German study finds xmrv

[Gerwyn]

She doesn't address it directly there, but instead outlines the WPI methods that include the activation/stimulation/biological amplification/culture steps.

However, she and Ruscetti state it directly in their recent response in Science magazine (bolds mine):

Sorry doc. you were right about culture.i misread it. fred getting unreliable.They were even cleverer than I first thought.They concentrated their sample by usinf reverse transcriptase PCR.you dont need actively replicating virus to pick up XMRV RNA.The WPI also used RT -PCR to hedge their bets in the run using fresh blood.The RT pcr would have amplified the viral RNA many orders of magnitude which could be then used to make sufficient cDNA to be detectible by nested PCR .they used Rt -PCR as their "culture " step.Very clever indeed and the basis of a very quick detection method! Why the english studies did noy use RT-PCR when looking for a RNA virus is beyond me!

 

[bullybeef]

In regards to the XMRV global prevalence, the ~6% is also mentioned on the National Cancer Institute website:

What is the prevalence XMRV infection in the general population?
The number of healthy people infected with XMRV, and what the distribution of infection is within the U.S. and worldwide, is unknown. The virus has been detected in about four percent of individuals from the same geographical region as infected patients with chronic fatigue syndrome. An analysis of more than 300 prostate tissue samples found that the virus was present in six percent of noncancerous prostates. These studies suggest that XMRV infection could be widespread, but there is no evidence for a new, increasing, or spreading XMRV infection. The NCI is developing molecular tools to determine the incidence of XMRV in the general population and is involved in coordinating a global effort to study the prevalence and distribution of XMRV.

http://www.cancer.gov/newscenter/pressreleases/XMRV_QandA

 

[Dr. Yes]

Do I get this right, that if XMRV writes itself over a 'start site' (one which contains genes that regulate or activate maybe 40 other genes - epigenetics?) you will get many more immune regulation and other problems and a much more serious condition?

Retroviruses don't 'write over' your DNA, but rather insert their sequences into it. If they are clean about it, they will only interrupt your sequence, but if they are 'messy' they can cause small mutations in the nearby human DNA. Of course the very act of inserting themselves is a mutation. Where they insert themselves is therefore critical; from Kim et al (2008) we know that at least in a prostate cell line, XMRV likes to integrate into parts of the genome associated with the regulation of transcription, such as transcription start sites and probably gene promoters. 'Start sites' are literally where transcription of a gene (ultimately to turn it into a protein) starts. Promoters are involved in regulating the transcription level of a gene, i.e. they are 'control knobs' for gene expression. Viral integration into such a site won't lead to defective proteins, but could well lead to too much or to little expression of them.

In other words, XMRV has shown a tendency to insert into the 'master switches' of genes. Some genes are themselves master switches of sorts, such as the CREBBP gene, which makes a protein that is essential to that CREB/CRE regulation Gerwyn is always yakking about Retro wink. The CREBBP gene happens to be one of the genes Kerr found to be dysregulated in CFS patients. Altering the expression of one such 'regulator' gene could indeed have a cascade effect on many other genes. It doesn't even have to be a part of the CREB/CRE group; many gene products such as IL-6 have multiple signalling roles in the body and genes for IL-6 were also found to be dysregulated by Kerr et al. However, XMRV insertions into those genes would have to take place in a lot cells in order to cause disease.

And is it also likely that the (last count I heard) 6 strains of XMRV discovered so far may prefer to bind to different sites?

Nobody knows exactly why retroviruses favor certain areas of the genome for insertion, but one reason may be an interaction between IN (the enzyme retroviruses use to integrate their DNA into ours) and the shape or the specific nucleotide sequence of particular sections of our genome. I don't know if any of the six strains prefer different binding sites; they differ by only a few nucleotides so unless those happen to be crucial to integration (the probability of that is obviously very low), I doubt they favor different sites. Technically retroviruses can insert themselves almost anywhere in the genome, by the way, but some sites are an order of magnitude or two more likely to be selected.

