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Lombardi et al 2011 - cytokine signature in CFS

Discussion in 'XMRV Research and Replication Studies' started by jace, May 13, 2011.

  1. jace

    jace Off the fence

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    in vivo 25: 307-314 (2011)

    Xenotropic Murine Leukemia Virus-related Virus-associated Chronic Fatigue Syndrome Reveals a Distinct Inflammatory Signature

    VINCENT C. LOMBARDI1, KATHRYN S. HAGEN1, KENNETH W. HUNTER4, JOHN W. DIAMOND2, JULIE SMITH-GAGEN3, WEI YANG3 and JUDY A. MIKOVITS1

    1Whittemore Peterson Institute, University of Nevada, Reno MS 0552, 1664 N. Virginia St., Reno, NV 89557, U.S.A.;
    2Triad Medical Center, 4600 Kietzke Lane M242,Reno, NV 89502, U.S.A.;
    3Nevada Center for Health Statistics and Informatics, University of Nevada, 1664 N. Virginia St., Reno, NV
    89557, U.S.A.;
    4University of Nevada Reno, Department of Microbiology and Immunology Applied Research Facility,1664 N. Virginia St., MS 199, Reno, NV 89557 U.S.A.

    Abstract.

    Background:

    The recent identification of xenotropic murine leukemia virus-related virus (XMRV) in the blood of patients with chronic fatigue syndrome (CFS) establishes that a retrovirus may play a role in the pathology in this disease.

    Knowledge of the immune response might lead to a better understanding of the role XMRV plays in this syndrome.

    Our objective was to investigate the cytokine and chemokine response in XMRV-associated CFS.

    Materials and Methods:

    Using Luminex multi-analyte profiling technology, we measured cytokine and chemokine values in the plasma of XMRV-infected CFS patients and compared these data to those of healthy controls.

    Analysis was performed using the Gene Expression Pattern Analysis Suite and the Random Forest tree classification algorithm.

    Results:

    This study identifies a signature of 10 cytokines and chemokines which correctly identifies XMRV/CFS patients with 93% specificity and 96% sensitivity.

    Conclusion:

    These data show, for the first time, an immunological pattern associated with XMRV/CFS.

    ````

    Introduction

    Chronic fatigue syndrome (CFS) is a poorly understood disease of unknown etiology, which is commonly characterized by innate immune defects, chronic immune activation and dysregulation, often leading to neurological maladies [reviewed in (1)].

    It can also involve other biological systems such as the musculoskeletal, gastrointestinal and endocrinological systems (2-4).

    Although several common symptoms are primarily reported and predominate, they may differ among individuals, are often intermittent and can persist for years, frequently resulting in substantial disability
    (5).

    Some of the most commonly reported physical symptoms include muscle weakness and pain, tender or swollen lymph nodes and chronic flu-like symptoms (6).

    Memory and concentration impairment, blurred vision, dizziness and sleep abnormalities represent some of the cognitive symptoms typically observed while immunological symptoms often manifest themselves through viral reactivation, RNase L dysregulation, decreased natural killer (NK) cell function and susceptibility to opportunistic infections (7-12).

    NK cell dysregulation may be associated with viral reactivation or viral persistence and may also lead to malignancy (13, 14).

    Indeed, clinical observations corroborate pathological manifestations in CFS as viral reactivations, particularly herpes virus such as cytomegalovirus (CMV), Epstein-Barr virus (EBV) and human herpes virus-6 (HHV-6), are common occurrences (15-17).

    Moreover, epidemiological studies have reported increased incidences of lymphoma associated with CFS outbreaks (18 ).

    These clinical observations suggest that a compromised innate immune system may play a role in CFS pathology.

    The completion of the human genome project enabled positional cloning studies to identify the RNASEL gene as the hereditary prostate cancer allele-1 (HPC1) (19).

    This discovery prompted Robert Silverman and his colleagues to search for a viral component to hereditary prostate cancer.

    Using a viral micro-array and tissue biopsies from individuals with hereditary prostate cancer they identified and sequenced the complete genome of a novel human gammaretrovirus, very similar in sequence to xenotropic murine leukemia virus and therefore termed the new virus xenotropic murine leukemia virus-related virus (XMRV) (20).

    Subsequent studies performed in our laboratory identified and isolated infectious XMRV in the blood of 67% of CFS patients (21).

    This work was performed using multiple techniques including PCR, electron microscopy showing budding viral particles, Western blot analysis of viral proteins and serology confirming that infected patients express antibodies to XMRV envelope proteins.

