Are Infections Just a Trigger of ME/CFS, or an Ongoing Cause of ME/CFS?

halcyon

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Did Dr Chia say why human DNA interferes with the PCR testing of enterovirus? I know nothing about the technicalities of PCR testing, but I am wondering if this interference is a common occurrence, or whether is it something specific to enteroviruses, or perhaps specific to intracellular infections (enterovirus of course can create long-term intracellular infections).
He seems to believe that the viral RNA is attaching itself to the DNA in the cell. He doesn't go into much detail about this but mentions that it's a phenomenon that he was aware of.

Did Dr Chia say which cell types of the brain tested positive for enterovirus, or wasn't this determined? In one of the two previous ME/CFS brain autopsies (the Richardson autopsy), it was found that the glial cells of the brain, plus the fibroblast cells around small vessels of the brain, stained positive for enteroviral VPI protein.
I don't believe this was determined. The samples were ground and kept in a medium. The samples were taken from the pontomedullary junction, medial temporal lobe, lateral frontal cortex, occipital lobe, cerebellum, and midbrain. I had a bit of trouble following this part of the talk, but it appears that they incubated these tissue samples with some of his serum and then did a western blot that showed reactivity in all six samples. For some reason, after applying DNAse, the RT-PCR only came back positive for enterovirus in the pontomedullary junction and frontal cortex.
 

alkt

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How would you interpret Dr John Chia's research which found significantly enteroviral infection in the gut tissues of ME/CFS patients, and when he treated these patients with IV interferon, many got full remission from ME/CFS, a remission lasting as long as 14 months in some cases, along with a simultaneous large reduction in enteroviral levels in their gut tissues.
the gut is always being infected whenever you eat or drink kiss someone put your hands fingers on your lips or inside your mouth. modern food processes do not get rid of whatever bacterials or viruse sthat have invaded the animals pre slaughter.common sense dictates some of these will survive for some time especially in the digestive systems of people on proton pump inhibitors or antacids.combined with run down immunity fighting abilities. and what exactly is their version of remission because if they do not become fully asymptomatic they are not in remission. we unfortunately know from experience this illness can take you to hell and back. even our good days are just a slight improvement and would be considered by most healthy people to be blood awful .
 

Hip

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So here is a summary of the ME/CFS brain autopsies so far, showing the areas of the brain in which enterovirus infections were detected:
Code:
——————————————————————————————————————————————————————————————————————————————————————
ME/CFS BRAIN       ENTEROVIRUS          PARTS OF BRAIN FOUND
AUTOPSY STUDY      TESTING METHOD       INFECTED WITH ENTEROVIRUS
——————————————————————————————————————————————————————————————————————————————————————

John Chia          Western blot         Pontomedullary junction (in the brain stem),
                                        medial temporal lobe, lateral frontal cortex,
2015                                    occipital lobe, cerebellum, midbrain.
                                 

                   RT-PCR               Frontal cortex, pontomedullary junction (in
                                        the brain stem).

——————————————————————————————————————————————————————————————————————————————————————

John Richardson    VP1 protein stain    In the cerebral cortex, a small fraction of
2001                                    the glial cells, and the fibroblasts in the
                                        adventitia of small blood vessels, stained
                                        positive for enterovirus.

——————————————————————————————————————————————————————————————————————————————————————

Mcgarry,Gow,Behan  PCR                  Hypothalamus and brain stem.
1994

——————————————————————————————————————————————————————————————————————————————————————
Mcgarry, Gow and Behan 1994 also tested the brains of 8 controls, and these were found to be enterovirus negative.

The full text of the John Richardson 2001 and Mcgarry, Gow and Behan 1994 studies are attached below as pdf documents.


Dr John Chia's 2015 ME/CFS brain autopsy is detailed here:
Chronic Enterovirus Infection in a Patient With Myalgic Encephalomyelitis / Chronic Fatigue Syndrome (ME/CFS) – Clinical, Virologic and Pathological Analysis

John Chia, David Wang, Andrew Chia, Rabiha El-Habbal, 2015

Presented at the 19th International Picornavirus Meeting, 2016

OBJECTIVES: A 23 y/o Caucasian male developed prolonged, recurrent gastrointestinal symptoms, followed by onset of severe ME/CFS (CDC criteria, ICC). At initial evaluation, Echovirus 11 antibody titer was ≥1:640 (normal <1:10); IgG and IgM antibody for EBV and HHV6 were negative, CMV IgG was positive. He failed to respond to a combination of α and γ interferon; and debilitating symptoms of the stomach and central nervous system were minimally alleviated by SSRI, benzodiazepine and acid-suppressants. Repeated MRI scans of brain and spinal cords showed normal results. The patient committed suicide 6 years after the onset of symptoms. Brain was harvested and frozen within 24 hours of death for evaluation of chronic viral infection.