This all seems well related to the existing body of research and to recent results: Dr Kerr, as I understand it, recently identified 80 genetic sequences which are abnormal in ME/CFS patients, and it could be that these are somehow related to the corruption of DNA by XMRV? (Not sure how that might work).

Abnormal gene expression does not necessarily mean mutation. Kerr et al looked for abnormal gene expression (upregulation or downregulation) and found it in 88 genes; to my knowledge they have not looked for mutations within those genes (which would by very interesting.. particularly to see if XMRV sequences can be found in there somewhere). The abnormal regulation of these genes could, hypothetically, be due to the insertion of a retrovirus that likes to integrate into the regulatory sites of genes, as XMRV does... or to the insertion of said retrovirus into 'regulator genes' like CREB or CREBBP.

 

[lansbergen]

Thanks Gerwyn and Dr. Yes for explaining.

 

[Dr. Yes]

Sorry doc. you were right about culture.i misread it. fred getting unreliable.

I'm afraid my trio of brain cells is a lot less reliable than Fred...



​

Why the english studies did noy use RT-PCR when looking for a RNA virus is beyond me!

See? Larry Curly and Moe forgot all about that. They can't figure why that step would be omitted either.

 

[Ecoclimber]

Wikipedia on XMRV and WPI has been updated with the latest information.

 

[bullybeef]

Yeah, I've noticed Wiki has changed some info on XMRV. But this new study is typically at the bottom of the page, whilst the 'dodgy' negative studies remain in the first paragraph.

 

[Cookie Monster]

But remember that they received transplant tissue or organs from healthy donors, so their chance of getting contaminated organs would have been around 3%.... meaning that would not account for 9% prevalence.

I still feel a more likely explanation that it was a long-standing xmrv infection in the first place that might have contributed to their organ failure, or them needing marrow transplant for whatever reason.

Hi Natasha,

Would it not be true that both statements are applicable to the 9.9% figure from this sub-set of the study? - i dont see myself that they are mutually exclusive options?

 

[natasa778]

Yes you are right Cookie Monster, they are not mutually exclusive.

Also your first theory that this subset of patients could have been infected through transplants would be strenghtened if we had data on those patients receiving blood transfusions etc before or after transplants, or if they had been in need of repeat procedures/transplants, from different donors. Each new exposure would raise chances their chances of xmrv infection by xxx whatever statistical number.... ?

 

[Cookie Monster]

Yes you are right Cookie Monster, they are not mutually exclusive.

Also your first theory that this subset of patients could have been infected through transplants would be strenghtened if we had data on those patients receiving blood transfusions etc before or after transplants, or if they had been in need of repeat procedures/transplants, from different donors. Each new exposure would raise chances their chances of xmrv infection by xxx whatever statistical number.... ?

Cheers for replying Natasha very much appreciated. I may throw out a quick question to the authors just out of curiosity.

Hopefully the WPI study on XMRV infection following blood transfusion should throw up some useful data down the line (now im already wondering if they themselves have differentiated those that simply had a blood transfusion, from those that had organ transplant+transfusion in their own study ).

 

[Cookie Monster]

I may throw out a quick question to the authors just out of curiosity.

would have helped if i had realised at the time that i cant write in german though

 

[anciendaze]

...Why the english studies did noy use RT-PCR when looking for a RNA virus is beyond me!

There's a confusing pair of letters here. RT is also used for real-time PCR, which the Groom study used, but not reverse-transcription.

I think the error comes from unconsciously relying on sloppy biological processes to leave a few copies lying around. If you have lots of virus, and lots of infected cells, you can get away with this. When you are pushing right to the limits of detection, sloppy thinking about sloppy biology does not work. More evidence scientists, like most people, spend more time operating on autopilot than thinking.

Another thing here is confirmation bias. This comes from confusion about confirmation of beliefs versus confirmation of hypotheses.

Using the parallel with black swans, the hypothesis that all swans are white corresponds to the hypothesis that all chronic fatigue syndrome is tied to psychological factors. Not finding a black swan confirms your belief that all swans are white, but is almost irrelevent to confirming the hypothesis.

