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Nguyen et al/NCNED: Calcium mobilisation in NK cells from CFS/ME patients is associated with...

Discussion in 'Latest ME/CFS Research' started by mango, Oct 12, 2016.

  1. mango

    mango Senior Member

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    Calcium mobilisation in natural killer cells from Chronic fatigue syndrome/Myalgic encephalomyelitis patients is associated with transient receptor potential melastatin 3 ion channels

    Nguyen T 1,2, Johnston S 3,4, Clarke L 3,4, Smith P 3, Staines D 3,4, Marshall-Gradisnik S 3,4.

    Author information
    1 The National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute, Griffith University, Gold Coast, Australia.
    2 School of Medical Science, Griffith University, Gold Coast, Australia.
    3 The National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute, Griffith University, Gold Coast, Australia.
    4 School of Medical Science, Griffith University, Gold Coast, Australia.

    Clin Exp Immunol. 2016 Oct 11. doi: 10.1111/cei.12882. [Epub ahead of print]

    Abstract
    Transient receptor potential melastatin subfamily 3 (TRPM3) ion channels play a role in calcium (Ca2+ ) cell signalling. Reduced TRPM3 has been identified in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) patients. However, the significance of TRPM3 and association with intracellular Ca2+ mobilisation has yet to be determined.

    Fifteen CFS/ME patients (mean age 48.82 ± 9.83 years) and 25 health controls (mean age 39.2 ± 12.12 years) were examined.

    Isolated NK cells were labelled with fluorescent antibodies to determine TRPM3, CD107a, and CD69 receptors on CD56Dim CD16+ NK cells and CD56Bright CD16Dim/- NK cells. Ca2+ flux and NK cytotoxicity activity was measured under various stimulants including PregS, Thapsigargin, 2APB and ionomycin.

    Unstimulated CD56Bright CD16Dim/- NK cells, significantly reduced TRPM3 receptors were seen in CFS/ME compared with HC. Ca2+ flux showed no significant difference between groups. Moreover PregS stimulated CD56Bright CD16Dim/- NK cells showed significant increase in Ca2+ flux in CFS/ME patients compared with HC.

    By comparison, unstimulated CD56Dim CD16+ NK cell showed no significant difference in both Ca2+ flux and TRPM3 expression. PregS stimulated CD56Dim CD16+ NK cells significantly increased TRPM3 expression in CFS/ME but this was not associated with a significant increase in Ca2+ flux.

    Furthermore, TG stimulated CD56Dim CD16+ NK cells increased K562 cell lysis prior to PregS stimulation in CFS/ME patients compared with healthy controls.

    Differential expression of TRPM3 and Ca2+ flux between NK cell sub-types may provide evidence for their role in the pathomechanism involving NK cell cytotoxicity activity in CFS/ME.

    https://www.ncbi.nlm.nih.gov/pubmed/27727448
     
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  2. Marky90

    Marky90 Science breeds knowledge, opinion breeds ignorance

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    Wut.

    Somebody care to lay it out in layman terms? :p
     
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  3. Never Give Up

    Never Give Up Collecting improvements, until there's a cure.

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    First they took some stuff, and did some stuff to it, and found, well, stuff.

    Someday I'll get around to studying all that calcium chanel signaling, but for now I'd just like to say Go Australian's!
     
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  4. M Paine

    M Paine Senior Member

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    Reduced NK Cell activity is associated with CFS/ME. NK Cells normally differ from NKT cells in that they don't express TCR or CD3 receptors, amongst other things. They usually express CD16 and CD56.

    NK cells are grouped into two main subsets, depending on the cell surface density of CD56 and CD16 expression. 90% of circulating NK cells are CD56dim CD16bright. The rest being CD56bright CD16neg/dim.

