New paper on hold

[JennyJenny]

This may or may not have happened, accidentally or on purpose.

Regardless, the blinded multi-lab study is the clincher to me. WPI couldn't reliably detect spiked positive controls, and inaccurately found negative controls as positive. And the most bizarre part to me is that Mikovits claimed to have no issues culturing virus all this time, then when it came time to actually put up or shut up, their culture samples mysteriously were contaminated with mycoplasma, and they for some reason couldn't re-run them?

What makes me crazy is that WPI then went and destroyed the notes. Just keep them. When an accounting department has a General Ledger that doesn't make sense they don't destroy it. They find out what happened, make a journal entry and re-run it. They keep the first GL, the journal entry and the second GL. I would think for a lab they would keep the notes/results, re-run tests and then have the second set of notes/results.

Am I wrong here?

 

[Hustler]

Nothing ever came from Silverman into the part of the lab where Mikovits did her Science paper work

 

[garnet10]

It appears that Dr. Mikovits stumbled on to an "inconvenient truth"--that research labs have been inadvertently creating novel retroviruses from mouse pro-viruses that can actually be aerosolized, contaminate labs, and then here's the great unknown: could they potentially cause disease in susceptible humans? Even if there was no risk to susceptible human beings--this knowledge that novel retroviruses are being created through scientific research is profoundly disturbing.

It is very interesting to note that the blood supply is now treated with Cerus' INTERCEPT after WPI and Cerus did research to show that INTERCEPT was able to inactivate retroviruses such as XMRV in the blood supply. So at the very least, the blood supply has been protected. It does make me wonder about other biologicals that are produced in the lab?



I understand that an XMRV-like retrovirus could not be found in CFS patients in several other studies, however:

In a public conference call with the CDC on September 10, 2013, Dr. Lipkin reported on his findings with Dr. Jose Montoya of Stanford University regarding a well-defined cohort of ME/CFS patients. Lipkin reported, "We found retroviruses in 85% of the sample pools. Again, it is very difficult to know whether or not this is clinically significant or not. And given the previous experience with retroviruses in chronic fatigue, I am going to be very clear in telling you, although I am reporting them in Professor Montoya's samples, neither he, nor we, have concluded that there is a relationship to disease."

I find this statement incredibly fascinating, and the fact there were no follow up studies done to be very curious. If the source of these retroviruses is 1) from our very own scientific research, and 2) it can be proven that they cause disease in a susceptible population of humans--well then that would be utterly tragic--a truly unacceptable public health disaster.

 

[garnet10]

This article was referenced by Dr. Mikovits in one of her lectures and looks interesting although their ending statement is very carefully worded, and rightfully so.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3801167/


And then there's this paper:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3302299/

which states, "Second, it is readily apparent that MuLV, a mouse leukemia virus, can infect human B cells."

It sounds like some exciting research is being done, just not much of it. And of course, research data can be interpreted in different ways--I did not critically look at any of these articles, I just found their conclusions possibly relevant and interesting. So far, as far as I know, neither study has been retracted.

 

[Vaporization]

I don't have enough coffee to get through this paper right now , but this part was very interesting.

Partial molecular cloning of the JHK retrovirus using gammaretrovirus consensus PCR primers

Clinical history

The IP was a 39-year-old woman with a 3-year history of an ill-defined, viral-like complex of symptoms. An initial, mild febrile illness was followed by a relatively acute onset of marked easy fatigability, and then by severe, delayed post exertional malaise and fatigue, somnolence requiring prolonged bed rest, impaired cognition and verbal expression, forgetfulness, lower extremity muscle weakness, myalgias and paresthesias, occasional arthralgias and drenching night sweats, lymphadenopathy, and immunologic anergy. She slowly regained full activity and function.

And then there's this paper:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3302299/

which states, "Second, it is readily apparent that MuLV, a mouse leukemia virus, can infect human B cells."

Hey, maybe we can find something that will wipe out human B cells and force the body to make new ones. Then we may have a treatment.

 

[garnet10]

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3801167/
Partial molecular cloning of the JHK retrovirus using gammaretrovirus consensus PCR primers

Unlike earlier reports, in which MLV-like sequences were identified in human source material, which may have been due to murine contamination, budding retrovirions were demonstrated repeatedly by electron microscopy in uncultivated lymphocytes of the index patient that were morphologically identical in their development to the virions in the JHK-3 cells, and immunological evidence was obtained that the index patient produced IgG antibodies that bound to the budding viral particles in patient PBMCs and in the JHK-3 cells.

These data indicate that the patient had been infected by JHKV, lending significance to the demonstration of JHKV amplicons in nucleic acids of the patient’s PBMCs. In future studies, the PCR primer sets described herein may expand the detection of an amplifiable subset of viruses related to MLV.


