Lipkin finds biomarkers not bugs

September 12, 2013

The CDC PCOCA telephone broadcast on 10 September 2013, featured a lengthy presentation from Dr Ian Lipkin who revealed some stunning initial results from the study that is primarily hunting for pathogens in ME/CFS. Simon McGrath and Russell Fleming (Firestormm) review this exciting and possibly game-changing news…

Lipkin - CFI

Dr Ian Lipkin
Lead Researcher
Chronic Fatigue Initiative (CFI)
Pathogen Discovery and Pathogenesis Study

Read the full Lipkin Transcript: Here.

Dr Ian Lipkin has been a human whirlwind in ME/CFS research since he became involved a few years back, and he’s just surprised us all by announcing the first results from the world’s largest ever biomedical ME/CFS study in a public broadcast!

He started off by lowering expectations: their results thus far do not implicate a single infectious agent and they have not discovered the cause of CFS. But he then wowed the hundreds listening to the conference call by revealing they had found strong evidence for immune overstimulation, both in blood plasma and in cerebro spinal fluid; and that they were now hard at work trying to identify what could be causing these abnormalities.

The big pathogen hunt draws a blank thus far

This study is looking exhaustively for any pathogen: viral, bacterial, fungal or parasitic, to see if a chronic infection could explain ME/CFS. Many of the results are in, but so far there is no clear sign of viruses at least. It is every bit a collaborative effort, with patients provided by long-established physicians including Dr Dan Peterson and Professor Jose Montoya.

The Key Players

Lipkin began by thanking the clinicians who had provided the all-important samples. Effectively it was a roll call of America’s top ME/CFS physicians:

Professors Jose Montoya, Anthony Komaroff and Nancy Klimas; and Drs Dan Peterson, Lucinda Bateman, Sue Levine and Donna Felsenstein.

The study itself was run by Dr Mady Hornig (CFI Principal Investigator) at the Centre for Infection and Immunity at Mailman School of Public Health; where Lipkin is also based.

He went on to acknowledged the very necessary support of the Chronic Fatigue Initiative, the Evans Foundation and an anonymous donor supporting Montoya’s work.

First the researchers looked at blood plasma for 285 CFS patients and 201 controls from Jose Montoya at Stanford (these are serious numbers). They searched for genetic evidence of infection using a panel that was able to detect 20 specific bacteria, viruses or parasites; including Epstein-Barr virus, Human Herpes Virus 6, enteroviruses, Influenza A and Borrelia bacteria.

Next they used high throughput sequencing, a method that was pioneered by Lipkin and colleagues that was used to discover over 500 viruses – so they felt fairly confident that if a virus was present, then they would have found it.

But they effectively drew a blank using both methods – only 1.4% of CFS patients were found to have HHV6 infection, but so did 1% of controls.

Other findings…

Lipkin said they also found Anelloviruses in 75% of those studied in plasma samples, but it was not specific for Chronic Fatigue Syndrome (we don’t currently have separate figures for patients and controls). Anellovirus is believed to cause widespread chronic infection in the human population, but has not yet been associated to any specific disease. “I really don’t know at this point what this finding means” said Lipkin.

They also found retroviruses in 85% of the sample pools. However, Lipkin expressed caution:

“It is very difficult at this point to know whether or not this is clinically significant. And given the previous experience with retroviruses in Chronic Fatigue, I am going to be very clear in telling you – although I am reporting this at present – in Professor Montoya’s samples neither he nor we have concluded that there is a relationship to disease …if I were to place bets and speculate, I would say that this is not going to pan out.”

Window on the brain: Cerebrospinal fluid samples

Both Lipkin and Hornig were clearly excited to have had access to the 60 cerebrospinal fluid samples from Peterson, and kindly donated by his ME/CFS patients. Cerebrospinal fluid bathes our brain and spinal cord so it gives access to what goes on in the brain, a prime site of interest to researchers, but hard to access; so these spinal tap samples are precious.

However, using the same molecular techniques to track down pathogens as they had applied to Jose Montoya’s samples, Lipkin’s team again drew a blank.

At the Invest in ME conference in May, Hornig said they had  tentative findings that there might just be a novel pathogen or pathogen candidate in these samples – but it was too early to say for sure. In reply to a question yesterday, Lipkin said there is no more to report:

we do not yet have a novel pathogen nor do we have a pathogen candidate in which I would have any confidence. So at this point we have no plans to publish anything.

Nonetheless, he also said they hadn’t finished all the DNA sequencing work on these samples, so this is not yet a dead end.

Immune stimulation and biomarkers

Up to this point we’d heard a lot of very impressive negative findings, based on large, well-defined samples of patients and controls, and using powerful research techniques. But what we most wanted to hear about was positive leads – and Ian Lipkin duly obliged:

plasma and blood

Plasma is the clear fluid left when blood is separated

They took plasma from the CFI cohort of 200 very well-defined ME/CFS cases and 200 controls collected by clinicians (See ‘Key Players’ above). Then they searched for cytokines and chemokines, which are regulators and controllers of the immune system.

The big finding was a decrease in levels of Interleukin’s IL-17, IL-2, IL-8 and in TNF-alpha – all of which have a role in pro-inflammatory responses. Lipkin believes these significant differences are worth follow-up in larger cohorts to better understand what they mean.

Professor Jonathan Edwards (who is an adviser to the planned UK Rituximab trial) commented on this forum yesterday, that it was a surprise to see TNF-alpha levels decrease, and suggested it could mean that ‘inflammation’ is too crude a concept for the process.

Lipkin and colleagues also found upregulation in Leptin, a hormone that plays a key role in regulating energy uptake and use, and Serpin, a protein family with multiple roles. More obscure cytokines, and and their cousins chemokines, were also affected but Lipkin didn’t give any further detail.

Two new patient types revealed

Ian Lipkin said they found something very surprising in their data: there appear to be substantial differences in profiles between those people who have had the disease for 3 years or less and those who have had the disease for more than 3 years. And that this is important because, “it could have implications for therapy as well as for diagnosis”.

In the ‘early group’, who have been ill for less than 3 years, there seem to be a number of markers suggestive of some sort of allergy aspect. For example there are often increased numbers of eosinophils in blood, with more differences in cytokines. But while this is tentative, he was more confident in the finding the IL-17 was elevated in these ‘early’ patients, compared with reduced levels in patients ill for over three years.

Lipkin said that he thought this 3-year division could be very important and that it hadn’t been something they were looking for; but would be looked at in any future work they did.

Cerebrospinal fluid: different differences

Lipkin’s team also found differences in cerebrospinal fluid biomarkers between patients and controls. I’m not sure from what he said if there were significant differences between the ‘early’ group and patients who had been ill for more than three years. However, they did find that patients had elevated levels of the TH-2 type cytokines IL-10 and IL-13 and elevated levels of four TH-1 cytokine: IL-1 beta, TNF-alpha, IL-5, and IL-17. Lipkin said this is compatible with a profile of some particular types of response that may provide insights into immunological dysregulation in Chronic Fatigue Syndrome.

So, overall they found clear differences between patients and controls in both plasma and spinal fluid. Many immune differences have been detected before, but findings are not very consistent and none had used such a large and robustly-defined patient cohort (meeting Fukuda and Canadian criteria), or with such carefully matched controls.

HEALTH WARNING

Dr Ian Lipkin again was very clear that he did NOT recommend anyone act on these provisional findings: he doesn’t want anyone to get a spinal tap, or measure the cytokine levels and hope to modulate them with drugs based on this research: it’s too early for that.

The lost years…

Back in 1999 Ian Lipkin published his first paper on CFS, which ruled out a connection between the illness and Borna Disease Virus (one of the many viruses he’s discovered). But crucially they found evidence of polyclonal B-cell reactivity. He commented that back then:

“there was a very strong sentiment in some portions of the scientific and clinical communities – not always and not everywhere – but in some portions of the community, that this was a psychological illness. What I said was that based on our findings we had very strong evidence that people with Chronic Fatigue Syndrome were truly ill with a physical illness and they deserved a “Deep Dive” to find out why they were ill.”

He noted that was a long time ago and dryly added “I am pleased to see that people are now paying much more attention to this disorder and what we can do about it”.

What next?

“I still believe the primary cause is likely to be an infectious agent.”

Having found clear signs of cytokine abnormalities, Lipkin wants to find out what is causing these changes; he currently thinks that an ongoing stimulus is causing immune system activation which might be resulting in the symptoms we associate with the disease. And interestingly, he still believes that  infection lies at the root of it all – despite not yet finding any infections.

The team – working closely with the clinicians he’s already mentioned – plan even more extensive work on a cohort of 200 CFI patients and controls. They will further test for viruses and crucially they will also look for bacteria and fungi by sequencing ribosomal RNA (which should detect unknown bacteria and fungi, as well as known ones).

Previous infection detection

The group has looked for current pathogens in the blood and spinal fluid samples they have – and has so far drawn a blank, though there is more work underway. However, what if the trigger for ME/CFS is an earlier infection that has since been cleared: the ‘hit and run’ scenario Hornig has described? Lipkin says they are looking for “shadows” of previous infections by searching for antibodies against specific infections, rather than the infections themselves (the body will continue to produce low levels of specific antibodies years after an infection has been cleared (the basis of much immunity)).

They will use a technique called LIPS to look for antibodies against particular known infection. In addition, as in their viral sequencing work, they will use an ‘unbiased’ system (using peptide arrays) that should detect all potential viruses – or rather antibodies against them. They don’t currently have an ‘unbiased’ system that works on bacteria or fungi.

Fecal future?

What excites Ian Lipkin most is the gut microbiome, the community of micro-organisms that live in our gut – which might seem odd given his focus so far on pathogens and the immune system. However, it turns out that the microbiome can have a profound influence on the immune system and may be important in many diseases. Hornig and Lipkin have already developed techniques to study the microbiome, and have shown it could be a factor in autism and even in the severity of colon cancer.

It turns out the best way to find out what lives in the microbiome is to measure what comes out of the gut: fecal matter, or ‘poop’ to you and me. Their ME/CFS microbiome project has started, but Lipkin felt there wasn’t enough material in the first attempt at gathering samples, so things have been delayed as they switch to a new technique (the ‘special’ collection cups).

The Big Ask: “we can’t do this without you”

Microbiome research is very expensive (analysis, not sample collection), and they currently only have the funds to do 10% of what’s needed. In fact, throughout the call, Lipkin was stressing the need for more funds, because good research costs and ME/CFS is woefully underfunded: there just isn’t enough money to do the work needed. The current science budget cuts (Sequestration) in the US makes the climate even harder.

“It is probably inappropriate for me to be passing the hat, but that’s precisely what I am doing.”

He urged patients to contact their representatives in Congress, demanding more funding for ME/CFS research – pointing out that in the early days of HIV, there was little funding until patients demanded it.  He also said that if patients were able to afford it, they should invest in research themselves.

Conclusion

ME/CFS research is coming of age. Ian Lipkin has reported these early – and soon-to-be published results – from a huge study, using 285 of Jose Montoya’s patients, the 200 patients from the Chronic Fatigue Initiative, plus the 60 ‘gold-dust’ spinal fluid samples from Dan Peterson’s patients. Using state-of-the-art techniques, they have largely ruled out a role for a specific pathogen – though that work is incomplete. And they have strong evidence of ongoing immune stimulation in ME/CFS patients, with abnormalities found in both the blood and spinal fluid. The next stage involves deeper searching for bacteria and fungi, looking for ‘shadows’ of previous infections and investigating the gut microbiome. These are exciting times!

