I know it's been a few years since this thread was active, but could somebody please answer just one question? Was reverse transcriptase ever found in either ME/CFIDS patients or autistic patients? Was it ever even looked for? If not, why not? And can we look for it?

After all the reading I've done, in the book, Plague, and in numerous newspaper and magazine and journal articles and papers -- this is the most important to me of many questions I still have about the whole XMRV saga.

Dear Dr. Miller:

I was gratified to receive your response, especially the first paragraph in which you said, "I appreciate your effort to understand the disease that affects your family, and to reach out to scientists who might have useful information. That said, I know very little about autism, including possible genetic or environmental components."

You and I have probably both reached the age where we're well aware of our strengths and weaknesses, and try to highlight the former, while minimizing the later. I take no offense and look upon it only as an opportunity to further connections between people who are interested in solving the medical problems which affect so many.

The problem with the little you know of autism is shared by so many. Considering that something like 50 kids came down with the measles in my state last year and it was treated as a public health emergency while at the same time approximately 5,000 were diagnosed with autism makes many of us wonder about the priorities of the public health system. Our community would feel so much better if upon a diagnosis of autism, a public health swat team swung into action giving every imaginable test to try and figure out what had gone wrong with that specific child.

You might be interested in this article I wrote in which the government confirms they are testing approximately 100 children with autism for the presence of the XMRV retrovirus. http://www.ageofautism.com/2011/06/government-confirms-testing-of-children-with-autism-for-presence-of-xmrv-retrovirus.html

I understand your frustration with the small amount of research funding presently going to XMRV research and offer an opportunity. Starting with the autism community, but branching out to other disease communities who feel they've been long neglected, including ME/CFS, Gulf War Syndrome, lyme disease, we are forming a new political party to advocate for more research dollars for these diseases.

It is called "The Canary Party" and you can access it here. http://www.canaryparty.org/ I would consider it an honor if you joined us.

All the best,
Kent Heckenlively

Dear Kent Heckenlively,

I appreciate your effort to understand the disease that affects your family, and to reach out to scientists who might have useful information. That said, I know very little about autism, including possible genetic or environmental components.

I should also say that I interpret the bulk of the scientific data to date to support the conclusion that XMRV is not present in humans. It's not that I want this to be the case. Indeed, I and many other retrovirologists were excited that we might use our knowledge of retroviruses to help understand and treat prostate cancer/CFS/autism.

So when you say your family members have tested positive for XMRV, I am prone not to believe the accuracy of the results, but would be glad to be convinced otherwise. Where were these assays done? If virus could be grown in culture from your wife and mother-in-law, what was the sequence of the virus(es)? These days, it is so easy to sequence viruses to establish their identity, that such sequencing would be required to document the nature of the virus detected. If the virus(es) very closely match known isolates of XMRV, such as VP62 or 22Rv1 XMRV, or known mouse retroviruses, then the finding may be the result of contamination. But if the viruses show some diversity, indicative of virus evolution in the new human hosts, then perhaps your family members are indeed infected. Have these assays been independently verified by a second lab?

Despite what I have said on this thread, I think your investigation of a potential link of autism to vaccination is a reasonable effort. I predict that XMRV will not be the culprit, but perhaps another virus is involved. As for me, while I am interested in the eventual resolution of the XMRV story, the small amount of funding that I have to study XMRV is about to disappear, and we are winding down our effort in this research area.

I wish you and your family the best, and hope you take no offense at anything that I have said.

Dusty Miller

Dear Dr. Miller:

I appreciate your continuing engagement with me on the question of whether infection by the XMRV retrovirus may be related to autism and the question of whether vaccines may play any role in this disorder.

I'm not sure if you've followed my writings on the question of the possible relationship of XMRV to autism so let me briefly recap the information known by my more regular readers.

My daughter with autism/seizures tested positive for XMRV through antibodies. My wife and my mother-in-law tested positive through culture. I have tested negative for the virus as has my father. My mother passed away fifteen years ago, but given the results with my father and I the more reasonable conclusion is that she would also have been negative.