This seems significant. In a patient with a CFS-like illness, these researchers isolated a novel gammaretrovirus in the Murine Leukemia Virus family. It makes one wonder if this is the same virus that Dr. Mikovits found in the blood of her CFS cohort? She definitely found something that was not related to Silverman's VP-62 plasmid (and when she sent him samples, he was unable to sequence her virus), and therefore it was not exactly XMRV. She also has an electron micrograph (albeit, from just one patient), of a virion budding from the cell of a CFS patient.


From Dr. Harvey Alter's now retracted paper:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2936598/
Detection of MLV-related virus gene sequences in blood of patients with chronic fatigue syndrome and healthy blood donors

We examined 41 PBMC-derived DNA samples from 37 patients meeting accepted diagnostic criteria for CFS and found MLV-like virus gag gene sequences in 32 of 37 (86.5%) compared with only 3 of 44 (6.8%) healthy volunteer blood donors. No evidence of mouse DNA contamination was detected in the PCR assay system or the clinical samples.





Dr. Elaine DeFreitas found evidence of a retrovirus in CFS almost 2 decades ago, but hers was similar to HTLV-II and her research could also not be replicated. Interestingly, she found also virus particles in mitochondria.

http://www.pnas.org/content/88/7/2922.short
Retroviral sequences related to human T-lymphotropic virus type II in patients with chronic fatigue immune dysfunction syndrome.

The majority of patients were positive for HTLV antibodies by Western blotting and for HTLV-II gag sequences by polymerase chain reaction and in situ hybridization. Twenty nonexposure healthy controls were negative in all assays.

The correlation between the presence of serum antibodies to HTLV and detectable' HTLV-II-like gag DNA or RNA in CFIDS patient blood was high but not universal. Of 13 pediatric patients positive by PCR, 10 were Western blot positive. Of 10 adult PCR-positive patients, 5 had positive Western blots. The lower frequency of antibody-positivity in adults is compatible with the reported incidence of cell-mediated anergy in adult CFIDS (27). Alternatively, antibody-negative patients may reflect latency of the retroviruslike gene. In fact, of 6 Western blot-negative CFIDS adults, HTLV-II gag mRNA could not be detected in 4. Conversely, of 6 Western blot-positive adult patients, 5 expressed HTLV-II gag mRNA.


I know that many researchers have been unable to replicate these retrovirus studies, but there certainly seems like there is some smoke there. And a viral infection would be consistent with the low NK cell function in CFS, as well as the "outbreaks" described, many of the symptoms of CFS, the waxing/waning nature of the symptoms etc. At least for some cases of CFS, a retrovirus makes sense. There are also some CFS patients who report they have been put into remission by anti-retrovirals.



In searching PubMed, I came across this article, likely done to assure the community that XMRV contamination did not work its way into vaccines:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3245253/

No Evidence of Murine Leukemia Virus-Related Viruses in Live Attenuated Human Vaccines

All eight vaccines were negative for XMRV and closely related MLV sequences using these two approaches. We found novel hamster genomic and retrovirus sequences in the JEV vaccine mostly likely originating from vaccine production in Syrian hamster cells. Our findings do not support the hypothesis that vaccines are a possible source of XMRV or MLV introduction into humans and are consistent with accumulating evidence on the absence of these viruses in humans.

Whoa!! So Japanese Encephalitis Vaccine contains "novel hamster genomic and retrovirus sequences"!

But what a relief to know that the other vaccines did not contain XMRV and closely-related MLV sequences.

 

[garnet10]

Regardless, the blinded multi-lab study is the clincher to me. WPI couldn't reliably detect spiked positive controls, and inaccurately found negative controls as positive. And the most bizarre part to me is that Mikovits claimed to have no issues culturing virus all this time, then when it came time to actually put up or shut up, their culture samples mysteriously were contaminated with mycoplasma, and they for some reason couldn't re-run them?

This is disconcerting, and unfortunately, I haven't been able to find any interview in which Dr. Mikovits, Dr. Ruscetti, Dr. Lo or Dr. Alter is given the opportunity to address this discrepancy. If anyone has any information on this, I would like to hear if they have an explanation.

Oddly enough, none of the labs were reliable:

Based on these findings, we conclude that currently available XMRV/P-MLV assays, including the assays employed by the three participating laboratories that previously reported positive results on samples from CFS patients and controls (2, 4), cannot reproducibly detect direct virus markers (RNA, DNA, or culture) or specific antibodies in blood samples from subjects previously characterized as XMRV/P-MLV positive (all but one with a diagnosis of CFS) or healthy blood donors.


It's so odd, because each of these 3 labs were being run by renowned, well-respected, experienced researchers. Hard to reconcile.


Finally, our findings are reassuring with respect to blood safety and indicate that routine blood donor screening for XMRV/P-MLV is not warranted at this time.

