Human Herpesvirus-6 Reactivation, Mitochondrial Fragmentation, Paper Pub. 4/1/20 - Dr. Prusty

raghav

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THE FIRST 31 POSTS IN THIS THREAD HAVE BEEN MOVED FROM Bhupesh Prusty: "we are on a perfect path for identifying potential transferable factors in ME/CFS blood that can cause mito dysfunction..." GoFundMe

https://www.immunohorizons.org/content/4/4/201

Human Herpesvirus-6 Reactivation, Mitochondrial Fragmentation, and the Coordination of Antiviral and Metabolic Phenotypes in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome
Philipp Schreiner, Thomas Harrer, Carmen Scheibenbogen, Stephanie Lamer, Andreas Schlosser, Robert K. Naviaux and Bhupesh K. Prusty
ImmunoHorizons April 1, 2020, 4 (4) 201-215; DOI: https://doi.org/10.4049/immunohorizons.2000006


Bhupesh's paper is now available online. Cheers. He just now mailed me.

 
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raghav

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Discussion
HHV-6 infection and reactivation alter host cell transcriptome, including the miRNAome (15, 37, 38). miRNAs regulate intraorganellar mechanisms, in which they perform the job of fine-tuning of the functions required to fulfill the metabolic demands of an organ or cell type. Our previous deep sequencing approach (15) revealed major changes in the expression of a large panel of human miRNAs upon HHV-6A reactivation that hints to a broad range of effects on mitochondria and associated metabolic functions upon viral activation. In this study, we studied quantitative changes in cellular as well as mitochondrial proteomics upon HHV-6A reactivation and found several significant alterations with close similarity to ME/CFS pathophysiology. PDP1 was downregulated in response to HHV-6A reactivation. PDP1 helps in reverting the negative effects of pyruvate dehydrogenase kinases on pyruvate dehydrogenase. A decrease in PDP1 suggests a potential decrease in functions of pyruvate dehydrogenase, which is also reported in ME/CFS patients (28). We also found a decrease in the SOD2 level that can lead to high amount of ROS within the cell. We have previously shown that HHV-6 infection induces ROS in host cells and alters expression of glutathione reductase (29). These observations strongly support a pathological link between HHV-6 reactivation and ME/CFS.

Hypometabolic state of blood cells is a characteristic feature of ME/CFS (39). Although we did not perform metabolic studies, our study on mitochondrial architecture in presence of CFS patient serum but not controls has revealed a transferrable activity in ME/CFS serum that fragments mitochondria and stimulates a coordinated antiviral state in naive responder cells assayed in vitro. This assay was highly sensitive (0.9–1.00; 95% CI = 0.6–1.0) and specific (0.8–1.0; 95% CI = 0.38–1.0) in identifying serum from patients with ME/CFS. In our studies, both viral reactivation as well as CFS patient serum treatment induced a typical M1 state of mitochondria (35, 36) in host cells, which was also accompanied by a strong proinflammatory state of the cell that protected the host cells against both incoming DNA and RNA viruses, consistent with oxidative shielding (27). There are many possible ways that a nearby healthy cell can sense potential infectious risk in the environment. Increased IFN response from the virus-transactivated cells can result in secreted IFN that can be sensed by the nearby cells, leading to a hypometabolic state (40). Several type I and type II IFNs, as well as IFN response genes, are reported to be high in the serum of ME/CFS patients (41, 42). However, we did not observe any increase in IFN-γ in PBMCs of a small group of ME/CFS patients (Supplemental Fig. 2B). In addition, we observed significant decrease in ISG expression upon treatment with ME/CFS serum. Hence, the strong antiviral phenotype in our assay system seems to be IFN-independent. Fragmented mitochondria can release broken mitochondrial DNA into the cytoplasm (43) and to the extracellular space that can induce TLR-mediated cytokine production in nearby cells (44). Newly synthesized, 8-hydroxyguanosine–containing mitochondrial DNA (ox-mtDNA) can directly activate assembly of the NLRP3 inflammasome (45). Interestingly excess of certain cytokines can also change mitochondrial metabolism (46) to prevent further pathogen growth. Certain complex disorders, such as amyotrophic lateral sclerosis, are associated with a hypometabolic state that is characterized by lower mitochondrial size and number (4749).

Autoimmunity is common in ME/CFS (50). Several autoantibodies are capable of inducing a proinflammatory state in target cells (51). Interestingly, HHV-6 is linked to several autoimmune diseases, including multiple sclerosis and Hashimoto thyroiditis (52). Autoantibodies against β2 adrenergic receptors (β2R) were found to be upregulated in a subset of patients with ME/CFS (53). Such Abs belong to a network of natural Abs against adrenergic, acetylcholine (cholinergic), and other GPCR receptors that were shown to be dysfunctional in various autoimmune diseases (54). Autonomic dysregulation is a hallmark of ME/CFS. There is evidence that the β2R-mediated regulation of cytokines by terbutaline is impaired in whole blood immune cells of CFS patients (55). A recent paper showed that influenza replication is inhibited by α2 adrenergic stimulation via cAMP inhibition (56). In contrast β2R stimulation is known to stimulate cAMP. Thus, a disbalance of adrenergic stimulation favoring cAMP downregulation might also be an explanation for our findings. However, similarities between adoptive transfer of HHV-6A reactivation culture supernatant and ME/CFS serum suggests a potential metabolic role in addition to other possibilities.