Thanks to the CDC for organizing the presentation and to Firestormm for providing a transcript of this talk, and above all to Dr Ian Lipkin for taking time out to talk to patients.

If you would like to donate to Ian Lipkin’s research, click here. In the comments box, put “for M.E/C.F.S Study” to make sure it goes to the right place.

 

Simon McGrath tweets on ME/CFS research:

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161 comments

{ 161 comments… read them below or add one }

Legendrew September 12, 2013 at 12:23 pm

Great article guys! Exciting to hear about the signs of immune activation and also interesting to me to hear about the increase in Leptin, i have to admit I overlooked it intially but reading the article it stand out to me as a very interesting upregulation to see. It makes a lot of sense to have upregulation of a molecule such as this given the evidence of mitochondrial and ATP production problems!

I think it's worth mentioning too that these findings given important evidence for both the autoimmune theory that Fluge and Mella are working on and indeed the pathogen theory that Lipkin himself believes. Hopefully some of these abnormalities hold up to further scrutiny!

Sasha September 12, 2013 at 12:57 pm

Well done, Simon – great article! And well done, Russ – great transcript!

Simon September 12, 2013 at 12:59 pm
Legendrew

Great article guys!

…interesting to me to hear about the increase in Leptin, i have to admit I overlooked it intially but reading the article it stand out to me as a very interesting upregulation to see. It makes a lot of sense to have upregulation of a molecule such as this given the evidence of mitochondrial and ATP production problems!

Thanks!

Re Leptin:

the blog

Lipkin and colleagues also found upregulation in Leptin, a hormone that plays a key role in regulating energy uptake and use

Yes, I was intrigued by that too, though I that line is pretty-much the extent of my knowledge of Leptin.

I think it's worth mentioning too that these findings given important evidence for both the autoimmune theory that Fluge and Mella are working on and indeed the pathogen theory that Lipkin himself believes. Hopefully some of these abnormalities hold up to further scrutiny!

Yes, and I think they key thing is replication, which is very much what Ian Lipkin plans to do. I have to say, these broad findings are probably consistent with many different theories so I will be very interested in what the future work turns out: certainly Ian Lipkin's main focus now seems to be to find what is responsible for these changes.

prioris September 12, 2013 at 1:38 pm

Overactive immune system has been known for at least 2 decades.

Antares in NYC September 12, 2013 at 1:53 pm

This is remarkable. I think it's interesting that this study makes ME/CFS somewhat similar to MS, particularly in the "hit and run scenario". A virus-like infection comes in shortly, disrupts the immune system, and it's all downhill from there.

Also, how do we pressure for more research funds? How do we organize and lobby to get these researchers what they need?

Legendrew September 12, 2013 at 2:04 pm
prioris

Overactive immune system has been known for at least 2 decades..

Indeed it has but we now seem to be getting to the bare bones of the issue with what exactly is wrong with strong evidence to support these initial claims – if nothing else these findings are good clues as of where to look next!

Sometimes finding nothing at all is the best outcome. In terms of viruses as a central causation, this somewhat conclusively shows little significance in the commonly held perpetrators which means we can move on to newer ideas without the baggage that these old ideas bring with them.

An analogy I made in a previous thread earlier today seems to also be relevant here I feel with regards to the 'hit and run' idea many people discuss. The reference to autoimmunity is however just a personal opinion I hold and for the purpose of this topic should be interchangeable with immune activation.

Legendrew

As an analogy I believe the genetic predisposition is like having petrol liberally doused over an area – it's invisible (but likely a few small hints that go unnoticed) – it then only takes a spark in the form of a viral or bacterial trigger to start the immune activation and through exploitation of the genetic predisposition and suddenly a whole fire breaks out – this is when the symptoms first appear and suddenly it becomes much more difficult to manage. There is little point in identifying the initial virus or bacterial 'spark' as this does nothing to ease the fire. The only option then comes in calming the aberrant immune response and trying to break the autoimmune cycle.

The interesting part here is the specifics of the immune activation – certainly not all of these will prove to be repeatable but if just a handful do we could be looking at a good trace of why the immune response has turned so aberrant.

Nielk September 12, 2013 at 2:11 pm

Thank you Simon and Firestormm for the hard work putting this clear review together and supplying a transcript for us. I had listened in to the meeting but, apparently missed a lot of the details.

It is clear to me that Lipkin found enough abnormalities/biomarkers for him to be excited to plunge in deeper for further studies. Any study showing differences between patients and controls is promising. It remains to be shown/proven what this will lead to.

Their finding of the three year marker showing differences in the results is intriguing. I wonder why three? As he stated though, they were not looking for that, it just has seemed to appear when comparing results.

I wish I could understand in layman's terms what these abnormalities in cytokines mean or what this Leptin issue means.
I almost wish I had gone through medical school in order to understand and follow these developments.:)

Waverunner September 12, 2013 at 2:33 pm

Great news! Thanks for the article. I truly hope that they advance the understanding of the disease mechanisms. It's fascinating that some inflammatory markers were even low. I also hope that microbiome analysis gets a lot cheaper through better implementation of computers and resulting automation.

Erik Johnson September 12, 2013 at 2:59 pm

In addition to regulating the hunger-response, Leptin controls the release of toxins from the fatty tissues.

Lipkins findings of Leptin is consistent with "the elusive biomarker for CFS" that we have known about for about fifteen years now.

Legendrew September 12, 2013 at 3:03 pm
Erik Johnson

In addition to regulating the hunger-response, Leptin controls the release of toxins from the fatty tissues.

Lipkins findings of Leptin is consistent with "the elusive biomarker for CFS" that we have known about for about fifteen years now.

With regard to Leptin, I think it is a very interesting find and it's more interesting that it has been found in the past. I'm not sure it will prove to be a causative agent in the disease pathology of ME but certainly it would be interesting to know why it appears to be getting up-regulated – I suspect it could be due to its role in metabolism considering the findings of dysfunctional mitochondria and Lactate build-up in the muscles – presumably through increased anaerobic respiration during trivial exertion.

roxie60 September 12, 2013 at 3:17 pm

Help in understanding, so is there an up- regulation in Leptin if you have CFS?

roxie60 September 12, 2013 at 3:27 pm

Checked SNPedia and there are Leptin related genes/snps for those who wish to travel down that rabbit hole.

Erik Johnson September 12, 2013 at 3:27 pm

As you might expect….It's complicated.
Leptin stimulates MSH "Melanocyte Stimulating Hormone"
Which in turn regulates the innate immune system.
High Leptin and low MSH is seen in illnesses where toxins are stored in the fatty tissues.
The high Leptin makes weight loss extremely difficult.

Since CFS researchers have yet to make the connection between this phenomenon and CFS, the connection remains to be explored. But for Lipkin to see it makes it more likely that CFS researchers will take an interest.

roxie60 September 12, 2013 at 3:41 pm

I wonder if the genes associated with Leptin will be determined to play any role. For now I checked three of the SNPs associated with Leptin and I have the common variety so they dont explain my expereince. I am one of the 'fighting this battle longer than three years' group. I still suspect when I had mono in 1995 that was the start of it all even though I did not recognize the long periods between crash and recovery as being related. The last 6-7 years I know realize the unmistakable relation. I am just coming out of another crash (6-7 weeks) and there is a difference, more function (although not sure I'll ever get back to 100%), not as sick all the time, symps not as severe. I wish I understood this crash and recovery element to whatever is causing this :confused:. Do others also experience this crash and recovery I am describing? There are some symps I have all the time but when I crash I am more disabled, other symps added and the ones I usually have seem worse then like a cloud lifting I enter a day or week that I realize I am much better. It is so friggen madding the inconsistency. I blame that most for the reason I lost my career, how can I or my employer count on me when I dont know one day to the next if I can function. :cry: Sorry for the derail I just want answers, to heal and get my life back!!!!

Erik Johnson September 12, 2013 at 3:58 pm

The variability was in my quote from Dr Ramsay.

Yes, I know what controls the crash phenomenon, and it is linked to leptin signaling.

acer2000 September 12, 2013 at 4:45 pm

It seems that he did find bugs. He reported that he found both anellovirus sequences and also retroviral sequences. He said he didn't know what it meant of if it would lead anywhere. Given the craziness that happened last time a researcher tried to associate a viral cause with CFS I don't blame him for being cautious and wanting more evidence before making any claims. But he did state he felt the cause still seemed like an infection and that all of the findings (bugs or not) should be followed up.

Also, I have a hard time reconciling the claims about IL-8. Several other studies have shown IL-8 to be significantly elevated in patients with CFS. My own IL-8 is one of the most consistently elevated markers, and I have participated in several of these studies through my CFS doctor (including this one). I'll admit though, I don't know the intricacies of testing for cytokines. Perhaps they vary based on time of day, or processing method or something.

Antares in NYC September 12, 2013 at 5:23 pm
acer2000

It seems that he did find bugs. He reported that he found both anellovirus sequences and also retroviral sequences. He said he didn't know what it meant of if it would lead anywhere. Given the craziness that happened last time a researcher tried to associate a viral cause with CFS I don't blame him for being cautious and wanting more evidence before making any claims. But he did state he felt the cause still seemed like an infection and that all of the findings (bugs or not) should be followed up.

Also, I have a hard time reconciling the claims about IL-8. Several other studies have shown IL-8 to be significantly elevated in patients with CFS. My own IL-8 is one of the most consistently elevated markers, and I have participated in several of these studies through my CFS doctor (including this one). I'll admit though, I don't know the intricacies of testing for cytokines. Perhaps they vary based on time of day, or processing method or something.

Acer2000:

You bring a very good point. I noticed he mentioned the anellovirus and traces of some retrovirus, but qualified it quickly as needeing more research. After the xmrv debacle, you can't blame any researcher for being utterly cautious.

I think the title is not necessarily misleading, but leaning heavily on Dr Lipkin's cautious approach to the findings.

Forbin September 12, 2013 at 6:17 pm

Back in 1999 Ian Lipkin published his first paper on CFS, which ruled out a connection between the illness and Borna Disease Virus (one of the many viruses he’s discovered). But crucially they found evidence of polyclonal B-cell reactivity. He commented that back then:

“there was a very strong sentiment in some portions of the scientific and clinical communities – not always and not everywhere – but in some portions of the community, that this was a psychological illness. What I said was that based on our findings we had very strong evidence that people with Chronic Fatigue Syndrome were truly ill with a physical illness and they deserved a “Deep Dive” to find out why they were ill.”

He noted that was a long time ago and dryly added “I am pleased to see that people are now paying much more attention to this disorder and what we can do about it”.

Dr. Lipkin is even more emphatic on this earlier in the call…

"My own interest in Chronic Fatigue Syndrome dates back to the mid-to-late 1990’s when I was working with Brigette Evangard, and one of the questions that came in earlier today had to do with polyclonal B cell activation. Those of you who may have read that paper that I wrote with Brigettea, which was ultimately published in 1999, was one of the first to really draw attention to the fact that people with Chronic Fatigue Syndrome who were diagnosed using the criteria at the time did in fact have a physical condition.

"Although we were unable to find evidence of infection with borna disease virus, which was the primary question at that point, we did demonstrate unequivocally that the vast majority of people who met these criteria had polyclonal B cell activation, so they clearly had some sort of immune activation. But the question was, why?"

