XMRV CFS UK study #II

[Kati]

Thank you Dr Judy! If you are reading this, you are my hero! Keep up the good work.

 

[bullybeef]

Another simple reason for the lack of in-depth research of this 2nd UK study is lack of funding. It is a common thing in the UK to be 'tight' or frugal. Having a national health service means all research has to be funded ‘on the cheap‘, and once the research has the facts, then the NHS/MRC/DOH etc will release the funds to further these studies. A good example is Wessely & co offering XMRV testing with a fee attached.

Maybe it explains why both UK studies were rushed and without rigour. Not to mention, as long as they can continue proving XMRV doesn't exist in ME, the purse strings can remain firmly closed, and ME remains ‘in the head‘.

 

[Adamskitutu]

All of the above suggests that the cohort-issue, whilst important in general terms, is actually a bit of a red-herring here, and takes us away from the key-issue which is methodology. Method, method, method.

Except the cohort under study IS method, method, method. studying an 'AIDS' cohort with no 'AIDS' patients - only 'fatigued with sore throat' would be a key problem methodologically - wouldn't happen because no one would dare. Yet this problem of methodology is happening in the XMRV/CFS studies. The cohort under study is NOT a red herring, it is a key problem. Really cannot be brushed off the table. Penguins and ostriches.

 

[oerganix]

Its always been method. You can't get zero results from large patient studies by choosing the wrong cohort- unless the WPI is super selective in their patient selection process - and they've stated that they haven't been. I think this is the key



Although honestly how the Retrovirology missed this I don't know. If they knew about it and ignored it they must have assumed that their techniques would have been able to pick up the vanishingly small levels of XMRV in unstimulated cells. That study cost alot of money!

You need to stimulate the cells first but check this out - they said that growing the white blood cells is usually required - not always. Since the UK study used what Dr. Vernon suggested were more sensitive techniques then you would have thought they would have picked up the virus at some point - since according to this statement it isn't always necessary to grow the white blood cells - but they obviously didn't. Perhaps the other factor the WPI listed came into play.

One the problems is that this appears to be an unusual bug - you can't do 'normal' work and find it. Somebody, either the DHHS group or the CDC or somebody else is going to follow the WPI's work to the letter and then we'll know.

Cort and others, I think Dr Yes put forth a plausible reason in his previous post where he compared research looking at/for HIV retrovirus. Put that with Gerwyn's ideas that you need more time to culture XMRV to a 'density' where it can be found consistantly and you have an explanation for why they looked but didn't find. Maybe they were using methods that work for an HIV type virus but won't work for XMRV. All the more reason researchers should be carefully studying and using WPI's methods totally.

 

[Advocate]

From Angela Kennedy:

PERMISSION TO REPOST

My response to "Absence of xenotropic murine leukaemia virus-related virus in UK patients with chronic fatigue syndrome" in 'Retrovirology 2010'

http://www.retrovirology.com/content/7/1/10

(This response is awaiting moderation by retrovirology - it was submitted 15 February 2010)

What were the characteristics of the patient cohort?

Comment:
Immediate questions that spring to mind relate to the patient cohort and whether they were similar enough to the patient cohort in the Lombardi et al project.

Wow. Who is Angela Kennedy? That is a really good letter she wrote.

 

[starryeyes]

this may have been posted..sorry if it has. its from the ME association.

"One small but important add on piece of research that The MEA is continuing to pursue is to see if some of those people in the UK who have tested positive for XMRV using the US test can now be retested by one of the UK groups. It would also be very interesting to see if a mutually agreed cohort of CFS blood samples and control samples can be tested by all three UK and US research groups to see if they produce the same XMRV results."

Yeah! Now they're talkin'. :sofa:

 

[creekfeet]

Thanks rebecca1995 for posting the full text. I'm still not able to access it on WPI's site.

Cort, or somebody, could the full text be posted on the home page as news, so that those who can't get it at WPI can easily find it even when this-here page is buried in further posts?

