Article: Dr. Mikovits on XMRV: The Sweden Talk

You can view the page at http://forums.aboutmecfs.org/content.php?313-Dr.-Mikovits-The-Sweden-Talk

 

They have a viral load test now? That is amazing news. Wonder if load will correlate with debilty?

 

If I understood it right they do.... I tried to confirm that but couldn't - but that's what I understood her to say.....

That would be one of the first questions I would guess

 

I wonder if there's any chance they'll be able to rush out a paper about the (possibly) blinded Dr. Bagni testing?

There's been a real lack of positive XMRV papers getting published. All this news could be great, but I'd love to have it in more detail and from a venue that other researchers would take note of. Are we at a strange point where it's easier to get negative papers published than positive? It's not surprising journals are being extra cautious, but if testing is being properly blinded, that should assuage some of the concerns about contamination... even if the conclusive proof disproving contamination is currently beyond us.

 

I would guess they would publish Bagni - since she is providing new evidence...I asked Dr. Mikovits about that - she just emailed me that their email server has been down for a couple of days - and she would get back to me when she can.

I'm sure the publication of a Bagni paper would be a big boost. .

 

Very interesting. Thanks for that Cort.

It would be an enormous advance if they have found a way to measure viral load, but I'm confused as to how they could, using existing knowledge and methodologies, considering the difficulties of detecting XMRV due to the apparently very low copy numbers in the blood.

Dr Bagni's study, using an XMRV-specific antibody test, would be extremely helpful if it was published... An XMRV-specific antibody test would be a huge step forwards.

Interesting to hear Judy talk about re-freezing samples in storage, and how it destroys XMRV... I wonder if this finding is linked to work done by the Blood Working Group.

Interesting that Judy is re-emphasising the importance of patient cohort... The emphasis seemed to have moved onto sample preparation and storage, in relation to the negative studies... I suppose it's important to focus on both issues, not just one or the other.

Very interesting that Alter's pMRV's might need a different cell line to grow in, which I suppose might lead to pMRV's being detected far more frequently once the methodology is advanced.

I didn't know that Frank Ruscetti discovered HTLV, so that's very interesting to learn.

I didn't know much about the relationship between Judy and the Ruscettis, so it's interesting to learn more about that.

I liked Dusty Miller's quote about the Hue study... It's something that I had thought about... I think Alter is said to have tested his samples for all known mouse DNA and retroviruses, so he would surely have tested for contamination by the mouse DNA that the Hue study looked at.
Also, how can Hue et al deny the existence of XMRV if they were not even attempting to detect or sequence XMRV, specifically, in their study? In their study, they got a positive reading with a test which wasn't specific to XMRV, and then said that the fact that their test wasn't specific for XMRV meant that their positive reading showed that XMRV doesn't exist! It seems like very woolly science. It's like holding a peach or a nectarine in your hand with your eyes shut, and saying that because I have detected a fruit in my hand but can't determine whether it is a peach or a nectarine, that peaches don't exist.

 

I would guess they would publish Bagni - since she is providing new evidence...I asked Dr. Mikovits about that - she just emailed me that their email server has been down for a couple of days - and she would get back to me when she can.

I'm sure the publication of a Bagni paper would be a big boost. .

Fingers crossed with it all. Thanks for keeping us informed. It's starting to play with my mind having the two opposing sides on XMRV moving furhter apart as time goes by. If the WPI has an XMRV test that can identify CFS patients with reasonable accuracy, why's it taking so long for them to prove this! (I'm not necessarily blaming them... I know they'll have to jump through hoops to get things published and so on... but even so). The last 18 months have been such strange time for us.

It would be an enormous advance if they have found a way to measure viral load, but I'm confused as to how they could, considering the difficulties of detecting XMRV due to the apparently very low copy numbers in the blood.

Gallons of blood! Suck 'em dry.

 

Cort, many thanks for this--nice to get some good news at this down time of the year! Really appreciated. Chris

 

Where the heck are the Singh/Bateman XMRV studies already?!!?

 

Where the heck are the Singh/Bateman XMRV studies already?!!?

