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Abstracts from the 1st XMRV Conference

Discussion in 'XMRV Research and Replication Studies' started by George, Sep 20, 2010.

  1. mojoey

    mojoey Senior Member

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    Not being able to copy and paste from the PDF is probably why this thread hasn't exploded.

    Abstract P_03 titled: XMRV productively infects primary antigen-presenting cells, authored by the NCI, gives more insight into reservoirs for the virus, suggesting that
    in contrast to the aforementioned study on EBV transformed B cell lines

    and concluding that
    The most interesting part of all this is perhaps the following comparison between MLV and XMRV:

  2. Lesley

    Lesley Senior Member

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    Anti-retroviral trial in patient with chronic lymphocytic leukemia.

    Here's an interesting one:

  3. Lesley

    Lesley Senior Member

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    Another interesting one is abstract P-19, involving families with neuroimmune diseases including CFS and Autism.

  4. mojoey

    mojoey Senior Member

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    Some more takeaways
    -------------------------------------------------------------------------------------------------------------------


    From Abstract P_04 by Cleveland Clinic Foundation:

    From Abstract P_10 by Functional Genetics Inc, R&D, Gaithersburg, USA

    Abstract P_18 from Spain:

  5. Otis

    Otis Señor Mumbler

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    WOW. My conclusion is that XMRV is prevalent in neuro-immune diseases and large-scale familial testing (perhaps including grand-parents, if possible) along the lines of this study is warranted.

    Did I say WOW?!
  6. jimm

    jimm

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    wild mice

    the infected wild mice had leukopenia...that really jumped out at me...i have leukopenia i.e low white cells accross the board.....people with HIV have similar I beleive, hence I was offered an HIV test by 3 docs now,
    that struck me as possibly very telling
  7. Otis

    Otis Señor Mumbler

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    Managed a quick and dirrty! OCR of the abstracts

    See the attachment.

    The formatting isn't great and the header and footers are embedded with the text, so definitely check against the original for errors.

    Was in Word but it was too big to upload so I had to convert back to PDF.

    Attached Files:

  8. mojoey

    mojoey Senior Member

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    Awesome Thanks a million Otis!
  9. urbantravels

    urbantravels disjecta membra

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    Many thanks Otis!!
  10. Otis

    Otis Señor Mumbler

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    My pleasure. I'm too PEMed to do any thinking so have at it!
  11. eric_s

    eric_s Senior Member

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    That's a lot of information!! I went out in the evening, so i'm a bit wasted and don't read much now.

    But one thing strikes me... the way i see it, there is quite a discrepancy between what i can read here now in the abstracts and the impression the Q&A session gave me.

    Why is this? And especially Dr. Coffin irritated me a bit in the Q&A (even though i respect him).

    My reaction to the information i can now see would be totally different than the message the Q&A was sending out. Do they want to keep the general public calm? And if so, it's really surprising how well they are succeeding in doing this. I'm not saying panic would be better, maybe things are good the way they are, but it's quite confusing.
    Anyway, it seems the science has already progressed further than i have thought and that's a good thing.

    Thanks to everybody who has worked on the documents in this thread.
  12. leela

    leela Slow But Hopeful

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    So much new info.....Brain exploding.....Eyes bleeding....
    (not to mention EBV and XMRV doing a dark ritual somewhere deep within, drawing the pentagram, sacrficing the goat...)
  13. eric_s

    eric_s Senior Member

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    I think we can be happy to have such smart people working for us now. I wonder if i would ever be able to do something as valuable as what they are doing.
    Lol... maybe they should switch to sacrificing weasels :tongue: but i don't want to joke about these things, i'm not catholic but now i feel like i should make the sign of a cross in front of myself.
  14. George

    George waitin' fer rabbits

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    Yep, yep and yep. The whole Q and A was about the politics not about the science. The Q and A will be watched by the media and can be dissected and writen up in nasty little sound bites. The Abstracts will most likely not be read by anyone in the media, too much work. (grins) plus what the heck is the fun of reporting on "science". (big grins)

    The PTB's are sweatin' bullets trying to keep the media out of this, especially with the holidays coming up. Man poor moms and dads, kids and ordinary people so sick they can hardly stand up, that's really good pre-holiday tear jerker stuff.

    So yeah, the science is way, way past a bunch of people trying to figure out if it's lab contamination. (roll eyes) At this point it more about methods of detection and pathagenisis. GOOOOOOO TEAM!
  15. leela

    leela Slow But Hopeful

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    really, my head has exploded, so not much intelligent thought can dribble out right now, and there's so much to say/think/feel... but that drivel in the Q&A...
    they're fighting about contamination and replication in blood, when they were presented with a mind-blowing paper from the macaque people that suggests blood isn't even the best place to look for XMRV? I mean, get the hell ON with it already!!

