Problems with Deisher’s Study— Part II: Biological Implausibility

My apologies for taking so long to get to Part II. Between losing one of the days I allotted for work to a non-stress test, grossly underestimating how much could be wrong in such a tiny section and trying to whittle it down, waiting for personal correspondence [updated 9/25/14], and, you know, life, it has taken a bit longer than I anticipated. I’m still going to plow through, though, because I am sure this won’t be last time this zombie hypothesis is given new life.

Deisher’s study* is incredibly, incredibly thin in the realm of biological plausibility. This is surprising (or not) because she is making some novel, extraordinary claims. Yes, she has a nice bibliography salad, but the studies she cites do not directly, or even indirectly at times, support her central hypothesis that DNA from fetal cell lines is a direct environmental cause for increasing autistic disorder (AD) diagnoses. Further, even her unpublished research that has been cited in newsletters and pro-life media means very little in terms of the hypothesis. So, where to begin? Perhaps, we should start with a very simplistic reminder of what DNA is.

DNA stands for deoxyribonucleic acid. It is our genetic “blueprint” located primarily in the nucleus of each cell in our body. The well-known double helix shape results from the base pairing of DNA nucleobases— adenine (A), guanine (G), cytosine (C), thymine (T)— with the deoxyribose (sugar) and phosphate groups forming the backbone. In the human genome, there are approximately 3 billion base pairs.
Pertinent to this conversation, it is important to remember that the DNA doesn’t “do” anything by itself anymore than the plans to a building builds the building. The DNA is “read” through a process called transcription. During transcription, an enzyme, RNA polymerase binds to the DNA at a promoter region just prior to the gene in question and produces messenger RNA (mRNA) after transcribing the termination sequence. The mRNA can then be decoded by the ribosome, an organelle, in a process called translation to assemble a protein. The protein-coding sequence of a gene is called an exon.

Now, let us consider the residual DNA that can be present in vaccines. In order to grow the viruses used in viral vaccines, you must use a cell line; viruses proliferate in cells. Some vaccines produce viruses in cell lines (WI-38, MRC-5) that were derived from the lung tissue of two fetuses who were the victims of unrelated elective abortions. Leaving aside the philosophical moral issues and our visceral reactions to the evil of abortion, from an objective, scientific standpoint, these are completely normal human cells. Their verifiable normality was the major reason for the development of the cell lines in the first place.

During the vaccine manufacturing process, the cells are destroyed to harvest the viruses. The product is also treated with the enzyme DNAse, which fragments the unprotected DNA at random. In total, the purification processes reduce the DNA to trace, nanogram(ng) levels. (A nanogram is one billionth of a gram.)

The first problem when looking at the biological plausibility of Deisher’s DNA hypothesis is what she is actually measuring and how accurate those measurements are. It should be noted that Deisher never tested MMRII (measles, mumps, rubella) or Varivax (varicella) vaccines; she only tested MeruvaxII (rubella) and Havrix (hepatitis A). It is unclear whether she can make any kind of valid assumption about the DNA content in the untested vaccines, especially as the 2 micrograms(µg) she uses as the DNA content of Varivax comes from an approval document that is over 20 years old. Further, Dr. David Gorski, MD, PhD, a published surgical oncologist and cancer researcher, noted under his nome de plume some potential problems in Deisher’s methods, including a lack of PCR (polymerase chain reaction) and the results potentially muddled when you combine the specificity of the Picogreen test with the insufficiency of degrading RNA by heating alone. You can read the details on his blog (though he is not charitable to the religious who promote bad science).

Like Gorski, I also noted something very curious when reading the study. If you’ve ever done experimentation in the sciences or engineering for review, you know that you will always have the evaluating party (professor or reviewer) expect to see some tables, pictures, graphs, etc. Yet, in the Deisher study, there is bupkis, other than not very detailed results. After some digging, I found an unpublished study of Deisher’s that follows the exact same methodology (ELISA kits–PicoGreen/OliGreen, SYBR gold staining after 4% agrose gel electrophoresis) on the exact same vaccines. These results should not give us much confidence with either the numbers published in the study nor in how they advance her hypothesis. You’ll notice the inconsistencies from Figure 2: some vials show both double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA); some have one or the other. The weights are not consistent. Only two base-pair measurements were taken with MeruvaxII and one is less than 200 base pairs. (That will be relevant when discussing FDA recommendations.)