[my bolding]

bel canto September 12, 2013 at 7:46 pm

Montoya's recently released results from a small (10 participants, I believe) study: They obtained blood draws for 25 consecutive days from these patients, who also, I believe, kept journals indicated their fatigue levels. The only finding that correlated directly with fatigue levels in most (not all) of these patients was the hormone leptin. I think that the overall levels were not abnormal, but the changes in leptin tracked the fatigue level changes.
I imagine the details of this are discussed somewhere in the forum, but I haven't had a chance to track it down yet.

Nielk September 12, 2013 at 7:59 pm
bel canto September 12, 2013 at 8:09 pm

Yes, thanks Neilk!

searcher September 12, 2013 at 9:25 pm

As bel canto said, what was interesting about the Montoya et. al study was that leptin wasn't particularly high– instead leptin levels correlated with fatigue in patients but not in controls. Leptin levels themselves do not appear to be a useful biomarker– they will be higher in overweight people even without fatigue.
From Montoya's paper:
"The relationship we observed between leptin and fatigue existed even though leptin levels were not abnormally elevated, and there was no statistical difference in leptin values between the CFS and control groups. "
So, although I think Shoemaker's work is interesting and relevant to CFS patients, I don't think he has made the same conclusion about leptin that Montoya and his team has.

The Montoya paper on leptin can be found at http://www.translational-medicine.com/content/pdf/1479-5876-11-93.pdf

HowToEscape? September 12, 2013 at 11:47 pm
Antares in NYC

This is remarkable. I think it's interesting that this study makes ME/CFS somewhat similar to MS, particularly in the "hit and run scenario". A virus-like infection comes in shortly, disrupts the immune system, and it's all downhill from there.

Also, how do we pressure for more research funds? How do we organize and lobby to get these researchers what they need?

By organizing.

There is a template for that, the AIDS folks were very well organized and targeted specific actions within a plan. If we just wait for "them" to "do something", we'll wait till we're old and eventually dead.

GcMAF Australia September 13, 2013 at 12:46 am

did he look at Lyme??

Simon September 13, 2013 at 2:43 am
Antares in NYC

This is remarkable. I think it's interesting that this study makes ME/CFS somewhat similar to MS, particularly in the "hit and run scenario". A virus-like infection comes in shortly, disrupts the immune system, and it's all downhill from there.

Also, how do we pressure for more research funds? How do we organize and lobby to get these researchers what they need?

I think it's worth stressing that the 'hit and run' scenario discussed by Mady Hornig is, at this point, only suggeseted as a possiblity for ME/CFS – and they don't yet have any resultf from their work looking for 'shadows' of previous infections.

I'm not in the US so don't have any idea of how to press for funds – but it might be worth posting something in the advocacy forum (I couldn't see anything there yet). There is probably discussion of it on the CDC phonecall thread too, but it's rather lengthy.

Nielk

Their finding of the three year marker showing differences in the results is intriguing. I wonder why three? As he stated though, they were not looking for that, it just has seemed to appear when comparing results.

That seems a pretty weird finding to me too: it almost implies some sort of shift in the illness after 3 years, which is hard to fathom. Hopefully things will be clearer when the new paper is published, which Ian Lipkin said would be 'very soon'.

acer2000

It seems that he did find bugs. He reported that he found both anellovirus sequences and also retroviral sequences. He said he didn't know what it meant of if it would lead anywhere. Given the craziness that happened last time a researcher tried to associate a viral cause with CFS I don't blame him for being cautious and wanting more evidence before making any claims. But he did state he felt the cause still seemed like an infection and that all of the findings (bugs or not) should be followed up.

Also, I have a hard time reconciling the claims about IL-8. Several other studies have shown IL-8 to be significantly elevated in patients with CFS. My own IL-8 is one of the most consistently elevated markers, and I have participated in several of these studies through my CFS doctor (including this one). I'll admit though, I don't know the intricacies of testing for cytokines. Perhaps they vary based on time of day, or processing method or something.

He did find some bugs, but said the Anellovirus was not specific to CFS (ie CFS were not greatly different from controls). It sounds like the retroviral thing was the same in that they found it in 'pooled samples'. Hopefully things will be clearer in the full paper.

Do you have any refs/links re the elevated IL8 findings elsewhere? Would be good to compare.

Antares in NYC

I think the title [Lipkin finds Biomarkers not Bugs is not necessarily misleading, but leaning heavily on Dr Lipkin's cautious approach to the findings.

I have to admit I took a slight liberty with the title, favouring snappiness over total accuracy – though I think it reflects the gist fo the talk.

Simon September 13, 2013 at 2:52 am
Nielk

I wish I could understand in layman's terms what these abnormalities in cytokines mean or what this Leptin issue means.

Wish I could too! Broadly, I think they are signs of immune activation. Intriguingly, when it comes to the cytokine findings in spinal fluid

Ian Lipkin

this is compatible with a profile of some particular types of response that may provide insights into immunological dysregulation in Chronic Fatigue Syndrome.

He didn't expand on this and I wish knew more, but clearly Ian Lipkin thinks this could be an important clue.

justy September 13, 2013 at 2:53 am

The three year marker difference is fascinating and when it is unraveled may explain why it is often said to those newly ill that the best chances of a full recovery are in the first 2-3 years. It is well known that after this (approximate) time period the illness can become chronic/relapsing/remitting.

Thanks Simon and Firestormm for the article and the transcript – very much appreciated :thumbsup:

Simon September 13, 2013 at 3:09 am

Leptin

Not surprsingly this has caused a lot of comment

bel canto

Montoya's recently released results from a small (10 participants, I believe) study: They obtained blood draws for 25 consecutive days from these patients, who also, I believe, kept journals indicated their fatigue levels. The only finding that correlated directly with fatigue levels in most (not all) of these patients was the hormone leptin. I think that the overall levels were not abnormal, but the changes in leptin tracked the fatigue level changes.
I imagine the details of this are discussed somewhere in the forum, but I haven't had a chance to track it down yet.

Leptin appears to have a role in signalling to the brain, is directly affected by the immune system and ties in with the HPA axis too (post by jeffrez). So it ticks all the boxes (but then so do many things).

The full text of Montoya's new paper (from earlier this year) is here: Daily cytokine fluctuations, driven by leptin, are associated with fatigue severity in chronic fatigue syndrome: evidence of inflammatory pathology

ABSTRACT

Background
…Studies have demonstrated elevated levels of inflammatory factors in patients with CFS, but findings are contradictory across studies and no biomarkers have been consistently supported. Single time-point approaches potentially overlook important features of CFS, such as fluctuations in fatigue severity. We have observed that individuals with CFS demonstrate significant day-to-day variability in their fatigue severity.

Methods
…we implemented a novel longitudinal study design to investigate the role of cytokines in CFS pathophysiology. Ten women meeting the Fukuda diagnostic criteria for CFS and ten healthy age- and body mass index (BMI)-matched women underwent 25 consecutive days of blood draws and self-reporting of symptom severity. A 51-plex cytokine panel via Luminex was performed for each of the 500 serum samples collected. Our primary hypothesis was that daily fatigue severity would be significantly correlated with the inflammatory adipokine leptin, in the women with CFS and not in the healthy control women. As a post-hoc analysis, a machine learning algorithm using all 51 cytokines was implemented to determine whether immune factors could distinguish high from low fatigue days.

Results
Self-reported fatigue severity was significantly correlated with leptin levels in six of the participants with CFS and one healthy control, supporting our primary hypothesis. The machine learning algorithm distinguished high from low fatigue days in the CFS group with 78.3% accuracy.

Conclusions
Our results support the role of cytokines in the pathophysiology of CFS.

The obvious problem with this is that it's a tiny sample. In any event 78% accuracy in separating high from low fatigue days in CFS isn't too great – and is tweaked for this specific sample so would probably be lower in an independent sample. And 6/10 isn't stunning either (though could be subgroups).

However, it is a really interesting approach and could be consistent with the Lipkin results. As ever, replication is needed.

PR thread discussing this paper

Firestormm September 13, 2013 at 4:38 am

Replication of Lipkin's preliminary work?

It is unclear at this point, the extent to which this might (at least in part), represent some means of replication for the Lipkin Study we reported on above; but the NIH grant to the UK Biobank team, led by Luis Nacul, does show some similarity to those areas studied by Lipkin's team:

It will be a 3 year longitudinal study I believe, ending in 2016, and a significant one at that – though only using plasma and no CSF but the controls will comprise MS patients (as well as healthy) and the ME patient cohort will include those in the severe category:

DESCRIPTION (provided by applicant):

A longitudinal study will be conducted of clinical presentation, immune phenotype, gene expression and virus infection among ME/CFS patients and MS and population controls frequency-matched by geographical area of residence, age-group (within 5 years), and sex.

Clinical samples will be collected for studies of NK cell function virology (herpesvirus infection), and gene expression and for banking as a resource for future ME/CFS research.

Hypothesis:

ME/CFS is associated with immune dysfunction, which results from – or predisposes to – herpesvirus infections. Immune dysfunction will present as alterations in NK cell function that may lead to, or result from, alterations in cytokine production and altere expression of diverse immune-associated genes.

Finally, we predict that the ME/CFS immune phenotype may fluctuate over time and in association with clinical presentation and that the majority of patients clinically characterized as having ME/CFS will show a biosignature distinct from that of controls, and that further alterations will be seen during episodes of clinical exacerbation.

Activities and objectives:

i) Collect clinical and other data and venous blood samples at baseline and 6 months or at another time during the 6-month follow-up period when there is a perceived "significant" deterioration in symptoms;

ii) Analyze blood samples for NK cell phenotype and function;

iii) Screen samples for evidence of herpesvirus infection and viral load, focusing on Epstein Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus-6 (HHV-6);

iv) Describe clinical phenotype and fluctuations over time;

v) Correlate the presence of symptoms and severity with markers of virus activity and immune function;

vi) Investigate gene expression profiles associated with ME/CFS and how they vary in relation to changes in disease severity, virus activity, NK cell function, and other markers of immune function;

and

vii) Securely store blood samples from patients and controls, anonymously linked to clinical and other data, as an open resource for researchers to conduct ethically-approved studies of ME/CFS.

Recruitment:

150 ME/CFS cases (50 severe, 100 non-severe), 75 MS controls, and 75 healthy controls will be recruited.

For immunology and virology, 100 cases, 50 MS controls, and 50 healthy controls will be sampled at 2 time points.

For gene expression, we will analyze 50 ME/CFS cases, 25 MS and 25 healthy controls, each at recruitment and one follow-up.

Cases will be selected from UK ME/CFS Disease Register and NHS ME/CFS specialty and primary care services in London and Norfolk, Suffolk, and Great Yarmouth and Waveney, UK; MS controls via the NHS; and healthy controls will be identified by ME/CFS patients (excluding blood relatives) or GPs.

Outcomes:

Identification of putative biomarkers for diagnosis, severity, and prognosis of ME/CFS, which can be evaluated in larger (ideally prospective) future studies.

In the long term, identification of robust biomarkers will allow clinicians to correlate ME/CFS phenotype (including clinical presentation, genetic, immune, and viral markers) with disease severity and prognosis and may reveal new options for interventions research.

Simon September 13, 2013 at 5:11 am

Thanks

Hope they take a good look at Lipkin's work first! He looked for several herpes viruses and found almost nothing. He was looking at plasma not NK cells, but if NK cells were infected I would have thought that some of the herpes virus would have been detectable in plasma. Lipkin also didn't include the severely-affected, but he has looked at 285 samples already with another 200 to go. IF they come back negative too that would surely be a concern for the biobank team?