To illustrate, I look out my window this fine morning, and do not see a black swan. This is consistent with the hypothesis that there are no black swans, and also with the much stronger hypothesis that all swans are white. (There may be gray or blue swans.) This may confirm my own belief, but any additional weight it adds to previous evidence is negligible.

 

[Rrrr]

judy mikovits comment to me about this german study: "it is great for all of us !!!"

 

[Rrrr]

does this german study essentially amount to a replication study of the original science paper study? (but maybe also surpasses a replication study's goals, in terms of knowledge gained?)

i mean, they found xmrv using a method similar to (better than!) the wpi study.

 

[Knackered]

does this german study essentially amount to a replication study of the original science paper study? (but maybe also surpasses a replication study's goals, in terms of knowledge gained?)

i mean, they found xmrv using a method similar to (better than!) the wpi study.

No, they didn't look at CCD CFS patients. Still good though, it proves XMRV is international.

 

[slayadragon]

Airplane, School and Medical Workers

In the Bible (Osler's Web) an early large survey in the US found that 90% of "CFS" patients were working in only one of five jobs at onset: doctors, nurses, flight attendants, pilots and teachers.

Someone brought this quote to my attention, so I'm going to insert a different explanation.

According to Dr. Ritchie Shoemaker, a high percentage of airplanes have very bad contamination problems with toxic mold.

Certainly, the airplanes I've been on since getting unmasked (and thus becoming able to easily identify the presence of mold by looking at how I feel rather than judging odors) have been extremely contaminated. And I always crashed big-time after every flight I took when my CFS was active. Rarely did other activities have as much of an effect on me.

Insofar as airplanes are contaminated, the effects would be magnified substantially due to lack of air circulation. Those who have read Osler's Web will remember how all the teachers except one got sick in the Truckee High School teachers' lounge, where the air circulation had been shut off. (The exception worked in his camper rather than the teachers' lounge during breaks).

Based on my own assessment of Truckee HS two years ago, the place has a mold problem. Erik Johnson, one of the Incline Village cohort who has recovered from mold avoidance, says that it was infinitely worse back when he was a student there and then during the epidemic.

Dr. Shoemaker reports that a high percentage of schools in general have mold problems. Some are remediated with great fanfare, but in most cases the debates about them go on for decades.

I'd guess with the doctors and nurses, this more may have been a pathogen issue. Certainly there is one or more pathogens (e.g. XMRV but maybe others such as Lyme or mycoplasma) involved as a root cause in CFS/whatever. The idea that this bug is usually transmitted casually or even through sex doesn't seem to have much epidemiological evidence behind it, as far as I've seen. The exception is what Erik writes of his own observations during the epidemic (below).

Possibly the pathogen always could be transmitted by blood though, and thus might have been a real issue earlier in the epidemic when health professionals weren't quite so cautious with needle sticks.

Just some food for thought....

Best, Lisa

*

I only know what I saw:

That in certain specific biotoxin-laden Sick Building Syndrome environments, the "whatever" was capable of being very infectious by casual contact, with a savagely quick incubation period of 24-48 hours.

After that initial phase, the capacity for transmission seemed to just disappear.

I asked Dr. Cheney about this at the time. He confirmed that many viruses are no longer transmissible after the initial phase, but he dismissed the environmental component as probably being just a consequence of the flu-like illness.

-Erik (2009)

 

[Gerwyn]

Someone brought this quote to my attention, so I'm going to insert a different explanation.

According to Dr. Ritchie Shoemaker, a high percentage of airplanes have very bad contamination problems with toxic mold.

Certainly, the airplanes I've been on since getting unmasked (and thus becoming able to easily identify the presence of mold by looking at how I feel rather than judging odors) have been extremely contaminated. And I always crashed big-time after every flight I took when my CFS was active. Rarely did other activities have as much of an effect on me.

Insofar as airplanes are contaminated, the effects would be magnified substantially due to lack of air circulation. Those who have read Osler's Web will remember how all the teachers except one got sick in the Truckee High School teachers' lounge, where the air circulation had been shut off. (The exception worked in his camper rather than the teachers' lounge during breaks).

