Testing cytokines outside of labs useless?!

Messages
2,284
Likes
7,537
Hi 🙋🏻‍♂️

I had a call with Dr. Pfeiffer (a lab doc and immunologist ) and he commented on my recent highly elevated results on IL-6 (388 pg/ml ref. <5.4) and IL-8 (6645 pg/ml ref. <62) that these are due to artefacts from the transport of the blood draw to the lab (hemolysis)...

I then called another lab (lab4more) and talked to a another lab doc and she said it's possible.

So: What is the benefit in testing cytokines when you are not in a hospital/lab? It seems to be absolutely useless.
She said to me the only reason why these results might be meaningful is because they are combined with other pro-inflammatory markers such as MIF (4566 pg/ml ref. <1000), MRP 8/14, Il-1b, very low IFN-g etc... But she said that testing cytokines and judge them alone when your not within a “testing-window” of approx. 4 hrs (!) doesn't mean anything really.
She also said cytokines and the whole cytokinins profile can change from one week to another.

So what's the thing with Patterson here who seems to look at one cytokine specifically (RANTES)?

I'm confused now. What's the matter with testing cytokines at all if you have to send them to a lab?

Thank you!

Martin
 

nerd

Senior Member
Messages
863
Likes
2,532
Hemolysis is a general issue with whole blood samples. The samples have to be delivered fresh and quickly, not refrigerated over night. The respective labs normally use courier drivers who collect and deliver the samples on the same day.

The typical indication for hemolysis would be elevated sodium and lowered potassium. If there's suspicion, labs can also identify hemolysis by other means, but most of the time, they don't check it and only the physician might raise the question.

However, the ME pathophysiology overlaps with this phenomenon. Abnormal sodium and potassium are part of it. It doesn't have to be hemolysis. The ME pathophysiology might also predispose blood samples to accelerated hemolysis.

This can be tested by looking at thrombocyte aggregation with from separate citrate, heparin, and EDTA samples from the same blood collection. These tests are generally inaccurate, so it's best to repeat the testing multiple times. If citrate and heparin don't increase or maintain the thrombocyte count but reduce it when compared to EDTA, it's pseudo-thrombocyte aggregation, meaning that the aggregation happened in the tube and not in the body. In this case, I'd expect hemolysis to happen more quickly in whole blood samples as well but electrolytes aren't a reliable indicator for ME. Moreover, thrombocyte count will always be inaccurate if not tested in fresh blood.

What could be tried to check if the lymphocytes are still intact is a lymphocyte transformation test to see if the unstimulated protein synthesis is still within reasonable boundaries and if a typical antigen mix can sufficiently stimulate the protein synthesis. 2-3 assays are necessary for that. If the baseline isn't too high or too low and if the stimulation index is high enough, the lymphocytes should be still intact and their contained cytokines shouldn't be too far off.
 
Messages
2,284
Likes
7,537
Unfortunate if true.

FYI I had to ship my samples to Patterson's lab overnight, on an ice pack.
The lab said I should not freeze it. Crazy, isn't it? Every lab has it's own rules, references etc. But my IL-8 for example is elevated on every test. And now increased by a hundred times. And everything this clown recommends is fish oil.
 

seamyb

Senior Member
Messages
560
Likes
1,312
So what's the thing with Patterson
I wouldn't be surprised if the Patterson hype machine was funded by 'Ivermectin "R" us'. He's the self-claimed leading researcher on long covid whether he gets results or not. Doesn't want to hear anything that isn't already in agreement with him. He's selling his brand, not trying to find answers.
 

nerd

Senior Member
Messages
863
Likes
2,532
This doesn't make sense. Hemolysis in serum isn't possible. RBCs are removed from it, so there can't be any aggregation. It's the very reason why this is done in the first place. The only possibility that they might mean is that the doctor's assistant centrifuged the sample too late, had your sample laying around for too long - basically a protocol violation. Maybe that's the case?

If assistants aren't trained properly, they might think that the normal rules apply for cytokines as well and that they can wait up to 24h if they refrigerate the sample. But I don't see any good reason why they would wait. It's bad practice.

The lab said I should not freeze it. Crazy, isn't it? Every lab has it's own rules, references etc.
It's generally true that most labs have different standards if they don't follow DIN standards but refrigeration and freezing are different things. Refrigeration should be fine, freezing can either preserve or destroy what you are looking for, it depends on the test.
 
Messages
2,284
Likes
7,537
If assistants aren't trained properly, they might think that the normal rules apply for cytokines as well and that they can wait up to 24h if they refrigerate the sample. But I don't see any good reason why they would wait. It's bad practice.
It was centrifuged 20 minutes after the blood draw. You're right: it doesn't make sense!
 
Messages
2,284
Likes
7,537
It's useless. And it explains what I have noticed so far: conflicting results. So the only way to be certain is: use multiple labs and repeat the test over time. If the values are reproducible you have a signature. But one result does not mean anything it seems. Thank you for this!
 

Wishful

Senior Member
Messages
4,514
Likes
8,249
Location
Alberta
I had a cytokine profile done. Whether it was done 'correctly' or not probably didn't matter. The interpretation of the results was: "There's something going on, but it's not a pattern I'm familiar with, so I can't help you." Unless you have a specific cytokine level to check, or something is drastically abnormal, I'm not sure what the value of the test is.
 

nerd

Senior Member
Messages
863
Likes
2,532
I had a cytokine profile done. Whether it was done 'correctly' or not probably didn't matter. The interpretation of the results was: "There's something going on, but it's not a pattern I'm familiar with, so I can't help you." Unless you have a specific cytokine level to check, or something is drastically abnormal, I'm not sure what the value of the test is.
It's use is mostly for personal research or to share it with researchers such as Dr. Patterson. I think the only cytokine that's actually of use is interferon gamma to see how much sense an interferon therapy makes. The interferon stimulus can also be checked.

I don't think that there's been sufficient research with lymphocyte stimulation in ME patients. This can be used to detect if the immune system has an encounter with pathogens or allergens such as mold, or if it is occupied by autoimmunity. Immune deficiency can also be detected. Moreover, it's possible to see if lymphocytes are already infected by viruses.
 
Messages
2,284
Likes
7,537
It's use is mostly for personal research or to share it with researchers such as Dr. Patterson. I think the only cytokine that's actually of use is interferon gamma to see how much sense an interferon therapy makes. The interferon stimulus can also be checked.

I don't think that there's been sufficient research with lymphocyte stimulation in ME patients. This can be used to detect if the immune system has an encounter with pathogens or allergens such as mold, or if it is occupied by autoimmunity. Immune deficiency can also be detected. Moreover, it's possible to see if lymphocytes are already infected by viruses.
You can also see chronic inflammation if that cytokine is almost gone while IL-4 is normal midrange
 
Messages
818
Likes
1,418
It's useless. And it explains what I have noticed so far: conflicting results. So the only way to be certain is: use multiple labs and repeat the test over time. If the values are reproducible you have a signature. But one result does not mean anything it seems. Thank you for this!
I'm due to have a "proper" IL-6 test soon (through an official pathology lab that does our normal blood tests). I'm going to see if it is at all similar to the result I have from the holistic medicine lab that did my full cytokine panel.

The first test was a few months ago now, but hopefully not too much has changed since then (although I have crashed since that time and used a few peptides.. So who knows).