XMRV CFS UK study #II

[garcia]

I was just reading your blog, Cort, and it brought up a question. I'm not sure if it has been asked before, but there hasn't been any great discussion on it.

What technique did the Japanese use? They looked at blood, and they did it the summer before the WPI study was published.

Has anyone else tried using their technique?

Does anybody even know what the technique was?

Here is a really good blog where the author lists all the XMRV papers and the various techniques they used, plus the results they got:

http://neuroskeptic.blogspot.com/2010/02/case-of-missing-retrovirus.html

Just scroll down to where it says: "Published Papers". You'll see that the Japanese study apparently used antibodies.

 

[Gerwyn]

This is from the original science paper:



Is this an unusual or usual step in PCR?

This is what they did with the T-cells






Do these statements indicate that you have to culture the wbc's first in a special way?

not unusual for looking for a latent virus activation is key concentrating your product is normal you need to culture Band T cells to have any chance of producing virus

 

[Cort]

Thanks Gerwyn,

If you're going to the trouble of culturing that essentially says this virus is hard to find (or latent) does it not? How can you ignore a signal like that?

That was a big study! That cost the UK government a ton of money of money. I just can't imagine. They're looking at hundreds of samples and every one is turning up negative - wouldn't they at some point culture a few to see if that was necessary? These are all very smart people. Their reputations are on the line as well. It really blows my mind. It makes me wonder if I'm missing something.

if that's true regarding the standard steps for retroviral amplification then shame and Gow and Kerr fpr participating in that study. How can a 'world-renowned retrovirologist' participate as well? Is that the state of research in the UK? Do they not dot their i's and cross their t's?

A problem with the cohort question is that the WPI has repeated said now that the patients in the study were typical patients and they were not characterized by the abnormalities stated in the Science paper. I believe the queston Dr. Vernon has regards the cohort concerns duration of illness (since that can effect viral presentaton), severity of illness (another potentially key factor regarding viral presentation) and treatment (since several of the patients have been identified as having lymphoma).

 

[Gerwyn]

Thanks Gerwyn,

If you're going to the trouble of culturing that essentially says this virus is hard to find (or latent) does it not? How can you ignore a signal like that?

That was a big study! That cost the UK government a ton of money of money. If they missed something as simple as that they should be FIRED! I just can't imagine. They're looking at hundreds of samples and every one is turning up negative - wouldn't they at some point culture a few to see if that was necessary? These are all smart people. It really blows my mind.

if that's true regarding the standard steps for retroviral amplification then shame and Gow and Kerr fpr participating in that study. How can a 'world-renowned retrovirologist' participate as well? Is that the state of research in the UK? Do they not dot their i's and cross their t's?

A problem with the cohort question is that the WPI has repeated said now that the patients in the study were typical patients and they were not characterized by the abnormalities stated in the Science paper. I believe the queston Dr. Vernon has regards the cohort concerns duration of illness (since that can effect viral presentaton), severity of illness (another potentially key factor regarding viral presentation) and treatment (since several of the patients have been identified as having lymphoma).

This virus would not have been found at all without the human genome project that is how difficult it is to find hidden since the dawn of virology!You are right about the points but they did conform to the canadian criterea it is easy to make the oxford look like feduka Its hard for people to appreciate the politics in the UK the big issue with bloods is whether the people who took the blood knew it was for virology purposes or not now the wpi is a centre for virus research but the uk people in the early samples cant have known what the blood was for do it wrong and no viable virus.All the british control group had one illness or other where there are high titres of endogenous viruses expressed making the test results impossible to authenticate i cant understand how a virologist who knew the implications would do this

 

[Angela Kennedy]

Thanks Gerwyn,

if
A problem with the cohort question is that the WPI has repeated said now that the patients in the study were typical patients and they were not characterized by the abnormalities stated in the Science paper. I believe the queston Dr. Vernon has regards the cohort concerns duration of illness (since that can effect viral presentaton), severity of illness (another potentially key factor regarding viral presentation) and treatment (since several of the patients have been identified as having lymphoma).

Cort,

Are you able to point to exactly where the above has been said? I've been going by the accompanying additional information to the Science paper, which was presented in PDF form (a copy of which I believe I've saved), which said the cohort fulfilled BOTH Canadian and Fukuda. This would be in order, because virtually all 'Canadian' patients would also fulfill the symptoms criteria for Fukuda.

Are you saying the Lombardi et al cohort did NOT fulfill Canadian criteria or the additional characteristics I cited above?