That should say “Tables from Figure 2”

For the sake of argument, let’s assume that her numbers are accurate. It isn’t necessary to dispute the numbers to demonstrate that the fear-uncertainty-doubt spread by this study is completely unjustified. In the long run, though, we should keep in mind that this might be a dubious assumption.

The mechanisms: Lack of evidence and plausibility
While Deisher admits that she and her coauthors “do not know the causal mechanism behind these new contaminants and autistic disorder,” she proposes two mechanisms: autoimmune reactions and/or genomic insertions and mutations. While she refers to these mechanisms as “known” and “clinically documented”, they are not and she does not back up that claim with sources.

We can basically rule out the idea of the trace, fragmented DNA by itself causing an autoimmune reaction. Unlike an allergic reaction, which involves hypersensitivity to an allergen, autoimmune reactions result when the immune system erroneously recognizes the body’s own cells as “foreign” and then attacks them. Deisher, herself, conceded in an expert testimony that there is no evidence for free DNA fragments causing an immune reaction: “I must note that Merck did investigate the possibility of an immune response to the contaminating DNA and found no response – which is one of the reasons we have focused on the theory of genomic insertion.” In the highly criticized Ratajczak study, citing Deisher as a source, the premise of vaccine-DNA-induced autoimmunity in the etiology of autism relies on genomic insertion. Truly, the only sources she cites in her study (Freimanis 2010, Tai 2008) refer to the potential genetic components of (completely unrelated) autoimmune diseases like rheumatoid arthritis and multiple sclerosis. “It is the proteins and not the DNA that generates the immune response in humans,” explains Rev. Dr. Nicanor Austriaco, OP. Therefore, we have to focus on the ridiculous implausibility of normal, human DNA inserting and expressing itself in such a way as to cause autism.

Think about all the things that would need to happen to make this hypothesis even possible. These normal, human fetal cell line DNA fragments would have to:

1. Enter the cells (these are presumptively neural cells in the brain, based on Deisher’s previous assertions)
2. Enter the nucleus of the cell
3. Insert or recombine into genome both behind/with a proper promoter region and have a termination sequence
4. Consistently code for all the genes associated with autism
   —or—

   Consistently code for some protein that triggers an autoimmune response to the cell (and that autoimmune response be responsible for autism)^#

5. Express that code in substantial quantity to cause systemic autism

Then, you would still have to demonstrate that this actually happens in autistic people. As you can see, this is a very, very, very tall order. To paraphrase Dr. Paul Offit, a well-published doctor and co-inventor of the rotavirus vaccine who is highly respected among his peers, the likelihood of all of this happening is roughly equivalent to a child flapping his arms and taking flight. But we can’t even get off the ground in step 1.

There are a great many barriers to getting even the trace amount of DNA from a vaccine to neural cells. The injection is given into the muscle or right under the skin— the DNA fragments would need to inexplicably bypass those cells and remain intact and functional through interstitial fluid and get into the blood stream. From there, the DNA fragments would have to inexplicably breach the blood-brain barrier, a highly selective barrier that impedes the influx of molecules into the brain from the blood. Despite what you may have heard, it is formed and functional at birth. “There is no clear mechanism to explain how DNA injected into a baby could ‘wreack havoc’ on the baby’s DNA,” says Fr. Austriaco, “especially since there is a blood brain barrier that prevents the movement of molecules from the blood into the brain.”