    In addition, gene sequencing and phylogenetic analysis confirmed these patients were indeed infected with XMRV that was >99% identical to previously published sequences but was obviously distinct from the only existing XMRV molecular clone, VP62 (20).

    Taken together, this work clearly rules out any possibility of gross contamination and additionally, represents the first identification and isolation of naturally occurring infectious XMRV.

    The connection between CFS and XMRV was further supported by the studies of Lo et al., who identified murine leukemia virus (MLV)-related sequences in the blood of 86% of CFS patients, further establishing a retroviral association with CFS (22).

    Presently, three families of retroviruses are known to infect humans; the human immunodeficiency viruses
    (HIV), the human T-cell leukemia viruses (HTLV) and now the human murine leukemia-related viruses.

    Both HIV and HTLV are known to dysregulate the innate immune system and promote the production of inflammatory cytokines and chemokines (23, 24).

    In light of the association between XMRV and CFS, it is not surprising that some of the most salient observations in CFS are the differences in cytokines and chemokines when compared to healthy controls (8 ).

    Previous reports, however, addressing the role of
    these molecules in CFS have produced conflicting
    results.

    Much of this emerges from such hindrances as small sample size, a limited number of cytokines surveyed at one time, insufficient patient population stratification, and insufficient negative control subjects.

    This has resulted in inconsistent reports in the literature for a number of cytokines including interleukins (IL) 6, 10 and 12.

    In spite of these conflicting results, a number of cytokines and chemokines have consistently been show to be associated with different subgroups of CFS.

    For instance, Natelson et al. showed elevated levels of IL-8 and IL-10 in the cerebral spinal fluid of patients with sudden, influenza-like onset CFS when compared to healthy controls (25).

    Additionally, Chao et al. have show neopterin and IL-6 to be up-regulated in subsets of CFS patients, indicative of a pro-inflammatory immune condition (26).

    However, these studies did not analyze the complex relationships between multiple cytokines and clinical disease.

    By applying conventional statistical analysis and 'machine logic' algorithms to the multiplex data, it is possible to identify cytokines and chemokines that are differentially expressed between two groups.

    To support this premise, we have used the xMap multi-analyte profiling technology that allows simultaneous measurements of multiple biomarkers in serum or plasma.

    In this study, a panel of 26 cytokines, chemokines and growth and angiogenic factors were analyzed in blood plasma of CFS patients and healthy control subjects.

    This study revealed a signature of 10 cytokines and chemokines, which showed a specificity of 93% and sensitivity of 96% in diagnosing XMRV-associated CFS in this patient cohort.

    Full text here http://www.megaupload.com/?d=V980UCHY

    If you are not a member of Megaupload, you have to wait for about a minute.
     
  2. Jemal

    Jemal Senior Member

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    Awesome, thanks for sharing. Looks like the WPI was finally able to publish something again?

    This seems to establish CFS (or XMRV related CFS as they call it) as an inflammatory disease. Which makes sense as anti-inflammatory medicine has helped me a lot.

    The bad news seems to be that we have a greater risk to get lymphoma...

    But now we have a test?
     
  3. pamb

    pamb Senior Member

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    It seems to me I should be jumping for joy? Is this getting much closer to proving a) XMRV is real and lots of ME/CFS people seem to have it b) leaning towards proving it is not a harmless bystander? I'm guessing much more is needed to prove it 'causes' something, but this certainly seems unlikely to be coincidence.

    I look forward to all the brainy science types explaining it soon :D
     
  4. Jemal

    Jemal Senior Member

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    I am almost sure this paper is not going to resolve the controversy. The good thing is that this paper doesn't look for XMRV in patients, but it looks for a certain signature in people that are XMRV+. XMRV is controversial, but I don't think there will be much controversy over the signature itself... I guess we'll see.

    For me it's more evidence that there's certainly something there.
     
  5. alex3619

    alex3619 Senior Member

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    Hi, I have a lot to learn about gamma delta T-cells (aka Dendritic Epidermal T-cells) as I know almost nothing about them. There are three main points I would like to raise at least until I know more. The first is they are half way between the innate and adaptive immune system, not completely part of either. Second, they reside in mucosal tissue (where Silverman has shown XMRV likes to grow), especially the gut but also the skin, lungs, tongue and reproductive epithelium. Third, they directly sense toxic bacterial products - they are able to detect bacterial infections based on the chemicals that bacterial produce.

    Bye
    Alex

    PS In case anyone is interested in why I discuss gamma delta T-cells, these are the ones upregulated in this study, and thought to produce many of the cytokines that were found upregulated.
     