I think we are all getting antsy about this but I rather she spend time doing the science to make her paper more solid and set a new benchmark instead of rushing to publication.

 

Thank you Dr Mikovits, very informative as always. Dr Bagni's study and Dr Singh's study very much need to be published. But good studies take time, why is it that the "contaminant theorists" can get their studies published with little more than a few weeks, or in the case of Simon Wessely 3 days of peer review! In seems there needs to be much more review of the claims made by contaminant theorists - for example if the following is true, it seems to me that the Hue/Towers paper should have been refused publication as does not support it's main finding.

"The main problem with Hue et al. is they propose that XMRV is a mouse virus, but I have looked at all of the mouse sequencing data available, and can find no copy of XMRV in the genome of any mouse sequenced to date. That includes the 129x1 strain mentioned in their paper. Sure one can find snippets of DNA that look similar to regions of XMRV, but nothing very extensive."

 

I think we are all getting antsy about this but I rather she spend time doing the science to make her paper more solid and set a new benchmark instead of rushing to publication.

That is my preference.

 

Unfortunately it isn't correct that in the UK study our blood was taken at our home. The UK Study consisted of 50 people who somehow got to a hospital just outside of London for the blood to be drawn. The blood was then flown out that day to the US. Also it isn't correct that everybody was housbound or bedbound. None of the participants would have been bedbound, these people are still waiting to have their blood drawn.

However it is true that all the patients have true ME and I guess this is what was meant by the importance of patient selection.

 

Finding a cell line that will grow a virus is often not easy but they got luckywhen it turned out that Dr. Mikovits hunch about a T-cell Lymphoma cell line defective in producing the interferons and that was susceptible to androgens which effect prostate tissues). Luckily she guessed right - its not always easy to find the right cell line - and the WPI was off and growing the virus. This is still the only cell line known that XMRV proliferates in.

I'm not sure how much this was 'lucky' or a 'hunch': the discovery of the XMRV-permissive LNCaP cell-line was apparently made by Dong in 2007, as referenced in the Lombardi Science paper!

 

"The main problem with Hue et al. is they propose that XMRV is a mouse virus, but I have looked at all of the mouse sequencing data available, and can find no copy of XMRV in the genome of any mouse sequenced to date. That includes the 129x1 strain mentioned in their paper. Sure one can find snippets of DNA that look similar to regions of XMRV, but nothing very extensive."

This Dusty Miller quote is indeed very interesting since it basically suggests that XMRV is not a mouse retrovirus at all (with just relics of it left in the mouse genome). Interestingly, the Hue phylogenetic tree included NO mouse XMRV sequences. This sort of work is Hue's specialism and if he couldn't find XMRV sequences either adds further weight to the view that XMRV is not an active murine virus.

However, the key thing about the Hue paper was that it found a very close link between XMRV from the human 22Rv1 cell line and the 2 complete XMRV sequences from CFS patients published in the Lombardi Science paper. In other words Hue is saying the contamination is from a human cell line, so you can ignore all the stuff about mouse DNA contamination. Apparently Judy Mikovits has tested her positive samples with the Switzer mouse mitochanodrail DNA assay and found nothing, but that obviously doesn't rule out contamination was from a human cell line.

Going back to the point that XMRV might not be an acitve mouse retrovirus, where did the XMRV in the human cell line 22Rv1 come from? It's been assumed XRMV was picked up when the cell line was xenografted onto mice (in 1999, I think), as part of the development of that cell line. But wait, 22Rv1 is a human PROSTATE CANCER cell line, so maybe the XMRV is of human not mouse origin, and was in the cell line all along? I'm pretty sure this has been speculated on in another forum (by Bob and Marco?) but didn't really take it in as it was over my head. Actually, my heads spinning so i'll leave it there.

 

I think we are all getting antsy about this but I rather she spend time doing the science to make her paper more solid and set a new benchmark instead of rushing to publication.

Well I just have the feeling that they are finished but that the journals won't publish them b/c of all the recent contamination papers. So that's making me concerned/anxious.

