1. The latest crowdfunder to support David Tuller's work has opened. To donate click here.
    Dismiss Notice
  2. Guest, the 'News in Brief' for the week beginning 18th October 2021 is here.
    Dismiss Notice
  3. Welcome! To read the Core Purpose and Values of our forum, click here.
    Dismiss Notice

Complement Component C1q as a Potential Diagnostic Tool for Myalgic Encephalomyelitis/Chronic Fatigue Syndrome Subtyping, 2021, Castro-Marrero et al

Discussion in 'BioMedical ME/CFS Research' started by Sly Saint, Sep 16, 2021.

  1. Sly Saint

    Sly Saint Senior Member (Voting Rights)

    Messages:
    6,861
    Location:
    UK
    Abstract
    Background: Routine blood analytics are systematically used in the clinic to diagnose disease or confirm individuals’ healthy status. For myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS), a disease relying exclusively on clinical symptoms for its diagnosis, blood analytics only serve to rule out underlying conditions leading to exerting fatigue. However, studies evaluating complete and large blood datasets by combinatorial approaches to evidence ME/CFS condition or detect/identify case subgroups are still scarce.

    Methods: This study used unbiased hierarchical cluster analysis of a large cohort of 250 carefully phenotyped female ME/CFS cases toward exploring this possibility.

    Results: The results show three symptom-based clusters, classified as severe, moderate, and mild, presenting significant differences (p < 0.05) in five blood parameters. Unexpectedly the study also revealed high levels of circulating complement factor C1q in 107/250 (43%) of the participants, placing C1q as a key molecule to identify an ME/CFS subtype/subgroup with more apparent pain symptoms.

    Conclusions: The results obtained have important implications for the research of ME/CFS etiology and, most likely, for the implementation of future diagnosis methods and treatments of ME/CFS in the clinic.

    https://www.mdpi.com/2077-0383/10/18/4171
     
    Michelle, Jan, diwa and 11 others like this.
  2. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    It's good that the article is open access, and that it's possible to read the feedback of reviewers. The study was partly funded by a patient group, although I bear in mind the previous studies by Castro-Marrero that were funded by a supplement manufacturer.

    The study was of a good size (250 people), all females.

    The paper is a bit frustrating in that the charts shown in the paper relate to efforts at subgrouping the sample and really don't tell us much, whereas the scatter plots are hidden away in a supplementary file. It's disappointing to see peer reviewers simply say 'Well done!' or focus on the fact that numbers under nine should be spelled out as text rather than given as a number, when the paper could have been improved a lot by better choice of figures.

    Vitamin D
    A high percentage of the patients were low in vitamin D. While it is likely that that is a result of a lot of time being spent indoors rather than being a cause of ME/CFS or shedding light on the pathology of the illness, it does suggest that people with ME/CFS may need to try to spend more time in the sun, or take supplements.

    While Vitamin D deficiency is very common in populations, references I found suggest levels should be over 40 ng/ml. That suggests this sample of people (living in Spain, a sunny country) really are quite deficient. Given the symptoms of Vitamin D deficiency do overlap substantially with the symptoms of ME/CFS, it would be good to see some more attention being paid to this. I'd like to see a well-conducted supplementation trial, and guidelines should note that people with ME/CFS are at risk of vitamin D deficiency.

    Screen Shot 2021-09-17 at 6.24.28 AM.png
     
    cfsandmore, Jan, Snow Leopard and 9 others like this.
  3. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    Complement factor C1Q
    The main finding of interest though was the high levels of C1q.

    Screen Shot 2021-09-17 at 6.45.41 AM.png
     
    Michelle, Jan, Snow Leopard and 5 others like this.
  4. Aslaug

    Aslaug Moderator Staff Member

    Messages:
    1,485
    Sorry but "need to try to spend more time in the sun" :laugh: I'm surprised at these numbers from Spain, but even in sunny parts of the world vitamin D deficiency is not uncommon due to clothing, sunscreen use and what time of day people are typically outdoors. Not least what time of year the samples were collected.

    Many of those patients look to be deficient, although reference ranges for vitamin D vary a lot by country (I assume the red lines are the range used in Spain). If one only cares about skeletal health, the reference range for sufficiency is >7.5 ng/ml (25 nmol/L), but most countries use >20 ng/ml (50 nmol/L) "to be on the safe side" while taking into account that vitamin D might have extraskeletal effects. 35-40 ng/ml (>75 nmol/L) are advocated by some specifically due to extraskeletal effects and I think @Mij mentioned it had been implemented in Canada. I think it has been also in other countries. In Norway it is seen as a bit fringe to advocate for people to have such high levels of vitamin D (our threshold for sufficiency is >16-20 ng/ml depending on what lab is used).

