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Deficient EBV-Specific B- and T-Cell Response in Patients with Chronic Fatigue Syndrome

Discussion in 'Latest ME/CFS Research' started by Ecoclimber, Jan 16, 2014.

  1. Kate_UK

    Kate_UK Senior Member

    The author of this paper Professor Carmen Scheibenbogen will be at the Invest in ME Conference in London, 30th May

    tickets are discounted prices until 12th March 2014
    Last edited: Jan 22, 2014
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  2. PDXhausted

    PDXhausted Senior Member

    NW US
    Ema likes this.
  3. This is EXACTLY me. I had mono, i was recovering normally, but i went back to training for my sport too soon. No one told me otherwise and i was too dumb to know on my own. Sure enough, start training, and this ungodly feeling of fatigue would take over after each workout, but it always went away. Until the day it didn't. July 3rd 2010, i came home feeling terrible, took a nap, but this time i woke up feeling worse then when i fell asleep. All down hill from there.

    My EBV titers are still off the charts three years, a round of valcyte and rituximab later.
    Fuschia likes this.
  4. MeSci

    MeSci ME/CFS since 1995; activity level 6?

    Cornwall, UK
    I was puzzled reading this for a moment. Do you mean modern/conventional medicine? Traditional medicine usually means the older type - what is commonly called 'alternative/complementary' these days.
  5. anciendaze

    anciendaze Senior Member

    I'm caught between two different groups using language in different ways, like the British and Americans "divided by a common language". In this case I meant mainstream medicine, though mainstream medicine doesn't call itself that. I did not mean traditional as in folk medicine. I could have done better in this case, but I'm afraid there is no general solution to the problem of writing for multiple audiences as divided as these.
    Otis likes this.
  6. anciendaze

    anciendaze Senior Member

    @Young&sick, the fact that your EBV titers remain high indicates your immune system is in better shape than many who have been ill for years. This research may explain the conflicting results from serology for EBV going back to the mid 1980s. You can't depend on the infected cells to tell you they are infected.

  7. Maybe. Its a common theme for, my titers are still high for EBV, CMV, and a type of pneumonia three plus years after my initial bout of mono that triggered all this.
  8. A.B.

    A.B. Senior Member

    Epstein Barr virus-specific immune defects in patients with persistent symptoms following infectious mononucleosis.

    This paper from 1986 describes similar EBV-specific immune defects.
    merylg and Kati like this.
  9. Dolphin

    Dolphin Senior Member

  10. Dolphin

    Dolphin Senior Member

    I haven't read the German paper yet.

    One paper that occurred to me was the following, where there was not good immune control of Parvovirus B19 in CFS. I don't have a good knowledge of biochemistry or immunology (nor the German paper, as I said) - are there any similarities?

    Simon likes this.
  11. Ecoclimber

    Ecoclimber Senior Member

    This is interesting on how EBY produces an enzyme BPLF1 to evade detection by TLR.

    PLoS ONE
    Epstein-Barr Virus Large Tegument Protein BPLF1 Contributes to Innate Immune Evasion through Interference with Toll-Like Receptor Signaling

    Michiel van Gent, Steven G. E. Braem, Annemieke de Jong, Nezira Delagic, Janneke G. C. Peeters, Ingrid G. J. Boer, Paul N. Moynagh, Elisabeth Kremmer, Emmanuel J. Wiertz, Huib Ovaa, Bryan D. Griffin, Maaike E. Ressing mail

    Published: February 20, 2014 DOI: 10.1371/journal.ppat.1003960

    Viral infection triggers an early host response through activation of pattern recognition receptors, including Toll-like receptors (TLR). TLR signaling cascades induce production of type I interferons and proinflammatory cytokines involved in establishing an anti-viral state as well as in orchestrating ensuing adaptive immunity. To allow infection, replication, and persistence, (herpes)viruses employ ingenious strategies to evade host immunity.

    The human gamma-herpesvirus Epstein-Barr virus (EBV) is a large, enveloped DNA virus persistently carried by more than 90% of adults worldwide. It is the causative agent of infectious mononucleosis and is associated with several malignant tumors. EBV activates TLRs, including TLR2, TLR3, and TLR9. Interestingly, both the expression of and signaling by TLRs is attenuated during productive EBV infection.

