Exosome-associated Mitochondrial DNA is Elevated in Patients with ME/CFS and Stimulates Human Cultured Microglia to Secrete IL-1β

Messages
92
Likes
176
Location
The Netherlands
Exosome-associated Mitochondrial DNA is Elevated in Patients with ME/CFS and Stimulates Human Cultured Microglia to Secrete IL-1β (Theoharides, 2021)

Abstract Background: Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a debilitating disease that presents with fatigue, sleep disturbances, malaise and cognitive problems. The pathogenesis of ME/CFS is presently unknown and serum levels of potential biomarkers have been inconsistent.

Methods: Exosomes were puried from serum obtained from patients with ME/CFS before and after exercise and their content of mitochondrial DNA (mtDNA) was determined by quantitative PCR. Exosomes from both patients and controls were incubated with cultured human microglia and release of interleukin-1beta (IL-1β) was measured by ELISA.

Results: Here we show that serum mtDNA, associated with exosomes, is increased in ME/CFS after exercise. Moreover, exosomes isolated from patients with ME/CFS stimulate signicant secretion of IL-1β from cultured human microglia.

Conclusion: These results provide evidence for a potential novel pathogenetic factor and target for treatment of ME/CFS.
 

pattismith

Senior Member
Messages
3,276
Likes
6,312
paper from Bardsen published september 2020 (maybe from the same study)

Interleukin-1β, heat shock protein 90α, and hypocretin-1 in chronic fatigue

Methods:
To explore mechanism of fatigue, a cohort of 71 patients with primary Sjögren’s syndrome were investigated.
CSF samples where available from 49 patients.
A method based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed for measurements of Hcrt1.
Hcrt1 was measured in CSF samples from 22 healthy subjects and 9 patients with narcolepsy type 1.

Results: Measures of HSP32, -60, -72, and -90α in plasma revealed that the concentrations of HSP90α were significantly higher in pSS patients with high fatigue versus low fatigue. A tendency toward higher concentrations of HSP72 was observed in patients with high fatigue compared to patients with low fatigue.
....
Analysis of IL-1β related proteins (IL-1Ra, IL-1RII, and S100B), IL-6, and Hcrt1 in CSF demonstrated that IL-1Ra showed significant association with fVAS scores together with the clinical variables BDI scores and pain scores. The relationship of the biochemical variables was explored in PCA, and two significant components appeared: Variables related to IL-1β activity dominated the first component while in the second component there was a negative association between IL-6 and Hcrt1. Fatigue was introduced as an additional variable in a second model. In this PCA, fVAS scores were associated with the first component as was the IL-1β related variables. In addition, the second PCA model revealed a third component that showed a negative relationship between Hcrt1 and fatigue.


Conclusions:


I) HSP90α and to a lesser degree HSP72 in blood may possibly be parts of a fatigue inducing mechanism.


II) The LC-MS/MS method with high selectivity and accuracy revealed considerably lower levels of Hcrt1 in CSF than previously reported.


III) IL-1β signaling is a primary driver in fatigue. Several other proteins and molecules interact with IL-1β in a complex network, in which several cell types (neurons, microglia, and astrocytes) probably participate.


IV) Hcrt1 also influences fatigue, but probably through another pathway than the IL-1􀁅 route.
 

pattismith

Senior Member
Messages
3,276
Likes
6,312
3. The Role of EV Cargo in Modulating Cytokine Production during Autoimmunity

At present there are number of reports that have shown an increase in circulating EV levels during autoimmune conditions [68,69].

Knijff-Dutmer et al. demonstrated that circulating levels of platelet-derived microparticles were significantly elevated in RA patients compared to those in healthy volunteers, and the relative abundance of these EVs correlated directly to disease activity [70].

In another study, Berckmans et al. reported that the number of platelet- and leukocyte-derived EVs was also elevated, specifically in the synovium of RA patients [71].

Others have shown similar systemic increases in EV levels during SLE, Sjogren’s syndrome, and MS [72,73].

Viñuela-Berni et al. also demonstrated an increase in circulating EVs in SLE and RA and that these EVs could potently stimulate the release of interleukin 17 (IL-17), TNF-α, and interleukin 1 (IL-1) in vitro [74].

Interestingly, a paradoxical decease in circulating microvesicles was observed by Sellam et al. in the most severe examples of autoimmune disease [75].
It is thought that this is likely due to increased phospholipase A2 secretion and activity, which amplifies EV uptake as seen by Duchez and colleagues in neutrophils [75,76].

Others have used preclinical models to show the pathophysiological role EVs play in autoimmune disease. For example, Kavian et al. used a murine model of systemic sclerosis in order to demonstrate that chemically inhibiting microvesicle release with pantethine decreased the extent of skin and lung fibrosis compared to mice that had not received the inhibitor of EV release [77].

In a similar fashion, Deng et al. have shown the benefit of inhibiting the release of receptor activator of nuclear factor kappa-B ligand (RANK-L)-positive EVs from osteoblasts in ovariectomized mice as a method of significantly reducing bone loss [78]. While the ovariectomized mouse does not represent a model of autoimmunity, the activity of RANK-L-positive EVs in the formation of mature, multi-nucleated osteoclasts, and indeed exaggerated bone loss, remains relevant to the study of RA [79,80]. EV cargo has been shown to modulate cytokine release during autoimmunity in a variety of ways [81].

.......

In the context of autoimmunity, Skriner et al. have previously shown the association of citrullinated proteins with synovial exosomes from individuals with RA [88].

Others have shown that synovial macrophage exosomes isolated from patients with juvenile idiopathic arthritis specifically contain the nuclear oncoprotein, and autoantigen, DEK which stimulates joint inflammation [89].