    <sigh>

    Thanks for this awesome thread. I shall tear myself away from the computer now and go put the pieces of my shattered head back together.

    love you all:hug:!!
  16. XAND

    XAND

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    Here goes the three posters of Irsi Caixa Spanish retrovirologist team, the ones that make the link with EBV and XMRV, and if I am not mistaken, I also understand that they DID find XMRV in CFS patients:

    Detection of XMRV sequences in EBV-transformed B cell lines
    J. Blanco1, J. Carrillo1, E. Garcia1, J. Areal2, B. Clotet1, C. Cabrera11
    IrsiCaixa Foundation, Retrovirology Lab, Barcelona, Spain;
    2HUGTIP, Urology Dept, Barcelona, Spain

    Background: XMRV infection has been found in humans linked with prostate cancer and chronic fatigue syndrome (CFS). XMRV is able to infect a wide range of cells and has been found in a variety of cell types both in Vitro and in vivo, including B cells and other immune cell types.

    Materials and methods:
    In order to determine the presence of XMRV sequences in B cells, we have screened 21 B cell lines available in our laboratory. These cells were generated by EBV immortalization of PBMC obtained from 11 CFS affected individuals (fulfilling both Fukuda and Canadian criteria), 5 healthy donors, 4 HIV infected individuals and 1 prostate cancer patient.
    DNA was extracted from dry cell pellets and XMRV sequences were amplified using a real-time PCR covering a 150bp pol sequence and using a nested approach in both gag and Env genes.

    Results:

    Envelope amplification yielded positive bands in 4 out of 21 individuals tested, 3 CFS affected individuals and 1 healthy donor.

    However, gag amplification yielded only 3 positive samples (1 CFS affected individual, 1 healthy donor and one HIV+ patient).

    In contrast, Real-time PCR of Pol fragment detected 7 positives samples in 14 individuals tested (4 SFC , 2 donors and 1 HIV+ individuals).

    To confirm the presence of XMRV sequences we performed sequence analyses of gag and env amplicons. The analysis of the three available gag sequences confirmed the XMRV characteristic 24-nt deletion, which is not found in any known exogenous MuLV. Sequences were 100% identical to reported XMRV sequences. Furthermore, envelope sequences were also homologous to previously described XMRV sequences. In this case, sequence variability was low or absent. Interestingly, most of changes observed corresponded to G to A mutations that were accumulated in one positive sample.

    Conclusions: Despite the discrepancies observed in the different PCR approaches using gag, pol or env sequences, our data suggest that EBV transformed B cell lines harbor XMRV specific sequences, and therefore this cell type may represent a reservoir for XMRV contributing to its potential pathogenesis.


    Pathogenesis

    Xenotropic murine leukemia virus-related infection of human lymphoid tissue ex vivo

    C. Cabrera1, M. Curriu1, J. Carrillo1, M. Massanella1, E. Garcia1, B. Clotet1, C. Carrato2, J. Blanco11
    Fundacio irsiCaixa, Retrovirology Laboratory, Badalona, Spain;
    2Hospital Universitari Germans Trias i Pujol,
    Departamento de Anatoma Patolgica, Badalona, Spain

    Background: The gammaretrovirus xenotropic murine leukemia virus-related virus (XMRV) has been recently associated with prostate cancer and chronic fatigue syndrome. In patients with both diseases, the virus has been found in a variety of cell types, including T and B cells. Moreover, in XMRV-infected rhesus macaques there is evidence of viral replication in lymphoid organs, suggesting that lymphocytes are a primary target for XMRV. Histocultures of tonsils support productive infection with various lymphotropic viruses, including HIV and HHV-6.

    In this study, ex vivo lymphoid tissue was used to investigate the pathogenic mechanisms of XMRV

    Materials and methods: Human tonsils from 2 healthy individuals undergoing tonsillectomy were collected in PBS and cultured in small pieces (2mm3) over gelfoam soaked in RPMI 1640 medium. Small tissue blocks were left uninfected or were infected ex vivo with a XMRV viral stock obtained from a 22Rv1 cell culture supernatant. Culture medium was replaced every 3 days. After 14 days in culture, uninfected and infected tissues were mechanically homogenized and cells were isolated. Viral infection was evaluated at different times in the cells migrating out of the tissue and at day 14 in tissue cells, by analyzing the presence of viral DNA by PCR. In addition, tissue cells were immunophenotyped and analyzed by flow cytometry

    Results: At day 7 post-infection cells migrating out of the tissue were positive for XMRV DNA. After 14 days of culture, tissue cells were also highly positive, confirming that XRMV (as other viruses) was able to infect human tonsil tissue fragments in the absence of exogenous stimulation. Despite this apparent efficient infection, cells showed similar percentages of T and B cells in uninfected and infected tissues. XMRV infection did not modify the percentage of CD3 (76 and 75% in XMRV+ and XMRV- tissue, respectively), CD4 (53% vs 52%), CD8 (39% vs 40%) or CD19 cells (3% vs 1%). A deeper analysis of T cell subsets showed that XMRV infection did not modify the nave/memory cell ratio, as evaluated by CD45RO staining, or immune activation markers, as evaluated by the expression of HLA-DR and CD38 in both CD4 and CD8 T cells

    Conclusions: The development of in vitro models to study XMRV pathogenicity is essential for understanding its role in human diseases. Our data show that histoculture of human lymphoid tissue is a suitable model for the analysis of XMRV pathogenesis. In this system we observed a infection by XMRV although this infection did not result in depletion of T or B cells nor an immune activation, suggesting the absence of evident depleting effect of XMRV in these cells in lymphoid tissue.


