ho, ISIS Report, October 2002
5."GM maize approved on bad science in the UK" by Mae-Wan Ho, Science in
Society 2002, 15, 10-25.
6.Kohli A.,Griffiths S, Palacios N, Twyman R, Vain P, Laurie D and
Christou P. Molecular characterization of transforming plasmid
rearrangements in transgenic rice reveals a recombination hot spot in
the CaMV 35S promoter and confirms the predominance of microhomology
mediated recombination" Plant.J. 1999, 17,591-601.
7.Ho MW, Ryan A and Cummins J. Cauliflower mosaic viral promoter – a
recipe for Disaster? Microbial Ecology in Health and Disease 1999 11,
194-7.
8.Ho MW, Ryan A and Cummins J. Hazards of transgenic plants with the
cauliflower mosaic viral promoter. Microbial Ecology in Health and
Disease 2000, 12, 6-11.
9.Ho MW, Ryan A and Cummins J. CaMV35S promoter fragmentation hotspot
confirmed and it is active in animals. Microbial Ecology in Health and
Disease 2000, 12, 189.
10.Quist D and Chapela IH. Transgenic DNA introgressed into traditional
maize landraces in Oaxaca, Mexico. Nature 2001, 414, 541-3, 2001.
11."Astonishing denial of transgenic contamination" by Mae-Wan Ho,
Science in Society 2002, 15, 13-14.
12.Netherwood T, Martin-Orue SM, O'Donnell AG, Gockling S, Gilbert HJ
and Mathers JC. Transgenes in genetically modified Soya survive passage
through the small bowel but are completely degraded in the colon.
Technical report on the Food Standards Agency project G010008
"Evaluating the risks associated with using GMOs in human foods"-
University of Newcastle.
13.Doerfler, W. and Schubbert, R. (1998). Uptake of foreign DNA from the
environment: the gastroinestinal tract and the placenta as portals of
entry, Wien Klin Wochenschr. 110, 40-44.p. 40.
14.Ferguson GC and Heinemann JA. Recent history of trans-kingdom
conjugation. In Horizontal GeneTransfer 2nd ed. (ed. M Syvanen & CI
Kado), pp 3-17, Academic Press, San Diego, 2002.
15.Ho MW. What's unspeakable in horizontal gene transfer? Heredity (in
press); "Averting sense for nonsense" by Mae-Wan Ho, Science in Society
2002, 16, 29-30.
16.McNicole et al (1997) The Possibility of Agrobacterium as a Vehicle
for Gene Escape. MAFF. R&D and Surveillance Report: 395.
17.Barrett et al (1997). A risk assessment study of plant genetic
transformation using Agrobacterium and implications for analysis of
trangenic plants. Plant Cell Tissue and Organ Culture 47: 135-144.
18.Sengelov G, Kristensen KJ, Sorensen AH, Kroer N, and Sorensen SJ.
Effect of genomic location on horizontal transfer of a recombinant gene
cassette between Pseudomonas strains in the rhizosphere and spermosphere
of barley seedlings. Current Microbiology 2001, 42, 160-7.
19. Kunik T, Tzfira T, Kapulnik Y, Gafni Y, Dingwall C, and Citovsky V.
Genetic transformation of HeLa cells by Agrobacterium. PNAS USA, 2001,
98, 1871-87; also, "Common plant vector injects genes into human cells",
ISIS News 2002, 11/12, p. 10
20."Stacking the odds against finding it" and "Averting sense for
nonsense" by Mae-Wan Ho. Science in Society 2002, 16, 28-30.

The questions first submitted to FSA 22 July 2002 were as follows:

1.Why were the transgenic soya samples so poorly characterised in terms
of GM content, structure of transgenic insert(s), states of degradation,
etc.?
2.Why was only one meal administered and monitored?
3.Why was only one small fragment of the entire insert subject to PCR
amplification, knowing that this would drastically underestimate the
presence of transgenic DNA?
4.Why did the researchers make what they know to be unjustified
assumption that transgenic DNA was absent in negative samples?
5.Why did the researchers not monitor for transgenic DNA in blood and
blood cells, when they are fully aware of previous research in mice
showing that transgenic DNA can indeed get into the blood, and from
there to other cells of the body?
6.Why was such a bad piece of research accepted by the FSA, and worse,
misinterpreted to indicate that GM foods are acceptable, when all the
indications are that the extent of horizontal transfer of transgenic DNA
is most likely to be much more extensive than the data indicate?



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