  6. Jemal

    Jemal Senior Member

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    So why would gamma delta T-cells be upregulated: is it a response by the immune system or is the virus doing that?
     
  7. gbmannc

    gbmannc

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    Meh, probably mouse contamination...
     
  8. gbmannc

    gbmannc

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    Is it going to resolve the controversy? Probably not. Is it a complete slap in the face to anyone that said it was contamination? It sure is. They showed a correlation to empirical data with these tests rather than to a list of symptoms. This is much harder to discredit, and also much easier to replicate I would imagine.
     
  9. currer

    currer Senior Member

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    Sounds interesting given that we have gut problems with bacterial overgrowth - ie our gut bacteria are not being properly controlled.
    Also the recent paper showing that XMRV replicates in mucosal tissue.
     
  10. heapsreal

    heapsreal iherb 10% discount code OPA989,

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    other info that i have read has said t-cells upregulate to compensate for poor nk function??? This describes my immune results.
     
  11. Forebearance

    Forebearance Senior Member

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    Finally! The WPI published something from the pile of data they have accumulated!

    Thank you for the info, Alex.
     
  12. gbmannc

    gbmannc

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    Hmmm, one of the few abnormal blood tests I've had is a almost nonexistent igA (Immunoglobulin A). igA is the antibody responsible for mucosal immunity. It exists in saliva, upper airway, GI tract etc. Coincidence? Anyone else test low igA?
     
  13. Jim

    Jim Senior Member

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    Which anti-inflammatories have helped you?

    Thanks.
     
  14. joshualevy

    joshualevy

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    Sorry, but this paper has nothing to say about XMRV; it is all about CFS. it does not have any data for or against the idea that XMRV is real, that CFS people have it, or that it has any effect on them.

    The paper compared people who have CFS and tested positive for XMRV, to people who were healthy. The healthy people were never tested for XMRV as part of this study. (Neither were the CFS patients, but they had previously been found XMRV positive as part of the previous study.)

    So this study can tell us about differences between CFS who also have XMRV and healthy people, but there is no way to know if what is seen has to do with being CFS+ or XMRV+ or both.

    If you ask me, this is awful experimental design. At a minimum, they should have had four groups (CFS+XMRV, CFS, XMRV, neither). That would have told us about both CFS and XMRV.

    Joshua Levy
     
  15. Jemal

    Jemal Senior Member

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    Ibuprofen, Doxycycline (antibiotic, but also anti-inflammatory), Amitriptyline (anti-depressant, but also anti-inflammatory)
     
  16. gbmannc

    gbmannc

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    They also only used patients that came down with acute illness as their initial presentation of cfs. I wonder if using other cfs+ xmrv+ cohorts would have different results.
     
  17. Sushi

    Sushi Moderator and Senior Member Albuquerque

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    Here is a Poster Paper from the XMRV Conference, Sept. 2010, entitled: Is the Mechanism of Systemic Immune Activation in XMRV Positive CFS Patients Similar to that in HIV? (Kenny De Meirleir, M.D. Ph.D, Vrije Universiteit, Brussel, Brussel, Belgium et al)

    This was over a year ago, preliminary, and with relatively few patients, but it might be interesting to compare.

    Sushi View attachment poster paper at XMRV conference 9-10.pdf
     
  18. Hope123

    Hope123 Senior Member

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    Yes, I agree on skimming the abstract. I think the WPI did the best it could with the data they had and I appreciate that. However, it is hard to "pin" the signature on XMRV without having and comparing cytokine findings in people who test XMRV neg. The upregulated cytokines could be a function of a group of people who have CFS and not those who are XMRV+. Interestingly though, some CFS studies have found NO upregulated cytokines in their subjects at all and that really makes me wonder about cohort selection.

    Anyhow, this does support the papers (like Klimas and even one of Peter White's papers) out there already that show some CFS patients have upregulated cytokines. And this paper opens up more areas for research.
     
  19. Angela Kennedy

    Angela Kennedy *****

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    I'm sorry - but - correct me if I'm wrong - did not the whole of the CFS cohort here have positive XMRV results? How do we know the controls were new controls, not those tested and found XMRV negative? (I'm genuinely asking here).

    Because, if we do have in this paper CFS sufferers with positive XMRV results at those high correlations, it seems rather silly to dismiss this paper has having 'nothing to say about XMRV'!
     
  20. toddm1960

    toddm1960 Senior Member

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    Josh I agree with you 100%, but it's just good to see WPI being published again. (which makes the scientific community look more like Jr. High than a body we're depending on for answers......but oh well) Maybe the next step is going to be connecting XMRV with these results.
     

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