me.cfs.forums: Congratulations to Dr Enlander on his new collaborative endeavour

We wish to congratulate Dr Enlander on his new collaborative endeavour.  The team put together by Dr Enlander looks impressive and we now have an opportunity to answer questions definitively. If the researches were willing to establish baseline immune profiles very much in the same manner as demonstrated by Dr Judy Mikovits we could establish unequivocally that ME is a neuroimmune disease.


Rituximab doesa lot more than deplete mature B cells. To begin, there is a molecule called IL-2 which is elevated in people examined by Dr. Mikovits who were ZMRVpositive. This molecule is a signal molecule and is pivotal in activating the immune system, and more importantly from our perspective, keeping it active. It  also controls the level of certain kinds of T regulator cellcalled FoxP3.   Without sufficient FoxP3 the T cells start to attack the immune system, and an over-high ratio of T Helper 17 (TH17) cells is created, as is found to be the case in auto-immune and chronic inflammatory diseases.  If IL-2 (an interleukin, a type of cytokine) remains high, then the T regs response is suppressed, and an autoimmune situation will arise.   Rituximab also acts as an antibody against IL-2 molecules and hence switches off or at least dampens down the activity ofthe immune system.


Rituximab could also be helping by removing TH17, a major source of autoimmune symptoms. It also reduces the level of a molecule called NF-kappaB. This is always high in people with an activated immune system and/or inpeople with activated microglia in the brain.  The activated microglia are the ultimate source of neurotoxicity and mitochondrial damage. Importantly cytokine levels are maintained by NF-kappaB as is the continued activation of microglia, yet another way in which Rituximab administration could aleviate the symptoms of someone with neuroimmune activation.

Much of the molecular biology surrounding HIV has been revealed using a mouse model of the effects of a gammaretroviral infection. The virus used is a gammaretrovirus like XMRV and ZMRV and thecondition has been dubbed Murine Aids.  This condition is caused by a replication defective retrovirus coding a protein, which causes B cells to expand once the retrovirus has infected the cell.  This is called clonal expansion.  The disease created is multisystemic likeME.  Obviously if the ME retroviruses are found in B cells, as theyare in Murine Aids, then reducing the number of these B cells will aleviate symptoms. Finally, the B cells first meet the antigens (bugproteins) produced by the Murine Aids virus in the spleen, lymph nodes and the payers patches in the gut.  Now, B cells tend to spend considerably more time in the compartments of the body where they first encountered anantigen than they do in the blood.  In fact only about 4% of preactivated CD20 B cells are present in the blood at any given time.  So looking for a ZMRV which inhabits CD20 B cells in the blood is not a very clever idea. 


If base line Th17  Nf-kappa B  and Tregs were established we could monitor the effects of rituximab directly and that would give the ammo for a specific licence application.


If then they are willing to use Dr Mikovits's/Dr Ruscetti's reverse transcriptase assay in its entirety, and Dr Lo's assay, we can establish the presence of ZMRVs once and for all.


Finally, there is good theoretical evidence that ZMRVs are not likely to be found in the blood at detectable copy numbers, hence a variation specifically looking for ZMRV sin gut tissue would blow the negative studies out of the water.


4 experts + onestudy = definitive answer.



Finally, for this blog we must comment on the increasing censorship endured by supporters of Dr Judy Mikovits and her research on patient forums. Wethink that hidden agendas should have no place on forums which are suppposed toserve the interests of patients rather than the intersts of individual administrators and moderators. We will endeavour to make this a safe place from which to receive unbiased information and a blog which unashamedly supports thecause of scientific research into our neurological disease and advocates for oppressed sufferers. The following is a post from Angela Kennedy a long term andhighly respected advocate.

I wanted to let people know that I have been prevented from posting on threads because I REPORTED OTHERS for blatantly breaking this forum's rules, and for them attacking other forum members, and using a campaign of intimidation and humilating tactics to silence, and called said others on what they were doing.

The moderator then said that what the person who started doing this on a particular thread, was 'fine', and has allowed it to stand.

This forum has been subject to abuse of moderator power for a long time, where moderators punish victims of abuse of forum rules, and use their power to advance their own partial positions, allowing others free rein to attack and intimidate others into silence and implied acquisance.

Free speech has been banished, and agent provocateurs of both the ideological and employed kind proliferate, spreading demoralisation and powerlessness.

Sadly forums have become one of the ways in which people who wish to advocate for this community to protect sufferers of ME/CFS against medical abuse. It is where people learn to mobilise, to discuss the issues, gather information. This forum is stopping that, and various other forums are on the way to this behaviour too.

I am speaking out because the level of abuse is blatant - I eventually recognised it for what it was. It took me a while because I actually do try and moderate my own behaviour to be reasonable. I'm no shrinking violet, and I don't have ME, so I can only imagine how dreadful this situation must be for people who are so ill - and find themselves powerless in the grip of such abuse of power and advancing of positions not in the interests of this community.

I empathise with those people on this forum - and I know there are enough of you because of discussions away from this forum- who have been intimidated and moderated into silence while others blatantly flout forum rules and intimidate others in order to advance points of view that are potentially and/or actually damaging to the interests of the ME community.

All I can hope is that this post is read before it is removed, and people will find some comfort in understanding the problem, and seek to get things changed. Whether that means leaving this forum and starting elsewhere, or having the courage to tell Cort and friends where they are going wrong has to be up to individuals.

