Biomarkers in the diagnostic algorithm of myalgic encephalomyelitis/chronic fatigue syndrome, 2022, Gravelsina et al

[SNT Gatchaman]

Biomarkers in the diagnostic algorithm of myalgic encephalomyelitis/chronic fatigue syndrome
Gravelsina Sabine, Vilmane Anda, Svirskis Simons, Rasa-Dzelzkaleja Santa, Nora-Krukle Zaiga, Vecvagare Katrine, Krumina Angelika, Leineman Iana, Shoenfeld Yehuda, Murovska Modra

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a complex disease that is mainly diagnosed based on its clinical symptoms. Biomarkers that could facilitate the diagnosis of ME/CFS are not yet available; therefore, reliable and clinically useful disease indicators are of high importance. The aim of this work was to analyze the association between ME/CFS clinical course severity, presence of HHV-6A/B infection markers, and plasma levels of autoantibodies against adrenergic and muscarinic acetylcholine receptors.

A total of 134 patients with ME/CFS and 33 healthy controls were analyzed for the presence of HHV-6A/B using PCRs, and antibodies against beta2-adrenergic receptors (β2AdR) and muscarinic acetylcholine receptors (M3 AChR and M4 AChR) using ELISAs. HHV-6A/B U3 genomic sequence in whole-blood DNA was detected in 19/31 patients with severe ME/CFS, in 18/73 moderate ME/CFS cases, and in 7/30 mild ME/CFS cases. Severity-related differences were found among those with a virus load of more than 1,000 copies/1,000,000 PBMCs.

Although no disease severity-related differences in anti-β2AdR levels were observed in ME/CFS patients, the median concentration of these antibodies in plasma samples of ME/CFS patients was 1.4 ng/ml, while in healthy controls, it was 0.81 ng/ml, with a statistically significant increased level in those with ME/CFS (p = 0.0103). A significant difference of antibodies against M4 AChR median concentration was found between ME/CFS patients (8.15 ng/ml) and healthy controls (6.45 ng/ml) (p = 0.0250). The levels of anti-M4 plotted against disease severity did not show any difference; however, increased viral load correlates with the increase in anti-M4 level.

ME/CFS patients with high HHV-6 load have a more severe course of the disease, thus confirming that the severity of the disease depends on the viral load—the course of the disease is more severe with a higher viral load. An increase in anti-M4 AchR and anti-β2AdR levels is detected in all ME/CFS patient groups in comparison to the control group not depending on ME/CFS clinical course severity. However, the increase in HHV-6 load correlates with the increase in anti-M4 level, and the increase in anti-M4 level, in turn, is associated with the increase in anti-β2AdR level. Elevated levels of antibodies against β2AdR and M4 receptors in ME/CFS patients support their usage as clinical biomarkers in the diagnostic algorithm of ME/CFS.

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[SNT Gatchaman]

Fukuda

 

[Andy]

"ME/CFS patients with high HHV-6 load have a more severe course of the disease, thus confirming that the severity of the disease depends on the viral load—the course of the disease is more severe with a higher viral load."

Or the immune system of someone who is less well will find it harder to control or eliminate infections that are potentially secondary.

 

[InitialConditions]

"ME/CFS patients with high HHV-6 load have a more severe course of the disease, thus confirming that the severity of the disease depends on the viral load—the course of the disease is more severe with a higher viral load."

Or the immune system of someone who is less well will find it harder to control or eliminate infections that are potentially secondary.

Indeed. Much of the science that gets posted here is shockingly bad, isn't it?

 

[Andy]

Indeed. Much of the science that gets posted here is shockingly bad, isn't it?

Sadly, yes. But if we, at least, don't point that out there is little counterpoint to the almost inevitable hype.

 

[BrightCandle]

Indeed. Much of the science that gets posted here is shockingly bad, isn't it?

I think its worse than just its bad. A lot of it is on already discredited approaches and the only way to get them to show a result is to set the study up with a lot of bias, looks like intentional fraud. The studies in progress don't look any better, NICE 2021 hasn't changed much.

 

[Creekside]

and in 7/30 mild ME/CFS cases.

If 23/30 mild cases didn't show the virus, then maybe it's not part of ME? My guess is that any immune system activation, from viral infections, tissue damage, or whatever else, will increase the severity of ME symptoms. It's just an extra load on top of ME's symptoms due to the immune system not working properly.

They should take the evidence that people can have ME without any of the viral infections that they keep studying (because it's easy to do and get funding for?) and stop funding ME research based on viral infection.

 

[Hutan]

The abstract doesn't tell us about the incidence of HHV-6A/B U3 in controls.