But still good to see so many chunky studies appearing, and I'm sure the biobank study will generate lots of valuable data and should confirm (or not) the Lipkin findings, even if they, like he, find no herpes.

Firestormm

Replication of Lipkin's preliminary work?

It is unclear at this point, the extent to which this might (at least in part), represent some means of replication for the Lipkin Study we reported on above; but the NIH grant to the UK Biobank team, led by Luis Nacul, does show some similarity to those areas studied by Lipkin's team:

It will be a 3 year longitudinal study I believe, ending in 2016, and a significant one at that – though only using plasma and no CSF but the controls will comprise MS patients (as well as healthy) and the ME patient cohort will include those in the severe category:

heapsreal September 13, 2013 at 5:31 am
Simon

Thanks

Hope they take a good look at Lipkin's work first! He looked for several herpes viruses and found almost nothing. He was looking at plasma not NK cells, but if NK cells were infected I would have thought that some of the herpes virus would have been detectable in plasma. Lipkin also didn't include the severely-affected, but he has looked at 285 samples already with another 200 to go. IF they come back negative too that would surely be a concern for the biobank team?

But still good to see so many chunky studies appearing, and I'm sure the biobank study will generate lots of valuable data and should confirm (or not) the Lipkin findings, even if they, like he, find no herpes.

I dont think thats correct in that he found almost none of the herpes viruses. From what i heard of the talk, hhv6 was mentioned but im not sure if he is referring to active infections or not.

Another thread people have said that its not 'the' cause, which is what he means. Im not even sure if he is referring to a particular test as in spinal fluid? herpes viruses do tend to favour nervous tissue? I think we need some clarrification on this as it does go against what alot of other cfs experts have found and although not everyone has had benefits from treating herpes viruses, it does seem to be a stand out sub set that seems to show some improvement with treatment.

I would also like to no if they tested for herpes viral particles and or lytic herpes viruses which are mentioned alot by dr lerner?? to me it makes sense that these viruses would be an issue if ones nk function is low, but??

The whole study hasnt really told us much other then they think theres some infectious cause and there are possible cytokine markers and they need to do alot more work and dont have the money to do it yet.

Firestormm September 13, 2013 at 5:36 am
Simon

Thanks

Hope they take a good look at Lipkin's work first! He looked for several herpes viruses and found almost nothing. He was looking at plasma not NK cells, but if NK cells were infected I would have thought that some of the herpes virus would have been detectable in plasma. Lipkin also didn't include the severely-affected, but he has looked at 285 samples already with another 200 to go. IF they come back negative too that would surely be a concern for the biobank team?

But still good to see so many chunky studies appearing, and I'm sure the biobank study will generate lots of valuable data and should confirm (or not) the Lipkin findings, even if they, like he, find no herpes.

Yeah, but if NK cells are not infected then it will serve to also rule out Herpes virus. So another confirmation of a poor theory – at least in so far as current infections go. Doesn't rule out this 'shadowing' or trigger concept of course.

And I think it is pretty safe to say that the UK Team are aware of Lipkin's presentation by now :)

If their study is really only 3 years (I took that figure from the NIH link but it isn't certain and is not mentioned in the actual text), it would be a shame; as they have said they will be looking to:

v) Correlate the presence of symptoms and severity with markers of virus activity and immune function;

And that could have resulted in a similar differential to what Lipkin was tentatively suggesting at the 3 year point. That's my presumption of course – because as you know I ain't no scientific nerd :alien: :D

heapsreal September 13, 2013 at 5:43 am

Serology, which is looking for evidence of previous infections, which I think much of what we need to do in the future must focus, is not part of this report that I am making to you today.

This is in relation to all the viral testing done by lipkin and his crew which might explain the low incidence of viruses??
Rereading the lipkin report it also says that the agent must be present in the plasma or spinal fluid as well as not being able to detect historical infections(whatever that means).

http://www.mecfsforums.com/wiki/Lipkin_presentation,_CDC_Conference_Call_9/10/2013

Firestormm September 13, 2013 at 5:45 am
heapsreal

I dont think thats correct in that he found almost none of the herpes viruses. From what i heard of the talk, hhv6 was mentioned but im not sure if he is referring to active infections or not.

Another thread people have said that its not 'the' cause, which is what he means. Im not even sure if he is referring to a particular test as in spinal fluid? herpes viruses do tend to favour nervous tissue? I think we need some clarrification on this as it does go against what alot of other cfs experts have found and although not everyone has had benefits from treating herpes viruses, it does seem to be a stand out sub set that seems to show some improvement with treatment.

I would also like to no if they tested for herpes viral particles and or lytic herpes viruses which are mentioned alot by dr lerner?? to me it makes sense that these viruses would be an issue if ones nk function is low, but??

The whole study hasnt really told us much other then they think theres some infectious cause and there are possible cytokine markers and they need to do alot more work and dont have the money to do it yet.

The point about HHV-6 was made very clear in both the transcript and Simon's review (see above).

From the transcript:

They found in the serum from ME patients and controls forwarded by Montoya:

So, when we look at the samples that we received and there we have 285 cases and 201 controls, that is to say 285 people with Chronic Fatigue; we found HHV-6 in 4 cases and in 2 controls – that is 1.4% of the cases and 1% of the controls. We then typed these HHV-6 samples and found that they were 4 for HHV-6 Type B, and 2 for HHV-6 Type A.

So, HHV-6 at least with respect to using the methods we have for direct detection of infectious agents, if it accounts for Chronic Fatigue, it will do so in a very small proportion. I should also add that Professor Montoya is a known expert in Human Herpes Virus 6 and there is the possibility that some people may have been preselected in some fashion. It’s a potential confound but in any evident I don’t think it is likely to account for much of what we see there.

The other samples that were assayed, using these methods, did not reveal any of the agents to which I have referred.

These same samples were studied using high throughput sequencing, which is a method that was really pioneered here – with which we have discovered over 500 viruses – so we feel fairly confident that to the extent of the technology’s capabilities at present; we would have detected everything that would have been present within these samples.

And in the CSF from Peterson:

Using the same approach as I have described (the multiplex panels I have already talked about with respect to Montoya’s work); we found no genetic evidence of microbial infection in those spinal fluid samples.

We had 1 patient who had HHV-6 B, but that’s a single patient out of 60 and I don’t consider that a statistically significant finding. Now it is possible that if we think that Chronic Fatigue has many different possible causes, that there may be a small subset of people who do have disease as a result of infection with single agents; but this is certainly not going to be a consistent theme that is going to be helpful to us in terms of diagnostics at this point.
We have not yet completed the ion tronic sequencing, so I can’t tell you what result we will obtain with that work.

That's the only references to HHV-6 in the presentation.

heapsreal September 13, 2013 at 5:56 am

I understand hhv6 but not much is mentioned of other herpes viruses. As mentioned in my last post it appears that he thinks theres more to learn from serology which they didn't do during these tests. Hhv6 may get a different result if serology testing was used??

Simon September 13, 2013 at 6:48 am
heapsreal

I dont think thats correct in that he found almost none of the herpes viruses. From what i heard of the talk, hhv6 was mentioned but im not sure if he is referring to active infections or not.

heapsreal

I understand hhv6 but not much is mentioned of other herpes viruses. As mentioned in my last post it appears that he thinks theres more to learn from serology which they didn't do during these tests. Hhv6 may get a different result if serology testing was used??

Hi heapsreal

Serology is testing for antibodies against a virus, 'looking for shadows', as Lipkin put it. Primarliy they are using this technique to find signs of prior infection – however, I think if an active infection is in a particular tissue, but not blood, the anti-virus antibodies (eh anti-HHV6) could still be in the blood. However, I'm not sure if herpes virus are generally not detectable in the blood – I'd just assumed they were.

Firestormm has already mentioned the low levels of HHV6 found in patients and controls (nb this was a very large and well-characterised cohort, probably much bigger than anything that has gone before). However, they also specifically tested for 4 other human herpes viruses: Cytomegalovirus, Herpes Simplex 1 & 2, and Epstein Barr virus (see transcript). HHV7 and Varicella zoser virus are the only other 2 human herpes viruses they didn't test for specifically, but the hightroughput 'unbiased' testing they did should have detected those if they were there (as I understand it).

Personally, I think we are learning a lot from this study because it is so large and robust – and there is much more to come, in the serology results and the unbiased search for fungi and bacteria.

alex3619 September 13, 2013 at 8:37 am
heapsreal

I understand hhv6 but not much is mentioned of other herpes viruses. As mentioned in my last post it appears that he thinks theres more to learn from serology which they didn't do during these tests. Hhv6 may get a different result if serology testing was used??

I think Lipkin looked for active infections for something like five herpes viruses, which are listed in his transcript. The point is he is looking for signs of acute infection. Serological evidence of past infection can be problematic, and typically this means looking for antibodies I think.

None of these address non-acute infections which are part of the lifecycles of some viruses, including herpes and enteroviral families. These are the tissue infections that keep being mentioned. From my current understanding, nearly all these types of viruses can infect B cells.

alex3619 September 13, 2013 at 8:48 am

On Leptin and Serpin I am starting to look into this more closely, though I looked at leptin more than a decade ago. There are many links here with ME pathophysiology. Two that intrigue me at the moment is that leptin can induce hypothalamic inflammation and insulin resistance. If you are obese, have high insulin, glucose or triglycerides; or have disordered stress response or sleep, then leptin could be involved. Its worth some more investigation.

Serpin is a family I think and not one hormone? It can be involved in regulation of other factors, including energy production. I have barely read anything on this though.

Gijs September 13, 2013 at 8:54 am

Business as usual. Nothing has been found. Sound familiar to me. I think the cause of this disease will never been found. It is to complicated. The cytokins Lipkin found are not consistent with earlier findings in other reports.

For example IL17 are not different by CFS patients compared with controls.

J Transl Med. (2012) 10(1): 191 Cytokine expression profiles of immune imbalance in post-mononucleosis chronic fatigue
Broderick G, Katz BZ, Fernandes H, Fletcher MA, Klimas NG, Smith FA, O’Gorman MR, Vernon SD, Taylor R

There are more reports with different outcomes then Lipkin e.a.

I do not think Lipkin has found a usefull biomarker or profile.

Sorry.

roxie60 September 13, 2013 at 9:12 am
alex3619

On Leptin and Serpin I am starting to look into this more closely, though I looked at leptin more than a decade ago. There are many links here with ME pathophysiology. Two that intrigue me at the moment is that leptin can induce hypothalamic inflammation and insulin resistance. If you are obese, have high insulin, glucose or triglycerides; or have disordered stress response or sleep, then leptin could be involved. Its worth some more investigation.

Serpin is a family I think and not one hormone? It can be involved in regulation of other factors, including energy production. I have barely read anything on this though.

Do you have links Alex to the Leptin info you reference? TIA

Sparrowhawk September 13, 2013 at 9:19 am

I thought Dr. Chia's main point is we're not going to find the viruses unless we're also testing a broad set of tissue samples, kind of to heapsreal's points above about the herpes family of viruses. Yes it's great they are testing cerrebrospinal and plasma, but if there is something we don't catch there, and it's already in all your nerve cells in various tissues, GI tract, etc. their "new happy home" then you may not find the usual viral counts or antibodies because the body can't fight them once they are in the nerve cells?

I do find the new direction on gut biome encouraging. If we can at least strengthen that area with some of their findings, that should do a lot to give the body a fighting chance to correct any other imbalances on its own.

