Passive transfer of fibromyalgia symptoms from patients to mice, 2021, Goebel et al

[SNT Gatchaman]

Unfortunately videos pem me too fast now, I can only read

A shame as I was going to suggest Dr Been's recently updated video series on immunology. Perhaps you might be able to listen to them, though the strength of his style is with the approachable cartoons he draws. He speaks softly and slowly which may well be appreciated by others here interested in this very complex subject. He's been making videos on many medical subjects for over 11 years. Here's the link to all of his lectures on immunology over the years.

The recent ones are —
Your Immune System (Part 1)
Your Immune System (Part 2)

It is a lifetime's learning getting to see all the ways antibodies can cause trouble.

I've taken the liberty of quoting Prof's earlier answer, and added some hopefully relevant Wikipedia links for us all.

Prof JE wrote earlier —

I would need a whole immunology textbook and the ones out there don't always make the simple things clear.

Antibodies bind antigen and can simply block its function without generating any inflammation.
To generate inflammation antibodies need to bind either complement or macrophage Fc receptors. They will do either or both if several antibody molecules are brought together - either bringing arms of complement C1q together or cell surface Fc receptors together.

Binding complement in the circulation is anti-inflammatory because the activated complement binds clearing receptors on red cells (CR1). Binding complement outside circulation releases C3a and C5a (chemotaxis) and binds to receptor CR3 leading to cell activation. Binding of Fc receptors leads to cytokine release - TNF and IL-1 - which produces much more destructive inflammation than complement.

The antibodies in RA are directed to immunoglobulin itself (Fc portion) and called rheumatoid factors. There are also antibodies to citrullinated peptide sites. The detail is very complicated but in simple terms the autoantibodies in RA are small enough to evade clearance by complement receptor 1 but just big enough to activate cytokines through Fc receptor IIIa. That receptor is preferentially expressed in joint lining - which is why it is a disease mostly of joints. Complexes of similar size contribute to lupus, which also often includes arthritis.

The complexes most typical of lupus are large. They cause damage because the clearing receptor CR1 is [not] operating effectively. That means that complexes can get big enough in the circulation to silt up in the kidney and brain with nephritis and encephalitis (never seen in RA where CR1 is working properly).

 

[Mister Person]

A shame as I was going to suggest Dr Been's recently updated video series on immunology. Perhaps you might be able to listen to them, though the strength of his style is with the approachable cartoons he draws. He speaks softly and slowly which may well be appreciated by others here interested in this very complex subject. He's been making videos on many medical subjects for over 11 years. Here's the link to all of his lectures on immunology over the years.

The recent ones are —
Your Immune System (Part 1)
Your Immune System (Part 2)



I've taken the liberty of quoting Prof's earlier answer, and added some hopefully relevant Wikipedia links for us all.

Prof JE wrote earlier —

I would need a whole immunology textbook and the ones out there don't always make the simple things clear.

Antibodies bind antigen and can simply block its function without generating any inflammation.
To generate inflammation antibodies need to bind either complement or macrophage Fc receptors. They will do either or both if several antibody molecules are brought together - either bringing arms of complement C1q together or cell surface Fc receptors together.

Binding complement in the circulation is anti-inflammatory because the activated complement binds clearing receptors on red cells (CR1). Binding complement outside circulation releases C3a and C5a (chemotaxis) and binds to receptor CR3 leading to cell activation. Binding of Fc receptors leads to cytokine release - TNF and IL-1 - which produces much more destructive inflammation than complement.

The antibodies in RA are directed to immunoglobulin itself (Fc portion) and called rheumatoid factors. There are also antibodies to citrullinated peptide sites. The detail is very complicated but in simple terms the autoantibodies in RA are small enough to evade clearance by complement receptor 1 but just big enough to activate cytokines through Fc receptor IIIa. That receptor is preferentially expressed in joint lining - which is why it is a disease mostly of joints. Complexes of similar size contribute to lupus, which also often includes arthritis.

The complexes most typical of lupus are large. They cause damage because the clearing receptor CR1 is [not] operating effectively. That means that complexes can get big enough in the circulation to silt up in the kidney and brain with nephritis and encephalitis (never seen in RA where CR1 is working properly).

Paki paki matua. There's a website called British society for immunology with byte sized immunology, also rather nice. https://www.immunology.org/public-information/bitesized-immunology
On that note I realise my pem isn't energy based in the same way cfs is... Whitney Dafoe can barely walk, or interact with humans but he can listen to music and watch TV all day. Yet I can't listen to music, watch TV, but I can get out of bed for more than 15 minutes and stay up for hours. Mine seems more mental/nerve based.

 

[JemPD]

Paki paki matua. Whitney Dafoe can barely walk, or interact with humans but he can listen to music and watch TV all day. Yet I can't listen to music, watch TV, but I can get out of bed for more than 15 minutes and stay up for hours. Mine seems more mental/nerve based.

I'm surprised to read that Witney can listen to music all day. Many many people with severe ME are hugely sensory sensitive & can barely tolerate any sound/visual movement at all. Lots of PwME say they get PEM from too much sensory input like sound/light etc so getting PEM from sensory input isnt 'non' ME.