Deisher previously has proposed a mechanism as to how the DNA bypasses the blood-brain barrier that is so far removed from reality, I can’t even come up with a cogent analogy for how absurd it is. In December 2009, Deisher claimed that “DNA injected into a muscle can be picked up by receptors on the nerve ending. Once DNA has been taken up in to the nerve endings, it can be transported back up the nerve’s axon to the brain.” Deisher might as well have claimed the DNA gets there by magic. If axons, the “nerve fibers” that conduct electrochemical impulses, could regularly transport unspecific, non-adapted macromolecules up to the brain, we would all be in seriously big trouble because that would mean the blood brain barrier is completely useless and the brain is an easy target for any toxin or pathogen that enters our bodies. Deisher says this occurs by retrograde axonal transport, but this makes no biological sense. Only select pathogens have adapted specifically to take advantage of retrograde transport to enter the nervous system; it is those specific adaptations that scientists are hoping to exploit for targeted gene therapy and Deisher knows this. (It’s right there in the sources she cites in the newsletter!) To imply that we should expect free, fragmented, non-adapted DNA to behave the same way is… seriously problematic.

Even if the DNA fragments somehow managed to get up to the brain, Deisher presents no evidence that they are capable of permeating the cell membrane. Even her own unpublished research, shows that not all cells will spontaneously uptake DNA into their cells. This was even after she incubated the cells with far, far higher concentrations of DNA than could exist in the body from a vaccine!

“Think about how hard it is to do gene therapy,” says Dr. Offit. In gene therapy, you have a specific, intact gene; you have a promoter region; you have a way to get it into the cells like a viral vector; you are injecting microgram(µg) quantities of this specific, intact, correctly coded DNA (1µg= 1000ng). Even then, it is very hard to get the genes expressed. “It would be the best news for gene therapy, ever. You can give random fragments of DNA and get the phenotypes of autism? Wow.”

Of course, no one, not even Deisher, has demonstrated that unprotected DNA fragments will enter normal human neural cells, enter their nuclei, and incorporate into the genome. In fact, people who work in biomedical research were the first ones to draw my attention to her unpublished research and its fatal flaws. Deisher tested 7 different cell lines, and only 2 showed DNA insertion into the host genome even with the high concentrations. Both NCCIT (teratocarcinoma) and U937 (lymphoma) are cancerous cell lines. She couldn’t get genomic insertion in any sort of naturally present, healthy cell. She could only get it in cancer cells isolated and cultured for research purposes in a lab. And, in case people haven’t taken note, cancer cells can do some weird things!

In fact, the “weirdness” of cancer cells is the entire reason the FDA has a recommendation of limiting residual DNA in vaccines to 10ng, but you would never know this from Deisher and SCPI. While not mentioned in the study, SCPI continues to exploit this recommendation to “prove” how “unsafe” the DNA levels are in fetal cell line vaccines. But reading the actual recommendation, to the contrary, shows just how quantifiably ridiculous it is to perceive residual fetal cell line DNA as a threat.

The 2005 draft presentation cited by SCPI, the 2008 update, and 2010 FDA guidance make it very clear that the concern is with limiting cancerous/continuous cell lines because of concerns about oncogenicity, the capability of the cells to induce tumors. This is not the case with established diploid cell lines like MRC-5 and WI-38. In the 2005 and 2008 presentation, you will see the term “no limit” for diploid cell lines; the 2010 guidance states, “For widely used human diploid cell strains, such as MRC-5 and WI-38 cells, measurement of residual DNA might be unnecessary because we do not consider residual DNA from these human diploid cells to be a safety issue.”

Another pertinent section in the 2010 guidance:

The risks of oncogenicity and infectivity of your cell-substrate DNA can be lessened by decreasing its biological activity. This can be accomplished by decreasing the amount of residual DNA and reducing the size of the DNA (e.g., by DNAse treatment or other methods) to below the size of a functional gene (based on current evidence, approximately 200 base pairs). Chemical inactivation can decrease both the size and biological activity of DNA.

This is actually important because, as you recall, in one of her unpublished studies, Deisher only found one vial of MeruvaxII with a bp size above that of what is considered a functional gene, even if we ignore the randomness and inconsistencies in the measurements. Further, in Victoria 2010, cited by Deisher in her study, the authors only focused on live-virus vaccines (not inactivated vaccines like Havrix for hepatitis A) “[b]ecause the chemical inactivation used in the manufacture of killed-virus vaccines is also likely to inactivate adventitious viruses.” (for brevity, think of this as the potentially problematic DNA sequences.) If Deisher were to try to say anything about the DNA in Havrix, she would have to prove that the current expectation regarding the effect of chemical inactivation on the DNA is wrong, too.