    Ubiquitination plays an important role in regulating TLR signaling and is controlled by ubiquitin ligases and deubiquitinases (DUBs). The EBV genome encodes three proteins reported to exert in vitro deubiquitinase activity.

    Using active site-directed probes, we show that one of these putative DUBs, the conserved herpesvirus large tegument protein BPLF1, acts as a functional DUB in EBV-producing B cells. The BPLF1 enzyme is expressed during the late phase of lytic EBV infection and is incorporated into viral particles. The N-terminal part of the large BPLF1 protein contains the catalytic site for DUB activity and suppresses TLR-mediated activation of NF-κB at, or downstream of, the TRAF6 signaling intermediate. A catalytically inactive mutant of this EBV protein did not reduce NF-κB activation, indicating that DUB activity is essential for attenuating TLR signal transduction. Our combined results show that EBV employs deubiquitination of signaling intermediates in the TLR cascade as a mechanism to counteract innate anti-viral immunity of infected hosts.

    Author Summary
    Epstein-Barr virus (EBV) is a human herpesvirus that persistently infects >90% of adults worldwide. One factor underlying the ability of EBV to establish such widespread and lifelong infections is its capacity to escape elimination by the human immune system.

    Among the first lines of defense against viral infection is the human Toll-like receptor (TLR) system. These receptors can detect the presence of viruses and initiate an intracellular protein signaling cascade that leads to the expression of immune response genes.

    The activation status of many proteins in this signaling cascade is regulated by the addition of ubiquitin tags. EBV has previously been reported to encode enzymes, called deubiquitinases (DUBs), which are capable of removing such ubiquitin tags from substrate proteins. In our study, we found that one of these enzymes, BPLF1, functions as an active DUB during EBV production in infected cells before being packaged into newly produced viral particles.

    Furthermore, our study provides insight into the way in which EBV can subvert the human immune response, as we show that BPLF1 can remove ubiquitin tags from proteins in the TLR signaling cascade. This inhibits TLR signaling and decreases the expression of immune response genes.

    ....Since EBV can activate TLR2, TLR3, TLR7, and TLR9, it would be of benefit if EBV were to evade innate immune activation. Here, we examined whether EBV DUB activity was present during productive infection in B cells and whether it could inhibit innate immune signaling....
    ...BPLF1 is expressed during the late phase of productive EBV infection and is incorporated into viral particles...

    ...During EBV infection, EBV components are sensed by host TLRs (a). These receptors activate signaling pathways through adaptor molecules MyD88 and TRIF, culminating in activation of transcription factor NF-κB (b). NF-κB induces production of proinflammatory cytokines (c) and obstructs viral replication. TLR signaling pathways are extensively regulated by ubiquitination, a process governed by cellular ubiquitin ligases and deubiquitinases (e.g. A20, d). The EBV-encoded DUB BPLF1 counteracts TLR-mediated NF-κB activation and can interfere with K63- and K48-linked ubiquitination of signaling intermediates, for example on TRAF6, NEMO, and IκBα (e). BPLF1 is expressed as a full-length protein during the late phase of productive EBV infection, is incorporated into the tegument of viral particles (f), and can subsequently be released into newly infected cells (g). It is as yet unclear where BPLF1 is processed to yield a shorter active fragment of ca. 280 aa (see Fig. 1). Thus, (processed) BPLF1 could exert its immunomodulatory functions towards TLR signaling not only in EBV-producing B cells

    Click on link above for full article:

    Raises the question on whether BRLF1 causes EBV to evade detection by the immune system to such a degree, that medical lab tests are not sensitive to accurately detect the extent of EBV infection in patients.
    Last edited: Mar 10, 2014
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  12. natasa778

    natasa778 Senior Member

  13. Ecoclimber

    Ecoclimber Senior Member

    J Virol. 2014 Mar 5. [Epub ahead of print]
    Analysis of host gene expression changes reveals distinct roles for the cytoplasmic domain of the Epstein-Barr virus receptor/CD21 in B-cell maturation, activation and initiation of virus infection.
    Arredouani MS1, Bhasin MK, Sage D, Dunn LK, Gill MB, Agnani D, Libermann TA, Fingeroth J.
    Author information

    Epstein-Barr Virus (EBV) attachment to human CD21 on the B-cell surface initiates infection. Whether CD21 is a simple tether, or conveys vital information to the cell interior for production of host factors that promote infection of primary B-cells is controversial, as the cytoplasmic fragment of CD21 is short, though highly conserved.