Based on my own assessment of Truckee HS two years ago, the place has a mold problem. Erik Johnson, one of the Incline Village cohort who has recovered from mold avoidance, says that it was infinitely worse back when he was a student there and then during the epidemic.

Dr. Shoemaker reports that a high percentage of schools in general have mold problems. Some are remediated with great fanfare, but in most cases the debates about them go on for decades.

I'd guess with the doctors and nurses, this more may have been a pathogen issue. Certainly there is one or more pathogens (e.g. XMRV but maybe others such as Lyme or mycoplasma) involved as a root cause in CFS/whatever. The idea that this bug is usually transmitted casually or even through sex doesn't seem to have much epidemiological evidence behind it, as far as I've seen. The exception is what Erik writes of his own observations during the epidemic (below).

Possibly the pathogen always could be transmitted by blood though, and thus might have been a real issue earlier in the epidemic when health professionals weren't quite so cautious with needle sticks.

Just some food for thought....

Best, Lisa

*

I only know what I saw:

That in certain specific biotoxin-laden Sick Building Syndrome environments, the "whatever" was capable of being very infectious by casual contact, with a savagely quick incubation period of 24-48 hours.

After that initial phase, the capacity for transmission seemed to just disappear.

I asked Dr. Cheney about this at the time. He confirmed that many viruses are no longer transmissible after the initial phase, but he dismissed the environmental component as probably being just a consequence of the flu-like illness.

-Erik (2009)

There is no known mammalian Mulv which is not sexually transferrable.XMRV would have to be totally unique if not transferrable by sexual intercourse.Dr Cheney needs to brush up on his virology>Try telling an AIDS patient that!

 

[eric_s]

Thanks CBS!

Eric had asked the following:

Does anyone know if the Bannert group could not find any XMRV at all? Or did they find the ~3% that would have to be there? If they couldn't find any, i think it's pretty safe to assume that they have used a different method compared to this latest study or did not work properly.
​

OK eric, here's what I found from the Bannert/Hohn paper provided by CBS:

Xenotropic Murine Leukemia Virus–related Gammaretrovirus in Respiratory Tract In total, 146 sera from prostate cancer patients and 5 healthy control individuals were tested negative for antibodies binding recombinant XMRV gp70 and Gag proteins in ELISA, although postive control immunized mouse sera reacted strongly (Fig. 5A and 5B). One patient serum that reacted strongly in ELISA against the recombinant pr65 protein was subsequently tested by immunofluorescence assay using HEK 293T cells expressing XMRV and cells expressing the gp70- or pr65 proteins alone. No XMRV specific binding was seen, indicating a non-specific ELISA reaction.

…As additional controls we tested the cell lines 22Rv1 (XMRV positive [17]) and DU145 (XMRV negative [9]). As expected, 22Rv1 was found to be strongly positive for RNA transcripts and for provirus (with In-For/Deletion-Rev primers), while DU145 was negative in both PCR approaches (data not shown).

From Figure 2: Nested-PCR screen of the first 16 QQ patients (lane 1-16) with the In-For and Deletion-Rev primer pair (upper panel) and In-For and In-Rev primer setup (lower panel); lane 17 = mouse tail DNA, lane 18 = water control outer PCR mix, lane 19 = water control inner PCR mix, lane 20 = pXMRV, lane 21 = pDG75, marker = 100 bp marker.In other words, the original Hohn/Bannert paper used water as a negative control, as well as cell line DU145 (XMRV negative). And they also tested "5 healthy control individuals".

Based on these low control numbers of actual humans, it would be impossible to determine prevalence in controls for comparison.

But did Hohn/Bannert in the original negative prostate paper use the same methods as Hohn did in his positive respiratory secretions paper????

Anyone??
​

Hi Parvo

Once again, this seems to be a situation, where it's very hard to make sense of the information we hear. Since the XMRV story started to make news, i try to figure out what's going on and just can't reach a conclusion. It seems when it's about CFS the world does not work in any logical way.