 

[Mister Person]

I see. Fibro = Cs, cs= glia, LDN = glia, fibro =LDN=glia. Why have I not been told this before. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6051899/
Cited 600 times. Mainstream paper that briefly touched on fibro

 

[RedFox]

I'm not terribly surprised personally. ME can skew towards physical or mental impairments. Unfortunately there's no statistics on this. I can walk for 30-45 minutes without getting PEM, but I have trouble taking care of myself due to cognitive impairment, so I can grasp that someone can only leave bed to use the bathroom but can listen to music all day.

 

[voner]

here is an follow on study.

Fibromyalgia patients with elevated levels of anti–satellite glia cell immunoglobulin G antibodies present with more severe symptoms, PAIN, Krock, Emerson, et al, Karolinska Institutet, Stockholm, Sweden

https://journals.lww.com/pain/Fulltext/9900/Fibromyalgia_patients_with_elevated_levels_of.274.aspx

open source. I have not read it yet, but I did pick out these two paragraphs. The first is about the use of pooled samples and IgG binding that has been discussed here:

In this study, we did not find a difference between FM IgG and HC IgG binding to neurons. We had previously reported elevated IgG binding to sensory neurons in vitro, but these experiments were done using pooled IgG preparations. When looking at the data points from single individuals in this study, many individuals have no or low levels of neuron binding IgG, but there are also individuals who have higher levels of IgG binding to neurons. It remains possible that a subset of FM patients have anti-neuron antibodies. Regardless, if autoantibodies binding SGCs are driving nociceptor hypersensitivity, then they must do so by indirectly activating SGCs or by activating neuronally expressed Fcγ receptors.5,48

This is their final paragraph in the Results section:

We found that anti-SGC IgG levels are elevated in 2 distinct FM cohorts. The level of these antibodies correlated with a more severe clinical presentation, while being unrelated to FM duration. Moreover, the level of anti-SGC antibodies detected in culture correlated with binding to human SGCs. These results, combined with our previous findings that FM IgG induces pain-like behaviour in mice, provide a possible answer to one mechanism of nociplastic pain in FM. Taken together, the data suggest that testing FM patients for elevated levels of anti-SGC IgG may be useful to stratify patients for therapies that interfere with antibody function such as B cell depleting drugs, IVIg, or plasmapheresis.

 

[Andy]

here is an follow on study.

Fibromyalgia patients with elevated levels of anti–satellite glia cell immunoglobulin G antibodies present with more severe symptoms, PAIN, Krock, Emerson, et al, Karolinska Institutet, Stockholm, Sweden

https://journals.lww.com/pain/Fulltext/9900/Fibromyalgia_patients_with_elevated_levels_of.274.aspx

open source. I have not read it yet, but I did pick out these two paragraphs. The first is about the use of pooled samples and IgG binding that has been discussed here:



This is their final paragraph in the Results section:

Discussion thread here, Fibromyalgia patients with elevated levels of anti–satellite glia cell immunoglobulin G antibodies present with more severe symptoms 2023 Krock et al

 

[Hutan]

A 2023 study failed to replicate the findings of the 2021 study:
Neutrophils infiltrate sensory ganglia and mediate chronic widespread pain in fibromyalgia, 2023, Sara Cavaria et al

 

[Mister Person]

https://classic.clinicaltrials.gov/...df&type=Intr&cond=Fibromyalgia&draw=3&rank=11
If this trial works, fibro will be autoimmune confirmed for some I think
Frcn inhibitor, as goebel described may help. Someone is running a trial, I wonder if he was involved

 

[Trish]

https://classic.clinicaltrials.gov/...df&type=Intr&cond=Fibromyalgia&draw=3&rank=11
If this trial works, fibro will be autoimmune confirmed for some I think
Frcn inhibitor, as goebel described may help. Someone is running a trial, I wonder if he was involved

Thread:
A Proof-of-concept Study to Evaluate the Efficacy and Safety of Rozanolixizumab to Treat [Adults] With Severe Fibromyalgia Syndrome, 2022-4, UK

 

[Sid]

I thought this had failed replication. Now they’re jumping from mice to interventional human trials?

 

[Mister Person]

I thought this had failed replication. Now they’re jumping from mice to interventional human trials?

Failed replication in a sample size of like 3? Was it? It was tiny and look at voners comment

 

[ME/CFS Skeptic]

Failed replication in a sample size of like 3? Was it?

This seems to be the data of the replication attempt, found in supplementary material of this paper: Neutrophils infiltrate sensory ganglia and mediate chronic widespread pain in fibromyalgia | PNAS


S5. Passive transfer of 8mg IgG derived from a fibromyalgia patient vs. pain free control into naïve recipient mice (n=4 mice per group). No differences in sensory thresholds were observed over a 7-day period to (i) punctate mechanical stimulation or (ii) noxious heat stimulation (P>0.05 for all time points 2 way RM ANOVA).