    TRPM3 (Transient receptor potential melastatin 3) encodes for a cation-selective channel, which forms part of calcium signalling and homeostasis. A previous study identified single nucleotide polymorphisms in TRPM3 genes as correlating to illness in CFS/ME.(Novel identification and characterisation of Transient receptor potential melastatin 3 ion channels on Natural Killer cells and B lymphocytes: effects on cell signalling in Chronic fatigue syndrome/Myalgic encephalomyelitis patients)

    This study used immunofluorescence to measure the expression of TRPM3, CD56, CD16 and a couple more receptors. They confirmed that TRPM3 expression is abnormal (low) in the less abundant subgroup of NK cells in CFS patients compared with controls. Calcium channel activity was measured, and was normal, until the cells were stimulated with PregS. That sort of suggests that it's not until these cells are 'activated', that the deficient calcium channel activity likely associated with TRPM3 expression is measurable.

    To further complicate matters, the more abundant subgroup of NK cells did express TRPM3 in a normal range, and increased expression when activated. This increase was not associated with an increase in calcium channel activity.

    I can't read the full paper, it's behind a pay-wall. Would really like to read the discussion if someone has access.
     
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  5. M Paine

    M Paine Senior Member

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    I think I had the PregS thing slightly wrong here... still trying to work out what's going on there. PregS may be activating the Calcium Channels in some way.

    "This current investigation showed inhibition of ER Ca2+ /ATPase pump and depletion of intracellular Ca2+ stores follow by PregS-activated TRPM3 increased cytotoxic activity in NK cells from CFS/ME patients"
     
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  6. adreno

    adreno PR activist

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    A connection between sphingolipids, TRPM3 and antibodies?

    https://www.ncbi.nlm.nih.gov/pubmed/20360246
     
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  7. CCC

    CCC Senior Member

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    Here's the conclusion:

     
  8. M Paine

    M Paine Senior Member

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    Finally got to read the full paper, thanks to the person who sent it through.

    PregS = pregnenolone sulphate

    From the paper,
    "TRPM3 ion channels are selectively activated by a potent neurosteroid, pregnenolone sulphate (PregS)"

    There are some really interesting/odd findings here. I find it a bit difficult to follow along with the two subgroups of NK cells. Not the easiest paper to read.

    For the sake of simplicity, it has been helpful to me thinking of CD56 bright CD16 dim/- as "immunosurveillance nk cells" (the 10%), and CD56 dim CD16+ as "cytotoxic nk cells" (the 90%).


    Immunosurveillance nk cells in CFS patients have lowered TRPM3 receptors. Ca2+ Flux normal. PregS activation of TRPM3 greatly increased Ca2+ Flux in these cells in CFS patients compared to controls. Odd that TRPM3 is low, but otherwise not much else to see here.

    On the other hand, Cytotoxic NK cells in CFS patients seem to have normal amounts of TRPM3 and Ca2+ flux, but when stimulated with PregS, they ramp up TRPM3 expression without increasing Ca2+ flux.


    From the discussion:

    "A possible rationale for this finding may be that TRPM3 ion channels expressed on CD56 dim CD16+ NK cells have impaired Ca2+ flux function, which is attempted to be compensated by
    increasing their surface expression."

    They also mention:

    "NK cells demonstrated significantly increased cytotoxic activity when stimulated with TG+PregS in CFS/ME compared with HC group. No significant differences between groups were seen with PregS, ionomycin and 2APB"

    I felt like this finding was very significant, but I can't quite work out the mechanics of TG + PregS exactly.

    "2-APB prevent the release of intracellular Ca2+ stores compared to TG which inhibit the replenishment of the Ca2+ into intracellular stores"

    At this point, I'm a bit lost about exactly now TG improves the Ca2+ flux situation. Nevertheless, it seems that the combination of PregS+TG impacts Ca2+ Flux in a way which improves the cytotoxicity of cytotoxic NK Cells in CFS patients.

    Seems very significant of an observation. I mean, decreased NK cytotoxicity would seem like an important cellular function for the immune system to put it lightly. Not only did they identify impairment of ion channel function, but they linked that directly to impaired cytotoxicity, taking it one step further and treating that problem in vitro with positive effect on cytotoxicity.

    @Jonathan Edwards... Do you have any comments about this paper?
     
