Phenotypic characteristics of peripheral immune cells of ME/CFS via transmission electron microscopy: A pilot study, Jahanbani et al, 2022

cassava7

cassava7

Senior Member (Voting Rights)
Fereshteh Jahanbani, Rajan D. Maynard, Justin Cyril Sing, Shaghayegh Jahanbani, John J. Perrino, Damek V. Spacek, Ronald W. Davis, Michael P. Snyder

Received: January 17, 2022; Accepted: July 25, 2022; Published: August 9, 2022

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Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a complex chronic multi-systemic disease characterized by extreme fatigue that is not improved by rest, and worsens after exertion, whether physical or mental. Previous studies have shown ME/CFS-associated alterations in the immune system and mitochondria.

We used transmission electron microscopy (TEM) to investigate the morphology and ultrastructure of unstimulated and stimulated ME/CFS immune cells and their intracellular organelles, including mitochondria. PBMCs from four participants were studied: a pair of identical twins discordant for moderate ME/CFS, as well as two age- and gender- matched unrelated subjects—one with an extremely severe form of ME/CFS and the other healthy.

TEM analysis of CD3/CD28-stimulated T cells suggested a significant increase in the levels of apoptotic and necrotic cell death in T cells from ME/CFS patients (over 2-fold). Stimulated Tcells of ME/CFS patients also had higher numbers of swollen mitochondria.

We also found a large increase in intracellular giant lipid droplet-like organelles in the stimulated PBMCs from the extremely severe ME/CFS patient potentially indicative of a lipid storage disorder.

Lastly, we observed a slight increase in platelet aggregation in stimulated cells, suggestive of a possible role of platelet activity in ME/CFS pathophysiology and disease severity.

These results indicate extensive morphological alterations in the cellular and mitochondrial phenotypes of ME/CFS patients’ immune cells and suggest new insights into ME/CFS biology.

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0272703
 
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It seems that lipid accumulation or abnormalities that suggest lipid accumulation might be occurring keep coming up in various studies. A patient here on the forum also had a muscle biopsy which showed increased lipids and was not definitive enough for another diagnosis.
 
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I am still reading this, but the whole excercise seems not scientifically valid, but I am not educated in these areas, so my question is can they validly make this claim? They are using so few data (2) points and the patients are so far apart on the illness spectrum. I have noticed a typo and some poor grammer usage, which always makes me suspicious also…

from the paper:

Interestingly, when compared to moderately affected twins, the percentage of stimulated T cells from the extremely severe ME/CFS carrying more than 6 abnormal mitochondria per cell was 2-fold higher (18% vs 38%, respectively) (S7 Table). These results suggest a positive correlation between disease severity and the extent of mitochondrial damage at single cell level after stimulation (S7 Table).
 
voner said:
my question is can they validly make this claim? They are using so few data (2) points and the patients are so far apart on the illness spectrum.
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I am treating it as an interesting initial study. It needs replication in a larger group, and probably a whole lot of other things. Yet such investigations have to start somewhere.

The implications if this can be validated in more patients with a greater range of severity are considerable.

I do want to remind people again that most such initial studies fail on follow up. Yet we need that follow up. Ignoring findings is not a good idea. We need to test them and see if they fail at some point.

I am amused that they are discussing lipid storage disease. My first proposed model of ME used Tarui disease (a glycogen storage disorder) as an analogy. Using what we know about lipid storage diseases we might be able to make guesses about what is happening.
 
Looking carefully at cells with an electron microscope seems like a good thing to do. There were a lot of preparatory steps, and with such a small sample (2 patients, 2 controls), I think there's a real risk that an inconsistency in those steps might result in observed differences e.g. more cells dying.