 

[Angela Kennedy]

Hi Angela,
One thing that I'm not clear on, and I wonder if anyone can clear this up for me?
The WPI study says they selected patients using the Fukuda and Canadian criteria...
does this mean that each patient has to fulfill both criteria,
or does it mean that that some patients fulfilled the Fukuda definition, and some the Canadian definition?
Although, I think that this question is a little redundant, because the WPI were testing for physical signs of organic illness anyway, so they were basically narrowing down the patients to an organic illness which has an effect on the body at the cellular level.
Bob

Hi Bob,
Many thanks for your warm welcome.

Regarding the cohort, the information I got was from the accompanying supporting online PDF to the Science article, which as I've said to Cort, I think I've saved. I did consult that PDF to reply to Cleare et al (co authors to the Erlwein et al paper) on Plosone in January. But today the link didn't work.

I have read that information as being both criteria were fulfilled by each patient . If one considers the additional abnormalities that 'characterised the cohort', and the likelihood of 'Canadians' also fulfilling Fukuda, it seems the most logical deduction. It should also be noted that Cleare et al referred to the Canadian Guidelines in order to explain why they did NOT use them in their study, indicating they understood the Canadian Guidelines identified the Cohort.

Having said that the devil is the linguistic detail (as with ALL 'CFS' criteria actually). This may need further clarification. As I think you're pointing out, the issue is specific organic abnormalities in the WPI cohort (something that is indicated in Canadian criteria, and confirmed in the additional abnormalities that characterised the cohort), whereas both Fukuda alone and Oxford are designed to weed out organic illness by the exclusionary clauses they employ linguistically.

By the way I don't think Canada is perfect, but this doesn't negate the need for a clearly defined patient cohort that follow their characteristics (as opposed to the almost diametrically different cohorts defined in Oxford and even Fukuda), especially in Britain, and it doesn't excuse those such as the UK projects ignoring their significance. That will go for PACE too, when the results are published.

Oh, and Peter Kemp's 'Penguins and Ostriches' rules ok!!! : )

 

[jackie]

Dumb "Q", I know...but is there ANY other way to find it?...in the gut, maybe..by way of endoscopy/etc....like ENT's? or tissue samples? ANYwhere else?

If I ruled the world...we'd start from scratch (with all on the SAME page).

Thanks!, (Obviously) no-scientist-jackie (this is so stupid I wish I could type it tiny...but I seriously want to know!)




BTW...pretty darned funny Anne!

 

[Gerwyn]

Dumb "Q", I know...but is there ANY other way to find it?...in the gut, maybe..by way of endoscopy/etc....like ENT's? or tissue samples? ANYwhere else?

If I ruled the world...we'd start from scratch (with all on the SAME page).

Thanks!, (Obviously) no-scientist-jackie (this is so stupid I wish I could type it tiny...but I seriously want to know!)




BTW...pretty darned funny Anne!

At the moment it can be found in epithelial cells of prostate tissue associated with cancer and with more difficulty in other types of prostate cells(stroma) apart from that it is in special cells of the immune system called B and T cells .We dont know of any other technique for isolating the virus from these cells apart from the one used by the WPI

 

[jackie]

Thanks Gerwyn! (I wish YOU were ruling our World....I bet we'd get to where we NEED to be a lot quicker!)



j

 

[oerganix]

• http://www.ifarablog.org/search/label/XMRV
• Mouse to Man? XMRV and Human Disease. Stephen Goff, Columbia Univ Coll of Physicians & Surgeons, New York, NY, US
• Virus-Host Interaction: HIV and XMRVOrgan and Cell Lineage Dissemination of XMRV in Rhesus Macaques during Acute andChronic Infection. Prachi Sharma, Yerkes Natl Primate Res Ctr, Emory Univ, Atlanta, GA, US
• XMRV: Examination of Viral Kinetics, Tissue Tropism and Serological Markers of Infection. John Hackett Jr, Abbott Diagnostics, Abbott Park, IL US
• SWINE FLU MEETS HIV 2009 Pandemic Influenza A (H1N1) Virus Infection in HIV-Infected Adults. Esteban Martinez. Hosp Clin, Barcelona, Spain

The first guy, Goff, speaking of XMRV in prostate cancer, says it 'easy' to cultivate. The second speaker, Charma, said they have infected macaques with XMRV and they found it in reproductive organs and .......I couldn't understand her, but she may have said lyphoid-something. The next guy, Hackett from Abbott spoke of their research, also around prostate cancer, and how they are looking for 'assays' to detect XMRV in blood, etc.