    I do with people would have their vitamin/mineral status checked (this will include a diet interview as there isn't a blood test for many nutrients), so deficiencies could be discovered and treated. Having those on top of ME feels so unnecessary :(
     
    Last edited: Sep 16, 2021
    Michelle, Jan, Caesar and 7 others like this.
  5. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    So C1Q -
    thanks to wikipedia, this is my understanding, which may be wrong and will certainly be incomplete, so do correct me:

    1. CIQ binds to molecules that suggest the immune system needs to do something
    C1Q can bind directly to bacterial and viral surface proteins, dying cells, and acute-phase proteins*. C1Q can also bind to [antibody (an IgM or IgG) + antigen] complexes.

    * Acute-phase proteins - proteins associated with inflammation which include CRP, and the mannose-binding lectin that another recent study looked at in ME/CFS).


    2. The binding sets a cascade of reactions off
    The binding activates the C1 complex which activates the 'classical complement pathway'.

    Classical complement pathway: That's a series of reactions that produces molecules that
    1. attract phagocytes;
    2. tags cells for phagocytosis;
    3. forms the Membrane Attack Complex that damages a cell membrane to kill the cell

    3. So, C1Q is a useful thing
    A lack of C1Q is therefore not a good thing, as the immune system won't work so well. A deficiency of C1Q apparently leads to lupus, where, Wikipedia says, there is an accumulation of autoantibodies and dying cells that aren't being cleaned up.

    4. But too much won't be good either
    Some of the molecules produced along in the cascade of reactions are called anaphylatoxins (fragments of C3,C4,C5 - namely C3a, C4a, C5a)
    A deficiency of C1-inhibitor can cause angioedema. C1-inhibitor defiency can be hereditary or acquired.

    So, it looks as though a substantial proportion of the women in this study had high levels of C1Q. I guess one question is, are the levels high enough to be contributing to their ME/CFS symptoms? Another one is 'why are the levels elevated?'
     
    Last edited: Sep 17, 2021
    Michelle, Jan, diwa and 10 others like this.
  6. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    Is the level of elevation of the C1Q significant?

    The unit of measure in this study is miligrams/dL. (1 mg/dL =10 ug/ml)

    This study suggested the normal range is 10 to 25 mg/dL.
    Mayo clinic's normal range is 12 -22 mg/dL. Using that range, most of this study population have high C1Q.
    Medscape suggests the normal level is 7 mg/dL, and the acute phase serum level is 13% higher.
    Clearly, there's variation in the ranges, maybe depending on the test method.

    It doesn't seem like the C1Q level in serum needs to be massively higher to suggest something is going on. C1Q levels can be much higher at the site where the action is taking place.

    (The Medscape article also talks about the lectin/ mannose-binding pathway, as an alternative to the classic pathway that involves C1Q. In that pathway, the acute phase serum level of MBL is 1000% of that of the normal level. So, you'd probably be wanting to see much higher levels in ME/CFS compared to controls in order to be convinced that higher levels of MBL are important.)
     
  7. Jonathan Edwards

    Jonathan Edwards Senior Member (Voting Rights)

    Messages:
    10,060
    The key question would be why a difference inC1q has not been picked up before. It is not measured routinely in the UK but probably would be in the US. I have not looked through the whole paper but I don't know if it gives a trawl of previous studies of C1q. Sorensen and colleagues did detailed studies on complement in CFS about ten years ago. The ME Biobank would have another cohort to study and may already have looked at C1q. Several groups looked for lupus-associated antibodies and are likely to have looked for C1q.
     
    FMMM1, Michelle, Skycloud and 8 others like this.
  8. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    I've made a thread for a Sorensen paper from 2003 here:
    Complement activation in a model of chronic fatigue syndrome, 2003, Sorensen et al

    It's an interesting paper with an exercise challenge. It looks as though they tested C1Q and did not find a significant difference - but I couldn't find any data reported for the C1Q comparison. The Medscape article suggested that the difference between normal and activated levels serum may not be big, and some of the articles I read suggested that levels change with age. So, perhaps the smallish sample size of the Sorensen study made it hard to get a significant difference given background noise.
     
  9. Aslaug

    Aslaug Moderator Staff Member

    Messages:
    1,485
    I don't think it is used routinely in Norway, although mine was checked before I was eventually diagnosed with ME (I apparently presented with lupus-like symptoms).

    The Wikipedia article (sans the lupus part, which I found a bit oversimplified) is in line with what we learned about the complement system in immunology @Hutan. A bit off topic, but recently some Norwegian researchers proved that a "truth" about the complement system and coagulation is in fact not true, and they discussed how it came to be a truth in the first place (bad data) https://www.jimmunol.org/content/207/6/1641 so I wonder if there are other things we don't know about these proteins.