The ending statement to this paper is interesting, because just a couple of years later, the FDA instituted Cerus's INTERCEPT system to remove retroviruses from the blood supply.

 

[garnet10]

So, what you're saying is that the conspiracy runs so deep that they were able to convince 5+ different labs, close to 60 authors (including Mikovits), and countless lab technicians to all lie and commit research fraud, publishing papers saying that they tested for MLVs when they never actually did?

And not only that, but they also convinced Maureen Hanson, who has a son ill with ME, to also lie on her study? But this time they convinced her to claim that she did find evidence of MLVs even though she didn't actually look, but then also convinced her to claim later that she couldn't rule out contamination? (ref).

What do you make of their reasoning why they found more antibody in CFS patients than controls? It seems like such a stretch to me--that because other labs could not replicate their findings, they had to come up with a reason:

While it is impossible to know whether or not contamination has occurred in another laboratory, one possible explanation for the much higher frequencies of positive patients versus controls in both the Lombardi et al. [1] and Lo et al. [9]studies could be the existence of reagent or environmental contamination at the time of assay of patient samples, but not when controls were assayed.

Really? That seems statistically improbable, doesn't it?

In the movie, What About ME?, there is a clip of Dr Alter expressing his disagreement that his results were due to contamination. But I cannot find anything after that.

Dr. Mikovits felt Lipkin's study design was flawed--that they eliminated the patients who would most likely have shown up positive, but who knows?

I just wish the door wasn't shut on this research. There are still so many unanswered questions.

 

[weyland]

What do you make of their reasoning why they found more antibody in CFS patients than controls?

From which study specifically?

In general, I think that antigen cross reactivity with HERVs is one possible explanation for finding increased titers of MLV reactive antibodies in CFS patients. People who are sick with an immune insult show increased HERV activity.

 

[garnet10]

From which study specifically?

In general, I think that antigen cross reactivity with HERVs is one possible explanation for finding increased titers of MLV reactive antibodies in CFS patients. People who are sick with an immune insult show increased HERV activity.

I was just confused--it was not antibody, it was actual sequences of a virus. Thank you for the explanation of a possible cause of finding elevated antibody titers as antigen cross-reactivity. Although that's an interesting thought about the immune insult and increased HERV activity--perhaps our own HERV's become opportunistic and turn on us?

Maureen Hanson in her follow up paper made this statement as a possible explanation for contamination of the samples in both the Lombardi et al and Lo et al studies:

While it is impossible to know whether or not contamination has occurred in another laboratory, one possible explanation for the much higher frequencies of positive patients versus controls in both the Lombardi et al. [1] and Lo et al. [9]studies could be the existence of reagent or environmental contamination at the time of assay of patient samples, but not when controls were assayed.

I thought this possibility seemed statistically unlikely. There is contamination only when testing the CFS patient samples, but not when testing the control samples? In two (actually three, if we include Hanson et al) separate studies?

 

[weyland]

Although that's an interesting thought about the immune insult and increased HERV activity--perhaps our own HERV's become opportunistic and turn on us?

It appears to be a bit of a double-edged sword. Yes, HERVs are activated by immune signalling and can probably cause additional inflammation/autoimmune disease/neoplasms, but they also help tune and potentiate immune response to other pathogens. It appears that their presence may be necessary for normal functioning of our immune system, which would make sense given that we've co-evolved with these viruses in our germline. I believe I posted this paper upthread on the topic. It's very dense, but the conclusion gives a good summary.

 

[Hustler]

Mikovits ISOLATED whole viruses.
The sequence was just done wrong.
That s all it is folks.
Scientific geeks playing with syntax and words who took advantage of the wrong sequence saying oh we replicated SCIENCE paper 'error'. Lame. Smart.....!

The only way to prove Mikovits right is to take antiretrovirals, then go smash down a few doors.

 

[garnet10]

Mikovits ISOLATED whole viruses.
The sequence was just done wrong.
That s all it is folks.
Scientific geeks playing with syntax and words who took advantage of the wrong sequence saying oh we replicated SCIENCE paper 'error'. Lame. Smart.....!

The only way to prove Mikovits right is to take antiretrovirals, then go smash down a few doors.

I've read Osler's Web and Plague and note the many other studies that were done by Grossberg, Martin, etc. and it appears that researchers keep finding retroviruses in a certain subset of CFS patients.

I wish there was a commercially available test that was like Wistar's PCR or C. V. Herst's OnCore PCR, or a test to find reverse transcriptase, that one could do, to suggest who might benefit from ARV's.

@Hustler, besides the debilitating fatigue, did you have any other markers of immune dysfunction such as very low CD4 counts or abnormal T helper to suppressor ratios? Elevated RNAaseL or elevated levels of interferon?