    Altered B, T and NK cell phenotype in chronic fatigue syndrome (CFS) individuals.

    M. Curriu1, M. Massanella1, J. Carrillo1, J. Puig2, J. Rigau1, B. Clotet1, C. Cabrera1, J. Blanco11
    Fundaci IrsiCaixa, Retrovirology laboratory, Badalona, Spain;
    2Hospital Universitari Germans Trias i Pujol, Fundaci Lluita contra la SIDA, Badalona, Spain

    Background: Several studies have reported controversial results on the dysfunction of the immune system in Chronic Fatigue Syndrome (CFS) affected individuals. Since the recently described Xenotropic murine leukemia virus- related virus (XMRV) can infect cells from the immune system (B, T or NK cells), we designed different panels of antibodies to deeply characterize the phenotype of B, T and NK lymphocytes populations in patients with CFS.

    Material and methods: We obtained blood samples from 12 individuals affected from CFS (fulfilling both Fukuda and Canadian criteria) and 15 healthy donors (HD). B, T and NK cell populations were phenotyped in fresh blood samples and analyzed by multicolour flow cytometry. Fresh PBMCs were obtained to characterize spontaneous ex vivo apoptosis and NK cytotoxic activity against EGFP-K562 cells, both analyzed by flow cytometry.

    Results: No changes in the percentage or absolute numbers were observed in principal populations of B (CD19+), T (CD3+, CD3+CD4+, CD3+CD8+) or NK (CD56+ CD16+) cells. However, significant differences were observed in various subsets of these populations.B lymphocytes from CFS individuals showed a decrease in marginal-zone marker CD1c, especially in memory IgG population. The percentage of memory IgG+ cells, effector memory CD38high IgG+ cells and plasmatic B cells (CD38high/CD27high) was also reduced in CFS individuals. In contrast, CFS individuals showed an increase in transitional B cells (IgD+CD38highCD5+CD10+), which present lower spontaneous ex-vivo apoptosis. T-cells from CFS individuals presented increased CD25 levels mainly within CD8+ population.

    Proliferation marker Ki67 was significantly diminished in CFS CD4 T-cells and a trend was also observed in CD8 T-cells. In addition, individuals with CFS showed increased CD5 levels within CD8 T-cells which could suggest an anergic state. No signs of altered senescence (CD57+ T cells) or activation (CD38+, HLA-DR+ or double positive cells) were observed in CD4 or CD8 subsets, although, CD8 T cells from CFS individuals showed higher expression of FAS and PD-1 and a slightly higher spontaneous ex-vivo apoptosis. NK cells showed a significant decrease in CD57+ expression and expressed higher levels of CD69 and activating NK Cytotoxic Receptors (NCR) NKp30, NKp44 and NKp46. Nevertheless, this altered phenotype did not impact function, as we did not observe differences in NK cytotoxic activity in CFS patients compared with HD.

    Conclusions: CFS patients showed qualitative but not quantitative alterations in all major immune cell types. B cells presented a phenotype partially similar to some autoimmune disorders or viral infections. Interestingly, an anergic phenotype observed in T cells from CFS individuals could be related with an impaired control of viral infections and could explain the lack of increased activation and senescence observed in our patients. Finally, altered NK phenotype did not seem to significantly modify cytotoxic activity. On the whole, the results suggest a global immune dysfunction with an unknown aetiology as potential contributor to the mechanism of CFS.
  17. Dolphin

    Dolphin Senior Member

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    For one interested in the "recovered" Lyndonville cases, perhaps have a look at this thread on a paper this year by David Bell and David Bell (sic)!: Definition of recovery in CFS (Bell & Bell, 2010) (inc follow-up of pediatric cohort) http://www.forums.aboutmecfs.org/sh...Bell-2010)-(inc-follow-up-of-pediatric-cohort)
  18. eric_s

    eric_s Senior Member

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    Thanks, XAND. They really already know a lot. Just those couple of lines above, wow...
  19. redo

    redo Senior Member

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    Please note that it says T and B cells in peripheral blood. XMRV is in little degree found in peripheral blood when the acute infection phase is done (see page 7).
  20. mojoey

    mojoey Senior Member

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    Great point redo. I was thinking that as I was reading the abstract, but got so tired typing all those sections out that I forgot to mention it ;)

    So researchers needs to look at B and T cells in lymph, bone marrow? Where else do they exist?

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