Dr. Bateman answers IOM questions from the community: Part 1
Clark Ellis brings us Part 1 of an interview with Dr. Lucinda Bateman, where she answered questions posed by the patient community ...
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Mouse retrovirus present in cell lines used for production of biologicals

Discussion in 'XMRV Testing, Treatment and Transmission' started by natasa778, Mar 15, 2010.

  1. MEKoan

    MEKoan Senior Member

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    That's really interesting, George!

    Leo Kanner, who coined the term Autism for that condition, said he began to see it in 1938 and published the first paper in 1943.

    Does anyone know how early vaccinations were made? Mice? There was intense pressure as far back as the turn of the 20th century, to mass vaccination against widespread and dangerous diseases.

    I've always been a fan of vaccination. All of us who work(ed) in international development were. The risk/benefit ratio seemed a no brainer. It may still be but it may be a more complex issue than we thought.

    I just don't know anything any more.
     
  2. MEKoan

    MEKoan Senior Member

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    From: The Wilbur A. Sawyer Papers

    The Yellow Fever Laboratory: Rockefeller Foundation, 1928-1937


    karma
     
  3. natasa778

    natasa778 Senior Member

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    Thanks for the lengthy explanation George, but few things...

    You used meat slicer analogy to describe contamination of cell lines, when here we are talking about something that is INTRINSIC to those cell lines. NOT something that would jump from those cutting blades, or something airborne. That ‘something’ would already come packaged integrated in the very genome of those cell lines.

    When you say radiation etc is used to decontaminate the cell lines, would those processes destroy/fragment any nucleic acid? Including cells’ own DNA? Wouldn’t that render those cells useless? Or if not, if cellular DNA is left mostly or at least partially intact, wouldn’t that mean that any/most retroviruses integrated in the genome would also 'survive'? A possibility?

    Also you say in the final stage the mice are checked… checked for what? How?
     
  4. natasa778

    natasa778 Senior Member

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    Limitations of the process to remove infectious viruses from cell lines

    www.emea.europa.eu/pdfs/human/bwp/026895en.pdf


    Note points 1.6 and 1.7 on page 3 – I could not copy and paste…

    On page 5 they say, under 3.2:
    "Results have shown that even small modifications in procedure or the particular laboratory strain of virus used can have a large effect on virus removal or inactivation"

    also note 3.5 "occasional cases have been reported" (what about unreported ones?)

    On page 6 they talk about rodent retroviruses. This guideline was written in 1994, what was the knowledge/awareness before then?

    Interesting discussion on page 9 on "Limitations of Validation Studies", i.e. numerous factors that may lead to INCORRECT ESTIMATE OF THE ABILITY OF THE PROCESS TO REMOVE NATURALLY OCCURRING VIRUS INFECTIVITY



    Exact science? Anything but, it seems ....
     

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  5. George

    George Guest

    You're right, my bad, post deleted.
     
  6. usedtobeperkytina

    usedtobeperkytina Senior Member

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    Clay, Alabama
    Holy S____!

    I don't understand it all, but this could be major.

    Tina
     
  7. starryeyes

    starryeyes Senior Member

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    This is the same information we came across back in October when we were trying to determine if DeFreitas's retrovirus CAV was the same as XMRV.

    I suspect that I got CFS from a vaccine as a kid but then EBV triggered it when I was 20.
     
  8. natasa778

    natasa778 Senior Member

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    Some further info and questions:

    FDA Letter to Viral Vaccine IND Sponsors - Use of PCR-based Reverse Transcriptase Assay (re detection of retroviruses in their products), dated 1998


    http://www.fda.gov/BiologicsBloodVaccines/SafetyAvailability/ucm105911.htm



    are recommended, but not legally required? What would infectivity studies be? Would those studies definitely rule out XMRV or any novel/still unknown exogenous infective retrovirus?


    does this leave room for in-lab contamination of cell lots via other cell lines?


    is it completely impossible that insect etc DNA could be a vector and contaminated?
     
  9. flybro

    flybro Senior Member

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    Is each vaccinated generation increasing potency of the and variety of virus'

    Is it possible that generation upon generation being vaccinated using disfferent strains of virus..

    increases the potency of the vaccine, and the variety of material available to the virus to mutate.
     