This evidence actually mitigates against a belief that current vaccines contain XMRV, although leaving out the question of how my mother-in-law came to acquire the virus.

To tell you about my wife, as a teenager she had psoriasis so severe it covered 90% of her body. Although she has been a speech therapist for more than 20 years in the rehabilitation unit of one of our best local hospitals, for the past few years she's noticed significant changes in her cognition. "I test people for brain injuries," she tells me, "and if you tested me you'd find significant deficits." She compensates for them, but strongly believes testing would reveal them. My mother-in-law was one of the first 1,000 people in the United States diagnosed with celiac sprue disease.

I must stand with my fellow autism parents, though, in my assertion that there was a change in my child after a vaccination. However, there were also many other complicating factors. My daughter had just gone off of breast milk and was on a cow's milk formula. She was only six months old so it was difficult to assess development as my wife and I were also late developers. At ten months of age she was diagnosed with a seizure disorder and at the age of three she was diagnosed with autism.

I believe the milk allergy plays into this disorder as for reasons unrelated to autism my son was kept on a hypo-allergenic milk formula when we switched him at fifteen months to save some money. At his eighteen month check-up the pediatrician ran him through a full developmental work-up, he passed with flying colors, and he recieved his shots without me giving it a second thought. A few days later my wife mentioned that she hadn't heard our son speak in a day or two. He'd had 15-20 words on the day of the visit to the pediatrician.

By chance I ran across the book, "Unraveling the Mystery of Autism" by Karyn Serrousi which thought gluten and casein were at the root of the problem. I put my son and daughter on the diet the next day. After 12 days my son said his first word again. it took him about a year to catch up in his language development, and about two years for his sensory problems to go away. The diet did nothing for my daughter and since that time I've always wondered what process we arrested in my son, but failed to do with my daughter.

After the diagnosis of my daughter with the XMRV retrovirus I contacted the UCSF pediatric AIDS unit to discuss the issue. I mentioned only that my daughter had been tested positive for this retrovirus, and that she had autism, and that there was a possibility that the two conditions could be linked. The representative with whom I spoke immediately said, "Well, that would explain why vaccines might trigger autism." I was a little stunned by this comment and asked him to explain further explain himself.

He said that all one needed to do was liken what is done with children who are born to HIV infected mothers with the situation I described. Babies born to HIV-infected mothers are given anti-retrovirals in order to lower the viral load prior to giving any vaccinations. As I'm sure you know, retroviruses like to hide out in the B and T cells of the immune system so that any stimulation of the immune system has the possibility of stimulating the virus. Although he was just the media representative, he felt fairly confident that this was a basic tenent of retrovirology. I must also note that an allergy, and my wife's family has been plagued with milk allergies, is also a condition which will produce an inflammatory response.

Considering that at a rate of 1 in 100 children with autism, this disease is more than 20 times more prevalent that polio was during any epidemic. A recent study suggests that autism is not genetic, as has long been the orthodoxy, but is more likely related to environmental factiors, of which prenatal infections may be a significant factor. http://archpsyc.ama-assn.org/cgi/content/full/archgenpsychiatry.2011.76

The question of whether vaccines at present contain XMRV is an open one, but nobody can doubt that vaccines are designed by their very nature to stimulate the immune system and produce a mild inflammatory response. Thus, regardless of the source of XMRV, a vaccination has the potential to stimulate XMRV, just as a vaccination will stimulate the HIV virus.

The finding of 10 times more viral particles of an unsuspected porcine circovirus than of the target virus in the rotarix vaccine raises the question of other unsuspected viruses circulating in biological materials given to humans. It is my hope that advances in technology, like deep sequencing, will enable us to answer these questions and make chronic diseases as rare as infectious diseases are today.

All the best,
Kent Heckenlively

Per...............

Is the asteroid bigger than a bread box?