METHOD: Using 5D8/1 and J2 monoclonal antibody, stomach and colon biopsies obtained 5 months after onset of illness were stained for viral capsid protein (VP1) and dsRNA, respectively, by immunoperoxidase technique. Blood drawn in Paxgene tube 3 years after illness was screened for enterovirus RNA by RT-PCR. 1 cm3 sample was taken from the ponto-medullary junction (PM), medial temporal lobe (MT), frontal lobe (FL), occipital lobe (OL), cerebellum (CL) and midbrain/hypothalamus area (MB) of brain. The brain samples were homogenized in 10 ml of serum-free medium. Aliquots were processed for viral cultures. Trizol-LS reagent was used for RNA and protein extraction, as well as other lysis agents. Protein samples were separated with Tris-Glycine and MES gels, wet and semi-dry transfer, then western blot was performed with Ibind (Life technology) using EV-, CMV- and HHV6-speci c mAbs, patient’s own serum and control serum samples. Viral culture was performed in WI-38 and BGMK-DAF cell lines. RT-PCR for conserved highly-conserved sequences of 5’ end and 3D polymerase sequence were performed on extracted RNA.

RESULTS: Stomach and colon biopsies stained positive for EV VP1 soon after initial infection documenting the initial viral infection; dsRNA was detected in the stomach biopsies. EV RNA was not detected in blood 3 years after illness. Initial culture of brain samples did not grow virus; 5’ EV RNA sequence was not detected by RT-PCR. Using 5 D8/1 mAb, western blot revealed 37K protein band in the brain samples, which corresponded to viral protein extracted from infected stomach biopsies and enterovirus culture, but not in brain biopsy samples taking from patients with brain tuberculoma and lymphoma. 3D pol gene was ampli ed from the DNase-treated RNA extracted from PM, MT and FL.

CONCLUSION: The analysis of the second brain specimen taken from a ME/CFS patient replicated the British findings published in 1994 (Ann. IM). The finding of viral protein and RNA in the brain specimens 6 years after documented acute enterovirus infection of the gastrointestinal tract is consistent with a chronic, persistent infection of the brain causing debilitating symptoms. EV is clearly one of the causes of ME/CFS, and antiviral therapy should be developed for chronic EV infection.

Source: 19th International Picornavirus Meeting, 2016

For a comprehensive list of studies that found enterovirus in ME/CFS, see this post.

An MEpedia article on ME/CFS postmortem brain autopsies.
 

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alkt

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I do think you're right that antibiotics alter the microbiome and that that may be an issue modern humans deal with regularly now. It's a reason antibiotic overuse is a bad idea. I don't think it's an ME trigger in particular, but who knows ... and there is such variability between people.

I agree that the root is genetic. It would be interesting to know if in fact the incidence really is increasing or it's just recognized more. It goes back to at least the 30's (the LA hospital outbreak), but probably earlier (neurasthenia). I don't know about how common it was though.
in england they date c f s back to the time of florence nightingale
 

barbc56

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I haven't had a chance to read the studies @Hip posted, but definitely plan to. But that's not a guarantee, I'll understand them.:rolleyes:

I do have some related questions. Since we've seen problems such as contamination with entrovirus studies, have there been any studies more recent that would take these into account and confirm these findings?.

I'm also wondering why they weren't followed up other than the usual factors hammpering me/cfs research.

Thanks for posting.

Barb
 

halcyon

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have there been any studies more recent that would take these into account and confirm these findings?
Autopsy findings or enterovirus research in general?

I'm also wondering why they weren't followed up other than the usual factors hammpering me/cfs research.
This is anyone's guess. It's not been replicated but it's not been definitely ruled out. For the last few years it's had to live in the shadow of the next big theory, first XMRV and now autoimmunity/rituximab. In theory Lipkin's microbiome project has the potential to look for these viruses though I'm not 100% sure they are going to. It sounds like Stanford has made some type of pathogen discovery and they will be bringing in a gastroenterologist to take GI tissue samples though I have no idea if they're looking for enteroviruses or something else. Somebody other than Dr. Chia needs to work in this area using his methodology or it's never going to get anywhere.
 