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  9. mango

    mango Senior Member

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    https://www.facebook.com/permalink.php?story_fbid=909726125826519&id=301252900007181
     
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  10. adreno

    adreno PR activist

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    What's TG?
     
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  11. lansbergen

    lansbergen Senior Member

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    Thapsigargin?
     
  12. anciendaze

    anciendaze Senior Member

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    Even if this does not work out in detail, I feel they are investigating the right kind of things. Calcium channels are involved in every situation where vesicles inside cells fuse with membranes to release their contents. This has obvious relevance to neurons and neurotransmitters, or neuromuscular junctions, but also plays very important roles in immune signalling which the medical profession scarcely worried about 15 years ago.

    (One example turns up in MCAS where mast cells release a range of biochemicals which mobilize immune response, or mislead it. We just saw the discovery that a significant percentage of the population with mysterious medical problems has multiple copies of the gene for alpha tryptase. Anyone who has been following the story of MCAS will pay attention to news involving tryptase.)

    A second aspect is the combination of an ion channel with ATP receptors. A similar combination shows up in K+ATP channels in mitochondria. Defects in such channels should be expected to cause problems in energy production or utilization and immune response.
     
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  13. eljefe19

    eljefe19

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    http://forums.phoenixrising.me/inde...n-in-chronic-fatigue-syndrome-patients.47443/

    Here's a thread on new research today on impaired energy metabolism in CFS but they found actually high levels of ATP but determined the mitochondria were not the source of much of it. It's all way over my head. Care to ELI5?
     
  14. anciendaze

    anciendaze Senior Member

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    @eljefe19,
    Not ready to provide a full explanation for anyone calling himself el jefe, my current goals are more modest.

    ATP plays many different roles, as does oxygen. I've been looking at purinergic signalling triggered by localized hypoxia for a while. One trigger for vasodilation comes when red blood cells encounter low oxygen levels they cannot correct by dropping oxygen they carry, and then dump ATP. This ATP is extracellular, and not manufactured by mitochondria within the receiving cell. ATP is also used to mark foreign or misfolded proteins within cytoplasm, and multiple copies are attached. Some are used to power cell machinery degrading proteins, others are released and serve as signals. Many biochemical signals require confirmation by purines like ATP to indicate the source of the signal is local and active.

    I'm still collecting information before committing to a theory.
     
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  15. Jonathan Edwards

    Jonathan Edwards "Gibberish"

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    I may be getting old but I cannot really follow what they are going on about.
     
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  16. Keith Geraghty

    Keith Geraghty Senior Member

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    from my basic level of cell biology I understand whats going on here but not whats causing this to go on - if there are polymorhps in nucelotides - that result in altered expression of Ca2+ channel proteins (channels) - whats causing the nucleotide shift - is this the big epigenes that MEGA hope to find. I suspect in ME/CFS immune alternations change many cells in the body, particularly immune cells - but if NK cells are losing cytotoxicity why? - I assume the levels arent reduced, its their ability to do what they do, which could be a clue - to either some sort of exhaustion or change in their activity as time dgoes on (similar to what the cytokine changes reported by Stanford).

    I'd like to know if macrophage activity and interferon levels are normal or not in these cases. The first phase immune response that changes as time goes by may be the reason we cant link standard viral pathway immune responses or bacterial pathway immune responses to things like NK cell activity and so on.

    I dont have the appropriate knowledge to even comment properly - however this Ca finding is intriguing.
     
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  17. BurnA

    BurnA Senior Member

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    What hope so does anyone else have I wonder!
     
  18. wastwater

    wastwater Senior Member

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    I bumped into another word of possible relevance,not sure what it is or if its of interest G-AlphaQ
     
  19. RogerBlack

    RogerBlack Senior Member

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    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5217865/
    This is a followup to the paper discussed in http://forums.phoenixrising.me/inde...-melastatin-3-ion-channels.44932/#post-730468

     
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  20. Rrrr

    Rrrr Senior Member

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    I'm not science smart, so maybe somebody can help me with this question I have. My calcium levels are always very high on my blood/lab work. Could this have something to do with it?
     

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