    Ubiquity of CD21 on normal B-cells, the diversity of this population, and the well-known resistance of primary B-cells to gene transfer technologies have all impeded resolution of this question. To uncover the role(s) of the CD21 cytoplasmic domain during infection initiation, the full--length receptor (CD21=CR), a mutant lacking the entire cytoplasmic tail (CT), and the control vector (NEO) were stably expressed in two pre B-cell lines that lack endogenous receptor. Genome-wide transcriptional analysis demonstrated that stable CD21 surface expression alone (either CR or CT) produced multiple independent changes in gene expression, though both dramatically decreased class I MAGE family RNAs and upregulated genes associated with B-cell differentiation (e.g. C2TA, HLA-II, IL21R, MIC2, CD48, PTPRCAP/CD45 associated protein). Temporal analysis spanning 72 hours revealed not only CR, but also CT expressing lines initiated latency. In spite of this, the number and spectrum of transcripts altered in CR compared with CT-bearing lines at one-hour after infection further diverged. Differential modulation of immediate early cellular transcripts (e.g. c-jun, multiple histones), both novel and previously linked to CD21-initiated signaling, as well as distinct results from pathway analyses support a separate role for the cytoplasmic domain in initiation of intracellular signals.


    Membrane proteins that mediate virus attachment tether virus particles to the cell surface initiating infection. In addition, upon virus interaction such proteins may transmit signals to the interior of the cell that support subsequent steps in the infection process.

    Herein, we show that expression of the Epstein-Barr virus B-cell attachment receptor, CD21, in B-cells that lack this receptor result in significant changes in gene expression, both before and rapidly following EBV-CD21 interaction. These changes translate into major signaling pathway alterations that are predicted to support stable infection.
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  14. Bob


    England (south coast)
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  15. heapsreal

    heapsreal iherb 10% discount code OPA989,

    australia (brisbane)
    Interesting, partially explains why ebv keeps popping up in the cfs/me arguement. I guess they cant say its not an issue in cfs/me but also cant specifically say it is an issue. Explains the t cell dysfunctions found in us as well as the nk dysfunctions.

    It might explain why some(cfsers) say they have never had ebv, their immune system cant produce antibodies to it, even though common information on ebv is that most adults have had ebv. When i hear someone (adult)with cfs/me hasnt had ebv, my thoughts are that maybe they have but their immune system isnt showing its so??
    rosie26 and Valentijn like this.
  16. anciendaze

    anciendaze Senior Member

    heapsreal, this is a very general problem in medicine, not confined to our disease. Measures of pathogen load are largely based on immune response to that pathogen, as measured in peripheral blood. This fails to cover cases where immune response is impaired, or where chronic disease has persisted long enough for immune tolerance to develop. Limitation to peripheral blood largely ignores processes going on primarily in tissues.

    One way around this is a provocative test in which an antigen is introduced, and failure to produce corresponding antibodies in the bloodstream then signals impaired response. There was such a test in the paper discussed. I can think of cases in which such a test might have saved lives of people suffering from known infectious diseases which crippled immune response. What happened instead was that lack of response was taken to imply lack of infection. This can cause doctors to miss big problems like TB or syphilis. Byron Hyde even found one such patient referred to him for CFS. In an unrelated case a patient treated for "depression" died of miliary TB.

    From one comment in that blog above, I would assume some of the people Cort talked to haven't given up the idea that replication of EBV genes and episomes must involve lytic replication of virions, which destroys host cells. This is another long-standing assumption which has not been thoroughly tested. The paper by the group in Berlin strongly suggests it is wrong.