I wonder if we should make a concerted effort to comment on the various articles reporting UK Study II, pointing out that they failed to replicate the WPI study. I'm seeing stuff like this: http://www.sciencedaily.com/releases/2010/02/100216142328.htm

]Further Doubt Cast on Virus Link to Chronic Fatigue Syndrome
ScienceDaily (Feb. 16, 2010) — Researchers investigating UK samples have found no association between the controversial xenotropic murine leukaemia virus-related virus (XMRV) and chronic fatigue syndrome (CFS).

A lot of people seeing that are going to feel so discouraged. I'd like to see us flood the comments with clarity and hope. Also, maybe we can prod the various editors toward a more reasonable editorial stance.

 

[gracenote]

As far as I can tell ScienceDaily has never posted on the original Science study. I kept watching for it after the study was published in October.

This is what comes up when I search ScienceDaily for XMRV and chronic fatigue syndrome.

Further Doubt Cast on Virus Link to Chronic Fatigue Syndrome
February 16, 2010 Researchers investigating UK samples have found no association between the controversial xenotropic murine leukemia virus-related virus and chronic fatigue syndrome. Their study calls into question a ...

New Virus Is Not Linked to Chronic Fatigue Syndrome, Suggests New Research
January 6, 2010 New research has not reproduced previous findings that suggested chronic fatigue syndrome may be linked to a recently discovered virus. The authors of the study say this means that anti-retroviral ...

Xenotropic Murine Leukemia Virus-Related Virus May Not Be Associated With Human Prostate Cancer
October 20, 2009 The xenotropic murine leukemia virus-related virus which has previously been linked to prostate cancer has been found to have a dramatically lower prevalence among German prostate cancer patients, if ...

First Evidence Of Virus In Malignant Prostate Cells: XMRV Retrovirus Linked To More Aggressive Tumors
September 8, 2009 In a finding with potentially major implications for identifying a viral cause of prostate cancer, researchers have reported that a type of virus known to cause leukemia and sarcomas in animals has . . .

creekfeet

I think your idea is great and we need to continue to do this. Maybe we can refine our "talking points" so we get a clear message across.

Quote from creekfeet: I wonder if we should make a concerted effort to comment on the various articles reporting UK Study II, pointing out that they failed to replicate the WPI study. I'm seeing stuff like this: http://www.sciencedaily.com/releases...0216142328.htm

A lot of people seeing that are going to feel so discouraged. I'd like to see us flood the comments with clarity and hope. Also, maybe we can prod the various editors toward a more reasonable editorial stance.

 

[Bob]

From Angela Kennedy:

PERMISSION TO REPOST

My response to "Absence of xenotropic murine leukaemia virus-related virus in UK patients with chronic fatigue syndrome" in 'Retrovirology 2010'

http://www.retrovirology.com/content/7/1/10

(This response is awaiting moderation by retrovirology - it was submitted 15 February 2010)

What were the characteristics of the patient cohort?

Comment:
Immediate questions that spring to mind relate to the patient cohort and whether they were similar enough to the patient cohort in the Lombardi et al project.

The Lombardi et al research cohort apparently met criteria for 'ME/CFS' as identified by Carruthers et al, in addition to Fukuda et al. which has been demonstrated to select patients with post-exertional malaise and fatigue, sleep dysfunction, pain, clinical neurocognitive, and clinical autonomic/ neuroimmunoendocrine symptoms (Jason et al).

Furthermore, the WPI give this information about their patient cohort in their supporting online material:

"Their diagnosis of CFS is based upon prolonged disabling fatigue and the presence of cognitive deficits and reproducible immunological abnormalities. These included but were not limited to peturbations of the 2-5A synthetase/RNase L antiviral pathway, low natural killer cell cytotoxicity (as measured by standard diagnostic assyas) and elevated cytokines particularly interleukin-6 and interleukin-8. In addition to these immunological abnormalities, the patients characteristically demonstrated impaired exercise performance with extremely low VO2 max measured on stress testing..." (www.sciencemag.org/cgi/content/full/117905/DC1)

The Erlwein et al project selected a rather different patient cohort:

""Patients in our CFS cohort had undergone medical screening to exclude detectable organic illness".

Advocate, I agree, this is great stuff...
It seems to be a major problem, in the UK, that in order for researchers to get funding from the MRC, the MRC doesn't allow the researchers to separate ME patients from Chronic Fatigue patients.
But, in the long term, I don't think it matters now, because I believe that everyone with the XMRV virus will eventually be tested and diagnosed as having XAND or some other name (XAND is the new name that the WPI gave to the disease caused by XMRV), and so the arguments about diagnostic criteria for ME will become irrelevant. That's if XMRV does turn out to be the cause of ME.