Lack of a strong HHV-6 and HHV-7 infection in ME/CFS patients in our study and several others has historically cast doubt on the involvement of these viruses in ME/CFS. However, in this study, we show that incomplete HHV-6 reactivation, even in a small fraction of latently infected cells, causes reactivated cells to secrete an activity that can be transferred in serum and produces mitochondrial fragmentation and coordinates a powerful antiviral program in responding cells. Our studies showed that only IE events, such as the transcription of several small noncoding viral RNAs, were needed to trigger the production and secretion of the mitochondrial fragmentation factor and transferrable antiviral state. No HHV-6 proteins are made during the incomplete reactivation events described in this paper. Specifically, no major change in HHV-6 replication was observed. This explains the failure of anti-herpesvirus drugs in a subset of patients because the HHV-6 polymerase is not expressed during an incomplete virus reactivation, and drugs that target the viral DNA polymerase would have no viral target.

The virus reactivation experiments described in this study show that an antiviral state is produced both in cells with unreactivated and reactivated HHV-6. This seems to be against the viral growth and hence fails to explain the short-term benefits of viral reactivation from the pathogen point of view, unless passive transmission of viral genome to daughter cells after mitosis plays a major role in HHV-6 genome propagation. Decreased IFN response in virus-reactivated cells might provide an advantage for survival of IE RNAs in the host cell cytoplasm. However, in this study, we have tested only the nonproductive transactivation state of the virus. Productive viral infection, with virion production and release, might bring in additional viral factor(s) that damage cellular ability to undergo a hypometabolic state to provide successful virus growth. Additionally, mitochondrial fragmentation often allows virus to acquire persistent or latent state under a hypometabolic state (57).

In this study, we found that none of 25 patients with ME/CFS had peripheral blood evidence of a fully reactivated HHV-6 or HHV-7 infection, and only 8 of 20 (40%; 95% CI = 0.19–0.64) had evidence of partial reactivation measured by FISH analysis of HHV-6 small noncoding RNA U14 in whole blood. However, using an in vitro reporter cell assay, we showed that serum from ME/CFS patients contained an activity that produced mitochondrial fragmentation, decreased mitochondrial ATP production, and induced a powerful antiviral state. In 2016, metabolomic analysis of patients with ME/CFS revealed a chemical signature that was similar to the evolutionarily conserved, hypometabolic state known as dauer (39). This dauer-like state was preserved by blocks to healing produced by abnormal persistence of the CDR (36). The CDR has been shown to be directly involved in both healing and the biology of aging (35). In this earlier work, it was hypothesized that the metabolic features of the CDR in ME/CFS patients could protect against certain kinds of infection, but no direct testing for antiviral activity was performed (39). Our current data show that only a small fraction of cells need to be latently infected with HHV-6 to trigger a secretory phenotype that is strongly protective against some of the RNA and DNA virus infection in neighboring and distant cells lacking HHV-6 DNA. The main conclusions of this study are illustrated in the graphical summary (Fig. 6). Larger multicohort studies involving ME/CFS patients from different age groups should be carried out in the future and should include methods for detecting and quantifying both productive and nonproductive (incomplete) viral reactivation events. Furthermore, potential factors affecting mitochondrial dynamics in ME/CFS patients should be systematically evaluated for their ability to induce a powerful antiviral state. Our mitochondrial reporter-based cell system will provide an opportunity to develop a diagnostic test for ME/CFS as well as provide a platform for further identification of potential factors that define ME/CFS pathophysiology.
 
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raghav

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1587686009838.png
1587686009838.png




FIGURE 6.
Graphical Summary.

(A) Mitochondria rapidly fragmented in cells containing integrated HHV-6 (iHHV-6) after partial virus reactivation in response to genetic and/or environmental stress. Mitochondrial fragmentation was associated with a decrease in cellular ATP production and a potent antiviral activity that was secreted into the supernatant. Genetic and environmental stressors were modeled by treatment with the histone deacetylase inhibitor TSA with or without substitution of the nonfermentable carbon source galactose for glucose in the medium. Partial iHHV-6 reactivation was characterized by the production of noncoding RNA but did not result in viral protein synthesis or replication. Red colored cells represent partial iHHV-6 reactivation. (B) When the supernatant of stressed iHHV-6 cells was transferred to responder cells, a potent antiviral activity was found. This secreted activity was distinct from IFN and TNF-α and was indistinguishable from the activity found in serum from patients with ME/CFS. The mitochondrial fragmentation, metabolic, and antiviral properties of this iHHV-6 cell–secreted and ME/CFS serum activity appeared to transfer together and could not be separated in this study. The antiviral activity had broad specificity. This activity prevented infection with either the RNA virus influenza-A or the DNA virus HSV-1. Purple cells represent cells with fragmented mitochondria, which, were resistant to virus infection. (C) Cells that lacked iHHV-6 could not be induced to secrete the antiviral and mitochondrial fragmentation activities found in (A). (D) When responder cells were exposed to serum from HC subjects or to supernatant from stressed cells that lacked iHHV-6, no antiviral or mitochondrial fragmentation activity was observed. Green colored cells represent virus-infected cells.
 
