Novel characterisation of mast cell phenotypes from peripheral blood mononuclear cells in ME/CFS

[Effi]

New study from the Marshall-Gradisnik team (NCNED).

http://www.ncbi.nlm.nih.gov/pubmed/27362406

BACKGROUND:
Mast cells (MCs) mediate inflammation through neuropeptides and cytokines, along with histamine and reactive oxygen species (ROS). Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is an illness characterized by an unexplained disabling fatigue with multiple physiological impairments as well as dysregulated cytokine profiles.

OBJECTIVE:
To determine mast cell phenotypes in isolated human PBMCs, in healthy controls and in CFS/ME patients. Second, determine receptor expression of RAGE and its ligand high mobility group box 1 protein (HMGB1).

METHOD:
Moderately severe CFS/ME patients (n=12, mean age 39.25±SD3.52 years), severe CFS/ME patients (n=6, mean age 43.00±SD4.02 years) and healthy controls (n=13, mean age 42.69±SD3.87 years) were included in this study. CFS/ME patients were classified according to the 2011 International Consensus Criteria. LSRFortessa X-20 Flow cytometry was used for the identification of phenotypic peripheral mast cell population in PBMCs using an exclusion marker Lin2 cocktail (anti-CD3, anti-CD14, anti-CD19, anti-CD20 and anti-CD56) and inclusion markers (CD117, CD34, FCεRI, chymase, HLA-DR and CD154) following comparative investigation. HMGB1 and soluble RAGE expression in plasma was measured by sandwich ELISA assay.

RESULTS:
There was a significant increase in CD117+CD34+FCεRI-chymase- mast cell populations in moderate and severe CFS/ME patients compared with healthy controls. There was a significant increase in CD40 ligand and MHC-II receptors on differentiated mast cell populations in the severe CFS/ME compared with healthy controls and moderate CFS/ME. There were no significant differences between groups for HMGB1 and sRAGE.

CONCLUSIONS:
This preliminary study investigates mast cell phenotypes from PBMCs in healthy controls. We report significant increase of naïve MCs in moderate and severe CFS/ME patients compared with healthy controls. Moreover, a significant increase in CD40 ligand and MHC-II receptors on differentiated mast cells in severe CFS/ME patients. Peripheral MCs may be present in CFS/ME pathology however, further investigation to determine their role is required.

 

[msf]

I just saw this and noticed that pretty much every other biomedical paper is from this group. It doesn´t seem like they have a theoretical framework for their research, but maybe it´s just too complicated for the likes of me!

 

[Marky90]

There are so many abnormalities everywhere, seemingly.. No doubt that whatever our disease process is, it causes systemic downward effects.

 

[Vasha]

This is potentially very exciting.
@Gingergrrl , @justy, @Strawberry -- other mast cell people--this is an attempt to draw a connection between mast cell problems and ME.

Good stuff: They used ICC (wondering why not CCC?) and have both _moderate and severe_ groups.
But: the groups are so small! 12, 6, 12.

Without the paper, it's hard to know what the stats look like, but it looks like there may be multiple comparisons, even on this small sample. Tagging @Valentijn .

I hope the paper is available soon. It's great that they are looking at this.

Vasha

 

[Bob]

For reference, and for search purposes...

Novel characterisation of mast cell phenotypes from peripheral blood mononuclear cells in chronic fatigue syndrome/myalgic encephalomyelitis patients.
Nguyen T, Johnston S, Chacko A, Gibson D, Cepon J, Smith P, Staines D, Marshall-Gradisnik S.
Asian Pac J Allergy Immunol. 2016.
June 30, 2016.
doi: 10.12932/AP0711.

Journal:
http://www.apjai-journal.org/

 

[A.B.]

Could someone explain what the findings mean (assuming that they are true), and to what degree they are consistent with what we know about ME/CFS?

 

[Bob]

I've only read the abstract. My (rudimentary) understanding of the abstract is that they've used flow cytometry to measure the variance in some types (phenotypes) of peripheral mast cells in the blood of ME patients compared to healthy controls. There were 18 patients and 13 controls. The study is described as a preliminary study (i.e. in my words, it's a small exploratory pilot study). No conclusions are made from the results: "peripheral [mast cells] may be present in CFS/ME pathology however, further investigation to determine their role is required."