And I agree with everyone else, that the negative results of UK research teams are purely down to methodology, which explains why not a single person has tested positive for XMRV in the UK yet... except for the private testing coming back from the VipDx labs, and Judy Mikovits' own testing of uk patients.

 

[omerbasket]

https://www.wpinstitute.org/news/news_current.html


February 18, 2010: WPI is aware of the recent UK study that was unable to detect the presence of XMRV in any CFS patient samples. Although researchers at the WPI were not involved in this project, our work in XMRV continues with researchers around the world. We look forward to the results of studies which replicate the methods used in the original research described in the journal Science in October, 2009.

Information Regarding XMRV Studies

1. The authors of the Science paper established the existence of XMRV as an infectious human blood borne retrovirus for the first time in blood of patients diagnosed with Chronic Fatigue Syndrome (CFS). Previous studies had established the presence of XMRV sequences and protein in human prostate tissue.

2. In the Science paper, the presence of XMRV in well-characterized patients with CFS was established using multiple technologies:

a) PCR on nucleic acids from un-stimulated and stimulated white blood cells;
b) XMRV protein expression from stimulated white blood cells;
c) Virus isolation on the LNCaP cell line; and
d) A specific antibody response to XMRV.

3. The authors of the two UK studies did not attempt to replicate the WPI study. Replication requires that the same technologies be employed. The WPI sent reagents and information to several groups of researchers in an effort to support their replication studies. Neither UK study requested positive control blood, plasma or nucleic acids from the WPI.

4. The collection, preparation and storage of DNA were completely different between the Science and UK papers. The latter studies do not show data on blood harvesting or storage. Nor do the studies disclose the quantity of isolated cells. Insufficient number of cells analyzed may result in failure to detect a low copy virus like XMRV, regardless of the sensitivity of the assay. Neither UK study provides detail to allow interpretation of how many white blood cells were analyzed.

5. Patient population selection may differ between studies.

6. The UK authors were unable to detect XMRV, even though 4% of healthy individuals were found to be infected in the US. Japanese scientists detected XMRV in 1.7% in healthy blood donors in Japan. The two previously identified human retroviruses have distinct geographical distributions.

7. Perhaps the most important issue to focus on is the low level of XMRV in the blood. XMRV is present in such a small percentage of white blood cells that it is highly unlikely that either UK studys PCR method could detect it using the methods described. Careful reading of the Science paper shows that increasing the amount of the virus by growing the white blood cells is usually required rather than using white blood cells directly purified from the body. When using PCR alone, the Science authors found that four samples needed to be taken at different times from the same patient in order for XMRV to be detected by PCR in freshly isolated white blood cells. More importantly, detection methods other than PCR showed that patients whose blood lacks sufficient amount of XMRV detectable by PCR are actually infected. This was proven by the isolation of viral proteins and the finding of infectious XMRV isolated from the indicator cell line LNCaP. The authors of the Retrovirology paper admit that their neutralization assay did not detect bacterially expressed XMRV gag and that positive control sera was needed to validate their assay. The WPIs monoclonal antibodies specifically and sensitively completed the immune response demonstrating the assays sensitivity and specificity for XMRV envelope.

Simply stated the only validated reliable methods for detecting XMRV in CFS patients, to date, are the methods described in Science. Failure to use these methods and validated reagents has resulted in the failure to detect XMRV. A failure to detect XMRV is not the same as absence of this virus in patients with CFS.

This is a very good response for the WPI, and it makes it clear that even the second UK study is not a proof of anything, because it didn't follow step by step the WPI's study. And beside that - why not cooperate with the WPI in order to make a replication, and the best replication possible?

I think that untill there are at least 3-4 studies of researchers that we don't know of a bad bias by them, that replicated the WPI's study and that the WPI confirms that their replication is good, we should not think that XMRV is not connected to ME/CFS. As mentioned in the WPI's message - they were the first to find XMRV in a blood example. As also mentioned there, XMRV is very hard to detect. Therefore, even a good scientist, that does not consult with the WPI and does not follow their study step by step, might not find XMRV even if it's really there.

