Failure to detect Xenotropic murine leukaemia virus-related virus in Chinese patients with chronic fatigue syndrome

Abstract:

BACKGROUND: Recent controversy has surrounded the question of whether xenotropic murine leukaemia virus-related virus (XMRV) contributes to the pathogenesis of chronic fatigue syndrome (CFS). To investigate the question in a Chinese population, 65 CFS patients and 85 blood donor controls were enrolled and multiplex real-time PCR or reverse transcriptase PCR (RT-PCR) was developed to analyze the XMRV infection status of the study participants. The assay was standardized by constructing plasmid DNAs and armored RNAs as XMRV standards and competitive internal controls (CICs), respectively.

RESULTS: The sensitivities of the multiplex real-time PCR and RT-PCR assays were 20 copies/reaction and 10 IU/ml, respectively, with 100% specificity. The within-run precision coefficient of variation (CV) ranged from 1.76% to 2.80% and 1.70% to 2.59%, while the between-run CV ranged from 1.07% to 2.56% and 1.06% to 2.74%. XMRV was not detected in the 65 CFS patients and 65 normal individuals out of 85 controls.

CONCLUSIONS: This study failed to show XMRV in peripheral blood mononuclear cells (PBMCs) and plasma of Chinese patients with CFS. The absence of XMRV nucleic acids does not support an association between XMRV infection and the development of CFS in Chinese.

 

Source: Hong P, Li J, Li Y. Failure to detect Xenotropic murine leukaemia virus-related virus in Chinese patients with chronic fatigue syndrome. Virol J. 2010 Sep 13;7:224. doi: 10.1186/1743-422X-7-224. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2945957/ (Full article)

 

Serological and virological investigation of the role of the herpesviruses EBV, CMV and HHV-6 in post-infective fatigue syndrome

Abstract:

Multiple previous studies have sought evidence for ongoing, active infection with, or reactivation of, Herpesviruses in patients with chronic fatigue syndrome (CFS), with conflicting results. This study aimed to clarify this by studying 20 patients enrolled in a well-characterized model of the onset and evolution of CFS, the prospective cohort of the Dubbo Infection Outcomes Study (DIOS).

The patients selected for examination included five CFS patients with primary Epstein-Barr virus (EBV) infection; five CFS patients with acute viral infection not caused by EBV; and 10 matched controls with prompt resolution of primary EBV infection. Serum samples from three timepoints were assayed using a comprehensive range of serological assays for EBV, HHV-6, and CMV. Viral genomes were assessed using quantitative PCR assays. All patients were seropositive for HHV-6, and 10 were seropositive for CMV at infection baseline (five patients and five controls). Low titer CMV IgM antibodies were found at infection baseline in two of these cases and three control patients. HHV-6 IgG antibody titers were highest at infection baseline but did not differ between the CFS cases and the control patients. There were increases in EBV IgG VCA p18, EBNA-1 IgG, and EA IgG titers over time, but these did not differ between CFS cases and control patients. EBV and HHV6 DNA levels were at control levels in a minority of samples, and CMV was undetectable in all samples. These data do not support the hypothesis of ongoing or reactivated EBV, HHV-6, or CMV infection in the pathogenesis of CFS.

 

Source: Cameron B, Flamand L, Juwana H, Middeldorp J, Naing Z, Rawlinson W, Ablashi D, Lloyd A. Serological and virological investigation of the role of the herpesviruses EBV, CMV and HHV-6 in post-infective fatigue syndrome. J Med Virol. 2010 Oct;82(10):1684-8. doi: 10.1002/jmv.21873. https://www.ncbi.nlm.nih.gov/pubmed/20827765

 

Review part 2: Human herpesvirus-6 in central nervous system diseases

Chronic fatigue syndrome (CFS) is a debilitating chronic illness [Fukuda et al., 1994] that often begins suddenly with a “flu-like” illness. Patients with CFS have great functional impairment [Komaroff et al., 1996]. The cost to the U.S. economy from lost productivity alone (not including medical care costs) is $9 billion annually [Reynolds et al., 2004].

While the pathogenesis of CFS is unknown, there is abundant evidence of an underlying biological process. In comparison to various health and disease control groups, patients with CFS have abnormal findings in the CNS and autonomic nervous system, evidence of chronic activation of various parts of the immune system, and disordered energy metabolism.