It appears this site is mainly about HIV. They will have a more complete conference at noon Pacific time, but I doubt there will be much on XMRV. I just thought it interesting that Goff says XMRV is easy to cultivate. And that there are studies being done on non-human primates going on right now.

 

[flex]

One thing is for sure regardless of scientific techniques - and that is, XMRV has been discovered in humans. To be honest from here on its a battle of politics and power. With the establishment and their years of denial- THEY HAVE GOT AN AWFUL LOT OF POO TO GET BACK IN THE HORSE!!

 

[Gerwyn]

• http://www.ifarablog.org/search/label/XMRV
• Mouse to Man? XMRV and Human Disease. Stephen Goff, Columbia Univ Coll of Physicians & Surgeons, New York, NY, US
• Virus-Host Interaction: HIV and XMRVOrgan and Cell Lineage Dissemination of XMRV in Rhesus Macaques during Acute andChronic Infection. Prachi Sharma, Yerkes Natl Primate Res Ctr, Emory Univ, Atlanta, GA, US
• XMRV: Examination of Viral Kinetics, Tissue Tropism and Serological Markers of Infection. John Hackett Jr, Abbott Diagnostics, Abbott Park, IL US
• SWINE FLU MEETS HIV 2009 Pandemic Influenza A (H1N1) Virus Infection in HIV-Infected Adults. Esteban Martinez. Hosp Clin, Barcelona, Spain

The first guy, Goff, speaking of XMRV in prostate cancer, says it 'easy' to cultivate. The second speaker, Charma, said they have infected macaques with XMRV and they found it in reproductive organs and .......I couldn't understand her, but she may have said lyphoid-something. The next guy, Hackett from Abbott spoke of their research, also around prostate cancer, and how they are looking for 'assays' to detect XMRV in blood, etc.

It appears this site is mainly about HIV. They will have a more complete conference at noon Pacific time, but I doubt there will be much on XMRV. I just thought it interesting that Goff says XMRV is easy to cultivate. And that there are studies being done on non-human primates going on right now.

could it have been macrophages and leucocytes? it isn t that hard to cultivate if you look in the right cells and the cells are numerous enough but the chances of finding it without "cultivation" techniques are remote

 

[parvofighter]

More on Coffin and Goff's Retrovirology Conference comments on XMRV

Hi Gerwyn and Oerganix, I transcribed some of the videos of this week's Conference on Retroviral Infections & Opportunistic Infections in this thread: http://forums.aboutmecfs.org/showth...etroviral-Infections-Opportunistic-Infections

The pi [FONT=&quot][/FONT]ce de r [FONT=&quot][/FONT]sistance: Coffin saying that there is no question that XMRV is real and infecting people. As CBS said in that thread, this really puts the first two UK studies in perspective.

Reason for cheer!:Retro smile::Retro smile::Retro smile::Retro smile:

 

[usedtobeperkytina]

And, a retrovirus that infects people, particularly in immune system cells has got to be one that causes illness.

Tina

 

[Gerwyn]

Hi Gerwyn and Oerganix, I transcribed some of the videos of this week's Conference on Retroviral Infections & Opportunistic Infections in this thread: http://forums.aboutmecfs.org/showth...etroviral-Infections-Opportunistic-Infections

The pi [FONT=&quot][/FONT]ce de r [FONT=&quot][/FONT]sistance: Coffin saying that there is no question that XMRV is real and infecting people. As CBS said in that thread, this really puts the first two UK studies in perspective.

Reason for cheer!:Retro smile::Retro smile::Retro smile::Retro smile:

thank you very much

 

[Dx Revision Watch]

One thing that I'm not clear on, and I wonder if anyone can clear this up for me?
The WPI study says they selected patients using the Fukuda and Canadian criteria...
does this mean that each patient has to fulfill both criteria, or does it mean that that some patients fulfilled the Fukuda definition, and some the Canadian definition?

Although, I think that this question is a little redundant, because the WPI were testing for physical signs of organic illness anyway, so they were basically narrowing down the patients to an organic illness which has an effect on the body at the cellular level.

Bob, I am still unclear on this point, too, that is, did all WPI patients fulfill both Fukuda and Canadian?

On 12 October I wrote to Dr Judy M and asked:

"I have received a query about the criteria used for this study.

I know you must be snowed under but I'd be very grateful if the criteria used for the study could be clarified, please.

(Our UK NHS site says: People with CFS had been diagnosed using standard criteria (1994 CDC Fukuda criteria and 2003 Canadian Consensus Criteria), and all had severe disability, prolonged disabling fatigue, cognitive defects and immune system abnormalities. They came from areas in the US with reported outbreaks of CFS.)