Of course, this is ultimately immaterial. Even with tumorigenic cell substrates (not normal diploid cells) a 10ng limit and reduction to < 200 bp, would provide a factor of safety greater than 10⁷! (10⁷ = 10,000,000). “It wouldn’t matter if it’s 10ng or 10µg,” Offit explains. “It isn’t going to do anything.”

Truly, Deisher should know just how much more DNA than what could be in a vaccine (DNA with verifiable oncogenes to boot) is required to produce an oncogenic event. SCPI declared,

As the FDA authors of a 2008 paper about the cancer dangers of residual human DNA state “Whether this residual cell-substrate DNA can induce tumors in vaccine recipients and thus represent a risk factor has been debated for over 50 years without resolution.”

First, you cannot conflate cancer vaccines and normal childhood vaccines. Even then, the article states clearly that it was only mice inoculated with two types of DNA each at 12.5µg who developed tumors. 12.5µg is the equivalent of 12,500ng and double that amount was needed, in total, to produce tumors. Compare that to the highest level of DNA Deisher claims is in fetal cell line vaccines… and, again, this is with known, complete cancer-causing DNA!

It’s seems if there is even a tangential mention of “DNA” and “vaccine” in the paper, Deisher is wholly uninterested if the concerns expressed are applicable to fetal cell line vaccines. In Mostovoy v. Sec’y HHS, in which Deisher was the expert for the petitioners, the Chief Special Master noted that the “petitioners’ claim that the FDA failed to meet its own safety standards when licensing vaccines containing human DNA is wholly unsupported by any part of the record.” This “record” includes several sources that have been cited by Deisher here and in the past. The Special Master decries things like a “misleading partial quotation” and “numerous unsupported assumptions and inconsistencies.” The summation in that case could very well sum up what we can say about Deisher’s current study: she and her coauthors “have claimed support for their position by relying on documents that furnish no such support.”

Lastly, let’s just remember the scale of things here. There are approximately 3 billion base pairs in the human genome, which exists in all the diploid cells of your body. There are an estimated 37.2 trillion cells in the human body. A nanogram is one billionth of a gram. A seven-pound newborn weighs 3.2 trillion nanogams.

One billion as a visual. One trillion is 1,000 billion.

This is why the scientific community would demand extraordinary, overwhelming evidence to take Deisher’s hypothesis seriously, not some David and Goliath conspiracy against the poor, persecuted pro-lifers: it is fantastically, mind-bogglingly implausible.

Really, I shouldn’t have had to say anything about this at all. As Rational Catholic Blog explained before, all current research into autism indicates a prenatal onset. The de novo mutations Deisher repeatedly invokes in her study are, by definition, germline mutations that occur in egg, sperm, or in the newly fertilized egg itself. You can’t get any more prenatal than that! “Further, these genes exert their effects in utero,” added Offit, meaning that they have been found to affect the development of the baby while he is still in the womb. Even with a hypothesis as fantastically implausible as Deisher’s DNA hypothesis, there is one hypothesis that I think we can declare impossible: time traveling vaccines.

Or are we to believe in some superstitious nonsense where WI-38 and MRC-5 cell lines are so infused with evil humors from the link to abortion that they can defy the laws of nature God wrote into creation?

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Stay tuned for Part III: Study design and Conclusions
See the previous posts on this study:
– Abortion, Autism and Immunization: The Danger of the Plausible Sounding Lie
– Problems with Deisher’s study— Part I: The numbers
– Looking a little closer at the numbers— A supplement to Part I

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* I will often conflate Deisher with her coauthors or Sound Choice Pharmaceutical Institute (SCPI). As her most ardent devotees do the same thing, I will do this for brevity.

# “It’s not at all clear whether the markers of inflammation sometimes reported in the brains of autistic children are a cause, a consequence, or merely an epiphenomenon of autism.” Dr. Gorski

Special thanks to Dr. Paul Offit and Fr. Nicanor Austriaco for their personal correspondence!