Bob September 13, 2013 at 10:58 am
Simon September 13, 2013 at 11:28 am
Gijs

Business as usual. Nothing has been found. Sound familiar to me. I think the cause of this disease will never been found. It is to complicated. The cytokins Lipkin found are not consistent with earlier findings in other reports.

For example IL17 are not different by CFS patients compared with controls.
[Cytokine expression profiles of immune imbalance in post-mononucleosis chronic fatigue]
Broderick G, Katz BZ, Fernandes H, Fletcher MA, Klimas NG, Smith FA, O’Gorman MR, Vernon SD, Taylor R

There are more reports with different outcomes then Lipkin e.a.

I do not think Lipkin has found a usefull biomarker or profile. Sorry.

Thanks. I do get the impression that these new cytokine findings don't quite fit with earlier ones. That said, generally cytokine findings are all over the place in this field.

Re that Broderick paper: it's very interesting in that it looks at cytokines over time in 300 adolescents who had glandular fever, and compares those that recover with those that get CFS. That's a strong experimental design – except there are not many CFS cases to go at. There were 22 CFS cases at 24 months and just 13 at 12 months – compare that with 285 for the Montoya sample in this study.(Though maybe those figures are not so different from the '3 years and under' subgroup in the Lipkin study?).

What I would particularly like to see is how consistent findings are between patients eg are patients consistently low for IL2,8 &17 – or are some low for 2, others for 8 etc. And also how big the differences are between patients and controls? Certainly for other studies there has been a high degree of overlap between patients and controls despite group differences. Will be good to see the paper when it's published.

Nielk September 13, 2013 at 11:31 am

These abnormalities could just be an outcome from the illness – not necessarily a causative agent.

Legendrew September 13, 2013 at 11:56 am
Nielk

These abnormalities could just be an outcome from the illness – not necessarily a causative agent.

I think there's little doubt that cytokines are not the causative agent – to me they stand out more as footsteps in the sand which are worth following. Abnormal levels of cytokines mean disruption of normal immune function and possibly abnormal and harmful immune loops. The true question is whether these loops are perpetuated by the continuing presence of a pathogen or are self-perpetuating as is seen in autoimmune activation.

voner September 13, 2013 at 11:57 am

Simon, Firestormm….. Thank you for that very well written article And the transcript. you are providing a valuable service to this community.

I Wonder how the Lipkin study fits in with the autoimmune/rituximab info? I guess we'll see what Dr. Edwards has to say about the transcript information….

i also wonder if Dr. Chia is ever going to get money to do his tissue sample studies. He is certainly a Lonewolf out there.

wastwater September 13, 2013 at 12:13 pm

I wonder if hhv6a aka human b cell lymphotropic virus or African swine fever virus was tested for

wastwater September 13, 2013 at 12:38 pm

I followed a link on anellovirus and it came up with torque teno virus,it is a very common virus

alex3619 September 13, 2013 at 12:47 pm
roxie60

Do you have links Alex to the Leptin info you reference? TIA

Its everywhere. I just gave a summary of a points on a few papers. Run a search and you will be overwhelmed with links. If I find really good review articles I will post those however.

One key word you might like to know is hyperleptinemia. High leptin has its own name.

PS Here is a paper that shows in mice some of the impact of hyperleptinemia can be reversed by resveratrol:

http://edrv.endojournals.org/cgi/content/meeting_abstract/33/03_MeetingAbstracts/SUN-146

PPS. My initial brief survey found no good reviews of leptin. Instead many papers will discuss it in their opening sections, and some will review a particular aspect of leptin. For example, the following paper discusses in its introduction the impact of leptin on the immune system, but the primary focus is breathing problems:

http://respiratory-research.com/content/pdf/1465-9921-11-152.pdf

There are so many links with possible ME issues that a word of caution is advised: these are all possibilities, but they require a lot of research to validate and test them. For example, hypoxia is known to induce increased leptin levels. However leptin also interferes with NO, inducing vasoconstriction. Its all very complicated.

wastwater September 13, 2013 at 12:59 pm

Another thing I noticed is Chromosomally integrated HHV-6 about the same as autism rates.

Firestormm September 13, 2013 at 1:53 pm

New study: http://www.ncbi.nlm.nih.gov/pubmed/24027260 "Cytokines do not change after exercise or sleep deprivation in chronic fatigue syndrome" Thread here. Made me wonder the circumstances under which the samples in Lipkin's study were taken. Of course we don't know them yet – but it might be prove interesting when we do.

————

Returning to the pathogen hunt, Lipkin said:

"And I will now tell you what the agents are that we can detect using this approach so that when I report to you these findings, you will have a better sense as to their significance.

So the agents that we can detect using this approach are:

  1. human herpes virus 6,
  2. cytomegalovirus,
  3. herpes simplex virus (type 1 and type 2),
  4. Epstein-Barr virus,
  5. Parvovirus,
  6. Borrelia species – normally associated with a number of other diseases that may look like Chronic Fatigue Syndrome,
  7. Black fever
  8. Chlamydia
  9. Enteroviruses
  10. Rhinoviruses
  11. Eastern equine encephalitis virus
  12. Porecco virus
  13. Polisen virus
  14. La Crosse virus
  15. Lymphocytic meningitis virus [?]
  16. Saint Louis Encephalitis virus
  17. West Nile virus
  18. Western equine encephalitis virus
  19. and influenza A virus

These particular tests are not capable of detecting historical infection, the agent must be present in the plasma or the spinal fluid, and in that case we examine[?] spinal fluid at the time that we do that assay.

The serology, which is looking for evidence of previous infections (which is where I think much of what we need to do in the future must focus), is not part of this first report that I am making to you today…

[Except for the HHV-6 we spoke about above]

The other samples that were assayed, using these methods, did not reveal any of the agents to which I have referred."

[And neither did the CSF samples either]

So then. Borrelia species bacteria. We hear an awful lot about people being told they have Lyme Disease and receiving treatment for it – or not; but it looks like there is not a role for this at least in this study.

Much more is said on the forums about various doctors testing folk for the bacteria and determining that this is the source of the CFS-like symptoms – how then can Borrelia not be a factor in Lipkin's results when those patients taking part were diagnosed using Fukuda and CCC?

Any thoughts? Or is this the beginning of the end for people with CFS being misdiagnosed with Lyme disease and receiving inappropriate treatments based on spurious testing?

Sasha September 13, 2013 at 2:31 pm

I thought Nancy Klimas said in Dr Peterson's pre-FDA workshop workshop (the audio went out as podcasts) that if she tests her patients, a minority are positive by PCR (blood, presumably) but by the time they've done three or four repeat tests, most are. I wonder how that ties up with Dr Lipkin's findings?

heapsreal September 13, 2013 at 3:01 pm

The serology, which is looking for evidence of previous infections (which is where I think much of what we need to do in the future must focus), is not part of this first report that I am making to you today

The above quote by lipkin is what many seem to be missing when i mention viruses etc. Serology might be about past infections but he seems very interested in this and thinks it maybe of importance. If serology is tested very high then dr's such as Lerner will say there is an active infection going on but lerner also mentions that these are viral particles, something that i think was missed in this study.

So at the moment i wouldnt say serology is dead and gone.

Sasha September 13, 2013 at 3:08 pm
heapsreal

The serology, which is looking for evidence of previous infections (which is where I think much of what we need to do in the future must focus), is not part of this first report that I am making to you today

The above quote by lipkin is what many seem to be missing when i mention viruses etc. Serology might be about past infections but he seems very interested in this and thinks it maybe of importance. If serology is tested very high then dr's such as Lerner will say there is an active infection going on but lerner also mentions that these are viral particles, something that i think was missed in this study.

So at the moment i wouldnt say serology is dead and gone.

I saw an infectious diseases specialist in the UK who told me my massive HHV-6 IgG titre didn't mean anything because the tests were enormously inaccurate, among other things. Given that almost everyone has been exposed to HHV-6 by adulthood, I wonder what Dr Lipkin can do with his serology tests that would be worth doing. Perhaps he's got better tests?

Firestormm September 13, 2013 at 3:14 pm
heapsreal

The serology, which is looking for evidence of previous infections (which is where I think much of what we need to do in the future must focus), is not part of this first report that I am making to you today

The above quote by lipkin is what many seem to be missing when i mention viruses etc. Serology might be about past infections but he seems very interested in this and thinks it maybe of importance. If serology is tested very high then dr's such as Lerner will say there is an active infection going on but lerner also mentions that these are viral particles, something that i think was missed in this study.

So at the moment i wouldnt say serology is dead and gone.

I am not ignoring this, far from it. What these 'shadows' will reveal is evidence of previous infections: possible triggers for the ME – if the same ones can perhaps be found across the cohort.

However, he did not detect them as 'active' infections and the theories that we have been discussing are in the main about previously 'dormant' viruses common to most people becoming 'reactivated' in people with ME.

If that had been the case: he would have detected them. So, in short, the shadows might reveal more about common pathogen triggers for ME – that might then tie-in with e.g. the over-stimulated immune system and/or 'looping' or B-cell influencing factors perhaps: but not infections that remain active.

Tally September 13, 2013 at 3:21 pm

I'm not too surprised about the differences in the disease after 3 years.

I can't say if it was exactly 3 years, but in the beginning I had horrible sore throats and didn't have muscle pain. Now, at fourth year I never get sore throat and pain is constantly present.

Does anyone know where we can donate money for the Lipkin's microbiome research?

SOC September 13, 2013 at 3:35 pm
Simon

Thanks

Hope they take a good look at Lipkin's work first! He looked for several herpes viruses and found almost nothing. He was looking at plasma not NK cells, but if NK cells were infected I would have thought that some of the herpes virus would have been detectable in plasma. Lipkin also didn't include the severely-affected, but he has looked at 285 samples already with another 200 to go. IF they come back negative too that would surely be a concern for the biobank team?

But still good to see so many chunky studies appearing, and I'm sure the biobank study will generate lots of valuable data and should confirm (or not) the Lipkin findings, even if they, like he, find no herpes.

Lipkin's team might have benefited from consulting the HHV-6 Foundation about HHV-6 testing:
[my bolding]

HHV-6 is never found in plasma or serum unless there is an active infection (or the individual has ciHHV-6). However, t

he absence of HHV-6 DNA in the plasma/serum does not mean that there isn’t a low-level persistent infection in the tissues (e.g. heart, thyroid, brain)

. HHV-6 DNA is not found in the plasma/serum except during the initial infection and transiently during an acute infection.

Firestormm September 13, 2013 at 3:45 pm
SOC

Lipkin's team might have benefited from consulting the HHV-6 Foundation about HHV-6 testing:
[my bolding]

Well I don't know if CSF counts as 'the brain'. What do you think?

Sasha September 13, 2013 at 3:48 pm
SOC

Lipkin's team might have benefited from consulting the HHV-6 Foundation about HHV-6 testing:
[my bolding]

Isn't Montoya connected with them? And Lipkin used Montoya's samples, I think.

Firestormm September 13, 2013 at 3:59 pm
Tally

Does anyone know where we can donate money for the Lipkin's microbiome research?

He wasn't asking specifically for funds – unless (my impression) you have access to a large trust fund – although the CFI is a registered charity: so I suppose you could ask them about establishing a means to donate smaller sums from the community perhaps – that might be a good idea.

Mainly he was talking about applying pressure to your political representatives. He said that Facui and (I forget) – the CDC and NIH – were receptive to his ideas and to ME research being pursued – but the implication was that of the funds that were available: we just don't have enough of a profile to gain access to what is needed.