From all the cohort, HHV-6A/B U3 genomic sequence in whole-blood DNA was detected in 9/33 (28.3%) control group individuals and in 44/134 (32.8%) patients with ME/CFS

When the incidence in both patients and controls is reported, HHV-6A/B U3 doesn't look that important. It was more common in people with severe ME/CFS, but actually substantially less common in people with moderate and mild ME/CFS. Of course, the diagnostic criteria make firm conclusions difficult, but, as @Andy says, it is looking more like HHV-6A/B U3 levels being a downstream consequence than a simple driver of the disease.

 

[Hutan]

The results showed that the median HHV-6 load in whole blood from patients with mild, moderate, and severe ME/CFS disease was 4,138 copies/106PBMCs (IQR: 19–8,244), 7,302 copies/106 PBMCs (IQR: 668–15,045), and 5,498 copies/106 PBMCs (IQR: 1,158–9,048), respectively, and no severity-related differences were found among these patients. Nevertheless, severity-related differences were found among those with a viral load of more than 1,000 copies/106PBMCs.
Proportion analysis showed that 84.2% of ME/CFS patients with severe clinical course, 72.2% with moderate clinical course, and 57.1% with mild clinical course had a viral load of >1,000 copies/106 PBMCs in comparison to the control group where only one of HHV-6-positive individuals (11.1%) had a viral load of >1,000 copies/106 PBMCs

That's some pretty fine slicing and dicing to get to a claim that there was a severity difference in HHV-6 load.

Actually, there was no relationship between viral load and severity. It was only when they looked at people with a viral load of more than 1000 copies that they found such a relationship.

 

[Hutan]

The median concentration of anti-muscarinic cholinergic receptor 3 (anti-M3) antibodies in ME/CFS patients’ plasma samples was 6.75 ng/ml, while in healthy donors, it was 2.29 ng/ml, without a significant difference between cases of ME/CFS and healthy controls (Figure 2A), as well as between female and male patients (Figure 2B), and without significant differences regarding disease severity (Figure 2C). ROC analysis did not reveal anti-M3 as a discriminating marker for ME/CFC (Figures 2D, E).

They found no evidence that concentrations of anti-muscarinic cholinergic 3 receptors are relevant to ME/CFS.

 

[Hutan]

Significant differences of anti-muscarinic cholinergic receptor 4 (anti-M4) antibody concentration was found between ME/CFS patients and healthy donors (p = 0.0029); the median concentration of anti-M4 in ME/CFS patients’ plasma samples was 8.95 ng/ml, while that in healthy donors was 6.45 ng/ml (Figure 3A). Anti-M4 levels did not differ significantly between the ME/CFS female patients and women from the donor group; however, significant differences (p = 0.0199) were found between the male groups (Figure 3B). Regarding the severity of disease, the median levels of anti-M4 (Figure 3C) were almost the same in all three subgroups of patients—9.64, 8.00, and 10.62 ng/ml for mild, moderate, and severe ME/CFS, respectively. ROC analysis showed that, in contrast to anti-M3, anti-M4 could serve as a good marker for ME/CFS in general (Figure 3D); however, it could not serve as a discriminating factor regarding ME/CFS severity (Figure 3E).

They did find a significant difference between the ME/CFS and control groups in concentrations of anti-muscarinic cholinergic 4 receptors. However, there was a lot of overlap and the differences aren't large. If anything, it looks like there is a relatively small subset of people labelled with ME/CFS with higher values than were found in the small control group. The study is not convincing evidence on its own.

 

[Hutan]

A significant (p < 0.0001) increase in anti-β2AdR levels was observed in ME/CFS patients (1.4 ng/ml) compared to healthy controls (0.81 ng/ml) (Figure 4A), and these statistical differences were also found in the respective female and male groups (Figure 4B). However, no differences in disease severity-related anti-β2AdR levels were observed in ME/CFS patients (Figure 4C). The ROC analysis showed that, in this study, anti-β2AdR has the best potential to be a good marker of ME/CFS in general (Figure 4D), but, as with anti-M4, cannot be a discriminating factor for the severity of ME/CFS (Figure 4E).

They found a significant difference between the ME/CFS and control groups in concentrations of anti-B2 adrenergic receptors. There actually is pretty convincing separation in this - it's worth having a look at the charts. I think there have been other studies on these receptors that have been less convincing, but I think this finding is one to note.

Although no disease severity-related differences in anti-β2AdR levels were observed in ME/CFS patients, the median concentration of these antibodies in plasma samples of ME/CFS patients was 1.4 ng/ml, while in healthy controls, it was 0.81 ng/ml, with a statistically significant increased level in those with ME/CFS (p = 0.0103).