Thanks a lot for your answers, here's what i think:
In the original Bannert study "Lack of evidence for xenotropic murine leukemia virus-related virus(XMRV) in German prostate cancer patients" he seems to have found 0 cases of XMRV infection. If i read your information correctly.
But as we can now see from "Xenotropic Murine Leukemia Virus-related Gammaretrovirus in Respiratory Tract" there seems to be a prevalence of XMRV in German healthy controls, people with RTI and people with RTI who have had transplantation. This is consistent with the Science study and the Japanese Red Cross values and so i think it's likely that those results are correct.
For that reason, i think that at least in "Lack of evidence for xenotropic murine leukemia virus-related virus(XMRV) in German prostate cancer patients" they have used a different method or did not apply their method correctly, because i don't see any reason why those German prostate cancer patients should all be XMRV negative, if XMRV is around in Germany and can be found in different groups of the population.
We can't say about the Bannert CFS study discussed in Prague because we don't know yet if they have found 0 cases there or if they have found some percentage to be positive.

Now what confuses me is this:
How long did the work on "Xenotropic Murine Leukemia Virus-related Gammaretrovirus in Respiratory Tract" take? It must have overlapped at least with the latest CFS study. Right?
I can hardly believe that Hohn is involved in all of those studies and while they can find XMRV in one study, as is to be expected at least in some one digit percentage of cases, he will not tell the Bannert group and they go on to publish a study where they couldn't find any.
So it would be very interesting to see what they have found in that study that they talked about in Prague. Because that one was only finished recently. So far i'm only sure that they did not find a high prevalence.
Nina said:

Next thing we know, Bannert's team present their findings at the Prague conference and say they still haven't found XMRV in CFS patients.

If that means they have not found any case of infection, then i don't understand what's going on. It could only mean that Hohn did not tell them about the other study's finding, that there they could find it, or that they just did not listen to him (not very intelligent) or that it was too late to not publish it. [Edit: Another possibility would be that the study only looked at such a small number of people that it would not pick up an ~5% prevalence, with some bad luck during the selection. I think now i can remember that it only involved 20 to 30 persons. But i'm not sure at all about that number.]

I really think all we can do is to either ask those people, which would be great if they answer, but i'm not too confident they will, or then to just wait for another study by them. I'm not German and i think the RKI and Charit are very good but i don't think it's their culture to answer emails from people outside of the scientific community (i mean not employed by a scientific institution) who they don't know. Probably that's much more common in America.

Ciao
Eric

 

[Martlet]

Okay - The exact circumstances of me getting ill. I'm giving these to try and find a way round all this, wondering whether any or all these factors came into play.

1. 12-hour flight from UK to Houston. Arrived so tired that we ordered room-service and I ate with my head resting on the table.
2. Next day, began the long drive to Alamogordo, NM, with two stops on the way, one in a not-so-savoury hotel in the Texas nowhere.
3. Five days in Alamogordo, visiting friends. Felt run-down. Wondered if there was some allergen causing this.
4. Drove to Colorado Springs, feeling somewhat better, but by the time we checked into our hotel, was so exhausted with flu-like symptoms that we had to go to the USAF base there to see a doctor. They ran some bloods while I slept, told me I had a viral infection and to (a) go to a lower altitude - even if it was only Denver - and (b) to stay in bed until I felt better. Checked into a hotel there and remember nothing for the next three days.
5. Felt well enough (but still not well) to continue our journey cross country to Cincinnati to visit friends. By the time we arrived in Cinci, my throat was so swollen and sore, glands up etc. that I had to see a doctor again. "Very bad tonsillitis" he said. Gave antibiotics.
6. Eight-hour flight back to UK and no recovery.

So, I was obviously exhausted. I'd obviously spent a long time on an airplane, exposed to goodness-knows-what. I met some sort of pathogen/allergen in the NM desert. I went down with a severe flu-like illness. I flew again, while ill.

I know it's only speculation, but it seems to me that there was enough in there to permit an opportunist infection such as XMRV may well be to have taken hold.

Any thoughts?

PS - One of my closest friends in UK already had ME and I saw her just before we flew to the US

 

[eric_s]

In regards to the XMRV global prevalence, the ~6% is also mentioned on the National Cancer Institute website:

Thanks bullybeef.