This sequestration that is being applied globally isn't helping: but if we can somehow raise the profile of ME in terms of solid research direction e.g. the kind of collaborative efforts on large scale that CFI is pursuing – and for things like (not my own preference) the microbiome – with some more background from Lipkin himself perhaps: you could try and pressure the government to increase funding for ME.

I think it needs to be specific. More specific than campaigns in the past. And personal – from you to your own Congressman etc. but it could be planned as part of a PR campaign or an even larger effort. We have – preliminary – biomarkers now: we'd need I think to wait for publication – but that's more credible now than ever before: so it gives us, well, some credibility and we'd not just be demanding funds as before.

We might also have more background detail from the publication of Lipkin's paper (whenever that will be). Be nice to hear something more solid (excuse the pun) about the microbiome and crap-shoot that he appeared so energised by. I think it could be an expensive waste: but I admit to knowing very little about the direction he wishes to take here.

In the transcript – and review – I provided a link I think to the Autism and microbiome paper he co-authored. Might be worth taking a look. I will when I get round to it.

Here's a general piece from Carl Zimmer (also there was another one (NYT I think) linked to in the transcript). You might find it interesting to read more about the possibilities:

A Living Drug Cocktail

by Carl Zimmer

And another recent one from Nat Geo:

When Our Microbes Chat, Dangerous Germs Are Eavesdropping

by Ed Yong

I think the Microbiome is fascinating, don't get me wrong, but I also think it can be a wild goose chase: unless you know what you are looking for it's very easy to get lost.

There are a hell of a lot of theories out there, and remedies for things that nobody can yet prove: but it would be good to get some hard science on the table in relation to PwME at last.

In a very mean spirited way, I am pleased that Lipkin and CFI are taking this challenge on. It is very expensive – as he said – and there's obviously no guarantees.

But I guess it has to be done by somebody – and I'd rather it was him and the incredible team he has assembled within CFI and elsewhere in collaboration.

snowathlete September 13, 2013 at 4:35 pm
Firestormm

New study: http://www.ncbi.nlm.nih.gov/pubmed/24027260 "Cytokines do not change after exercise or sleep deprivation in chronic fatigue syndrome" Thread here. Made me wonder the circumstances under which the samples in Lipkin's study were taken. Of course we don't know them yet – but it might be prove interesting when we do.

————

Returning to the pathogen hunt, Lipkin said:

So then. Borrelia species bacteria. We hear an awful lot about people being told they have Lyme Disease and receiving treatment for it – or not; but it looks like there is not a role for this at least in this study.

Much more is said on the forums about various doctors testing folk for the bacteria and determining that this is the source of the CFS-like symptoms – how then can Borrelia not be a factor in Lipkin's results when those patients taking part were diagnosed using Fukuda and CCC?

Any thoughts? Or is this the beginning of the end for people with CFS being misdiagnosed with Lyme disease and receiving inappropriate treatments based on spurious testing?

One possible explaination for the zero cases in this study might be that anyone with Lyme disease did not make it into the study in the first place, as the people picking the partisipants are good ME/CFS docs who can spot the difference and if I remember correctly Borrelia is in the CCC under conditions that should be excluded before diagnosis.

Which is why there are still plenty of people with Lyme disease who get misdiagnosed with ME/CFS without Lyme having been ruled out by their non-expert doctors, and subsequently ME/CFS docs find a fair number of patients that come to them with diagnosed ME/CFS actually have Lyme.

Firestormm September 13, 2013 at 4:36 pm

From MECFS Forums:

Re: CDC Conference Call Tues. Sept.10 3:00pm

« Reply #43 on: Today at 09:44:32 PM »
Wildaisy said:
I asked Dr. Lipkin about the retroviral sequences and this was his reply:

Quote

"The sequences were not specific for CFS. We find them in many people with and without disease."​

I don't believe the forthcoming paper will provide any further detail about these sequences. Not sure when we might discover more.

snowathlete September 13, 2013 at 4:41 pm
Tally

I'm not too surprised about the differences in the disease after 3 years.

I can't say if it was exactly 3 years, but in the beginning I had horrible sore throats and didn't have muscle pain. Now, at fourth year I never get sore throat and pain is constantly present.

Same here. I can't say when it happened exactly, but I was ill for quite a while with sore throats etc, still working etc. trying to find out what was wrong. Then BAM, something changed, the sore throats disappeared but I got much more ill.

snowathlete September 13, 2013 at 4:48 pm

Great article Simon, thank you!
And thanks Russ for your work on all this too!

Some really interesting stuff there. Lots of questions spring to mind! But I agree that one of the reasons why there is so much value to these studies is the large and robust cohort.

A nice surprise to hear about some of these results up front before publication. I guess that helps speed the process up of getting the funding he needs to keep things going, else it might take even longer.

So if I understand correctly, the sequencing for bacteria and fungi is yet to be completed. Would it be a surprise if he found something here? I'd have thought not as bacterias can be just as nasty as viruses and cause cytokine changes. Perhaps though, they have started and not found anything yet, so although they can't confirm it's not bacteria at this stage, they are fairly confident that it's not going to be?

If anything, it seems to me that Lipkin is even more convinced now of an infectious agent being involved, with these immune results, than he was before.

Regarding cytokine results not matching other studies, as I understood it, processing has a major impact on the result, so the important thing would be consistancy within Lipkin's tests (which I am sure will be the case) and any subsequent validation tests by someone else.

PDXhausted September 13, 2013 at 7:32 pm

Has anyone ever done research looking for pathogens in ear/nose/throat tissue? It seems that many people that have a post-viral onset do so from mono or a mono-like illness where the symptoms concentrate in those areas. I'm under the impression that viruses that park in those areas don't always infect the blood or spinal fluid.

For me personally, one of my first symptoms was tinnitus. It just popped up one day, and progressively got worse, and then I crashed a few weeks later and developed the rest of my symptoms. Along with that I developed excessive earwax and get ear infections that come and go. I think the microbiome is interesting and important, but I'd love for someone to virus-hunt in my ears.

Firestormm September 13, 2013 at 8:24 pm
snowathlete

One possible explaination for the zero cases in this study might be that anyone with Lyme disease did not make it into the study in the first place, as the people picking the partisipants are good ME/CFS docs who can spot the difference and if I remember correctly Borrelia is in the CCC under conditions that should be excluded before diagnosis.

Which is why there are still plenty of people with Lyme disease who get misdiagnosed with ME/CFS without Lyme having been ruled out by their non-expert doctors, and subsequently ME/CFS docs find a fair number of patients that come to them with diagnosed ME/CFS actually have Lyme.

I tend to agree and Lyme Disease is an exclusion under CCC. This result though does rule out Borellia as am infectious agent involved in CFS it would seem which would endorse the view that CFS is a distinct disease.

I'd be pretty darn careful who tested me for Borellia before being ruled out and treated – because there's a lot of uncertainty surrounding some of the tests and treatments – but we needn't get into any of that here.

There was one virus that was omitted which I did think about: Q Fever. I think from my early days with ME is was one of those that was considered a trigger at least. I guess it could show up for some people as a 'shadow'.

Made me wonder how Mady and Lipkin might have arrived at their choice of pathogens?

And finally, here is someone who is evidently not impressed with Lipkin's work:

Dr J Deckoff-Jones and her Bats in the Belfry :zippit:

alex3619 September 13, 2013 at 9:09 pm

Firestormm , Q fever would be part of the bacterial studies, not viral studies. As a result we have yet to hear the verdict on that. Q fever can lead to Post Q Fever, and some of those become indistinguishable from ME. There are parts of the world where outbreaks of this are still common, including in Europe.

Let me reiterate again, the current results from Lipkin are about acute viral infection and immune markers. Viruses are still not ruled out, just acute viral infection of likely viral pathogens.

alex3619 September 13, 2013 at 9:17 pm

Just to clarify what I think Lipkin found (or did not find): he did not find evidence of acute viral infection, but did find immune markers. HHV-6 etc. do not always present as acute viral infections, and are not ruled out except as acute viral infections. In other words, the classic viral model, the one doctors learn in med school, does not fit ME. Is that somehow a surprise?

Firestormm September 13, 2013 at 9:22 pm
alex3619

Firestormm , Q fever would be part of the bacterial studies, not viral studies. As a result we have yet to hear the verdict on that. Q fever can lead to Post Q Fever, and some of those become indistinguishable from ME. There are parts of the world where outbreaks of this are still common, including in Europe.

Let me reiterate again, the current results from Lipkin are about acute viral infection and immune markers. Viruses are still not ruled out, just acute viral infection of likely viral pathogens.

Thanks Alex. Isn't Borellia a bacteria? This was what he said, prior to listing the agents he could test for:

Let me begin first by talking about the work that we have done with Jose Montoya of Stanford. Now, our role here was to try to look specifically for infectious agents and as many of you will know, Dr Montoya is a rigorous clinician who has been working in this field now for a decade or more; and he sent us plasma samples [that] we characterised using a method that allows us to detect genetic evidence of infection [from] a wide variety of bacteria, viruses and parasites.

I know he later – when talking about what they are also doing at present – he says:

In addition we specifically look for bacteria using a method called 16S ribosomal RNA sequencing, which is a method that allows us to amplify by polymerase chain reaction (PCR) a region that we can then sequence and look for variability in the presence of specific bacteria. We do the same thing by using a method called 18S ribosomal RNA to look at fungi and I know that many people are interested in not only viruses but also bacteria and fungi.

And it's that part that obviously hasn't been reported on. But the results he did talk about were for active infections as per the list I cited above.

Not sure if these bacteria and fungi searches will be part of the microbiome and poo-hunt or not: I'd presume so – what do you think?

alex3619 September 13, 2013 at 9:25 pm
roxie60

Do you have links Alex to the Leptin info you reference? TIA

You might find this abstract interesting. Be warned its only an abstract. Many of us have circadian issues, particularly long term patients, which I have noted before I start to see after year 3 … which matches the timing of the shift in cytokines Lipkin observed.

http://jap.physiology.org/content/early/2013/07/16/japplphysiol.00630.2013.abstract

Diminished leptin signaling can alter circadian rhythm of metabolic activity and feeding

Leptin, a hormone mainly produced by fat cells, shows cell-specific effects to regulate feeding and metabolic activities. We propose that an important feature of metabolic dysregulation resulting in obesity is the loss of the circadian rhythm of biopotentials.

Then there is this:

http://onlinelibrary.wiley.com/doi/10.1111/cpf.12030/abstract

The associations between peak O2 consumption and leptin in 10- to 12-year-old boys

We can conclude that leptin first of all correlated negatively with relative peak O2 consumption. Absolute VO2peak correlated with leptin only in total group.

I wonder if our leptin goes up on day two of a repeat CPET test? It would be interesting to find out.

taniaaust1 September 13, 2013 at 9:33 pm
PDXhausted

Has anyone ever done research looking for pathogens in ear/nose/throat tissue? It seems that many people that have a post-viral onset do so from mono or a mono-like illness where the symptoms concentrate in those areas. I'm under the impression that viruses that park in those areas don't always infect the blood or spinal fluid.

For me personally, one of my first symptoms was tinnitus. It just popped up one day, and progressively got worse, and then I crashed a few weeks later and developed the rest of my symptoms. Along with that I developed excessive earwax and get ear infections that come and go. I think the microbiome is interesting and important, but I'd love for someone to virus-hunt in my ears.

