Dr. O'Leary. Mr. Chairman and members of the committee, the
purpose of my testimony is to report the scientific results in
a series of children with autistic enterocolitis that has just
been described to you. Nothing in my testimony should or must
be construed as anti-vaccine; rather, it encourages safe
vaccination strategies. The opinions that I am expressing in
the text and in my presentation are those of my own.
These studies were undertaken following an approach made to
me by Dr. Andrew Wakefield, who has just submitted independent
testimony. The studies represent a transnational,
multidisciplinary research program aimed at elucidating the
causes and pathogenesis of inflammatory bowel diseases in
association with developmental disorders of childhood.
Next slide.
Dr. Wakefield has alluded to measles virus as a potential
associated factor in the pathogenesis of autistic
enterocolitis, and he came to our laboratory to seek molecular
confirmation.
Next slide, please.
He posed some questions. The first question: Was measles
virus present in gut biopsies from these children?
Next slide, please.
Where was it located? How much was there? Could it be
sequenced--could we actually confirm there was measles virus?
And finally, could different molecular technologies actually
independently confirm the presence of measles virus?
May I inform you that I am a pathologist and a molecular
biologist. My area of diagnostic expertise is histopathology,
which is understanding and examining the cellular basis of
disease.
Next slide, please.
The blinded study included 46 cases; 25 children had a
diagnosis of autistic enterocolitis, and 21 controls were
included in the study, including 15 normal children who did not
have a developmental disorder, four children with Crohn's
disease, which is a chronic inflammatory bowel disease
condition, and two children with ulcerative colitis, a very
similar condition but one which is histologically different.
We examined terminal ileal biopsies, and from the blocks of
tissue that were given to us, we looked at four to six serial
sections on the one case. What we wanted to do was to see if we
could identify the virus; could we replicate it on multiple
sections from one patient. Where available, fresh tissue was
examined.
Next slide.
The positive control materials in this study were
transvected measles virus-infected viral cells which contained
measles virus genome, and measles virus-infected brain tissue.
As negative controls, we screened a panel of other virally
infected cells lines.
Next slide.
To confirm the presence of measles virus, we adopt a five-
hit strategy. The first was to localize and identify the
presence of measles virus RNA in the tissue sections from these
patients.
The second was to quantify using a technology called TaqMan
real-time quantitative PCR. I have worked in this technology
for the last 6 years, and it is approximately 1,000 times more
sensitive than existing PCR-based technologies.
Finally, we wanted to confirm the sequence of the virus, so
we employed capillary fluorescent-based sequencing.
Next slide.
Let me summarize what in-cell PCR is. I know this is
technical, but I think it is extremely important. Measles virus
is an RNA virus, and RNA when it is removed from the body is
extremely easily degradable. PCR basically is like a
photocopying reaction. We can make multiple copies of a
particular gene that we are interested in looking for.
In-cell PCR allows us to demonstrate these genetic
sequences in cells and without cells, looking down the
microscope.
Next slide.
TaqMan quantitative PCR basically is a revolutionary PCR-
based technology which is sequence detection-specific. You will
only get a positive result in this assay if the desired gene of
interest that you are looking for is present in the tissue
section.
Next slide.
In our laboratory, we are one of the few laboratories in
the world that actually perform RNA inhibition assays. This is
effectively to test how degraded the RNA in our samples is, and
No. 2, is there anything in the environment of the laboratory
that is contributing to the breakdown of RNA in tissue samples
of cells.
Next slide.
I think it is extremely important in relation to the
detection of low-copy viral infections, and I think our
laboratory has a reputation for the detection of low-viral copy
numbers, that we have strict anti-contamination measures. What
I mean here is that we do not want to generate false-positive
results.
So we have separate, isolated extraction and PCR areas.
This is in a newly built, custom-designed molecular biology
facility. We have two independent laboratory sites, and for the
purpose of this investigation, we employed in situ
hybridization, which is basically a way of taking a cloned or a
fragment of DNA or RNA that you have in your laboratory and
looking for the presence of that in a tissue section or cell.
We can use the technology of in-cell PCR that I have described,
solution phase PCR, and TaqMan PCR, which are complementary
technologies, but TaqMan PCR is 1,000 times more sensitive than
standard solution phase PCR technologies, and then sequencing.