This community has been under siege for too long. Friendly fire has ALWAYS been a problem, and the level of friendly fire has intensified with the advent of Mikovits's research. Sadly this is far more serious an issue than silly spats on a forum.

No evidence that a strain of XMRV was created in a cell line in the 1990s

Update on the paper Paprotka et al.

The argument in Paprotka et al. (a study led by Pathak and Coffin) is that the sequence isolated by Silverman, which he claimed came from patient VP-62, is a result of a unique recombination event. This unique recombination event occurred between a virus, found in many species of mice, and a hitherto undiscovered replication defective erv, when the xenograft which was involved in the creation of the 22Rv1 line was passed through mice.

They claim, but cannot prove that these mice were NU/NU and or Hsd mice, as they contain the viruses or at least the virus fragments they deem to be necessary to further their hypothesis.

They argue that their assays did not amplify XMRV in the early xenografts, but after several passages of the cells through the proposed mice their assays were able to amplify XMRV. Thus XMRV must have been absent from the early xenografts and created by this unique recombination event.

So we have to judge between two explanations. One is that the VP-62 sequence was generated by a unique recombination event, or that the PCR assays used lacked the clinical sensitivity needed to detect low copy VP-62 sequences in the early xenografts.

Given that a unique event is so very unlikely they must provide very strong evidence that VP-62 equences were indeed absent from the early xenografts and that their apparent absence was not caused by the failure of their PCR assays. Absence of evidence is not evidence of absence.

If one focuses on the initial quantitative PCR assay using one envelope primer it becomes clear that there is no evidence provided regarding the copy number of VP-62 sequences per 100 cells in the NU/NU or Hsd mice, later xenografts or early xenografts. This is because the PCR amplified what were known to be VP-62 sequences from the DNA taken from 22Rv1 cell line and the CWR-R1 cell line. The problem here is that the copy number of VP-62 in these cells is huge, being 2000 copies per 100 cells and 3000 copies per 100 cells respectively. We do know that this assay could detect ERV sequences in the mice and the early xenografts, but we don’t know what the copy number of ERV sequences is compared to the copy numbers of VP-62. Thus it is quite possible for VP-62 sequences to be present but undetectable, due to a lower copy number for example. There are other more complex reasons. In any event absence of evidence is not evidence of absence. We also have the problem that this assay with the env primer was not used to screen other species of lab mice or wild derived mice. Had this assay being able to detect either XMRV or ERV sequences in these mice the paper would have collapsed. Certain vested interests have argued that it would be silly to use this assay with this primer to screen the other mice because the primer could amplify both VP-62 and ERV sequences. In that case why use that primer to screen NU/NU and Hsd mice, and indeed why use that primer at all. I wonder what the defense is for not using this primer and assay to any screen wild mice. Wild derived mice are not wild mice.

There is no evidence that this assay can amplify XMRV specific env sequences other than in the DNA taken from 22Rv1 and CWR-R1 cells. There is no evidence that this assay with this primer could detect VP-62 sequences in the DNA extracted from lab mice, the early xenografts or the later xenografts. The later xenografts were not screened using this assay. In fact figure S3 in the SOM (Supporting Online Material) shows that provided this primer was used at all to screen the later xenografts using the second single round PCR, then that PCR assay could not amplify any product. As this primer was used to screen the early xenografts with the second PCR assay (Figure S3), not to use it to screen the later xenografts would be a serious departure from the scientific method.

Next we examine the second PCR single round assay with a gag primer, which is only capable of detecting VP-62 gag sequences. Figure 2D and 2E suggest that this assay, with this primer, was used to screen the early xenografts. This is however in direct conflict with the text, which states that all the primers used with the single round assay were capable of amplifying both VP-62 and ERV sequences. This is prima facie evidence of misfeasance and at the very least should be investigated. In any event, it seriously weakens if not invalidated their claim that gag sequences were not present in the early xenografts. Indeed we once again have the scenario that there is no evidence that this single round assay can detect VP-62 gag sequences in anything other than the DNA taken from the 22Rv1 or CWR-R1 cell line either. So they cannot provide evidence that their assay could detect VP-62 GAG sequences in mice or later xenografts let alone the early xenografts.

Thus I will leave it to those without vested interests to consider whether these authors have provided convincing evidence that VP-62 sequences are absent from the early xenografts, bearing in mind that absence of evidence is not evidence of absence and that the alternative explanation is a unique recombination event, which cannot have happened in the past and its future occurrence is a virtual impossibility.

Finally the paper does not fulfill the minimum requirements of surviving a peer review process. There is not enough evidence to enable the scientific merit of the study to be judged or enough data in the methodology section to enable replication. The claim that it is custom and practice among a certain group of retrovirologists not to include such details unless they deem it necessary merely means that none of their studies should have passed the Science peer review process or indeed any rigorous peer review process. Contrary to statements made by certain vested interests the Science magazine is not published merely for the perusal of certain retrovirologists but also other scientists from different disciplines. I would remind everyone what the requirements of science are:

"Data and materials availability
All data necessary to understand, assess, and extend the conclusions of the manuscript must be available to any reader of Science."

Paprotka et al., ‘Recombinant Origin of the Retrovirus XMRV’ does not meet the requirements as specified by Science. 

Permission to repost 

Thank you to Pumpkin for the post.