Unintended spread of a biosafety level 2 recombinant retrovirus & XMRV

[cold_taste_of_tears]

Found this information on another site, and no one answered the person posting it. There are many genius minds here (sadly I am not of them!).
So I thought this was the ideal place to ask you guys - what you think of this.

Should the WPI know about this?
We already have the incredible Elaine De Freitas link to CFIDS associated Retrovirus. How about this?
It was only published 8 weeks ago.


Journal of Retrovirology, Sept 2009
Stang A, et al
Department of Molecular and Medical Virology, Ruhr-University Bochum, Germany
Available online at: http://www.retrovirology.com/content/6/1/86

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Unintended spread of a biosafety level 2 recombinant retrovirus

abstract:

“Gene fragments for both viruses could be detected in a broad range of permissive cell lines from multiple species. Furthermore, experimental infections of cells negative for these viruses showed that both viruses replicate rapidly to high loads. We decided to further analyze the genomic sequence of the MuLV-like contaminant virus. Surprisingly it was neither identical to MuLV nor to the novel xenotropic MuLV related retrovirus (XMRV) but showed 99% identity to a synthetic retrovirus which was engineered in the 1980s.”


In detail:

''It has to be mentioned that each of both viruses was found in at least one aliquot of all cell lines tested. Thus, we cannot report a single cell line which is not permissive for one of both viruses. Furthermore, experimental infections of retrovirus-negative aliquots of selected cell lines showed that both viruses are highly infectious and propagate to high viral loads as determined by viral RNA copy numbers in the supernatants and proviral genome copies of extracted cellular DNA. In the early stage of infection even some cytopathic effects (CPE) could be observed. In contrast, no CPE was seen in persistently infected cultures possibly due to adaptation of cells to the retroviral infection.”

2 - “In summary, there have been numerous publications about retroviral contaminations like recent reports of ecotropic murine leukemia virus in various cell lines. The most frequent retrovirus found in this context is squirrel monkey retrovirus (SMRV). One study even reported the detection of SMRV related sequences in commercial interferon preparations in 1998. Although the sequences were found only as DNA and therefore rather derived from cellular DNA carrying proviral genomes than viral particles, it clearly demonstrated the contamination of the interferon producing cell line with SMRV. Germany's Central Commission of Biosafety (ZKBS) recently reported that SMRV was detectable in 128 samples of 4279 cell cultures from different laboratories throughout the country.

“The present report extents these studies by identifying for the first time a presumably synthetic chimeric retrovirus as a contaminant. This gene-modified organism seems to have replicated and spread intensely in a broad set of cell lines for several years without being noticed. This hybrid amphotropic/Moloney murine leukemia virus was engineered in the 1980s and neither the virus itself nor the plasmid (pAMS) containing its proviral genome were ever used in our laboratory. Although the precise source for the contamination could not be traced back, sharing cell lines with other laboratories seems the most likely explanation.”
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Further reading:

Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene
(1985)

Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production,(1986).

 

[greybeh]

I sent a link to the WP Institute. I don't know what this means but I think they should be made aware of it.

 

[fresh_eyes]

Thanks so much for posting this, cold_taste. I know they do lots of gene manipulation on MuLV...yikes.

 

[imready]

This is over my head

cold_taste_of_tears, your should post this on the WPI facebook page. I read this artical and have no idea what it means, can somebody explain it to me.

 

[Marylib]

Wpi

cold_taste_of_tears, your should post this on the WPI facebook page. I read this artical and have no idea what it means, can somebody explain it to me.

I don't think you can post anymore on the WPI facebook page, except in the "fans" section which seems to be a kind of general forum.

I could be wrong -- someone could try.

 

[kurt]

It means engineered MLV strains have 'escaped'

cold_taste_of_tears, your should post this on the WPI facebook page. I read this artical and have no idea what it means, can somebody explain it to me.

I asked a friend who is a biomedical engineer. Here is what he says:

"That article provides proof of principle for genetic engineering vectors, presumed to be uninfectious, actually getting out of the lab. If it is showing up unexpectedly in cell culture, then where else could that virus be (especially if it is infecting human cells in cell culture)? If that virus can do it, then how many more engineered retroviruses can do it? Scientists have engineered hundreds of these things over the past 30 years, many of them coming from the MLV strains…"

Basically, an engineered virus escaped from its lab in the 1980s. The virus came from an MLV strain. That virus is now appearing in some cell culture lines, these are the 'media' used in labs to help grow specimens they are studying. Once a cell line is contaminated, its offspring remain contaminated. How this line became contaminated will probably not be found out, but the fact that this type of line has been contaminated by an escaped, engineered MLV derived strain is worthy of note.