PDXhausted. Some time ago, Newcastle University (Australia) did a study on the bacteria Staph in the nose and found that those who have ME/CFS tend to carry a toxin producing kind of Staph there (25% of healthies carry Staph there but not the kind we tend to do).

This of cause isnt causing our ME/CFS but seems to be one of those things which can go with it and just one added thing knocking at our health. (So you may want to consider having a nasal swab done and then treating Staph if found there).

taniaaust1 September 13, 2013 at 9:55 pm

For anyone interested in the Anellovirus finding.

"Anellovirus infections are highly prevalent in the general population. A study in Japan found that 75–100% of patients tested were infected with at least one of the three human anelloviruses, and many were infected with multiple species "

So it seems that finding of the percentage found in us has no meaning at all (unless of cause the tiers were far higher then normal??).

acer2000 September 14, 2013 at 1:11 am

I think the issue with sequencing blood and CSF is that some of these agents don't really inhabit either. For example, Lyme testing via PCR in blood and in CSF is specific, but has very low yield. Its mainly a tissue based infection. Some other pathogens might be the opposite. Its really hard to make hard conclusions as to what extent a pathogen is involved unless you know where it primarily lives and those tissues are sampled. So its important that they also look for the antibodies as well.

Tally September 14, 2013 at 3:17 am
Firestormm

Mainly he was talking about applying pressure to your political representatives. He said that Facui and (I forget) – the CDC and NIH – were receptive to his ideas and to ME research being pursued – but the implication was that of the funds that were available: we just don't have enough of a profile to gain access to what is needed.

This sequestration that is being applied globally isn't helping: but if we can somehow raise the profile of ME in terms of solid research direction e.g. the kind of collaborative efforts on large scale that CFI is pursuing – and for things like (not my own preference) the microbiome – with some more background from Lipkin himself perhaps: you could try and pressure the government to increase funding for ME.

I think it needs to be specific. More specific than campaigns in the past. And personal – from you to your own Congressman etc. but it could be planned as part of a PR campaign or an even larger effort.

Thank you for the elaborate reply.

Unfortunately I'm not from the US, so there's nothing I can do about the government. The best I can do is donate a bit every month.

Simon September 14, 2013 at 3:42 am
SOC

Lipkin's team might have benefited from consulting the HHV-6 Foundation about HHV-6 testing [re problems detecting in blood]

Sasha

Isn't Montoya connected with them? And Lipkin used Montoya's samples, I think.

Interesting point. Montoya is an HHV6 expert, and the HHV6 foundation are funding his pilot study:
Valganciclovir shows promise during first randomized clinical trial for treatment of HHV-6 & EBV in CFS patients | HHV-6 Foundation
So would be interesting to hear what Montoya would say on that issue (and yes, he did supply the samples for this study)

Gijs September 14, 2013 at 4:38 am

The news is that there is no news. Lipkin did find nothing special. Only the CSF findings are interesting as a marker because they are consistent. This need to be validated in a larger population. This can proof 'infection' in the nervous system. Which is the cause of this disease, i believe.

alex3619 September 14, 2013 at 4:54 am

I think we might find leptin can be used as a treatment biomarker, but will be useless diagnostically. In other words, leptin might tell a doctor if a treatment is more likely to be helping than harming.

Leptin cannot be used diagnostically for two reasons: its found to be high in many conditions including obesity, diabetes and the metabolic syndrome; and its individually variable. However as a treatment biomarker the comparison will always be your own leptin levels.

Legendrew September 14, 2013 at 4:57 am
Gijs

The news is that there is no news. Lipkin did find nothing special. Only the CSF findings are interesting as a marker because they are consistent. This need to be validated in a larger population. This can proof 'infection' in the nervous system. Which is the cause of this disease, i believe.

I'd argue the point that CSF markers serve as proof of an infection, viral or otherwise, of the nervous system. They certainly hint at a form of immune dysregulation within the brain and perhaps entire central nervous system but are not even close to showing an infectious causation, certainly that could be one cause of such a dysregulation but there are numerous others to further explore too.

heapsreal September 14, 2013 at 5:19 am
Legendrew

I'd argue the point that CSF markers serve as proof of an infection, viral or otherwise, of the nervous system. They certainly hint at a form of immune dysregulation within the brain and perhaps entire central nervous system but are not even close to showing an infectious causation, certainly that could be one cause of such a dysregulation but there are numerous others to further explore too.

Many viruses like the nervous system, so its possible that those markers found in csf are from viruses. Meningittis is one that comes to find and a recent study i read somewhere and dont have a link, maybe someone can help out, but with some type of new testing ebv was found to be a major cause of viral meningittis where the older testing methods couldnt detect which infection was the issue. chickenpox/shingles loves the nervous system.

All comes down to which types of tests are the best for which viruses.

I still cant get past many of us not having chronic viral infections when most of us have low nk function, who's job it is to kill viruses. if its herpes virus or some other virus, if they cant find them then they probably havent looked in the right places or dont have the right tests.

A quick search of other auto immune illnesses too show viral infections play apart in some of them. There is even some with MS who have improved on valtrex. Maybe when all this is sorted they will possibly find viral infections implicated in other illnesses that they dont have the final answer in like many auto immune illnesses.

Thats my opinion anyway.

Legendrew September 14, 2013 at 5:38 am
heapsreal

Many viruses like the nervous system, so its possible that those markers found in csf are from viruses. Meningittis is one that comes to find and a recent study i read somewhere and dont have a link, maybe someone can help out, but with some type of new testing ebv was found to be a major cause of viral meningittis where the older testing methods couldnt detect which infection was the issue. chickenpox/shingles loves the nervous system.

All comes down to which types of tests are the best for which viruses.

I still cant get past many of us not having chronic viral infections when most of us have low nk function, who's job it is to kill viruses. if its herpes virus or some other virus, if they cant find them then they probably haven't looked in the right places or don't have the right tests.

A quick search of other auto immune illnesses too show viral infections play apart in some of them. There is even some with MS who have improved on valtrex. Maybe when all this is sorted they will possibly find viral infections implicated in other illnesses that they dont have the final answer in like many auto immune illnesses.

That's my opinion anyway.

A few good points here: NK cells however play a more major role in innate immunity – the job of clearing up viruses is generally down to the adaptive immune system with T-cells and B-cells + the antibodies the B-cells produce.

To me it seems a little strange that despite these results people still hold strongly onto the viral causation. I understand the argument of tissue-specific viruses but I would expect some clues to still be present in these samples. There has to come a point where we come to accept that viruses aren't the major cause of pathology in ME and I think we're fast approaching that – certainly it is worth testing these things to the fullest of our abilities but if the evidence isn't there you have to move on to other hypotheses. I think there is little argument that they are the most common triggering factor, likely due to the immune response they initiate, but if results searching for them keep turning up a blank we need to explore other areas and I think autoimmunity is both the most associated with the viral ideas and also probably the most exciting area at the minute.

Snow Leopard September 14, 2013 at 5:42 am

The failure to replicate virus results is not a recent phenomena, it is a consistent one over 20 years. Viruses seem to be key triggers for onset of symptoms, but evidence for chronic infection has remained elusive for a long time, and there are many other good hypotheses out there…

globalpilot September 14, 2013 at 6:08 am
Legendrew

A few good points here: NK cells however play a more major role in innate immunity – the job of clearing up viruses is generally down to the adaptive immune system with T-cells and B-cells + the antibodies the B-cells produce.

To me it seems a little strange that despite these results people still hold strongly onto the viral causation. I understand the argument of tissue-specific viruses but I would expect some clues to still be present in these samples. There has to come a point where we come to accept that viruses aren't the major cause of pathology in ME and I think we're fast approaching that – certainly it is worth testing these things to the fullest of our abilities but if the evidence isn't there you have to move on to other hypotheses. I think there is little argument that they are the most common triggering factor, likely due to the immune response they initiate, but if results searching for them keep turning up a blank we need to explore other areas and I think autoimmunity is both the most associated with the viral ideas and also probably the most exciting area at the minute.

The reason I think viruses may still be involved (not necessarily causal, but involved) is because Dr Chia found enterovirus in about 2/3 of my stomach biopsy cells. Unless the sample was contaminated I just can't see how this cannot be significant. I hope to hear from Dr Chia on these issue – ie lack of enteroviral RNA in the plasma and CSF.

I do agree, however, it is puzzling there is no RNA of these viruses in the blood if they are involved.

Firestormm September 14, 2013 at 6:14 am
globalpilot

The reason I think viruses may still be involved (not necessarily causal, but involved) is because Dr Chia found enterovirus in about 2/3 of my stomach biopsy cells. Unless the sample was contaminated I just can't see how this cannot be significant. I hope to hear from Dr Chia on these issue – ie lack of enteroviral RNA in the plasma and CSF.

I do agree, however, it is puzzling there is no RNA of these viruses in the blood if they are involved.

I admit to knowing very little about enteroviruses. I do know that it's possible these widespread infections are not always the cause of disease. I would though like to hear from Dr Chia with regard to Lipkin's work here. Maybe once it's published he will be willing to review? I am sure many doctors involved in ME work will be reviewing the data and hopefully talking to their patients – even if only to reassure.

heapsreal September 14, 2013 at 6:20 am

Definitely should be sub typing. I think many hold onto the virus theory as there is a group of cfsers who improve on antivirals. Myself dont believe it is the cause but treating them can help alot. Nk cells are there for a reason and work with other t cells and b cells. No reason why this dysfunction can reduce e the immune systems ability to contrl viral infections.
The autoimmune theory so far hasn't had alot of success at all that has been posted by people here?

Some recent studies show not only nk but cd8 t cells have low function. I believe its these abnormalities that allow one to get a variety of infections or maybe hit and run. I dont know what they would call this, an immune deficiency or dysfunction.

heapsreal September 14, 2013 at 6:25 am
Snow Leopard

The failure to replicate virus results is not a recent phenomena, it is a consistent one over 20 years. Viruses seem to be key triggers for onset of symptoms, but evidence for chronic infection has remained elusive for a long time, and there are many other good hypotheses out there…

Proper accurate testing as well as well defined cfs study participants could account for there inability to find viruses. Doesn't seem to be a problem for lerner Peterson klimas, as well as being able to treat and improve many of them.

lansbergen September 14, 2013 at 6:29 am

I refuse to believe the immunesystem attacks self for no reason. I always have and likely always will.

heapsreal September 14, 2013 at 6:30 am
Firestormm

I admit to knowing very little about enteroviruses. I do know that it's possible these widespread infections are not always the cause of disease. I would though like to hear from Dr Chia with regard to Lipkin's work here. Maybe once it's published he will be willing to review? I am sure many doctors involved in ME work will be reviewing the data and hopefully talking to their patients – even if only to reassure.

I dont think lipkins current work has alot to offer. All it seems to be doing is giving us more questions then answers. We will have to wait until its finished.
Montoya is doing a separate pathogen study?? Do we know when this will be finished??

globalpilot September 14, 2013 at 6:52 am
lansbergen

I refuse to believe the immunesystem attacks self for no reason. I always have and likely always will.

Same here. It makes no sense whatsoever.

Legendrew September 14, 2013 at 7:05 am
lansbergen

I refuse to believe the immunesystem attacks self for no reason. I always have and likely always will.