    Vitamin D can regulate the immune system, so vitamin D status may be important to see certain differences? This is where the threshold for >75 nmol/L may be important. Since endothelium permeability was mentioned, vitamin D may play a role in that too. Vitamin D has been found to regulate epithelium permeability in response to various pathogens, but only when above a certain threshold (in cell culture and animal models, I've also seen it in human studies but they are often of poor quality. Such as the one that gave supplements, said "vitamin D levels and epithelium integrity improved" but did not provide any measurements on vitamin D serum levels pre/post supplementation...). I'm not 100% sure but I think some of the proteins between epithelium cells that can be regulated by vitamin D is also found between endothelium cells.
     
    Michelle, Jan, Skycloud and 7 others like this.
  10. Grigor

    Grigor Senior Member (Voting Rights)

    Messages:
    342
    This is from Prusty's talk:

     
    sebaaa, Aslaug, ukxmrv and 1 other person like this.
  11. Jonathan Edwards

    Jonathan Edwards Senior Member (Voting Rights)

    Messages:
    10,060
    I cannot work out what Prusty's slide is supposed to indicate.It doesn't seem to have anything to do with high C1q levels.

    C1q contributes to formation of C1 complex, not the other way round.
     
  12. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    It's not clear to me what that slide is suggesting. That exercise and anti-C1q antibodies reduce levels of functioning C1q? So C1q can't do it's job helping to defeat pathogens and clear the rubbish up?
    But, I don't think we have any evidence of high levels of Anti-C1Q antibodies in ME/CFS (unlike the situation with Lupus). I mean, if we had high levels of Anti-C1q antibodies, wouldn't we have been diagnosed with Lupus?
     
    sebaaa, Michelle and Caesar like this.
  13. Caesar

    Caesar Senior Member (Voting Rights)

    Messages:
    119
    I have this rare illness Hereditary Angioeadema (HAE). Type II with normal C1-INH, but very low function.

    I used Danazol for many years as a treatment but had to stop due to long-term side effects.

    During the discontinuation period, I went from being healthy to getting many different acute serious symptoms.
    Everything from unusual muscle contractions and muscle twitches, pain, histamine and food problems, inflammation and severe fatigue.
    Before I after a year finally was diagnosed with ME/CFS.

    I'm now using a pure C1-INH treatment for my HAE, which does not affect my ME/CFS.

    A close family member developed Lupus shortly after the same Danazol treatment.

    It is not uncommon for someone with HAE to also have another autoimmune disease.

    Several other studies have drawn the relevance of Bradykinin to ME/CFS too, and Bradykinin is the very essence of HAE with low or non-functioning C1 proteins.

    I'm not quite sure if I understand the significance of this study and the relationship C1 to C1q, but I think it's an exciting field to explore.
    In a broad perspective across different diseases. But of course I'm no expert. :)
     
    Last edited: Sep 21, 2021
    cfsandmore, sebaaa, Michelle and 9 others like this.
  14. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    Thank you @Caesar, that's really interesting. My son and I get generalised swelling when in PEM - I don't wear rings, and lately now don't wear my watch either for that reason. There is facial swelling too, around the eyes. My son has had a couple of episodes of very obvious facial swelling when exhausted. And I know some other members here have problems with oedema too.

    So, you have normal levels of the C1 inhibitor but it isn't functioning well? How is that tested for? Does it mean that you have high levels of C1Q? From what I read, problems with C1 inhibitor don't seem to automatically mean high levels of C1Q, and in fact levels of C1Q can be low in people with angioedema.

    It's certainly complicated.
     
    sebaaa, Wyva, Aslaug and 5 others like this.
  15. Caesar

    Caesar Senior Member (Voting Rights)

    Messages:
    119
    @Hutan
    Have you had the swellings examined with blood tests? Tested C1-INH, C3 and C4?

    Due to the C1-INH treatment, I now have high levels with C1, due to the fact that C1 has low function. But originally, my C1-INH is normal, yes.

    Blood tests reveal this. C3, C4, C1-QUAN, C1-FUNC.

    I'm not sure about this C1Q. It is beyond my knowledge. It is, as yoy say, complicated. But I found this online:

    [​IMG]
    https://www.researchgate.net/figure/Algorithm-of-HAE-diagnosis_fig3_324737619
     
    Starlight and Hutan like this.
  16. Hutan

    Hutan Moderator Staff Member

    Messages:
    16,579
    Location:
    New Zealand
    Doctors don't seem to be interested. My record of lab tests doesn't have any complement-related tests, but I've been slack about copying results into it these last couple of years. I don't recall any complement-related tests.
     
    sebaaa, Wyva, Skycloud and 3 others like this.
  17. Snow Leopard

    Snow Leopard Senior Member (Voting Rights)

    Messages:
    3,207
    Location:
    Australia
    Given the absence of other excessive inflammation markers in circulation in most patients, I wonder about the other functions C1Q has (non classical complement cascade), and why it might be of high abundance in the first place...