CNS abnormalities have been found using MRI [Buchwald et al., 1992; Schwartz et al., 1994a; Lange et al., 2001; de Lange et al., 2005], functional MRI [Tanaka et al., 2006], SPECT [Schwartz et al., 1994b; Schmaling et al., 2003], and positron-emission tomography (PET) [Yamamoto et al., 2004]. Neuroendocrine studies reveal hypofunction of corticotropin releasing hormone (CRH) neurons in the hypothalamus [Demitrack et al., 1991], disruption of both serotonergic and noradrenergic hypothalamic pathways [Demitrack et al., 1992; Cleare et al., 1995], and of growth hormone secretion [Moorkens et al., 2000]. Typically, these abnormalities are in patterns opposite to those seen in major depression. Cognitive testing has revealed abnormalities [Tiersky et al., 1997; Daly et al., 2001; Deluca et al., 2004] that are not explained by concomitant mood disorders [Marcel et al., 1996]. Autonomic nervous system testing has found abnormalities—particularly postural orthostatic tachycardia syndrome, neurally mediated hypotension, and heart rate variability during head-up tilt testing [Bou-Holaigah et al., 1995; Freeman and Komaroff, 1997; Stewart, 2000; Naschitz et al., 2002].

The immunological findings described most commonly in CFS are impaired function of natural killer cells, increased numbers of CD8+ cytotoxic T cells that bear antigenic markers of activation on their cell surface, and increased production of various pro-inflammatory and TH2 cytokines [Komaroff, 2006]. Many of these cytokines can produce symptoms characteristic of CFS: fatigue, fevers, adenopathy, myalgias, arthralgias, sleep disorders, cognitive impairment, and mood disorders.

Many recent studies of patients with CFS have identified disorders of energy metabolism [Myhill et al., 2009], increased allostatic load [Maloney et al., 2009], and increased oxidative and nitrosative stress [Maes and Leunis, 2008].

Cases of CFS can follow in the wake of well-documented infection with several infectious agents, and may be more likely when the symptoms of acute infection were most severe [Hickie et al., 2006]. The first large study on the possible role of HHV-6 in CFS included 259 patients with a “CFS-like” illness (the case definition had not yet been developed) and age- and gender-matched healthy control subjects. Primary culture of lymphocytes showed active replication of HHV-6 in 70% of the patients versus 20% of the control subjects (P < 10 −8) [Buchwald et al., 1992].

Some subsequent studies have employed only serological techniques that do not distinguish active from latent infection. The results have been mixed: a slight preponderance has showed an association between CFS and HHV-6 infection [Ablashi et al., 2000; Reeves et al., 2000; Hickie et al., 2006].

In contrast, other studies have employed assays that can detect active infection: PCR of serum or plasma, IgM early antigen antibodies, and primary cell culture. Most of these studies have shown an association between CFS and active HHV-6 infection [Patnaik et al., 1995; Secchiero et al., 1995; Wagner et al., 1996; Zorzenon et al., 1996; Ablashi et al., 2000; Nicolson et al., 2003], whereas a few have not [Koelle et al., 2002; Reeves et al., 2000]. The number of patients in the studies that have found an association between CFS and active HHV-6 infection (N = 717) is much larger than the number in studies that have failed to find an association (N = 48).

Several observations, summarized above, together suggest that active infection with HHV-6 may cause some cases of CFS. First, active infection with HHV-6 is present in a substantial fraction of patients with CFS. Second, HHV-6 is tropic for the nervous system and immune system cells, and CFS is characterized by neurological and immunological abnormalities. Clinical studies with antiviral drugs that have in vitro activity against HHV-6 could provide strong evidence in favor of, or against, the hypothesis that HHV-6 may trigger and perpetuate some cases of CFS.

You can read the full article here: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4758195/

 

Source: Yao K, Crawford JR, Komaroff AL, Ablashi DV, Jacobson S. Review part 2: Human herpesvirus-6 in central nervous system diseases.J Med Virol. 2010 Oct;82(10):1669-78. doi: 10.1002/jmv.21861. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4758195/ (Full article)

 

Mouse retroviruses and chronic fatigue syndrome: Does X (or P) mark the spot?

Five years ago, a retrovirus resembling a murine leukemia virus (MLV) was found in patients with prostate cancer (1), and last year, a similar gammaretrovirus was identified in patients with chronic fatigue syndrome (CFS) (2). The agent was named xenotropic MLV-related virus (XMRV), because its env gene was nearly identical to that of xenotropic MLV, an infectious endogenous MLV that preferentially infects cells from foreign species, including humans (Fig. 1A) (3). The two reports struck a common chord, because the viral sequences found in prostate cancer and CFS were nearly identical. A second common theme emerged in reports from Europe that XMRV was rarely found, if at all, in prostate-cancer samples or patients with CFS; however, other investigators confirmed the presence of XMRV in prostate-cancer samples from North America (4). Although a recent report from the Centers for Disease Control and Prevention (CDC) did not find a link between XMRV and CFS (5), distinct MLV-related sequences are found in serial samples collected from the mid-1990s to 2010 from patients with CFS reported in the study by Lo et al. (6) in PNAS. However, the reasons for the current geographical restriction and the source of the infection are baffling.