 

The factor she keyed in on was the number of times a sample has been frozen and thawed and then refrozen. She said “if you're looking for a retrovirus and a sample has been thawed and refrozen that will break up the nucleic acids and you won't be held to find the retrovirus again”. It appears that freezing and thawing and then testing is okay but freezing and thawing and then freezing and thawing and retesting again is not okay.

This gives me an uncomfortable sense of dj vu. I remember that Elaine DeFreitas said the same thing about her retrovirus. She said that when samples were frozen and thawed, the strands of DNA would begin to break into tiny pieces, and then PCR wouldn't work. You couldn't amplify it. The CDC kept freezing their samples, even after Dr DeFreitas told them they shouldn't. The CDC said it would only be a problem if the virus were found in very low copy numbers.

So here we are 20 years later, dealing with a retrovirus found in very low copy numbers. We've got a lab that's consistantly finding this retrovirus in CFS patients, and a CDC that is consistantly not finding it, and now the researcher who is finding it starts saying it's important not to freeze & thaw the samples. This is beginning to feel like a recurrent nightmare.

And STILL no one has done a true replication study, using the WPI's techniques...

 

This gives me an uncomfortable sense of dj vu. I remember that Elaine DeFreitas said the same thing about her retrovirus. She said that when samples were frozen and thawed, the strands of DNA would begin to break into tiny pieces, and then PCR wouldn't work. You couldn't amplify it. The CDC kept freezing their samples, even after Dr DeFreitas told them they shouldn't. The CDC said it would only be a problem if the virus were found in very low copy numbers.

So here we are 20 years later, dealing with a retrovirus found in very low copy numbers. We've got a lab that's consistantly finding this retrovirus in CFS patients, and a CDC that is consistantly not finding it, and now the researcher who is finding it starts saying it's important not to freeze & thaw the samples. This is beginning to feel like a recurrent nightmare.

And STILL no one has done a true replication study, using the WPI's techniques...

'

I don't know. I would think retrovirologists know all about and freezing and thawing samples and the effects on viruses but this is a very rare virus and a new one and who knows?

From what I remember Folks at the CDC later said they acceded to all her requests over time and they never got another positive results. Its a shame that her virus never got the study that XMRV is getting. Then again she was never able to do the things the WPI was able to do--grow the virus, show that its able to infect cells, antibody tests......they were way ahead of the game from the get go.

 

That is my preference.

Mine, too.

The reason these lousy papers are getting out so quickly is that they're being published in journals that do poor reviewing. Several weeks (or 3 days ROFLMAO!) is nowhere near enough for a thorough, rigorous review.

I hope the papers we want to see aren't out in a hurry because the intent is to have them published in important, reliable journals -- and that takes time. Part of why it takes time is that the reviewers are knowledgeable, busy researchers who take the time to read and analyze the paper, bring up potential questions, and make suggestions for improvement. Then there's the rewrites to strengthen the paper, then the re-review.... Then the paper has to get into the publication cycle of a journal that all the best researchers want to be published it in. That doesn't happen in [snigger] 3 days.

I'm impatient, but I'll wait for the good stuff.

 

Going back to the point that XMRV might not be an acitve mouse retrovirus, where did the XMRV in the human cell line 22Rv1 come from? It's been assumed XRMV was picked up when the cell line was xenografted onto mice (in 1999, I think), as part of the development of that cell line. But wait, 22Rv1 is a human PROSTATE CANCER cell line, so maybe the XMRV is of human not mouse origin, and was in the cell line all along? I'm pretty sure this has been speculated on in another forum (by Bob and Marco?) but didn't really take it in as it was over my head. Actually, my heads spinning so i'll leave it there.

Yeah thats exactly what I said. Perhaps XMRV is the reason 22Rv1 was cancerous to begin with. Also, I am pretty sure LNCaP and 22Rv1 are different lines. So just because they found XMRV in 22Rv1, doesn't mean the same is true of the LNCaP line used by WPI for their study. And I'd hope that regardless, they tested the cell line for XMRV before they tried to combine it with blood samples in culture. You have to make sure your culture medium isn't already infected before you run the culture - otherwise it kind of defeats the purpose of culturing. They did this right?