  10. natasa778

    natasa778 Senior Member

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    ok I'm answering one of my own questions here, sort of:


    http://jvi.asm.org/cgi/content/abstract/73/7/5843

    Evidence of Avian Leukosis Virus Subgroup E and Endogenous Avian Virus in Measles and Mumps Vaccines Derived from Chicken Cells: Investigation of Transmission to Vaccine Recipients

    Reverse transcriptase (RT) activity has been detected recently in all chicken cell-derived measles and mumps vaccines. A study of a vaccine manufactured in Europe indicated that the RT is associated with particles containing endogenous avian retrovirus (EAV-0) RNA and originates from the chicken embryonic fibroblasts (CEF) used as a substrate for propagation of the vaccine. We investigated the origin of RT in measles and mumps vaccines from a U.S. manufacturer and confirm the presence of RT and EAV RNA. Additionally, we provide new evidence for the presence of avian leukosis virus (ALV) in both CEF supernatants and vaccines. ALV pol sequences were first identified in particle-associated RNA by amplification with degenerate retroviral pol primers. ALV RNA sequences from both the gag and env regions were also detected. Analysis of hypervariable region 2 of env revealed a subgroup E sequence, an endogenous-type ALV. Both CEF- and vaccine-derived RT activity could be blocked by antibodies to ALV RT. Release of ALV-like virus particles from uninoculated CEF was also documented by electron microscopy. Nonetheless, infectivity studies on susceptible 15B1 chicken cells gave no evidence of infectious ALV, which is consistent with the phenotypes of the ev loci identified in the CEF. PCR analysis of ALV and EAV proviral sequences in peripheral blood mononuclear cells from 33 children after measles and mumps vaccination yielded negative results. Our data indicate that the sources of RT activity in all RT-positive measles and mumps vaccines may not be similar and depend on the particular endogenous retroviral loci present in the chicken cell substrate used. The present data do not support transmission of either ALV or EAV to recipients of the U.S.-made vaccine and provide reassurance for current immunization policies.


    Re "infectivity studies on susceptible 15B1 chicken cells gave no evidence of infectious ALV" - how reliable are those??

    This does not inspire that much confidence, or does it? No details on PCR preparation of cells etc. Why not a culture study? Electron microscope? Why only 33?
     
  11. Gemini

    Gemini Senior Member

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    Koan,

    You asked "Does anyone know how early vaccines were made?"

    How about the 1950's polio vaccines? Byron Hyde's list of 63 suspected ME/CFS outbreaks by decade:
    1930-39...... 5
    1940-49...... 4
    1950-59...... 29
    1960-69...... 6
    1970-79...... 6
    1980-89...... 12
    1990...... 1

    shows a cluster in the 1950's.

    Gemini
     
  12. Gerwyn

    Gerwyn Guest

     
  13. MEKoan

    MEKoan Senior Member

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  14. Gerwyn

    Gerwyn Guest

     
  15. natasa778

    natasa778 Senior Member

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    btw that study was done by CDC ...
     
  16. natasa778

    natasa778 Senior Member

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    There is more....

    Re retroviral contamination of chicken embryo-grown products there were a few studies in the late 90s indicating that there may be a problem. Have a look at this one by the Swiss group

    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC191429/?tool=pubmed.

    ... read the first and second paragraphs of discussion. They detected RT activity in all vaccines, but WHO experts concluded that it poses no danger to humans. Then in the next sentence we find out that they haven't a faintest clue what causes that RT activity. But they do suspect retroviruses are involved. No worries then.

    Shortly (very shortly!) after that study there were 3 negative studies, 2done by CDC, and 1 by FDA, saying there is no problem with retro contamination, everything is hunky dory, nothing to see, go home, book is closed.

    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2631681/?tool=pubmed

    http://jcm.asm.org/cgi/content/full/39/2/675?view=long&pmid=11158127

    http://jvi.asm.org/cgi/content/full/73/7/5843?view=long&pmid=10364336

    case closed!



    then this one comes along in 2008:

    Molecular characterization of three recombinant isolates of avian leukosis virus obtained from contaminated Marek's disease vaccines.