Is this correct?"​

On 30 November, (with an apology for the delay) I received this response:

"This statement is correct with the exception that the patients came from all areas of the US including but NOT limited to reported outbreaks of CFS"​

which doesn't answer the point, per se, because I should have asked specifically for that to have been clarified.


For the Swedish Gottfries study, co-funders, ME Research UK, have said, on their website:

http://www.meresearch.org.uk/research/projects/xmrvsweden.html

"The researchers will retrospectively test previously
stored samples from 3 groups of patients (20
Fukuda-defined ME/CFS, 20 fibromyalgia, 20 irritable
bowel) and 20 controls.

In addition, they will prospectively test samples from
120 ME/CFS patients (defined on the Fukuda 1994
and the Canadian 2003 criteria, similar to patients in
the original 2009 report in Science), who will also
have functional assessments."​

I have a Swedish contact who is participating in this study and has given blood who has told me that they, themselves, fulfill both Fukuda and Canadian. But again, it is unclear to me whether the Swedish study is testing samples from patients who all fulfill both or whether some will meet Fukuda, only.

I did enquire whether patient information sheets in relation to this study were given out, but apparently not.

Suzy

 

[natasa778]

We dont know of any other technique for isolating the virus from these cells apart from the one used by the WPI

Gerwyn, what do you think about investigating actual tissue by FISH or similar techniques? Judy mentioned microglia as a xmrv target (dead on imo), not sure about CFS but there are certainly available autism brain banks to do something like that. Are there any CFS tissue repositories out there and what type of tissue would they contain? Is FISH something that would be relatively easy to create and employ for xmrv?

 

[Dx Revision Watch]

Thanks Gerwyn,

If you're going to the trouble of culturing that essentially says this virus is hard to find (or latent) does it not? How can you ignore a signal like that?

That was a big study! That cost the UK government a ton of money of money. I just can't imagine...

Cort, I'm coming to this thread late and forgive me if I've missed something in earlier posts, but which "UK government" funded study are you referring to here, please?

The reason I ask is because if you mean the

Groom, Boucherit, Makinson, Randal, Baptista, Hagan, Gow, Mattes, Breuer, Kerr, Stoye and Bishop

study, it was joint funded by the MRC (file reference (KB) U117592729 and (JS) U117512710); The Wellcome Trust (grant ID 084955) and the CFS Research Foundation, UK, with an acknowledgement to The Cunningham Trust for funding to SH.

Incidently, Prof Stephen Holgate who chairs the MRC "CFS/ME Expert Panel" is a member of the Research Committee of the CFS Research Foundation which has funded much of Jonathan Kerr's gene work (for which Holgate has been a collaborator).

Suzy

 

[Angela Kennedy]

Looking at the information Suzy has supplied here, together with the supporting online material for the Lombardi et al Science paper (I found the pdf!), I would say on face value that it would appear all patients fulfilled both, especially as no discussion of stratification between both criteria sets is given in the paper.

However, I believe it does need clarifying by the WPI. I guess it needs to be established whether ALL patients in the sample met the Canadian criteria (by definition those sort of patients SHOULD meet Fukuda symptoms criteria anyway).

I also wanted to say that I also think there are various fundamental problems with the testing for XMRV methodology. Technical mircrobiological testing methodology is not my area of knowledge, hence my not pressing that issue. But I understand it's vital importance.

Nevertheless, the issue of patient cohort is also crucial, not just in regard to XMRV, but for future 'CFS' research. Tom Kindlon kindly put up a paper on co-cure today showing that the Canadian criteria is starting to be engaged with in research projects. Leonard Jason led a good validation project of Canadian criteria in 2005. I have been involved in endeavouring to ensure that PACE trial authors do not get to ignore the Canadian guidelines (without being called on frequently and publicly it at least) since the trial was in planning stages and since. So the issue of patient cohorts is a hot one.

I wonder - might it be useful to consider starting a new thread on establishing Canadian Cohorts in 'CFS' research?

 

[Gerwyn]

Gerwyn, what do you think about investigating actual tissue by FISH or similar techniques? Judy mentioned microglia as a xmrv target (dead on imo), not sure about CFS but there are certainly available autism brain banks to do something like that. Are there any CFS tissue repositories out there and what type of tissue would they contain? Is FISH something that would be relatively easy to create and employ for xmrv?

if the uique sequence in the env rna sequence is big enough and the cells are actively replicating in theory yes I would have to check the fine detail s to give a better answer.I would go for the tissue of the reproductive organs or possibly semen of males known to be CFS as properly diagnosed becuae of its accelerative properties on XMRV replication