You can read the rest of this comment here: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2936611/

Comment on: Detection of MLV-related virus gene sequences in blood of patients with chronic fatigue syndrome and healthy blood donors. [Proc Natl Acad Sci U S A. 2010]

 

Source: Courgnaud V1, Battini JL, Sitbon M, Mason AL. Mouse retroviruses and chronic fatigue syndrome: Does X (or P) mark the spot? Proc Natl Acad Sci U S A. 2010 Sep 7;107(36):15666-7. doi: 10.1073/pnas.1007944107. Epub 2010 Aug 23. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2936611/ (Full article)

 

XMRV: does this virus hold the key to myalgic encephalomyelitis/CFS?

Abstract:

In October 2009 a team of researchers from the Whittemore Peterson Institute, in association with the National Cancer Institute and the Cleveland Clinic in the USA, made a discovery that could potentially open the door to useful treatments for myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). The researchers, led by Judy Mikovits, discovered a significant correlation between ME/CFS and an infectious retrovirus called xenotropic murine leukaemia virus-related virus (XMRV). XMRV is classified as a gammaretrovirus, belonging to the same broad family as HIV, but more closely related to a group of viruses that cause cancers such as leukaemia. XMRV is the first member of the gammaretrovirus genus of retroviruses to be found in humans; the research to fully understand the connection between ME/CFS and XMRV, as well as what it means to have the virus, is ongoing. The disastrous impact of AIDS on human health has significantly raised the profile of retroviruses as human pathogens, and XMRV has potentially serious health implications not only for patients, but also for those caring for people with ME/CFS.

 

Source: Crowhurst G. XMRV: does this virus hold the key to myalgic encephalomyelitis/CFS? Br J Nurs. 2010 Jul 22-Aug 11;19(14):919-22. https://www.ncbi.nlm.nih.gov/pubmed/20647985

 

Absence of evidence of xenotropic murine leukemia virus-related virus infection in persons with chronic fatigue syndrome and healthy controls in the United States

Abstract:

BACKGROUND: XMRV, a xenotropic murine leukemia virus (MuLV)-related virus, was recently identified by PCR testing in 67% of persons with chronic fatigue syndrome (CFS) and in 3.7% of healthy persons from the United States. To investigate the association of XMRV with CFS we tested blood specimens from 51 persons with CFS and 56 healthy persons from the US for evidence of XMRV infection by using serologic and molecular assays. Blinded PCR and serologic testing were performed at the US Centers for Disease Control and Prevention (CDC) and at two additional laboratories.

RESULTS: Archived blood specimens were tested from persons with CFS defined by the 1994 international research case definition and matched healthy controls from Wichita, Kansas and metropolitan, urban, and rural Georgia populations. Serologic testing at CDC utilized a Western blot (WB) assay that showed excellent sensitivity to MuLV and XMRV polyclonal or monoclonal antibodies, and no reactivity on sera from 121 US blood donors or 26 HTLV-and HIV-infected sera. Plasma from 51 CFS cases and plasma from 53 controls were all WB negative. Additional blinded screening of the 51 cases and 53 controls at the Robert Koch Institute using an ELISA employing recombinant Gag and Env XMRV proteins identified weak seroreactivity in one CFS case and a healthy control, which was not confirmed by immunofluorescence. PCR testing at CDC employed a gag and a pol nested PCR assay with a detection threshold of 10 copies in 1 ug of human DNA. DNA specimens from 50 CFS patients and 56 controls and 41 US blood donors were all PCR-negative. Blinded testing by a second nested gag PCR assay at the Blood Systems Research Institute was also negative for DNA specimens from the 50 CFS cases and 56 controls.

CONCLUSIONS: We did not find any evidence of infection with XMRV in our U.S. study population of CFS patients or healthy controls by using multiple molecular and serologic assays. These data do not support an association of XMRV with CFS.

 

Source: Switzer WM, Jia H, Hohn O, Zheng H, Tang S, Shankar A, Bannert N, Simmons G, Hendry RM, Falkenberg VR, Reeves WC, Heneine W. Absence of evidence of xenotropic murine leukemia virus-related virus infection in persons with chronic fatigue syndrome and healthy controls in the United States. Retrovirology. 2010 Jul 1;7:57. doi: 10.1186/1742-4690-7-57. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2908559/ (Full article)

 

Comment on “Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome

Abstract:

Lombardi et al. (Reports, 23 October 2009, p. 585) reported an association between the human gammaretrovirus XMRV and chronic fatigue syndrome. However, their results may be misleading because of various potential sources of bias and confounding. If real, the association may lack generalizability because of the specific characteristics of the cases studied and could be due to reverse causality.