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Just read the comments on s4me - awful and so negative. Especially from one person in particular. Talk about kicking you down 🙄 And when asked to explain, told that it would be impossible as we unscientific folk would never understand (paraphrased).... sigh and double sigh!!
Could you share a link? I didn't exactly find that...yet....
 

Badpack

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I was asking this one other doc if he could explain his extreme negativity, because he just said its bad and Prusty should feel bad for doing it without any reason whats pretty distasteful over your colleagues. Because how he speaks he clearly was claiming this right during his time as a scientist. But the mods just deleted me. Strange times.
 
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I looked at some of this.

The person posting the negative comments is entitled to their opinion I suppose. I am not in a position to refute or argue with it.

I do agree with this comment generally:" I am afraid I could go on for pages but basically I learnt to discount papers where the abstract shows no logical relation between results and conclusion - and includes no data. I was in biomedical science for thirty years and in the early days an abstract had to be easily intelligible and contain hard facts. Sadly in recent years anything seems to be publishable. But good science stays the same - an intelligible abstract with hard data." (quote is from that thread)


Abstracts here lately seem to do a poor job of- conveying what they discovered.

"The results are discussed"...is NOT a helpful summary.

But this is trivial compared to whether this research is- informative, valid etc.
 
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So if HHV-6 is some KEY virus...and given these statements at the HHV-6 web site, I again ask

WHY is there no antiviral therapy for people wiht ME? Why is this just all TALK.

And having been an Eppstein Barr victim my whole life: my brain is not wrapping cognitively around another virus that supposedly EVERYBODY is carrying.
 

sb4

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So if HHV-6 is some KEY virus...and given these statements at the HHV-6 web site, I again ask

WHY is there no antiviral therapy for people wiht ME? Why is this just all TALK.

And having been an Eppstein Barr victim my whole life: my brain is not wrapping cognitively around another virus that supposedly EVERYBODY is carrying.
I have no real idea so take this with a cup of salt but my understanding is that it can be any virus that partially activates, so that could be also why the antivirals might not work. If the antivirals are meant to target fully replicating viruses.
 

MonkeyMan

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Just read the comments on s4me - awful and so negative. Especially from one person in particular. Talk about kicking you down 🙄 And when asked to explain, told that it would be impossible as we unscientific folk would never understand (paraphrased).... sigh and double sigh!!
The guy who made these negative comments admits on Page 2 of the thread that he didn't read the whole paper. That strikes me as arrogant, closed-minded, and unscientific.
 
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I'm thinking ALL these herpes viruses- may be skilled in similar arenas.

Perhaps all these herpes viruses are capable of fragmenting mitochondria under the right set of circumstances.

Something is definately not going smoothly in my hippocampus (like any form of short term memory and moving that to long term storage)...something is WRONG there.

And this antiviral situation- feels like maybe in my case, this benefit may have hit when I GOT MUCH WORSE three years ago.

I seem to ONLY GET this weird awful STOMACH FLU-like thing that shuts down my intestines and is a near death experience. 5 times in two years. And I am very isolated: hence I don't even think sometimes it really IS a flu.
 
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I have no real idea so take this with a cup of salt but my understanding is that it can be any virus that partially activates, so that could be also why the antivirals might not work. If the antivirals are meant to target fully replicating viruses.
I don't have scientific clue neither but that makes sense to me...
And antivirals are very toxic stuff in the long term.

I think we can't beat those small f..ing bugs... So I think the treatment should be telling the cells to stop the CDR instead.

Maybe finally the CBT will be the answer! We have to sit down with our cells and ask them please to behave well haha :rofl: (just a joke, hoping not being misundetstood).

About the criticism of "that" person in "that" other place... Maybe sometimes, some people, live from the memories of the past glories?

Maybe it is easy to criticize the work of young and smart people who do their best without the money they should deserve because the governments don't give us a shit.... While you sit there and remember what you did (or not) for the ME.

Just a thought.
 

junkcrap50

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Just read the comments on s4me - awful and so negative. Especially from one person in particular. Talk about kicking you down 🙄 And when asked to explain, told that it would be impossible as we unscientific folk would never understand (paraphrased).... sigh and double sigh!!
He's the reason I don't go over there. Everybody seems to be sycophants for him there.
The guy who made these negative comments admits on Page 2 of the thread that he didn't read the whole paper. That strikes me as arrogant, closed-minded, and unscientific.
Bingo.