It's an incredibly complex topic and one I plan to explore more in a possible article in the future. Suffice it to say that there is generally a trigger in autoimmune diseases which moves it from a silent (ie non symptomatic) state into a seemingly active and debilitating state. These triggers can include non-self things such as viruses or bacterial infections – but the immune response then appears to be self perpetuating.

alex3619 September 14, 2013 at 8:08 am

Serpins are a superfamily of proteins, and their function is critically reliant on proper folding. We already know we have increased protein misfolding (KDM finding?) and that we have a mechanism for decreased folding: glutathione deficiency. So are septins, while abundant in quantity, really abundant functionally or deficient? I don't know.

http://www.ncbi.nlm.nih.gov/pubmed/21781239

Here is a recent review on serpins:

http://genomebiology.com/2006/7/5/216

Serpins are involved in regulating proteases, but also as transport molecules, involving both corticosteroids and thyroid hormone. Its worth noting that one of these, corticosteroid binding hormone, may be involved in producing CFS-like symptoms.

Have a gander at this table: http://genomebiology.com/2006/7/5/216/table/T1

In order to make sense of Lipkin's finding we need to know which serpins are involved. Until then the range of possible impact is very open to wild speculation.

alex3619 September 14, 2013 at 8:21 am
globalpilot

The reason I think viruses may still be involved (not necessarily causal, but involved) is because Dr Chia found enterovirus in about 2/3 of my stomach biopsy cells. Unless the sample was contaminated I just can't see how this cannot be significant. I hope to hear from Dr Chia on these issue – ie lack of enteroviral RNA in the plasma and CSF.

I do agree, however, it is puzzling there is no RNA of these viruses in the blood if they are involved.

If the biospy included staining of cells to be viewed under a microscope, the possibility of contamination is unlikely. If its just an assay to detect presence of the cells, then contamination is possible. I don't recall exactly, but haven't both been done and the biopsies found to be positive?

alex3619 September 14, 2013 at 8:27 am
lansbergen

I refuse to believe the immunesystem attacks self for no reason. I always have and likely always will.

One theory of autoimmune is due to how the antibody immune system works. The B cells randomly reshuffle some of their DNA to make antibodies … which have random targets. Any that match native proteins near where this occurs should be eliminated. A few years back there was a paper showing that proteins that are only expressed far from where the B cells are adapting may not lead to elimination of these B cells.

When the immune cells are activated by something its due to a protein or protein complex match. If there is sufficient activation then the activated cells multiply – clonal expansion. This is when we develop resistance to pathogens – we have sufficient antibodies to fight them. This is what Rituximab targets – it kills B cells. This is what Lipkin found in the earlier paper. Our immune systems are reacting to something.

B cells might also create autoantibodies in numbers due to cross-reactivity. The cells are responding to something else, but the antibodies have sufficient affinity to self-proteins that they get weakly attacked.

Gijs September 14, 2013 at 8:47 am

(neuro)toxines -produced by microflora- can trigger the immunesystem. I think this is the next step to investigate.

Legendrew September 14, 2013 at 8:47 am
alex3619

One theory of autoimmune is due to how the antibody immune system works. The B cells randomly reshuffle some of their DNA to make antibodies … which have random targets. Any that match native proteins near where this occurs should be eliminated. A few years back there was a paper showing that proteins that are only expressed far from where the B cells are adapting may not lead to elimination of these B cells.

When the immune cells are activated by something its due to a protein or protein complex match. If there is sufficient activation then the activated cells multiply – clonal expansion. This is when we develop resistance to pathogens – we have sufficient antibodies to fight them. This is what Rituximab targets – it kills B cells. This is what Lipkin found in the earlier paper. Our immune systems are reacting to something.

B cells might also create autoantibodies in numbers due to cross-reactivity. The cells are responding to something else, but the antibodies have sufficient affinity to self-proteins that they get weakly attacked.

Yep – I think it is also worth mentioning that a small degree of autoimmunity with regards to the production of autoantibodies is a good thing. The argument that autoimmunity needs an external stimulus is a little silly when you consider the benefits that autoimmunity has with regard to detecting and successfully fighting cancerous cells. If autoimmunity was a bad things then it would likely no longer exist in the human (and other organisms) population as the genes and traits responsible for it would have not been selected for during evolution.
Autoimmunity only becomes a bad thing when the immune response gets stuck in cycles over-expressing randomly created autoantibodies and the response that ensues therein.

alex3619 September 14, 2013 at 10:39 am

http://www.jimmunol.org/content/174/6/3137.full

Leptin in Immunology
Abstract

Leptin is an adipokine which conveys information on energy availability. In humans, leptin influences energy homeostasis and regulates neuroendocrine function primarily in states of energy deficiency. As a cytokine, leptin also affects thymic homeostasis and, similar to other proinflammatory cytokines, leptin promotes Th1 cell differentiation and cytokine production. We review herein recent advances on the role of leptin in the pathophysiology of immune responses

This is an old paper, so take what it says too seriously. I am citing it to introduce a point. In obesity research high leptin from fat cells is thought to not make it to the brain at a high enough rate, and so does not send proper signals to the hypothalamus.

Leptin sends a signal to the brain that there is plenty of energy available, crank up the metabolic rate. What if there isn't, if the leptin is derived some other way? If you have low energy capacity.with high leptin, what happens? What happens if the signal is distorted, as found in obesity, and the hypothalamus is not getting the message that there is too much energy?

Its also worth noting that women make more leptin than men. Is this related to why more women get ME than men?

Marco September 14, 2013 at 10:40 am
Legendrew
alex3619

One theory of autoimmune is due to how the antibody immune system works. The B cells randomly reshuffle some of their DNA to make antibodies … which have random targets. Any that match native proteins near where this occurs should be eliminated. A few years back there was a paper showing that proteins that are only expressed far from where the B cells are adapting may not lead to elimination of these B cells.

When the immune cells are activated by something its due to a protein or protein complex match. If there is sufficient activation then the activated cells multiply – clonal expansion. This is when we develop resistance to pathogens – we have sufficient antibodies to fight them. This is what Rituximab targets – it kills B cells. This is what Lipkin found in the earlier paper. Our immune systems are reacting to something.

B cells might also create autoantibodies in numbers due to cross-reactivity. The cells are responding to something else, but the antibodies have sufficient affinity to self-proteins that they get weakly attacked.

Yep – I think it is also worth mentioning that a small degree of autoimmunity with regards to the production of autoantibodies is a good thing. The argument that autoimmunity needs an external stimulus is a little silly when you consider the benefits that autoimmunity has with regard to detecting and successfully fighting cancerous cells. If autoimmunity was a bad things then it would likely no longer exist in the human (and other organisms) population as the genes and traits responsible for it would have not been selected for during evolution.
Autoimmunity only becomes a bad thing when the immune response gets stuck in cycles over-expressing randomly created autoantibodies and the response that ensues therein.

I quite like the theory of the immune system as a cognitive network that arises out of the Santiago theory of cognition.

No need to discriminate between self and non-self or to consider autoimmunity as necessarily a problem except in the (relatively) rare case of a massive infection.

In this theory the role of the immune network is not to seek out and kill but to regulate both
self and non-self within the parameters necessary to maintain the organism's physiological integrity. Therefore a certain level of foreign organisms is tolerated and regulated. Otherwise how could we tolerate our own beneficial bacteria and how would symbiosis (eg mitochondria) have evolved.

http://combusem.com/CAPRA4.HTM

Quite possibly bollocks though ;)

alex3619 September 14, 2013 at 10:54 am

Marco Maturana's theory is complicated and requires careful reading and analysis. I met Humberto Maturana, the founder of much of modern systems biology, as I went to one of his two day seminars. Systems that survive are ones that maintain balance, but that does not mean its seeking balance. Its just mechanisms that tend to maintain balance. Systems that adapt or evolve to maintain balance better have an evolutionary advantage.

Systems view always have to take details into account, and environment, and mechanism and … Its one reason why so many try to avoid them. Its not easy taking a systems view, nor even easy to know where to put the boundaries for the system analysis.

What it does imply is that the current state of the system is also a reflection of how it has changed during its history. In other words, its something dynamic. The immune system is definitely dynamic. I would like to see more work on treating the immune system as a dynamic system over time, but alas that is not where the science is going. The research is usually heavily reductionistic, which is what makes it more amenable to scientific enquiry.

Marco September 14, 2013 at 11:12 am
alex3619

Marco Maturana's theory is complicated and requires careful reading and analysis. I met Humberto Maturana, the founder of much of modern systems biology, as I went to one of his two day seminars. Systems that survive are ones that maintain balance, but that does not mean its seeking balance. Its just mechanisms that tend to maintain balance. Systems that adapt or evolve to maintain balance better have an evolutionary advantage.

Systems view always have to take details into account, and environment, and mechanism and … Its one reason why so many try to avoid them. Its not easy taking a systems view, nor even easy to know where to put the boundaries for the system analysis.

What it does imply is that the current state of the system is also a reflection of how it has changed during its history. In other words, its something dynamic. The immune system is definitely dynamic. I would like to see more work on treating the immune system as a dynamic system over time, but alas that is not where the science is going. The research is usually heavily reductionistic, which is what makes it more amenable to scientific enquiry.

Hi Alex

Must have been an interesting seminar.

I feel there's a lot in the theory but as you know its getting on a bit now – I don't know if mainstream medicine has embraced any of it – plus while these theories have been highly influential (and influenced by) cybernetics and chaos theory – Varela and Maturana may have over-extended the theory into areas where they've less expertise.

I do like the idea though that autoimmunity may involve a failure of connectivity in the immune network (Broderick I believe suggested something with regard to immune findings in ME/CFS).

There's also the possibility that complex systems (as the immune system must be) may be prone to rapid flips to another stable (but dysregulated) state.

All intellectually satisfying but unlikely to be of any practical help in the near future.

snowathlete September 14, 2013 at 11:26 am
acer2000

I think the issue with sequencing blood and CSF is that some of these agents don't really inhabit either. For example, Lyme testing via PCR in blood and in CSF is specific, but has very low yield. Its mainly a tissue based infection. Some other pathogens might be the opposite. Its really hard to make hard conclusions as to what extent a pathogen is involved unless you know where it primarily lives and those tissues are sampled. So its important that they also look for the antibodies as well.

It does seem like some pathogens favour tissue but many, including Borrelia, are often enough found in the blood. If any had been missed in the patient exclusion phase then with as many subjects as this (260?) and with the best techniques available being used, you'd have thought if present then at least some cases would have been picked up, but instead we have zero. Borrelia as a cause therefore looks pretty unlikely now, though some other bacteria being responsible is yet to be ruled out as i understand it. It'll be interesting to see if they find anything, or not.

Legendrew September 14, 2013 at 11:58 am
snowathlete

It does seem like some pathogens favour tissue but many, including Borrelia, are often enough found in the blood. If any had been missed in the patient exclusion phase then with as many subjects as this (260?) and with the best techniques available being used, you'd have thought if present then at least some cases would have been picked up, but instead we have zero. Borrelia as a cause therefore looks pretty unlikely now, though some other bacteria being responsible is yet to be ruled out as i understand it. It'll be interesting to see if they find anything, or not.

As far as i'm aware Borrelia is an exclusionary criteria from ME anyway although I guess that depends upon which diagnostic criteria the physician is using. This extensive search is certainly worthwhile but I think it's unwise to pin hopes on this given the history pathogens have had in terms of ME research. From a personal standpoint I doubt any infectious agent lies at the heart – perhaps they simply serve to exacerbate the problem in those who have them.

I agree that even if these pathogens are hiding away in tissues there should be some evidence of the active infection. As far as i'm concerned if the pathogen is inert in these tissues there should be no symptoms so I feel that line of thought should be throw out instantly.

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