    Historical evidence of how long it takes medical researchers to discard a convenient assumption which is wrong are not encouraging.
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  17. Annesse

    Annesse Senior Member

    Following is some information on this I will be putting up on my Facebook page tonight

    In a new study from Germany (link to study below) researchers found that patients with chronic fatigue syndrome had increased levels of EBV and a deficient immune response to the virus.
    Research has also shown that patients with lupus, multiple sclerosis, scleroderma, rheumatoid arthritis, Sjogren's syndrome, and other autoimmune diseases have increased levels of EBV and a deficient immune response to the virus (link to study below).
    So, if these diseases originate with missing enzymes that digest proteins (protease and DNase 1), how would a lack of these enzymes explain these study findings?
    EBV is a member of the herpesvirus family and according to the National Institutes of Health nearly 95 percent of all people between the ages of 35 and 40 have been infected by the virus.
    The EBV is responsible for the viral infection known as mononucleosis (or “mono”). Although the symptoms of mono, which include a fever, sore throat, and swollen lymph glands eventually go away, EBV takes up permanent residence in "B" cells in the immune system where it can lie dormant for years. Therefore, "persistent B cell activation" would drive the reactivation of EBV. For instance, in the following study the researchers concluded that B cell activation during flares drives frequent EBV reactivation in lupus.
    Exhausted cytotoxic control of Epstein-Barr virus in human lupus.
    Larsen M, Sauce D, et al. 2011. PLoS Pathog. 7(10):e1002328. doi: 10.1371/journal.ppat.1002328. Epub 2011 Oct 20.
    “Systemic Lupus Erythematosus (SLE) pathology has long been associated with an increased Epstein-Barr Virus (EBV) seropositivity…EBV reactivation appears to be an aggravating consequence rather than a cause of SLE immunopathology. We therefore propose that autoimmune B cell activation during flares drives frequent EBV reactivation, which contributes in a vicious circle to the perpetuation of immune activation in SLE patients.”
    Persistent B cell activation and the subsequent reactivation of EBV could also eventually deplete the T cells that attack virus infected cells--CD8+T cells. In the following study published in the Journal of Virology the researchers stated that chronic viral infections often result in ineffective CD8+T cell responses due to functional exhaustion or physical deletion.
    Viral persistence alters CD8 T-cell immunodominance and tissue distribution and results in distinct stages of functional impairment.
    Wherry EJ, Blattman JN, Murali-Krishna K, van der Most R, Ahmed R. 2003. J Virol. 77(8):4911-27.
    “Chronic viral infections often result in ineffective CD8 T-cell responses due to functional exhaustion or physical deletion of virus-specific T cells…Persistent infection led to a disruption of the normal immunodominance hierarchy of CD8 T-cell responses seen following acute infection and dramatically altered the tissue distribution of LCMV-specific CD8 T cells in lymphoid and nonlymphoid tissues…antigen appeared to be the driving force for this loss of function, since a strong correlation existed between the viral load and the level of exhaustion. Further, epitopes presented at higher levels in vivo resulted in physical deletion, while those presented at lower levels induced functional exhaustion. A model is proposed in which antigen levels drive the hierarchical loss of different CD8 T-cell effector functions during chronic infection, leading to distinct stages of functional impairment and eventually to physical deletion of virus-specific T cells. These results have implications for the study of human chronic infections, where similar T-cell deletion and functional dysregulation has been observed.”

    Take home message:
    The EBV takes up permanent residence in "B" cells. Persistent B cell activation would drive the reactivation of the EBV. Chronic EBV reactivation would result in the functional exhaustion or physical deletion of EBV virus-specific immune cells.
    Persistent B cell activation would explain why patients with chronic fatigue syndrome and autoimmune disease have increased levels of EBV and a deficient immune response to the virus.
    So, the question we need answered is: "What would cause persistent B cell activation in patients with chronic fatigue syndrome and autoimmune disease and how would this be directly related to the missing enzymes protease and DNase 1?"
    We will answer this question in our next post.
    Last edited by a moderator: Apr 28, 2015
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  18. Ecoclimber

    Ecoclimber Senior Member

    PLoS One. 2014 Mar 10;9(3):e90855. doi: 10.1371/journal.pone.0090855. eCollection 2014.
    Direct Infection of Dendritic Cells during Chronic Viral Infection Suppresses Antiviral T Cell Proliferation and Induces IL-10 Expression in CD4 T Cells.
    Baca Jones C, Filippi C, Sachithanantham S, Rodriguez-Calvo T, Ehrhardt K, von Herrath M.
    Author information

    Elevated levels of systemic IL-10 have been associated with several chronic viral infections, including HCV, EBV, HCMV and LCMV. In the chronic LCMV infection model, both elevated IL-10 and enhanced infection of dendritic cells (DCs) are important for viral persistence.

