Deep Sequencing of BCR Heavy Chain Repertoires in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome, 2025, Ryback et al

I’m really liking Audrey Ryback’s work. Especially how she hedges her claims and doesn’t overstate findings. I really hope she stays in the field.

Edit: I just realised she has a bluesky account.
https://bsky.app/profile/aryback.bsky.social

 

(I think this is a testament to how much impact Chris Ponting jumping into the field has made, having a very renowned researcher like him in the field winning big grants attracts talented PhD students and postdocs to work with him who then get experience in the field.)

 

I came across this tweet from the ME/CFS Biobank. In the linked video Dr Jaqueline Cliff presented data from her teams mostly NIH funded study on HHV6B saliva vs T-cell markers and found correlation between the two in Mild/Moderate ME/CFS and not in severe (slide 40mins in). I thought it was interesting in light of the IGHV30 finding being relevant only in Mild/Moderate in this study.
https://twitter.com/user/status/1899478703148535982

 

Are we aware of any consensus in these areas in ME/CFS (no matter how small?) What do we think about the TGF-β angle in 2025

 

Aha!
How about a consensus of 2 - Jo Cambridge and myself?
We are trying to put together a specific story to go out soon.

I think we can claim that TNF alpha, IL-1 and IL-6 are not major players, just from the negative findings on tissue change. A role for TGF beta and or GDF15 (probably suppressive) seems plausible especially if largely localised to lymphoid tissue and maybe muscle. There must be something activating at a very local cellular level that is not showing up in serum. We have some thoughts that but need to clarify them a bit.

 

THF-beta was mentioned here (I have no idea about the relevance): https://www.s4me.info/threads/tgfβ-...yndrome-in-children-2025-goetzke-et-al.43084/

 

One of the largest cytokine studies in ME/CFS was the Montoya Stanford study which showed consistent elevation of TGF-B across severities and strong p-value. The p-values were
0.0179 : Mild
0.0126 : Moderate
0.0089 : Severe

N = 192 ME/CFS, 392 Healthy control

That 2017 paper mentioned TGF-B was elevated in 5 of 8 studies. I don't see a thread for the paper.
https://pmc.ncbi.nlm.nih.gov/articles/PMC5576836/

 

Made a thread here:

Cytokine signature associated with disease severity in chronic fatigue syndrome patients, 2017, Montoya et al

 

Two leading experts sounds good to me.

I look forward to whenever your ideas are out there so that they can inform my own research ideas in a similar vein

When we both have the ideas solidified we should have a future, detailed chat (long overdue) -

- If it's something we can feasibly sample and assay from people I am open to discussing potential project ideas down the line if capability is needed

 

Uh-uh, I would want to be thought of as a 'leading expert'!

We are hoping to put something out quite soon, then everyone can chat about it. I am hoping that the DecodeME results will make it easier to assess models based on these existing ideas. Again, that should be soon.

One possible relevant assay might be Elispot - looking at cell-cell signalling in freshly obtained blood mononuclear cells.

 

I’m very late to this paper and thread, and not well enough to read through it all in detail. Does anyone know if anyone know if there is a good lay summary anywhere? If not, would anyone like to post one?

 

I think this comment by @forestglip is really good:
https://www.s4me.info/threads/seque...aria-infection-2024-ryback.37615/#post-593268

 

FYI - looks like there are some pitfalls with Elispot relating to sample processing time. Something to be aware of.
Optimization of Peripheral Blood Mononuclear Cell Processing for Improved Clinical ELISpot Assay Performance

 

Yes I am well aware of these. We worked Elispot technology early on around 1990. I am actually thinking of co-culture experiments where this sort of sample delay would not be that relevant.

 

I've seen @Jonathan Edwards in other contexts talk about how lymphocytes in the blood aren't expected to be very interesting, which makes sense to me, since as I understand it, those are mostly the naive ones still bouncing around, waiting to see their matching antigen.

In the case of B cells, especially where we've already seen differences in the BCRs here, it seems like one logical next step is doing a similar analysis of the differences in antibody gene segments, since the antibodies provide information about the more "interesting", activated B cells.

 

I am not sure that you can do that. If you purify IgG from ME/CFS patients you will get a preparation that contains maybe 100,000 different protein sequences all mixed together. In theory you could separate thereby electrophoresis but in practice nobody has ever got any clear bands doing that - you just get a smear of 100,000 overlapping indistinguishable bands.

in order to analyse sequences in proteins you need to so something like MALDI-TOF mass spectrometry and that only works if you have a single molecular species in your sample. If you are just looking for usage of something like VH3-30 or VH4-34 you can get some idea of the proportion of IgG using the gene product but it isn't that simple.

Maybe the most significant finding of this study, as far as I can see, is that it does not support the idea of a specific antigen-driven antibody response with full affinity maturation. To that extent it does not support a classical autoantibody response.

 

Interesting, thanks, I didn't know about any of those technical limitations.

 

Time for some more wild conjecture from someone with no medical training and little understanding…

Maybe all people with a susceptibility to ME/CFS have the imbalance (maybe not the right word) seen in mild/moderate and something about severe causes things to become balanced again?

Could there be a response in the body which kicks in and attempts to rebalance the imbalance seen and that is part of the disease mechanism? A part which has negative impacts elsewhere causing what we see as ME/CFS, and this part ‘wins’ in more severe ME?

 

Yes, and we have @Simon M to thank for Chris jumping in: https://theconversation.com/ignored...he-origins-of-a-modern-medical-scandal-241149 (Thread here)

 

One thing I’ve particularly enjoyed is papers which I’ve gone off and learned about the background and science involved in more detail. It can take many months. Picking up bits and revisiting and re-listening to papers but it’s really interesting.

I’m just starting to understand bits about B cells hence the focus of my questions/thoughts. But am also now starting to see where the complement system fits in and can see why there’s been interest here too. But that’s my next avenue of exploration. So much to learn!

So forgive me if I revisit things, I’ve been through the whole thread again too, but I have some questions/thoughts/notes which cropped up while doing so… (will post in a couple of posts later)