They've detected some statistically significant differences in phenotypes (not necessarily anything other than a small or modest difference) using very small numbers of subjects.

In moderate and severe ME patients there is an increase in naive mast cells, and an increase in CD117+CD34+FCεRI-chymase mast cell populations, compared to healthy controls. And there is an increase in CD40 ligand (also known as CD154) and MHC-II receptors on differentiated mast cells in severe ME patients compared to both moderate ME patients and controls.

What do the results mean? I don't think they tell us anything at this stage, except that there may be some measure of difference in immune cell activity between moderately affected patients, severely affected patients and controls. This study is so small that the findings may not be replicable.

I'm not sure if I've read any mast cell theories in relation to ME, or if I've read any substantial ME research involving mast cells.

Beyond that very rudimentary understanding, I'll pass it over to others to explain what's going on...

 

[Valentijn]

Without the paper, it's hard to know what the stats look like, but it looks like there may be multiple comparisons, even on this small sample. Tagging @Valentijn .

Hard to guess, without seeing the full paper. It doesn't seem to be published yet.

Could someone explain what the findings mean (assuming that they are true), and to what degree they are consistent with what we know about ME/CFS?

Increased mast cells and/or mast cell activity might cause inflammatory reactions to a lot of things. But if these differences are small, or not corrected for multiple comparisons, they might not be meaningful.

If the findings hold up, it might show that mast cell dysfunction is causative of the disease to some extent, or it might just be a downstream effect of the disease, only causing specific symptoms. It would be interesting to see how much overlap there is between values in patients and controls ... if there is some overlap, it would be a less interesting result. Looking at the gene(s) for our mast cells also might help in determining if dysfunction is the cause of the disease or a result of the diseases.

What I do like is that they included severe patients and evaluated them separately. But this weakens the statistical power, especially with a small number of patients and controls, so again, the results might not be meaningful.

It also might be fun to look into values post-exercise, etc, if associations in this study are looking a bit weak.

 

[Kyla]

I'm glad to see this. I hope someone does a study looking for Mast cell mediators as well ( methyl-histamine, prostaglandin, leukotrienes etc).

I don't know if Mast cell activation is "the" answer, ( or a subset, or represents people with Primary MCAD or mastocytosis who have been misdiagnosed ) but I know anecdotally that mast-cell treatments have been more effective for me than anything else.

 

[Strawberry]

Would this be a blood test that is as definitive as the bone marrow test that Dr Afrin performs?

I'd be willing to donate blood to them, as I all ready have to OMI. I'm tired of how slowly a diagnosis moves with this disease. Allergy wise I am finally starting to feel better, but I still can only stand a few minutes before I am visibly shaking... I want a 100% diagnosis, not an "I think you have this." I know, I ask too much....

 

[Vasha]

Could someone explain what the findings mean (assuming that they are true), and to what degree they are consistent with what we know about ME/CFS?

@A.B. Sorry for not saying more about why I will be excited if this pans out over time...@Valentijn covered much of what I was thinking:

Hard to guess, without seeing the full paper. It doesn't seem to be published yet.
If the findings hold up, it might show that mast cell dysfunction is causative of the disease to some extent, or it might just be a downstream effect of the disease, only causing specific symptoms. It would be interesting to see how much overlap there is between values in patients and controls ... if there is some overlap, it would be a less interesting result. Looking at the gene(s) for our mast cells also might help in determining if dysfunction is the cause of the disease or a result of the diseases.

Connecting mast cell issues to ME would provide more evidence of immune dysfunction, and more information about how the disease works.

Obviously, this is a very small, nascent study. But if mast cells turn out to be important, then I would be very excited about the practical benefits:

-If mast cell dysfunction turns out to be a feature of the disease (maybe in a subset, maybe more), then we'd have something that is testable, that can and should be tested for in all patients, and that . . .

-can be TREATED. Like so many things we deal with, treatment is trial and error, and imperfect, but it is treatable.
Treating MCAS has made a big difference in my quality of life, and it would be great if that were true for others!