    This report highlights the relationship between enhanced viral tropism for DCs and the induction of IL-10 in CD4 T cells, which we identify as the most frequent IL-10-expressing cell type in chronic LCMV infection.

    Here we report that infected CD8αneg DCs express elevated IL-10, induce IL-10 expression in LCMV specific CD4 T cells, and suppress LCMV-specific T cell proliferation. DCs exposed in vivo to persistent LCMV retain the capacity to stimulate CD4 T cell proliferation but induce IL-10 production by both polyclonal and LCMV-specific CD4 T cells.

    Our study delineates the unique effects of direct infection versus viral exposure on DCs. Collectively these data point to enhanced infection of DCs as a key trigger of the IL-10 induction cascade resulting in maintenance of elevated IL-10 expression in CD4 T cells and inhibition of LCMV-specific CD4 and CD8 T cell proliferation.

    The host-pathogen relationship in chronic viral infections requires the establishment of equilibrium between the host immune response and viral replication. While this balance of competing interests aids in protecting the host from the immunopathologic consequences of continuous inflammation, such a truce can also result in the prolonged persistence of the virus within its host. Studies over the last decade have identified several characteristics common to multiple persistent viral infections including elevated levels of systemic IL-10 and T cell exhaustion [1][8].......

    Click on link above for full free abstract:


    PLoS One. 2013; 8(4): e62850.
    Published online Apr 24, 2013. doi: 10.1371/journal.pone.0062850
    PMCID: PMC3634751
    Epstein Barr Virus and Helicobacter pylori Co-Infection Are Positively Associated with Severe Gastritis in Pediatric Patients
    María G. Cárdenas-Mondragón, Ricardo Carreón-Talavera, Margarita Camorlinga-Ponce, Alejandro Gomez-Delgado, Javier Torres, and Ezequiel M. Fuentes-Pananá*
    Yoshio Yamaoka, Editor


    H. pylori infection is acquired during childhood and causes a chronic inflammatory response in the gastric mucosa, which is considered the main risk factor to acquire gastric cancer (GC) later in life.
    More recently, infection by Epstein-Barr virus (EBV) have also been associated with GC. The role of EBV in early inflammatory responses and its relationship with H. pylori infection remains poorly studied.
    Here, we assessed whether EBV infection in children correlated with the stage of gastritis and whether co-infection with H. pylori affected the severity of inflammation.

    Methodology/Principal Findings
    333 pediatric patients with chronic abdominal pain were studied. From them, gastric biopsies were taken and inflammation graded according to the Sydney system; peripheral blood was drawn and antibodies against EBV (IgG and IgM anti-VCA) and H. pylori (IgG anti-whole bacteria and anti-CagA) were measured in sera. We found that children infected only by EBV presented mild mononuclear (MN) and none polymorphonuclear (PMN) cell infiltration, while those infected by H. pylori presented moderate MN and mild PMN.

    In contrast, patients co-infected with both pathogens were significantly associated with severe gastritis. Importantly, co-infection of H. pylori CagA+/EBV+ had a stronger association with severe MN (PR 3.0) and PMN (PR 7.2) cells than cases with single H. pylori CagA+ infection.


    Co-infection with EBV and H. pylori in pediatric patients is associated with severe gastritis. Even single infections with H. pylori CagA+ strains are associated with mild to moderate infiltration arguing for a cooperative effect of H. pylori and EBV in the gastric mucosa and revealing a critical role for EBV previously un-appreciated.

    This study points out the need to study both pathogens to understand the mechanism behind severe damage of the gastric mucosa, which could identified children with increased risk to present more serious lesions later in life.