 

[Gerwyn]

This is a very good response for the WPI, and it makes it clear that even the second UK study is not a proof of anything, because it didn't follow step by step the WPI's study. And beside that - why not cooperate with the WPI in order to make a replication, and the best replication possible?

I think that untill there are at least 3-4 studies of researchers that we don't know of a bad bias by them, that replicated the WPI's study and that the WPI confirms that their replication is good, we should not think that XMRV is not connected to ME/CFS. As mentioned in the WPI's message - they were the first to find XMRV in a blood example. As also mentioned there, XMRV is very hard to detect. Therefore, even a good scientist, that does not consult with the WPI and does not follow their study step by step, might not find XMRV even if it's really there.

I think the point is that a good scientist following normal scientific protocol would consult the WPI a poor or coerced scientist would not

 

[cfs since 1998]

But the probability of this is so infinitesimally small, it is just not worth considering. With an almost random sampling of possible CFS patients being 50%+, it seems impossible that a cohort picked according to any definition would be ZERO in two separate studies. Really, it's not even worth considering. I'm no statistician, but I feel comfortable guessing that it has less than .001% probability.

Assume that XMRV really is present at a rate of only 1.7% in the general population, which is the rate found in Japan. That means the probability of a random person NOT having XMRV would be 0.983 (98.3%). Further assume they had completely the wrong cohort and might as well have selected people randomly. They did PCR on a total of 299 people, and the probability of 299 random people not having XMRV would 0.983^299 which is 0.0059, or 0.59%. Usually in science something is considered not attributable to random chance if the probability is less than 5%, or sometimes 1%.

 

[julius]

Assume that XMRV really is present at a rate of only 1.7% in the general population, which is the rate found in Japan. That means the probability of a random person NOT having XMRV would be 0.983 (98.3%). Further assume they had completely the wrong cohort and might as well have selected people randomly. They did PCR on a total of 299 people, and the probability of 299 random people not having XMRV would 0.983^299 which is 0.0059, or 0.59%. Usually in science something is considered not attributable to random chance if the probability is less than 5%, or sometimes 1%.

I wasn't referring to the 1-4% in general population. Could you try to do the number for this problem;

Let's assume that the selection process for the patient cohort in the UK studies was the same as the 'selection process' for the VIP commercial tests (ie not very stringent controls, non specialist GP diagnosis, throw dart at dartboard etc).

If the VIP commercial test turned up 50% pos. (in UK) then what is the chance of having 0 in the patients in both these studies?

 

[Hope123]

I was reading an article Kurt recommended about the search for retroviruses in various illnesses. (See my HERV and XMRV thread in this subforum.)

The odd thing about this study is that Kurt's article talks about how people with various autoimmune/ inflammatory conditions commonly produce antibodies to a retroviral/ HERV fragments leading to false-positive antibody results. I think CFS, whatever the cause, is a chronic inflammatory state (supported by some prior studies). If so, the odd thing is the ABSENCE of antibodies in the whole CFS group (except one person) in this study. I would expect more CFS subjects to have antibodies, even false-positive antibodies. Or, as discussed earlier, if by entirely random chance, the same percentage of antibodies as the controls.

Which leads me to think about the one sentence the researchers give about CFS subjects unable to produce antibodies due to poor immune status. Here's a truly wild thought............they did mention a very small number of controls having neutralizing antibodies specific for XMRV. What if they could characterize that antibody more and produce it as vaccine therapy for CFS sufferers? I'm skipping a whole bunch of steps here but this makes me wonder.........................

 

[julius]

Hope123

You find this answers some of your questions;
http://forums.aboutmecfs.org/showth...escape-virulence-factor-encrypted-in-envelope

Basically, research found that XMRV has the ability to suppress the immune system. and there is mention of the possibility to 'turn off' this ability, allowing the immune system to produce the necessary antibodies.

 

[cfs since 1998]

I wasn't referring to the 1-4% in general population. Could you try to do the number for this problem;

Let's assume that the selection process for the patient cohort in the UK studies was the same as the 'selection process' for the VIP commercial tests (ie not very stringent controls, non specialist GP diagnosis, throw dart at dartboard etc).