Posted by: Media Scholar | July 17, 2011 at 02:30 AM

I hope the asteroid is as big as North and South American in mass/size and is headed directly for the Earth. A bread box would burn up in our atmosphere and do nothing. I want to see something that would put the earth out of commission. No more vaccines, no more nothin'

Oh, I feel so much better that the manufacturers and the FDA are going to the greatest lengths to making sure that the vaccines are not contaminated with other retroviruses. Thank you Dr. Miller. Oh, wait a minute. Did not the porcine retrovirus sneak in and in large quantities in the rotavirus vaccines? Yes, they did miss that one. It was only because some independent lab with "new" technology found it that we know now that the vaccine was contaminated. So they pulled one vaccine off the market and kept giving the other to small babies. Then, when they finally discovered that the other vaccine was contaminated with both versions of the porcine retrovirus, they decide to put both vaccines back on the market knowing full well that they contain the porcine retrovirus. Nice.

P.S.

regarding your comment: "And certainly, any cell line infected with XMRV would not pass testing required for use of the cells in vaccine manufacture. Part of the reason is because XMRV replicates so well in many cell lines that it is easy to detect by standard reverse transcriptase assays"

Are you forgetting that a virus may well be latent and not actively replicating? Are you forgetting that xmrv was discovered decades after vaccines and other biologicals have been in use? Also worth remembering instances when RT activity was detected in the past in cell lines used for production of biologicals, but was deemed (by regulators and manufacturers) to "present no danger". Even though they would not have a clue as to its source...

---------

FDA Letter to Viral Vaccine IND Sponsors - Use of PCR-based Reverse Transcriptase Assay (re detection of retroviruses in their products), dated 1998

http://www.fda.gov/BiologicsBloodVac.../ucm105911.htm

… Whichever PBRT test you choose to employ, its performance should be validated, especially with regards to the lower limit of detection, the specificity (generality for retroviral RTs, negative for non-retroviral polymerases with RT activity), and the reproducibility of the assay. The lower limit of detection ("sensitivity") should be comparable with the published literature. ... it is possible that the results will be positive, which may require further characterization of the source of the positive activity. In the latter case, IND sponsors are recommended to contact CBER for further guidance in this regard, prior to undertaking any additional testing. …. If positive results are obtained, infectivity studies may be requested to demonstrate that the source of the positive activity is not an infectious retrovirus.

“are recommended”, but not legally required? What would “infectivity studies” be? Would those studies definitely rule out XMRV or any novel/still unknown exogenous infective retrovirus? Infectious ALV HAS BEEN detected in vaccines, and deemed to present no danger? Reassuring? What about such testing in 30's, 40's, 50's?

D Miller,

There isn't anything wrong in your reply on the surface, but the reality is while that yes vaccine makers and the FDA do go to great lenghts to detect viral contamination (I should know- I worked in vaccine development and manufacture for more than a decade and know this first hand) you can't test for what you don't know about. These tests are specific for certain known viral contaminants. So, no matter how hard you try there is no certainty that you have a culture free of viral contaminants.

And yes, with good laboratory practices you should easily control contamination. But all it takes is one time and your culture is screwed. Working all day long everyday I'm willing to bet some 24 year old lab tech in the middle of switching playlists on his ipod will inadvertantly mix up spent media with fresh media. It happens. I've seen it happen again and again. Humans are careless.

Here are my comments about Heckenlively's post:

Re: XMRV -Science Journals
Scarier than Science Fiction

This piece by Kent Heckenlively confirms that a little
bit of knowledge can be dangerous!

The problem with the piece is that he got the wrong
definition of "xenotransplantation".

Xenotransplantation refers to transplantation of cells
between species, not just from mice to humans, as
Heckenlively quotes.

In fact, the report he discusses deals with
xenotransplantation of human cells into mice, not the
reverse.

This procedure is used to coax human cancer cells to
grow outside of humans for further study.

Surprisingly, human cancer cells are often difficult to
grow in cell culture, outside of a human, but are more
easily grown in nude mice.

While mouse cells have been transplanted into a few
humans, this procedure is very rare and could in no
way explain chronic fatigue syndrome or autism.

A. Dusty Miller, PhD

Heckenlively responds with the comment:

"I understand a Dr. Dusty Miller from the Fred Hutchison Cancer Center has raised some questions as to my suggestion that mouse tissue which harbors XMRV may possibly end up in a vaccine."

Nowhere did I mention vaccines. Neither did the paper Heckenlively finds so scary.