It also might be fun to look into values post-exercise, etc, if associations in this study are looking a bit weak.

Yes! Also to see if mast cell degranulation/activation contributes to PEM.

So... can't get the cart before the horse, but I do hope that this team and/or others keep pulling on this thread to see where it goes.

Vasha

 

[justy]

@paolo - have you seen this thread?

Thanks fr tagging me in @Vasha - this is my dilemma currently. I have MCAS, Lyme, EDS and M.E - my M.E dr says MCAS is because of Lyme (known through research to cause mast cell degranulation). EDS is also more common in people with MCAD'S.

Afrin believes MCAS is always a primary dx - in his book he says he doesn't go along with the view that it is secondary. My Lyme dx is clinical. Do I reall have Lyme or ME? or do I have primary MCAS? with no Dr in the UK capable of dx and treating as per Afrin then their is no hope to ever find out.

 

[Vasha]

@paolo - have you seen this thread?

Thanks fr tagging me in @Vasha - this is my dilemma currently. I have MCAS, Lyme, EDS and M.E - my M.E dr says MCAS is because of Lyme (known through research to cause mast cell degranulation). EDS is also more common in people with MCAD'S.

Afrin believes MCAS is always a primary dx - in his book he says he doesn't go along with the view that it is secondary. My Lyme dx is clinical. Do I reall have Lyme or ME? or do I have primary MCAS? with no Dr in the UK capable of dx and treating as per Afrin then their is no hope to ever find out.

Yes, relate to this--it's so confusing, and then hard to know what to prioritize.

For treatment purposes, I've mostly given up on trying to figure out what's "primary" and what's "secondary" because I think it's going to be a long time before we know (and it may vary a lot by patient).

Dr. Afrin's book is great. I do think, however, that experts can see a lot of nails to hit with whatever hammer they favor--natural human instinct for him to see what he discovered as primary. E.g., maybe there's a genetic weakness that predicts both EDS and MCADs? Of course, if something is truly primary, then it has to be fixed for everything else to be fixed.

Vasha
P.S. My "like" doesn't apply to the "no Dr in the UK" part!--I'm sorry, @justy.

 

[paolo]

@paolo - have you seen this thread?

No, I didn't know about this study. I'm reading.

By the way, I've wrote this post about a possible link between post-exertional malaise and mast cells mediators release.

 

[justy]

No, I didn't know about this study. I'm reading.

By the way, I've wrote this post about a possible link between post-exertional malaise and mast cells mediators release.

I read your post about this earlier, but right now I cant get that link to work.

 

[paolo]

I read your post about this earlier, but right now I cant get that link to work.

Now the link should work! Thanks.

 

[justy]

Now the link should work! Thanks.

yes that's great - working now, so more people can read it.

I am surprised more people on this forum aren't interested in Mast cell issues - they are quite common it seems in PWME, people with untreated Lyme, people with EDS - there is quite an overlap.

Having hung around on masto/MCAD sites for a while now I am struck at how similar some of their symptoms and issues are to people with M.E - including severe ME. They have patients who are bedbound and cant do any exercise etc. I don't feel at all out of place there.

 

[ahmo]

Thx @paolo Great image on your blog.

 

[paolo]

I am surprised more people on this forum aren't interested in Mast cell issues - they are quite common it seems in PWME, people with untreated Lyme, people with EDS - there is quite an overlap.

I think that mast cells and their mediators will be studied in ME/CFS in the following months/years. The paper by Griffith University is the very first one which provides experimental data on mast cells in CFS patients. But I've read somewhere that dr. Hornig is now interested in histamine, too. Her hypotesis is that histamine may be one possible root cause for orthostatic intollerance in CFS, I've read. Moreover, the NIH study also includes serum tryptase mesures. So, there is now some interest in the ME/CFS scientific comunity for mast cells and their mediators.

 

[Bob]

I am surprised more people on this forum aren't interested in Mast cell issues - they are quite common it seems in PWME, people with untreated Lyme, people with EDS - there is quite an overlap.

I think it's just a case of there not being much or any (?) research on mast cells in ME/CFS patients.