Next slide, please.
Just to reiterate again the point: Our laboratories go to
desperate lengths, No. 1, to prove that we do not have RNA
inhibition, but second to prove that we do not have
contamination. We purposely in all of our plates set up
contamination. This is effectively to outrule the possible
generation of false-positive results.
Again, I would appeal to the non-scientific members of the
committee that this is an extremely important control that must
be included if you are looking for low-copy viral gene
detections.
Next slide, please.
And of course, to confirm the presence of measles virus,
the gold standard is to confirm the sequence of the virus, to
say yes, this is measles virus RNA that we have got in the
tissue sections.
Next slide, please.
Let us look at the results now. We carry out an extensive
set of optimization reactions where we had measles virus
clones, looking at several regions of the measles virus genome,
F, N, and H. The technical detail of this is not important, but
just to show you that we could look at several genes within the
measles virus genome, and we could reproducibly make copies of
these or amplify them.
The bottom panel shows you an experiment which allows us to
detect the measles virus in viral cells, showing what the
optimal concentration of the probe that we require to detect
the virus is, and you can see clearly that the optimum
concentration is 1 to 1.5 micrograms per ml.
Next slide, please.
This slide shows the results of in-cell PCR by two
different technologies. One is in-cell solution phase PCR, the
other is in-cell TaqMan PCR, which produces a green signal in
the right-hand panels, top and bottom. Again it demonstrates
that we can clearly identify measles virus in these transvected
cells.
Next slide, please.
This is a patient with subacute sclerosing panencephalitis.
Measles virus is the cause of this condition. So of course, if
we are to reliably detect measles virus in biopsies of the
children that Dr. Wakefield provided to us, we should of course
be able to identify measles virus in brain biopsies from
patients with SSPE.
The left panel and the middle panel show measles virus
presence in these biopsies, and the right-hand panel is the
negative control.
Next slide, please.
This is a case of an autistic child No. 1. You can see the
top left-hand panel is a microscopic appearance of what the gut
looks like--and this is not normal gut epithelium from a child;
it is heavily inflamed. The bottom left-hand panel show a
spidery, black deposit, outer width of the cells that are in
green. This is actually measles virus RNA. The top right-hand
panel again in a different field shows a heavier deposit of
black, which again is measles virus, and again, I need to point
out to you that this is outer width of the cells that are
surrounding it.
Measles virus was located in the tonsils, in the lymphoid
hyperplastic areas that Dr. Wakefield described on endoscopy.
And the negative control is absolutely clean.
Next slide, please.
This is another case, a second autistic child, again
showing a very similar findings--a black deposit, which is
measles virus, outside of inflammatory cells, associating the
fibrillary matrix, and the negative controls here for mumps are
absolutely clean.
Next slide, please.
I think this slide is extremely important. If you look at
the left-hand panel, this is measles virus for the nucleocapsid
gene. It is a black signal. It again is the outer width of the
cells, but it has a spidery, cobwebby morphology. There are
little processes and tentacles coming out of this region. If
you look at the top right-hand panel, the follicular dendritic
cell has processes, dendrites, which form the cell matrix,
which is very, very similar to what we see on the left-hand
panel. And the immunocytic chemical staining analysis which Dr.
Wakefield alluded to gives exactly the same results.
Next slide, please.
By solution phase PCR, by what we call fairly standard
laboratory protocols, we can detect the measles virus in gut
biopsies from these children, and the negative controls are
appropriately negative.
Next slide, please.
Even in a paraffin-embedded tissue section that has
probably been sitting around in wax for 4 to 5 years, we can
detect measles virus. That is an astounding finding in view of
the fact that wax and fixation by itself breaks down RNA. This
graph shows in real time, we can look at the accumulation of
the PCR product that we are making, confirming that measles
virus is actually present in this biopsy.
Next slide.
And of course, we can sequence it, and we can say that this
is measles virus RNA present in the biopsies of these children.
To do this properly, you need to do it in a forward and
negative strand. I started off by saying that measles virus is
an RNA virus. When we make copies of the RNA virus, we make
copies in two-strand forms. So if we are to sequence it, we
should sequence in a forward and reverse strand.
Looking at that electrophoretogram, the sequences in both
directions are exactly the same.