    Persistent infections often lead to chronic inflammation, a well documented cancer risk factor. Gastric cancer (GC) generally starts with an inflammatory process mainly associated with infection by Helicobacter pylori (H. pylori) [1].

    GC is the fourth most common type of cancer and the second cause of death by cancer world-wide, affecting particularly Asian and Latin American countries [2]. More recently, GC has also been associated with Epstein-Barr virus (EBV) but the role of the viral infection in early inflammatory gastric responses remains poorly studied.

    H. pylori infects over 50% of the world population, with a higher prevalence in developing countries. Infection is usually acquired early in life; in Mexico, about 50% of children are infected by the age of 10 [3].

    Inflammation after infection in children is usually associated with a low level of polymorphonuclear (PMN) and mononuclear (MN) cells infiltrating the gastric mucosa [4]. It has been suggested than the earlier the infection, the greater the risk to present GC later in life, conceivable because of a long lasting (decades) chronic inflammatory reaction to the infection [5].

    Only a fraction of H. pylori infected individuals develop gastroduodenal disease: <15% peptic ulcer, <3% GC and <1% MALT lymphoma [6]. The outcome of H. pylori infection depends also on environmental, host and bacterial factors.

    Among the most important bacterial virulence factors is the pathogenicity island (CagPAI), which encodes a type IV secretion system (T4SS) that translocates the effector protein CagA into epithelial cells [7]. CagA activates multiple signaling pathways triggering cellular phenotypes associated with oncogenic transformation [7]. Moreover, transgenic mice expressing CagA develop adenocarcinomas of the digestive tract. Based on these data, CagA has been recognized as the first known bacterial oncoprotein [8], [9].

    EBV infection has been consistently associated with several types of lymphoma, nasopharyngeal carcinoma (NPC) [10], [11] and more recently to GC [12], [13], [14].

    EBV infection also occurs early in childhood and usually persists in B cells, with most infected individuals carrying the virus asymptomatically in a latent stage in these cells.

    It is not clear when EBV infects the gastric mucosa and whether infection induces an inflammatory reaction, as observed with H. pylori.

    EBV reactivation from infected B cells has been proposed to facilitate infection of the epithelial basolateral face [15]. In that scenario, the titer of anti-EBV antibodies against structural proteins has been proposed to correlate with the level of viral reactivation and as a prognostic marker in NPC [10], [16], [17].

    To our knowledge, no studies have previously addressed whether EBV infection in children is associated with inflammation in the gastric mucosa or whether there exists a cooperative effect between EBV and H. pylori correlating with the severity of the inflammatory reaction.

    In this study, we analyzed antibodies against EBV and H. pylori in sera of pediatric patients with chronic abdominal pain.

    Our results strongly suggest that single infection by either EBV or H. pylori is associated with a mild to moderate inflammatory response in the gastric mucosa; however, co-infection with both pathogens is significantly associated with severe gastritis.

    Even infection with H. pylori cagA+ strains is not associated with severe inflammatory responses in the absence of EBV. These data argue for a previously unknown critical role of EBV infection in the induction of an inflammatory response in the gastric mucosa of children.

    Click on link for above full abtract
    Last edited: Mar 21, 2014
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  19. anciendaze

    anciendaze Senior Member

    Notice that this kind of pathology violates Koch's postulates. Neither EBV nor h. pylori alone is responsible.
  20. Ecoclimber

    Ecoclimber Senior Member

    Because of the incredible and rapid technological advances over the last few decades (DNA sequencing..etc.), our understanding and knowledge of biological and human systems and their processes do not stand up necessarily to postulates formed in later part of the 19th Century. But, then that is my thought process. Koch's postulates have been controversially applied to conclude that "HIV does not cause AIDS (in support of HIV/AIDS denialism) and that oncoviruses do not cause cancers"(Wiki). I also have a problem with Occam's Razor applying to or explaining all phenomenon...the key operative, all in the fields of virology and molecular biology. I believe Bradford Hill criteria has replaced to some degree Koch's postulates. Suffice to say, as Shakespeare once stated in Hamlet, "There are more things in heaven and earth, Horatio, than are dreamt of in your philosophy."
    Last edited: Mar 22, 2014

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