But what about vaccines? Yes, there is the possibility of undetected viruses in vaccines, but vaccine manufacturers and the FDA go to great lengths to avoid this possibility. And certainly, any cell line infected with XMRV would not pass testing required for use of the cells in vaccine manufacture. Part of the reason is because XMRV replicates so well in many cell lines that it is easy to detect by standard reverse transcriptase assays, which measure an enzyme that is present in all retroviruses.

What about retrovirus spread in the lab? XMRV is not like the ghouls from the Night of the Living Dead, able to be shot and still pop up to attack. XMRV and other retroviruses are fragile, such that treatment of these viruses for a few seconds with detergent or 70% ethanol, common laboratory disinfectants, kills them very effectively. However, culture medium from infected cells can contain over ten million infectious virus particles per milliliter, so one does need to be careful to prevent virus spread. For example, feeding infected and uninfected cultures with the same pipette can easily spread the virus. But with proper care, it is simple to keep XMRV and other retroviruses under control.

In the 1990's I use to show my students a video were a top scientist that worked for the CDC would fly all over the United States to look for the hantavirus. He would come in looking like a great hunter/warrior/ doctor wearing his protective gear hmmmmmmm made the girl's in my class hearts all go pitter patter, and the boys green with envy.

So, where might this great warrior/hunter/doctor of mouse viruses from the CDC at now????

the truth will never ever be told, it has been a complete cover-up since day one...anytime any researcher got close to any answers the carpet was pulled from under them...there is no doubt in my mind that 45% of the hiv virus envelope will be a major step to the truth...in 1969 25 million u.s. dollars changed hands between the dept of science and technology and the u.s. military and low and behold the hiv epidemic started years later and the exact words used in that meeting were immuno-deficient and immuno-compromised where one will kill the host and the second will incapacitate the individual...we are all led to believe that m.e. has been around for a century but that is an outright lie, cfs is a new entity created in a u.s. lab...hiv virus is not naturally occuring, it is a synthetic creation...xmrv is a total waste of time and is would only stand to cover-up the full truth from emerging...we know that u.s. troops were vaccinated before iraq and we also know the threat imposed by suddam if they were to invade iraq and the u.s government knows what suddam had as a weapon because it was the u.s who armed suddam with these weapons prior to iraq invasion...this is exactly why the cdc/nih have stalled for decades on cfs and including autism and they will continue to do so...they will alwys come out and say they care and in the end it is all part of the military bio-warfare that was created...i will never believe in all the hype that comes from especially cdc/nih...they are ruthless.....sincerely and always the truth aidan walsh southampton, u.k p.s. xmrv is probably a result of a piece of the aids virus.....

The best-laid schemes o' mice an' men
Gang aft agley,
An' leave us nought but grief an' pain,
For promis'd joy!

Burns, R (1785) "To a Mouse, on Turning Her Up in Her Nest with the Plough" http://en.wikipedia.org/wiki/To_a_Mouse

Isn't it *odd* that Vincent Racaniello chose to kill off his mice after the xenograft study was released online?

"Transgenic mice susceptible to poliovirus
Posted: 12 Jul 2011 07:32 AM PDT
Yesterday I terminated the last remaining mice in my small colony, including the line of poliovirus receptor transgenic mice that we established here in 1990. Remarkably, I had never written about this animal model for poliomyelitis which has played an important role in the work done in my laboratory.

While I was still working on poliovirus as a postdoctoral fellow with David Baltimore, I became interested in how the virus causes disease. There were no convenient animal models to study poliovirus pathogenesis, so I began to think about the cellular receptor for the virus and how it could be used to make a mouse model for infection. When I moved to Columbia University Medical Center in 1982, I decided to identify the cellular gene for the poliovirus receptor. This work was carried out by the second graduate student in my lab, Cathy Mendelsohn. She identified a gene from human cells that encoded a protein which we believed to be the cellular receptor for poliovirus. When this human gene was expressed in mouse cells, it made them susceptible* to poliovirus infection (the mouse cells were already permissive for poliovirus replication). The gene encodes a transmembrane glycoprotein (illustrated) that we called the poliovirus receptor (PVR), later renamed CD155. Over the years we worked extensively on PVR, with the goals of understanding its interaction with poliovirus during entry into the cell. In one project we collaborated with Jim Hogle, David Belnap, and Alasdair Steven to solve the structure of poliovirus bound to a soluble form of PVR. The image of that complex decorates the banner at virology blog and twiv.tv.