    Three natural recombinant avian leukosis viruses (ALV; PDRC-1039, PDRC-3246, and PDRC-3249) expressing a subgroup A gp85 envelope protein and containing long terminal repeats (LTR) of endogenous ALV-E viruses were isolated from contaminated commercial Marek's disease vaccines, cloned, and completely sequenced. Their full genomes were analyzed and compared with representative strains of ALV. The proviral DNA of all three isolates displayed 99.3% identity to each other, suggesting a possible common ancestor, even though the contaminating viruses were obtained from three separate vaccine serials produced by two different vaccine manufacturing companies. The contaminating viruses have a genetic organization typical of replication-competent alpharetroviruses. The proviral genomes of PDRC-1039 and PDRC-3246 are 7497 bp long, and the PDRC-3249 is three base pairs shorter because of a deletion of a threonine residue within the gp85 coding region. The LTR, gag, pol, and the transmembrane (TM) region (gp37) of the env gene of all three viruses displayed high identity to endogenous counterpart sequences (>98%). Only the surface (SU) region (gp85) of the env gene displayed high identity with exogenous ALV-A (98.7%). Locus-specific polymerase chain reaction (PCR) analysis for ALV endogenous sequences (ev loci) in the chicken embryo fibroblasts used to produce the original vaccine vials identified the presence of ev-1, ev-2, ev-3, ev-4, and ev-6 in all three vaccines. Homologous recombination most likely took place to involve the SU region of the env gene because the recombinant viruses only differ in this particular region from the consensus ALV-E. These results suggest that the contaminating ALV isolates probably emerged by recombination of ALV-A with endogenous virus sequences before vaccine preparation. Barbosa T, et al Avian Dis. 2008 Jun;52(2):245-52.


    wt* ???

    Can we sleep soundly as WHO says we should. Or toss and turn and reopen the book?

    Could these be the same lab/s that were involved with De Freitas follow up research, those that could not find anything: HIV and Retrovirology Branch, Division of AIDS, STD, and TB Laboratory Research, Centers for Disease Control ?

    are these labs linked to the branch of CDC that is now in charge of CFS??
     
  17. Gerwyn

    Gerwyn Guest

    the other study did not have a clue what produced rt activity either they were making stuff up

    as to your other vcomments in my view yes yes yes
     
  18. natasa778

    natasa778 Senior Member

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    Gerwyn do you mean one (or all 3) of CDC/FDA ones?
     
  19. Athene

    Athene ihateticks.me

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    OMG this is HUGE! XMRV in vaccines. Natasa you are amazing.

    I found this web page about the 1950s polio vaccine, which was made using monkey organs and was contaminated with "simian virus - 40" from Rhesus monkeys.
    This virus causes various types of cancers. People infected with it via vaccines can transmit it to others sexually and through blood transfusions.

    http://209.85.129.132/search?q=cach...accine 1950s made by&cd=1&hl=it&ct=clnk&gl=it

    When they realised the mistake they started using Green Monkeys instead. Anyone heard of Green monkey disease?

    And finally, a quote from the web page:
    "Dr. Urnovitz revealed significant evidence that human immunodeficiency virus type 1 (HIV-1) is a monkey hybrid virus which was produced when 320,000 Africans were injected with polio virus contaminated with live simian immunodeficiency virus (SIV) in the late 1950's. Apparently, viral fragments combine easily with other viruses to produce these hybrids called "chimeras." Prior to this revelation, health officials were blaming AIDS on the habit of certain Africans to consume monkey flesh. What can be done now? "Make it in anything but animals," said Barbara Loe Fisher of the National Vaccine Information Center, which criticizes vaccine safety."

    As far as I understand it, this would be an exact parallel/precedent for similar contamination of vaccines cultivated on rodent organs with XMRV. Could this be why some people with XMRV are well and others have CFS and others cancer? Perhaps the reaction depends on which other viruses you are infected with and thus which "hybrids" are produced in your body.

    Who had heard of this polio vaccine contamination? How many people know that Guillain Barre Syndrome is caused by vaccine damage? They're very good at hushing these things up.
     
  20. natasa778

    natasa778 Senior Member

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    Thanks Athene, none of this is about xmrv though. So far.
     

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