Comment on: Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. [Science. 2009]

You can read the rest of this comment here: http://science.sciencemag.org/content/328/5980/825.1.full

 

Source: Sudlow C, Macleod M, Al-Shahi Salman R, Stone J. Comment on “Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome”. Science. 2010 May 14;328(5980):825; author reply 825. doi: 10.1126/science.1183545. http://science.sciencemag.org/content/328/5980/825.1.full (Full article)

 

Comment on “Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome”

Abstract:

Lombardi et al. (Reports, 23 October 2009, p. 585) reported a significant association between the human retrovirus XMRV and chronic fatigue syndrome (CFS). However, the cases with CFS and the control subjects in their study are poorly described and unlikely to be representative. Independent replication is a critical first step before accepting the validity of this finding.

Comment on: Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. [Science. 2009]

You can read the full comment here: http://science.sciencemag.org/content/328/5980/825.2.full

 

Source: Lloyd A, White P, Wessely S, Sharpe M, Buchwald D. Comment on “Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome”. Science. 2010 May 14;328(5980):825; author reply 825. doi: 10.1126/science.1183706. http://science.sciencemag.org/content/328/5980/825.2.full (Full article)

 

Comment on “Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome”

Abstract:

Lombardi et al. (Reports, 23 October 2009, p. 585) reported detection of the human gammaretrovirus XMRV in the blood cells of patients with chronic fatigue syndrome (CFS). However, the patient description provided was incomplete. The inclusion of patients from a “CFS outbreak” previously linked with a viral infection, without confirmation in sporadic CFS cases, casts doubt on the role of XMRV in the pathogenesis of CFS.

You can read the full comment herehttp://science.sciencemag.org/content/328/5980/825.3.full

Comment on: Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. [Science. 2009]

 

Source: van der Meer JW, Netea MG, Galama JM, van Kuppeveld FJ. Comment on “Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome”. Science. 2010 May 14;328(5980):825; author reply 825. doi: 10.1126/science.1183906. http://science.sciencemag.org/content/328/5980/825.3.long (Full article)

 

Prevalence of xenotropic murine leukaemia virus-related virus in patients with chronic fatigue syndrome in the Netherlands: retrospective analysis of samples from an established cohort

Abstract:

OBJECTIVE: The presence of the retrovirus xenotropic murine leukaemia virus-related virus (XMRV) has been reported in peripheral blood mononuclear cells of patients with chronic fatigue syndrome. Considering the potentially great medical and social relevance of such a discovery, we investigated whether this finding could be confirmed in an independent European cohort of patients with chronic fatigue syndrome.

DESIGN: Analysis of a well defined cohort of patients and matched neighbourhood controls by polymerase chain reaction.

SETTING: Certified (ISO 15189) laboratory of clinical virology in a university hospital in the Netherlands. Population Between December 1991 and April 1992, peripheral blood mononuclear cells were isolated from 76 patients and 69 matched neighbourhood controls. In this study we tested cells from 32 patients and 43 controls from whom original cryopreserved phials were still available.

MAIN OUTCOME MEASURES: Detection of XMRV in peripheral blood mononuclear cells by real time polymerase chain reaction assay targeting the XMRV integrase gene and/or a nested polymerase chain reaction assay targeting the XMRV gag gene.

RESULTS: We detected no XMRV sequences in any of the patients or controls in either of the assays, in which relevant positive and negative isolation controls and polymerase chain reaction controls were included. Spiking experiments showed that we were able to detect at least 10 copies of XMRV sequences per 10(5) peripheral blood mononuclear cells by real time as well as by nested polymerase chain reaction, demonstrating high sensitivity of both assays.

CONCLUSIONS: This study failed to show the presence of XMRV in peripheral blood mononuclear cells of patients with chronic fatigue syndrome from a Dutch cohort. These data cast doubt on the claim that XMRV is associated with chronic fatigue syndrome in the majority of patients.

Comment in: Chronic fatigue syndrome and human retrovirus XMRV. [BMJ. 2010]

 

Source: van Kuppeveld FJ, de Jong AS, Lanke KH, Verhaegh GW, Melchers WJ, Swanink CM, Bleijenberg G, Netea MG, Galama JM, van der Meer JW. Prevalence of xenotropic murine leukaemia virus-related virus in patients with chronic fatigue syndrome in the Netherlands: retrospective analysis of samples from an established cohort. BMJ. 2010 Feb 25;340:c1018. doi: 10.1136/bmj.c1018. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2829122/ (Full article)