 

[ME/CFS Skeptic]

https://classic.clinicaltrials.gov/...df&type=Intr&cond=Fibromyalgia&draw=3&rank=11
If this trial works, fibro will be autoimmune confirmed for some I think
Frcn inhibitor, as goebel described may help. Someone is running a trial, I wonder if he was involved

Looks like the initiative came from the Belgian company UCB Biopharma, the producer of the drug. More info here: Rozanolixizumab by UCB for Fibromyalgia (Fibromyalgia Syndrome): Likelihood of Approval (pharmaceutical-technology.com)

So unsure if there was a connection to the mice study of Goebel.

 

[chillier]

Failed replication in a sample size of like 3? Was it? It was tiny and look at voners comment

This seems to be the data of the replication attempt, found in supplementary material of this paper: Neutrophils infiltrate sensory ganglia and mediate chronic widespread pain in fibromyalgia | PNAS

View attachment 19792
S5. Passive transfer of 8mg IgG derived from a fibromyalgia patient vs. pain free control into naïve recipient mice (n=4 mice per group). No differences in sensory thresholds were observed over a 7-day period to (i) punctate mechanical stimulation or (ii) noxious heat stimulation (P>0.05 for all time points 2 way RM ANOVA).

This n=4 value is referring to the number of mice and not the number of patients so they are effectively technical replicates. Looking through the paper it's weirdly difficult to find out how many patients and controls they're using it's not even mentioned in the methods. They mention it only in the figure 5 legend when they do adoptive transfer of neutrophils (in bold).

Adoptive transfer of neutrophils from primed mice confers widespread pain in naïve mice. (A) Neutrophils were isolated from patients with fibromyalgia (n = 9) and healthy control subjects (n = 5) and administered into naïve male and female mice (1 × 106 cells i.v. in PBS suspension). Cells from each subject were injected into four mice. (B) Pain visual analogue scores (VAS) of healthy control subjects and subjects with fibromyalgia. (C) Mechanical hypersensitivity to von Frey stimulation of mice following i.v. adoptive transfer of neutrophils (*P < 0.05, ***P < 0.001 two-way RM ANOVA control vs. patient cells). (D) Latency to nociceptive behavior in the hot plate assay (two-way RM ANOVA primed vs. control). (E) Evoked action potential firing of neurons in the deep dorsal horn of naïve mice (n = 16 cells) before (open circles) and 1 h after (red circles) administration of neutrophils from patients. Total number of action potentials fired over 10 s stimulation with (i) innocuous brush, (ii) noxious thermal stimuli, (iii) punctate von Frey stimuli, and (iv) noxious mechanical stimuli (*P < 0.05, **P < 0.01 paired t test). (F, i) Ex vivo imaging of neutrophil cell counts in L4 DRG following adoptive transfer of neutrophils from patient (n = 4 mice) or control blood (n = 3 mice). White line illustrates 50 µm. (*P < 0.05 unpaired t test). (ii) Representative of mean images for (F, i).

The only context we get for the igG adoptive transfer is a throwaway line in the discussion,

Recent work indicates an immunological basis for fibromyalgia symptoms as passive IgG transfer from patients to mice can confer mechanical pain behavior (48), although we were unable to reproduce these findings (SI Appendix, Fig. S5).

and their igG extraction protocol in the methods (no mention of numbers or pooling)

The concentration of IgG present after dialysis was determined using a BCA assay and adjusted by dilution with PBS. The IgG solution was stored at 4 °C until i.p. administration (8 mg) into naïve mice.

Unless I missed something, it could just be a single patient and control, or my guess would be that they took n = 9 patients and n = 5 controls as they did in figure 5, and then pooled them.

 

[chillier]

The figure to compare it to in the Goebel & Andersson paper would be figure 3 panels C and D:

These show mechanical sensitivity and heat sensitivity as in the corresponding figure @me-cfs-skeptic just posted - but it is using a cold plate rather than a hot plate. The data here correspond to day 4 in that figure. Each of these dots is a technical replicate (a mouse) from a pooled cohort. They show two cohorts here and they're about the same size as the cohort (I think is) used above (Pool 1, n = 8 FMS and n = 12 HC; Pool 2, n = 14 FMS and n = 10 HC).

As above, they inject with 8mg purified igG. In contrast however they do this 4 times on 4 consecutive days in the Goebel paper. So I think a crucial difference may be that they are using 4x less igG in the replication than in the Goebel paper.

Then again, they are vague about what they're doing in the replication so it could be they do this too (but they certainly don't say so)

 

[Mister Person]

Hey PEJE. 'This latter model of increasing insular glutamate was recently reverse translated to show that findings of “small fiber neuropathy” could be induced simply by increasing CNS glutamate, as is known to occur in FM' this mean anything? Re sfn talk a while back

 

[Mister Person]

@Jonathan Edwards also, am I right on this a small trial with a negative outcome is treated as that it is true, but a small trial with a positive outcome is treated as if it's false. Why do we do this? Do we assume that if something is effective it should show through in the small sample?

 

[SNT Gatchaman]

See follow-up study Postacute COVID-19 syndrome and fibromyalgia syndrome are associated with anti-satellite glial cell IgG serum autoantibodies but only fibromyalgia syndrome serum-IgG is pronociceptive (2025, PAIN)