    Emerging and novel functions of complement protein C1q
    https://www.frontiersin.org/articles/10.3389/fimmu.2015.00317/full

    C1q: A fresh look upon an old molecule
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5582005/

    C1q as an autocrine and paracrine regulator of cellular functions
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5366276/

    Endothelial cells in particular can secrete significant amounts of C1q in certain circumstances.
     
    sebaaa, FMMM1, Michelle and 5 others like this.
  18. Caesar

    Caesar Senior Member (Voting Rights)

    Messages:
    119
    I understand. These are unusual blood tests and not all doctors know about it. I know that well. So HAE is underdiagnosed.

    By the way, I was not aware that oedema could occur during PEM in ME / CFS, so thanks for the information.
     
    FMMM1, Aslaug, Hutan and 1 other person like this.
  19. Samuel

    Samuel Senior Member (Voting Rights)

    Messages:
    398
    i haven't been able to determine if angioedema is associated with m.e. i have been assuming it isn't really, but really want to know.


    this algorithm alone is enough to confuse me. but that's me.

    it is not clear to me if "ae with normal c1-inh" is a broad category or a distinct entity. i will assume broad category. but why does the flowchart stop there?

    i do not understand what 95% refers to.

    in my case, if you take my c4 as low [is low-normal], then i pass through to the quantitative, which is normal, then thus to function, which i was told was normal. there is no entity attached to that. the flowchart stops there. my case is a non-entity yet again.

    i have technical issues tring to get access to the paper. might have brain issues reading it. perhaps it clarifies.


    i have not had c1q tested. i did try 4-6x normal amounts of cetirizine for many months, with no apparent effect on angioedema. that is an uptodate protocol. thus, it is non-histaminergic. if it is histaminergic, you pay for cetirizine. if it is not, you pay 400,000+ usd per year.

    i do have a note that some of these tests, including c1q, might not be reliable.

    > C1-INH functional assays were available to 93% of
    responding CHAEN physicians. However, confidence in this
    assay was relatively low; 41% said they did not trust the
    results, citing concerns about the handling and storage of
    samples, as well as the sensitivity and specificity (the
    chance of getting a false positive or negative,
    respectively) of different assays. ---
    https://angioedemanews.com/2019/01/10/canadian-dcotors-need-more-info-hereditary-angioedema-tests/
     
    Last edited: Sep 22, 2021
    Caesar likes this.
  20. Caesar

    Caesar Senior Member (Voting Rights)

    Messages:
    119
    I don't think angioedema is directly associated with ME/CFS.
    But the interaction between C4, C1-INH, C1q, estrogen, Bradykinin and Kallikrein I think may possibly create an indirect association to autoimmune diseases, and is ME/CFS it?

    Autoimmune diseases are common in HAE patients. So, what's going on here?
    We can somehow not prove anything, but the question is worth asking.

    It is also interesting that some ME/CFS patients develop non-allergic swellings during PEM.
    So is there an individual complicated and isolated indirect event involved? Maybe, maybe not... but interesting, and worth looking at.

    Regarding the flawchart I referred to to demonstrate C1-inh vs C1q, it was clearly deficient. It did not take into account high C1-INH and high C1q. So something significant is missing there.
    It also did not address HAE type III. Angioedema with normal values of C4, C1-Quan and C1-Func, but with the same non-allergic edema as type I and II. Type III is not fully understood.
    https://www.annallergy.org/article/S1081-1206(12)00505-4/fulltext

    HAE attacks occur in outbreaks over a few hours to up to 5-6 days. Sometimes the outbreak comes quickly, other times it builds up over days. The whole body is affected, including internal organs, and it is random where the edema develops. The gastrointestinal tract, groin, genitals, joints, hands, feet and face are classic places where edema attacks starts. And the outbreaks affect patients differently on an individual basis.

    HAE attack occurs as a result of stress, mental and physical strain and overexertion, incorrect load and pressure on the body, unusual bodily movements, temperature changes, some foods, infections, virus, bacteria, allergies etc.
    HAE attacks have same triggers as PEM in ME/CFS, actually.

    It is also worth mentioning that a special transient fatigue (HAE fatigue) occurs some time before and during an HAE attack. Because of this, it is often known many hours in advance that an attack is underway. No matter what the trigger was.

    I have not taken a C1q blood sample. I have to ask my HAE doctor about this.
     
    Last edited: Sep 21, 2021
    sebaaa, Samuel, Wyva and 2 others like this.

Share This Page