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The differences in mitochondria look interesting (A - healthy mitochondria; B and C, not healthy). It's not that healthy controls didn't have some disordered looking mitochondria, it's that the ratios of healthy to unhealthy mitochondria are quite different.
However, ME/CFS stimulated T cells showed substantially higher level of swollen and abnormal mitochondria (vesicular/ compartmentalized and/or swollen mitochondria) (1.9- and 1.8-fold increase with a p-value = 0.00004 and p-value = 0.003, respectively)
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the number of stimulated T cells carrying more than 3 swollen mitochondria per cell was significantly higher in the ME/CFS group, (25% of the ME/CFS vs 5.3% healthy control cells contained more than 3 swollen mitochondria per cell) (p-value = 0.001)
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I thought the discussion and conclusion were thoughtfully written, and think they are worth reading. I hope this team will be funded to do similar studies on a larger sample and that other researchers will attempt to replicate and build on the findings.
 
4 people is too small to be a study per se. The authors are like, "We screwed around with an electron microscope and here's what we saw." It sounds like something a 19th-century scientist would do. Not to discount this technique--our knowledge of ME is stuck in the 19th century, and screwing around will generate a lot of hypotheses to test rigorously.

The burning question: Is this finding striking enough to attempt a larger experiment?
 
Looking at the cells of two people with ME is too little to draw even the barest conclusion. It all just feels so anecdotal, whether or not it meets some official definition of a study. Maybe I don't know what I'm talking about.

Jaybee00 said:
Do you think they would have secured funding to examine 100 patients without having preliminary data?
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Certainly not. This is precisely why such research is valuable--to generate a hypothesis and attract the funds to test it.
 
Much good quality information can be developed from in-depth studies of even one subject. One of the particular strengths of this paper is that they were able to look at a monozygotic twin pair: one with, one without ME. That removes a lot of potential uncertainty. And they could compare those findings on a scale of healthy to very severe (with the affected twin as moderate).
 
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RedFox said:
Looking at the cells of two people with ME is too little to draw even the barest conclusion. It all just feels so anecdotal, whether or not it meets some official definition of a study. Maybe I don't know what I'm talking about.


Certainly not. This is precisely why such research is valuable--to generate a hypothesis and attract the funds to test it.
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Agreed re: N=2. We do know that we are extremely heterogenous and missed diagnosis happen. We certainly cannot draw any conclusion from this paper or generalize to our community.

Further comments:
The 2 subjects investigated are men and within ME women are over-representated by a 4 to one ratio. Moreover, it has been established that men and women have different metabolic findings.

Also: There are very few details about the participants, other than age, sex and the fact that they were successfully employed prior to illness. It is unclear whether they had an infectious or gradual onset and does it matter? (I would want to know) Then one would like to know especially for the very severe, his nutritional status (BMI, feeding type: oral, gastric or j-tube- or parenteral) and whether that matters.
 
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Jaybee00 said:

Not sure what this means. This is a pilot study—see title “Phenotypic characteristics of peripheral immune cells of ME/CFS via transmission electron microscopy: A pilot study”

Do you think they would have secured funding to examine 100 patients without having preliminary data?
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The problem with this is that Ron Davis has been touting for the last 6 years that they want to fast track research and find, and i quote from back in 2018, "hopefully a cure within months". This has allowed them to raise more than 30 million dollars from patients.

I get it that it's a pilot study but they could have done at least a bunch more patients.
 
Milo said:
this work, particularly the electron microscopy is detailed and labor intensive, and not something you'd do by the hundreds.
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Its probably only useful to establish an hypothesis, but as more and more are studied with similar findings then it might be considered a stronger hypothesis. What I suspect they might try is to find potential biochemical signatures. They can mass test those using more developed methods.

First see what might be wrong in a small sample. Then develop a further hypothesis that is easily testable. Then major studies can begin.
 
cassava7 said:
TEM analysis of CD3/CD28-stimulated T cells suggested a significant increase in the levels of apoptotic and necrotic cell death in T cells from ME/CFS patients (over 2-fold). Stimulated Tcells of ME/CFS patients also had higher numbers of swollen mitochondria.

We also found a large increase in intracellular giant lipid droplet-like organelles in the stimulated PBMCs from the extremely severe ME/CFS patient potentially indicative of a lipid storage disorder.
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@Cohen, maybe things to explore with Professor Tate, who will have access to the electron microscope technology?
 
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