Shortly after identifying PVR as the cellular receptor for poliovirus, a new student, Ruibao Ren, joined my lab. For his project I suggested he create transgenic mice with the human gene for PVR. We already knew that synthesis of PVR in mouse cells allowed the complete poliovirus replication cycle. Together with Frank Costantini and JJ Lee, Ruibao produced PVR transgenic mice and showed that they were susceptible to poliovirus infection. The illustration at top left shows a PVR transgenic mouse with a paralyzed left hind limb after poliovirus inoculation.

Poliovirus transgenic mice were used for many years in my laboratory to study how the virus causes disease, and to identify the mutations that attenuate the neurovirulence of the Sabin vaccine strains. A good summary of this work can be found in my review, ‘One hundred years of poliovirus pathogenesis‘. But there is a dark side of this story that I wish to briefly recount. When we first developed PVR transgenic mice, my employer decided to patent the animals. Until the patent issued, we could not share the transgenic mice with other researchers. As a consequence, others developed their own lines of PVR transgenic mice. One of these lines has been qualified by the World Health Organization to determine the neurovirulence of the Sabin vaccine strains. However, Columbia University realized little income from the PVR transgenic mice – such animals cannot be patented in Europe. By patenting the mice, we simply delayed research progress. Because of this experience I am personally very wary about patenting biological discoveries.

There are several reasons why I decided to stop doing research with mice. The cost of housing and breeding mice is very high, nearly $1.00 US per cage per day, and I simply don’t have the funds to support such work. More importantly, no one in my laboratory has any interest in working with mice: the last student to do mouse work left years ago. Although there are many interesting experiments to be done using viruses and mice, that line of work ended for the Racaniello lab on 11 July 2011.

*A susceptible cell bears the receptor for the virus; a permissive cell allows viral replication. A susceptible and permissive cell allows the complete viral replication cycle.

Ren, R., Costantini, F., Gorgacz, E., Lee, J., & Racaniello, V. (1990). Transgenic mice expressing a human poliovirus receptor: A new model for poliomyelitis Cell, 63 (2), 353-362 DOI: 10.1016/0092-8674(90)90168-E"

The International ME Association has also posted an answer to Dusty Miller's response at http://www.mecfsforums.com/index.php/topic,8102.msg98678.html#msg98678

"Dr Miller demonstrates the danger of an overreliance on metaphors in the guise of definitions. The study (Zhang, 2011) demonstrates how recombinant MLV viruses have spread from mice into cell lines and tissue cultures. The study further demonstrates that X-MLVs can enter other cell lines being held in the laboratory, at the same time. These recombinant X-MLVs are highly infectious and hence the process of xenografting can easily explain the entry of a number of related but different X-MLVs into the human population. The presence of these viruses could in every way explain the aetiology of ME/cfs and autism"

An excellent report, thank you.

Bear in mind that the US and UK government Health services have been actively blocking any research into ME/CFS since 1985. A retrovirus was found in us in the early 1990s and that work was suppressed.

Please help us by calling our illness ME/CFS. The other way around is what the psychs are using in an attempt to hang on to the derogatory name of CFS.

witches brew now takes on a new meaning. Green that vaccine? Not possible. One or two ingredients, is not greening a vaccine. Especially if they still use animal and human cell lines. I am now wondering if the simean CMV virus found in polio has helped the retrovirus cauldron even more? Since I am positive per WPI, and my kids with autism, the bigger question is, how do we get out of this fine mess? HIV children often have neurological problems very close to autism, hmmm?

Postscript:

Which means to them that the autism epidemic -- having humans with communication problems is perhaps not such a bad thing.

Our scientific arrogance has now reached an all time high. A sure sign we are overdue for an asteroid impact.