If the VIP commercial test turned up 50% pos. (in UK) then what is the chance of having 0 in the patients in both these studies?

I know, but I was saying what the worst case scenario would be. To calculate what you are asking you have to assume that WPI/VIP's tests are correct, they haven't been confirmed, meanwhile the general population figures of 1.7% to 4% have been found by three different labs to my understanding, so it is a more reasonable assumption.

The first UK study used 186 CFS patients and the second 170 CFS patients. IF it is *assumed* that 50% of randomly selected CFS patients have XMRV, the probability that you randomly select 186+170 negative CFS patients is 0.5^356 or ... 6.8 10^-108. That is 0.(107zeroes)68, or 0.(105zeroes)68%, or "impossible" for short.

 

[flex]

The UK government have issues around ME locked away from the public until 2071. They recently extended this period from 2030. So what is it they know that they are trying to hide until we are all dead. They have whitewashed this whole matter for decades now. So cover up, fraud and inhuman unethical treatment would not be ill described as a "conspiracy theory". When do they intend to stop this cover up? It would appear at least 2071!!

So back to the question, "what do they know, how long have they known it and what are they hiding"? Well, how do the studies here seem to find not one single person with a trace of XMRV when other geographical studies have shown a prevalence of up to 4% even in healthy controls. The question of cohorts and pre screening of organic illness has come up a number of times.

What exactly does the the pre screening of organic illness mean? They know things that we don't know and they have these issues locked away until 2071. I really wonder who made the first connection to XMRV!! This is not the first retro virus connection - that was made back in the early nineties. Defreitas was run out of town. What did they do with the evidence she provided. Did they really scrap it or will we find out in 2071?

 

[Hope123]

Thanks, Julius. Yeah, I read that thread prior and it was interesting.

This probably comes down to technique again and which antibodies the WPI vs. other groups are looking at but if that thread was entirely true, why is it the WPI had claimed previously that 95% of their CFS subjects were antibody positive? This was unpublished information but repeated by various press releases and even Dr. M or Peterson at one of their presentations. Also, you have 19/33 PCR negative CFS patients - from Science study at WPI - and another small group at NCI (separate group - 8/15) - exhibiting antibodies. (This from CFSAC.)

I'm not expecting an answer from you or anyone at this point but just putting my thoughts out there trying, like everyone else, to reconcile different pieces of data.

 

[julius]

I know, but I was saying what the worst case scenario would be. To calculate what you are asking you have to assume that WPI/VIP's tests are correct, they haven't been confirmed, meanwhile the general population figures of 1.7% to 4% have been found by three different labs to my understanding, so it is a more reasonable assumption.

The first UK study used 186 CFS patients and the second 170 CFS patients. IF it is *assumed* that 50% of randomly selected CFS patients have XMRV, the probability that you randomly select 186+170 negative CFS patients is 0.5^356 or ... 6.8 10^-108. That is 0.(107zeroes)68, or 0.(105zeroes)68%, or "impossible" for short.

Yeah...that's even smaller than I thought. Thanks so much. I have trouble making change for a twenty.

About the confirmation of VIP/WPI tests, I feel that they have been confirmed. The original paper had two replication studies built in to it (CC, NCI) It's too bad they couldn't publish those seperately.
Also, I think it is safe to say VIP/WPI are finding 'something'. And the UK studies aren't finding that 'something'.

 

[julius]

Thanks, Julius. Yeah, I read that thread prior and it was interesting.

This probably comes down to technique again and which antibodies the WPI vs. other groups are looking at but if that thread was entirely true, why is it the WPI had claimed previously that 95% of their CFS subjects were antibody positive? This was unpublished information but repeated by various press releases and even Dr. M or Peterson at one of their presentations. Also, you have 19/33 PCR negative CFS patients - from Science study at WPI - and another small group at NCI (separate group - 8/15) - exhibiting antibodies. (This from CFSAC.)

I'm not expecting an answer from you or anyone at this point but just putting my thoughts out there trying, like everyone else, to reconcile different pieces of data.

The anitbody tests were not done with the same reagents.

<edit> Oops, misread your post. Scratch that response.