Next slide, please.
In summary, then, in terms of the association that Andrew
Wakefield alluded to, I can confirm that his hypothesis is
correct--24 out of 25 children--that is 96 percent of the
children's biopsies that he sent to my laboratory, blinded--
children with autistic enterocolitis harbor measles virus
genomes.
Three out of four patients with Crohn's disease--these are
children with Crohn's disease--and one out of two children with
ulcerative colitis also contain measles virus. That is an
interesting biological fact.
Finally, 1 of 15, 6.6 percent, of control children harbored
measles virus genome. I think it does not take greater
statistical analysis to work out that there is a significant
difference between 24 out of 25 and 1 out of 15.
Next slide.
Solution phase RT-PCR, which is standard laboratory-based
technology, was positive in all children with autistic
enterocolitis for different regions of the measles virus
genome. That is extremely important. And we can sequence
measles virus isolates from these children.
Final slide, please.
However, the infection that is present in these children is
at extremely low copy, at about 5 to 30 transcripts per
approximately 2,000 cells.
Now, again, I want to add some technical overlay on this.
The way that we quantify RNA in our laboratory is by a method
called copy RNA. Again, we are one of the few laboratories
worldwide that would do this as the most accurate way of
actually measuring RNA. Indeed, industry now is accepting this
as a standard way of quantifying RNA in cells of tissue
sections.
Finally, the presence of measles virus was identified by in
situ hybridization, by solution phase PCR, by TaqMan PCR, by
in-cell PCR, by sequencing. This really, in terms of
technology, is as good as it gets right now.
I hope that I have given you evidence here which is
compelling in relation to the presence of measles virus in
children with autistic enterocolitis.
Thank you for your time and attention.
Mr. Burton. Thank you, Professor, and when we get to the
question-and-answer phase, I am sure that we will try to ask
questions in layman's terms and try to get some answers so that
we can understand everything that both you and Dr. Wakefield
said--but I think we got the gist of it.
Note/Warning:
Autistic people have fought the inclusion of ABA in therapy for us since before Autism Speaks, and other non-Autistic-led autism organizations, started lobbying legislation to get it covered by insurances and Medicaid.
ABA is a myth originally sold to parents that it would keep their Autistic child out of an institution. Today, parents are told that with early intervention therapy their child will either be less Autistic or no longer Autistic by elementary school, and can be mainstreamed in typical education classes. ABA is very expensive to pay out of pocket. Essentially, Autism Speaks has justified the big price tag up front will offset the overall burden on resources for an Autistic’s lifetime. The recommendation for this therapy is 40 hours a week for children and toddlers.
The original study that showed the success rate of ABA to be at 50% has never been replicated. In fact, the study of ABA by United States Department of Defense was denounced as a failure. Not just once, but multiple times. Simply stated: ABA doesn’t work. In study after repeated study: ABA (conversion therapy) doesn’t work.
What more recent studies do show: Autistics who experienced ABA therapy are at high risk to develop PTSD and other lifelong trauma-related conditions. Historically, the autism organizations promoting ABA as a cure or solution have silenced Autistic advocates’ opposition. ABA is also known as gay conversion therapy.
The ‘cure’ for Autistics not born yet is the prevention of birth.
The ‘cure’ is a choice to terminate a pregnancy based on ‘autism risk.’ The cure is abortion. This is the same ‘cure’ society has for Down Syndrome.
This is eugenics 2021. Instead of killing Autistics and disabled children in gas chambers or ‘mercy killings’ like in Aktion T4, it’ll happen at the doctor’s office, quietly, one Autistic baby at a time. Different approaches yes, but still eugenics and the extinction of an entire minority group of people.
Fact: You can’t cure Autistics from being Autistic.
Fact: You can’t recover an Autistic from being Autistic.
Fact: You can groom an Autistic to mask and hide their traits. Somewhat. … however, this comes at the expense of the Autistic child, promotes Autistic Burnout (this should not be confused with typical burnout, Autistic Burnout can kill Autistics), and places the Autistic child at high risk for PTSD and other lifelong trauma-related conditions.
[Note: Autism is NOT a disease, but a neurodevelopmental difference and disability.]
Fact: Vaccines Do Not Cause Autism.
International Badass Activists 