XMRV is closely related to a host of engineered viruses from MLV, including those used in gene therapy (that has been halted because of patients getting leukemia from the viruses). Supposedly these viruses used in genetic engineering are designed to NOT be pathogenic to humans. So if they are out there, don't think that automatically means they are making us sick. In fact some day we may even be customers, who knows, maybe gene therapy could help some PWC in the future.

I don't know if this angle of XMRV has been discussed yet here, there are so many threads now it is hard to keep track. But no doubt the researchers at WPI and in the replication studies are well aware of the possibility that we harbor those types of retroviruses as well. If it becomes relevant we will probably hear about it eventually.

 

[jenbooks]

I wondered about this right at the start--inadvertant human-engineered contamination ie we created our own monster, considering how long we've engineered MLV and the fact that we work so exclusively with lab mice when testing anything. There was also one interesting study (I don't feel like finding the citation at the moment but it's somewhere on Cort's site and we discussed) where MLV increased infectivity of XMRV and researchers warned therefore that if you were working with cell lines you did not know were infected with XMRV you could have trouble.

MLV does have promoter sequences that were only recently removed and in gene therapy trials on bubble babies caused cancer. That's because the virus which was used as a vector (with seemingly potentially harmful or infectious sequences removed) to insert a healthy gene into stem cells occasionally landed and integrated next to oncogenes. Then the promoter sequences swiched on the oncogenes and those cells replicated in an uncontrolled fashion as cancer does and therefore gained a competitive advantage over other cells leading eventually to cancers that had to be treated in the bubble babies (who'd been cured of bubble baby disease thru gene therapy).

So there is a chance these murine viruses could theoretically potentiate cancer that way if the contamination got into the blood supply--who knows. It's too upsetting to me to actually look into in detail.

 

[starryeyes]

I'm glad you answered Jen, I was thinking of you when I read this thread. Also it's interesting to hear from the biomedical engineer, Kurt.

This is turning out to look like what we've all been discussing for many years. Doesn't this fit in well with the Cover-up theory too?

tee

 

[jenbooks]

I don't know teekjay (coverup). I don't know enough about whether an infected cell line, which is being used to study genetic manipulation or in vitro cell effects, could somehow get into human beings working with those cell lines. I guess I'd rather not speculate.

 

[George]

rate of spread

I don't know teekjay (coverup). I don't know enough about whether an infected cell line, which is being used to study genetic manipulation or in vitro cell effects, could somehow get into human beings working with those cell lines. I guess I'd rather not speculate.

Good catch, this is contamination of germ lines. Millions of dollars of research material pretty much down the tubes. It's important that researchers know this before doing work and getting screwy results. Worse case senerio would be that cell lines used by WPI come out as contaminated. But I doubt it. However this study regarding the contamination could be used to muddy the waters.

I don't think any thing as recent as 1980 could have time to reach 17 million people in 30 years or less. Most Phd'ers take this back to 1934 for first cases so I don't see how it would play.

 

[jenbooks]

George

I wanted to thank you for that URL with the interesting history on HIV.

I guess one thing I will say is that--as they admit in the world of gene therapy for instance--we underestimate viruses. They've been evolving for quite a long time. They have many tricks up their sleeves. They are adaptable. We manipulate things in labs as if we are truly gods--when in fact sometimes we're pawns .

 

[George]

Welcome

I wanted to thank you for that URL with the interesting history on HIV.

I guess one thing I will say is that--as they admit in the world of gene therapy for instance--we underestimate viruses. They've been evolving for quite a long time. They have many tricks up their sleeves. They are adaptable. We manipulate things in labs as if we are truly gods--when in fact sometimes we're pawns .

Yeah, it's inevitable that our inattention and lack respect will come back and bite us in the butt sooner or later.

The smoking gun in HIV isn't very. . .well smoking. (grin) I think it may be a little more identifiable for XMRV. I've been reading up about mouse virus and zoonosis. We've live side by side with mice since the days of the cave, in the 1600 the Chinese started breeding mice as pets. Fancy mice became a big deal in Victorian England in the late 1800's and people breed them, white, silver, gray, big eared, little eared and everything in between. Yet no retro-viral epidemics or evidence of viral zoonosis in all that time.

The only correlation between event and illness that works well for the accepted time frame that I have been able to find is the first inbreeding of mice by Clarence Cook Little. He inbreed the strains to 99% identical type. The first breed that was sent out to Hospitals and research facilities was called "dba" and is still being used in research today. That was around 1920 then, we get our first out break of CFS in 1934 LA General Hospital which was attached to the teaching school (including labratory) at USC. Many of the early outbreaks were affiliated with hospitals and people who worked in hospitals.