Similarly, Hasilo et al. have reported the presence of diabetes autoantigens, such as glutamate decarboxylase and glucose transporter 2, in exosomes isolated from pancreatic islet cells [90].

....
. Regulatory T-cells are known to attenuate inflammation, in part, through the secretion of anti-inflammatory cytokines such as IL-10; however, the relative abundance of these cells, and therefore, their anti-inflammatory secretome is decreased during autoimmunity.
.....
. Despite the understanding of the individual roles that both EVs and cytokines exert during autoimmunity, the impact that cytokines associated with EVs may have in disease states remains widely underappreciated.
etc: cf full text
 

pattismith

Senior Member
Messages
3,276
Likes
6,312
In the same 2020 paper about IL-1beta:

Currently, one of the most well-studied cytokines in relation to its association with EVs is IL-1β.

On average, across several biological systems, the abundance of IL-1β has been shown to be equally distributed between EVs and free, soluble levels [105].

Unlike most other cytokines, IL-1β lacks a signal sequence and therefore has a non-conventional secretion pathway in association with EV release [108].

Previously, IL-1β has been shown to be released associated with exosomes from dendritic cells in patients with lupus [109].

IL-1β synthesis and release via EVs is highly regulated and dependent on the activation of the NOD-like receptor family pyrin domain containing 3 inflammasome.

The inflammasome is a multiprotein complex, which directs inflammatory signalling in a range of cells and its activity has been shown to be a key driver in a range of autoimmune conditions, including RA and T1D [110,111].

It has been demonstrated that non-classical secretion of IL-1β is mediated by microvesicle shedding in monocytes, macrophages, dendritic cells, and microglia and following the activation of purinergic receptors on the surface of EVs,

IL-1β is released into the extracellular space [112–114].

Increased purinergic receptor expression and signalling has been reported in the inflamed synovial tissue of arthritic rats and has been implicated in the pathogenesis of SLE [115,116].

Activation of synovial fibroblasts with IL-1β induces an arthritic phenotype, increasing cartilage degrading enzymes as well as IL-6 and vascular endothelial growth factor [117].

The pathogenic role of helper T-cells (Th cells) in autoimmunity has been well described; however in recent years, the role and impact of IL-17-secreting T-cells during autoimmune conditions such as RA, psoriasis, and SLE have been reported [118].

Hebel et al. have shown that IL-1β activates CD4+ T-cells, in conjunction with CD3 and CD28 stimulation, causing the release of IL-17.

Sustained IL-1β signalling in combination with TGF-β and/or IL-6 causes committal of T-cell differentiation into a Th-17 fate [119].

Interestingly, the authors showed that IL-1β stimulation also induced the release of IFN-γ, and others have shown that IFN-γ induces the increased shedding of EVs by increasing the activity of EV-packaging machinery, such as interferon-stimulated gene 15 [119,120].

Ultimately, IFN-γ stimulates further inflammasome activation, therefore inducing further IL-1β synthesis and release via EVs in chronic inflammation.

Stimulation by IL-1 family members has been seen to induce the release of IL-6-containing EVs from mast cells in a manner independent of de-granulation [121].

While the involvement of mast cells during autoimmunity is debated by some, data exists to support their pathogenic role in RA and multiple sclerosis [122].

In recent years, an autoimmune component has been implicated in the pathogenesis of amyotrophic lateral sclerosis and the release of EVs containing IL-6 from astrocytes is thought to contribute to disease pathogenesis and activity [123,124]
 

pattismith

Senior Member
Messages
3,276
Likes
6,312
I have extremly high IL-1b found in my serum as well.
Would there be any recommendable IL-1B blocker to try?

Or Interleukin 1Beta rezeptor Antagonists?
Anakinra trial failed to improved fatigue in a CFS/ME trial so I don't know what to answer.

Do you have POTS? OI?
I don't have them and respond to yellow mustard (Sinapis Alba). Do I respond because of it's anti-inflammatory properties? vascular modifying properties?
Too much questions and not enough answers remain in our disease...
 

Pyrrhus

Senior Member
Messages
2,632
Likes
7,155
Location
U.S., Earth
The two main points that I get from this study are:
  • The amount of mitochondrial DNA in exosomes from the blood of ME patients increases after exercise.
  • Protein content of exosomes was lower in ME patients compared to controls and was even lower after exercise.
 

pattismith

Senior Member
Messages
3,276
Likes
6,312
Polymorphism of interleukin-1β and interleukin-1 receptor antagonist genes in children with autism spectrum disorders 2020
https://doi.org/10.1016/j.pnpbp.2020.109999

Our data show alterations in the IL-1β system, with abnormally increased serum levels of IL-1β and IL-1RA in the children with ASD.
Further, polymorphisms in the IL-1β-511 and IL-1RA genotype variants correlated positively with autism severity and behavioral abnormalities. IL-1β-511 and IL-1RA gene polymorphisms could impact ASD risk and may be used as potential biomarkers of ASD.
Variations in the IL-1β and IL-1RA systems may have a role in the pathophysiology of ASD.
Another study in Turquie (2021) makes the link between autism and polymorphism in the IL-1 beta rs1143634 allele T
 
Messages
20
Likes
60
"Hebel et al. have shown that IL-1β activates CD4+ T-cells, in conjunction with CD3 and CD28 stimulation, causing the release of IL-17. "

For what it is worth, this seems to tie in with the findings of Drs. Liisa Selin and Anna Gil, which is in a different thread.
IL17, in particular, was elevated in males.
Cosentyx is an IL-17 inhibitor, but I do not know of any off-label use for CFS.