Hip

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Since we've seen problems such as contamination with entrovirus studies, have there been any studies more recent that would take these into account and confirm these findings?.

I'm also wondering why they weren't followed up other than the usual factors hammpering me/cfs research.
I have not heard of contamination problems in the context of enterovirus studies; usually the main issue with enterovirus is detection sensitivity. Chronic enterovirus infections can lead to significant intracellular infections, yet with very few viral particles in the blood, making these chronic infections more difficult to detect than your average virus.

There have been a significant number of enterovirus studies over the decades (a list of some of them is given in this post), many with strong positive results, and it is something of a mystery as to why, with this evidence base, there has been so little further research in this area. Dr Chia's studies, which were published 10 years ago now, and have not yet been replicated, and I know of no other ME/CFS enterovirus studies that have been conducted since then.

I don't really understand why this strong evidence has not be acted upon.


As for ME/CFS brain autopsies, there definitely needs to be more of these.
 

barbc56

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I have not heard of contamination problems in the context of enterovirus studies; usually the main issue with enterovirus is detection sensitivity. Chronic enterovirus infections can lead to significant intracellular infections, yet with very few viral particles in the blood, making these chronic infections more difficult to detect than your average virus
Thanks. I must have misread entrovirus as retrovirus?

Confusion reigns at the moment.

Barb
 

acer2000

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It sounds like Stanford has made some type of pathogen discovery and they will be bringing in a gastroenterologist to take GI tissue samples though I have no idea if they're looking for enteroviruses or something else.
Do you have any more info on this?
 

halcyon

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Do you have any more info on this?
Only what was in their spring newsletter:

High Throughput Sequencing/Pathogen Discovery: Through our continued partnership with Holden Maecker PhD at Stanford and W. Ian Lipkin MD and Mady Hornig MA, MD at Columbia University, our effort of looking for pathogens present or abundant in ME/CFS patients has yielded exciting results. We are in the process of preparing a manuscript for submission to a peer-reviewed journal.

Universal Pathogen Discovery: Building upon preliminary data we collected from our GEISD-Pathogen Discovery study, we are planning to, in addition to blood, sample cerebrospinal fluid, lymph nodes, bone marrow, and NK cell compartments, in a partnership with bioengineer Stephen Quake DPhil. In collaboration with Linda Nguyen MD, gastroenterologist and motility expert at Stanford, we will also collect gastrointestinal biopsies. This study will be the first comprehensive effort to search for pathogens in sites never attempted before.
 

MeSci

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Building upon preliminary data we collected from our GEISD-Pathogen Discovery study, we are planning to, in addition to blood, sample cerebrospinal fluid, lymph nodes, bone marrow, and NK cell compartments, in a partnership with bioengineer Stephen Quake DPhil. In collaboration with Linda Nguyen MD, gastroenterologist and motility expert at Stanford, we will also collect gastrointestinal biopsies. This study will be the first comprehensive effort to search for pathogens in sites never attempted before.
I wonder where they will get control samples, as the findings won't be much use without controls. Who would volunteer to provide CSF, bone marrow or gastrointestinal biopsies? @Jonathan Edwards has previously said that students may be keen to do this - rather them than me!
 

msf

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Yes, it seems like the 2nd study is going to go ahead now that they have secured funding. I wonder what the infection-sceptics make of this? It must seem like rather a waste of money to them...
 

Izola

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the gut is always being infected whenever you eat or drink kiss someone put your hands fingers on your lips or inside your mouth. modern food processes do not get rid of whatever bacterials or viruse sthat have invaded the animals pre slaughter.common sense dictates some of these will survive for some time especially in the digestive systems of people on proton pump inhibitors or antacids.combined with run down immunity fighting abilities. and what exactly is their version of remission because if they do not become fully asymptomatic they are not in remission. we unfortunately know from experience this illness can take you to hell and back. even our good days are just a slight improvement and would be considered by most healthy people to be blood awful .
Alkt: Thanks for all your good posts. You make things more understandable to my really broken brain. :confused::aghhh::)

You said, in part, "This illness can take you to hell and back." :devil: When do we get to the "and back." part?"

You did make the curtain open for a moment, though! Thanks :):D iz
 

Izola

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You weren't lost for long--or maybe I was. Am I talking to myself? Broke arm. med techs didn't give a poop. I do. Hurts. Can't see. Have to have that looked at. ha ha
I am really lost and talking to self. Deep pain does that?