I'm still looking into connections and I haven't heard back from any of the e-mails I've sent so far but the correlation really pops for me.

Sorry for going over old stuff but I'm just really digging into this. Makes me feel useful for the first time in a long time.

 

[jenbooks]

It's very very interesting and I hope you continue to post your thoughts about it. I doubt you will get on the record speculation from within the field as it's too risky for them.

 

[George]

Might as well doing something I'm good at while I wait!

It's very very interesting and I hope you continue to post your thoughts about it. I doubt you will get on the record speculation from within the field as it's too risky for them.

Oh, no I didn't even think about e-mailing the researchers. I've got e-mails out to librarians in LA and Florida and Jackson Laboratory. I'm interested in old records that would put certain inbreed mouse strains in those areas with a 10 year pre-outbreak window.

In other words it keeps my mind busy while I wait for more news. It beats screaming (sheepish grin). Course now I'm hanging out here and it's keeping me nicely distracted too.

 

[jenbooks]

What a great idea.

I did look back at the early work on MLV in the 70s when they published work demonstrating it was not infectious in human cell lines. Of course I am mindful of the Jurassic Park line, Life will find a way. Add that to 'adapt or die.' Well anyway I'm very curious to see what you turn up. I had no idea for instance that we kept mice as pets a few centuries ago.

 

[Mark]

Good catch, this is contamination of germ lines. Millions of dollars of research material pretty much down the tubes. It's important that researchers know this before doing work and getting screwy results. Worse case senerio would be that cell lines used by WPI come out as contaminated. But I doubt it. However this study regarding the contamination could be used to muddy the waters.

I don't think any thing as recent as 1980 could have time to reach 17 million people in 30 years or less. Most Phd'ers take this back to 1934 for first cases so I don't see how it would play.

Wouldn't that depend on the transmission vector George? If it had some better way of spreading more rapidly, wouldn't that make it infect more people quicker. I mean, look at H1N1! What sort of transmission model might fall in between airborne and HIV-vectors and reach 17 million people (or rather, 5% of the population) in 30 years? Also there are the outbreaks themselves to explain: they are quite tricky to explain with the more limited transmission vectors, the best theory yet is as XMRV as passenger in other viruses.

We might even be able to figure out the transmission vectors from all this info, if we assume the lab-escape theory and it turns out that's correct.

I know quite a few people feel that the previous historical outbreaks rule out a virus appearing in the 1980s, but I'm not convinced of that yet. Those previous variants could be, for example, some other, very similar condition (possibly endogenous) that's now being activated by XMRV along with other conditions. Or something...needs more thought this one...

This question's a really crucial one to ponder because the genetically-engineered theory is pretty compelling to me...and if it were true, having to then explain the 4% population reach within that time, and also the historical outbreaks, would yield a lot more...um...tenuous but interesting conclusions.

 

[flybro]

Unintended spread of a biosafety level 2 recombinant retrovirus

This link has info on it mentioning XMRV and a synthetic virus.

http://www.retrovirology.com/content/6/1/86

Does anyone think this is related to XMRV in the WPI studies. Or Dr Defriatas (spelling sorry) studies.

From the link

The first hint for the presence of contaminating retroviruses in one of our cell lines was obtained by electron microscopy of exosome-like vesicles released from the supernatants of transfected 293T cells. Random amplification of particle associated RNAs (PAN-PCR) from supernatant of contaminated 293T cells and sequencing of the amplicons revealed several nucleotide sequences showing highest similarity to either murine leukemia virus (MuLV) or squirrel monkey retrovirus (SMRV). Subsequent mass spectrometry analysis confirmed our findings, since we could identify several peptide sequences originating from monkey and murine retroviral proteins. Quantitative PCRs were established for both viruses to test currently cultured cell lines as well as liquid nitrogen frozen cell stocks. Gene fragments for both viruses could be detected in a broad range of permissive cell lines from multiple species. Furthermore, experimental infections of cells negative for these viruses showed that both viruses replicate rapidly to high loads. We decided to further analyze the genomic sequence of the MuLV-like contaminant virus. Surprisingly it was neither identical to MuLV nor to the novel xenotropic MuLV related retrovirus (XMRV) but showed 99% identity to a synthetic retrovirus which was engineered in the 1980s.




Thanks for looking.

 

[_Kim_]

Hi flybro,

There is another discussion in progress about this article.

http://forums.aboutmecfs.org/showthread.php?t=1117&highlight=biosafety

Cheers,
Kim