Mar 5, 2022·edited Mar 5, 2022Liked by Modern Discontent
Total biology newb here, but my naive understanding was the lymph drains to the nodes from there into the blood stream. When the original "it stays at the injection site" claims were made, I was puzzled as that's not how I understand the lymph system to work from my studying of exercise physiology. I could be totally wrong, but my understanding was when you exercise you create lots of bits and pieces of detritus (damaged muscle cells, etc) and something like massage is great as it flushes all that "exterior to the damaged cells" gunk up to the lymph nodes and then out to the blood stream to be processed and expelled as necessary. (I remember an admonition to "massage towards the heart" for that reason).
So I recently started reading up on the immune system. In my reading on the immune system I see macrophages et al grab bits and pieces of pathogen (also vaccine), travel to the lymph nodes. Here they can present to T and B cells, get the adaptive response started, etc, etc.
If the lymph nodes drain into the blood, what's to stop the various bits and pieces of uneaten but successfully transported to the lymph node vaccine components (or generated spike) from being piped back into the blood system and thence around the body to wherever they can latch on?
Is it that only the cells with the right receptors (or what have you) can leave the lymph into the blood stream? Or is it more it's impossible to not get eaten by neutrophils, macrophages, etc on the way to the lymph node?
Apologies if this is really dumb. I am trying to learn :D
Never attribute your learning process to dumbness! Many of us are learning this as we go along. It's sometimes about the process of learning more than it is to always getting things down to a T.
But you're thinking is correct I believe. My reading of immunology is very basic it's one of the topics I really struggle with but I believe the pathway is to activate the immune system so that cells can travel to lymph nodes, ramp up immunity to then go out and target the pathogen.
I think one way to steelman the concerns over vaccine debris is to examine if similar issues may occur with viral infections. If viral debris can make their way into the bloodstream this may be a great concern with these spike proteins and really any prior vaccination regimen.
Now, the actual activities of the spike granting abilities to enter the bloodstream may be an issue. But one of the main factors here is the mRNA being found within the lymph node and that should be looked at in closer detail.
Otherwise there's so much here that I am still learning so much about.
The main issue with this study though is the presence of luciferase within the liver which certainly falls outside of the idea of "staying within the injection site".
Mar 5, 2022·edited Mar 5, 2022Liked by Modern Discontent
It would be generous to call the official narrative of what happens after injection "coherent." Does the mRNA payload go to the lymph nodes, or just spike (after transfected muscle cells express it)? But we know that the official narrative would be wrong in either case since the LNPs go straight to the ovaries in the Pfizer study.
Lymph-transported spike would likely dump into the bloodstream, though pre-existing antibodies may form a complex with loose spike which results in better interception. The LNPs are not clear-cut, but I think they would be intercepted in the Subcapsular Sinus floor which normally screens bacteria and viruses (though with the caveat that what we call an "infection" usually implies that the SCS has been breached, and so innate immunity has been overcome) - https://www.frontiersin.org/articles/10.3389/fimmu.2019.00347/full - Figure 2 is a good illustration. Excitingly, the authors tease the idea that the SCS can "grow" pathogens to generate more attention from the immune cells underneath. This could account for how memory immunity doesn't always have to wait for innate immunity to fail before developing tools against a specific bug. (SCS uniformity may be much more relevant to immunodeficiency/AIDS than lymphocytes but I still haven't looked into the whole AIDS thing at all.) However, once again the LNP dispersion suggests that this stuff isn't being intercepted at all, so I think that suggests it is just leaking into the blood from the injection site rather than evading the SCS. In this quote the last sentence answers your question about the bits and pieces.
"The “flypaper” function of SCS macrophages is also applicable to lymph-borne bacteria. Fluorescently labeled Pseudomonas aeruginosa, an extracellular bacterium, was found in the LN parenchyma and blood 8 h post-injection when the macrophages were depleted, while bacteria were limited to the SCS when the macrophage layer was intact (36). More specifically, lipid antigens, such as lipopolysaccharide found on bacteria, has also been shown to localize with the SCS macrophages (37). This further defines the “flypaper” function of the SCS macrophages as it is not only preventing a systemic spread, but specifically limits pathogens to the SCS in the LN. Restricting pathogens to the SCS is at least partially achieved by the expression of CD169 on the macrophages, as CD169 interacts with α2,3-linked sialic acids expressed on the surface of cells or microbes. Biotinylated exosomes specifically bound to SCS macrophages on tissue sections while biotinylated bovine serum did not, suggesting the CD169+ macrophages retain extracellular vesicles and microbes rather than free flow proteins at the sinus "
Your explanation of the % encapsulation part helps me think up a rationale why an "inferior" Acuitas LNP5 was included to begin with - perhaps as a benchmark for the in-house LNP12 formulation. Presumably BioNTech was hopeful but not confident that their own version was ready for prime-time.
Thank you for the kind remarks and links.
Speaking of LNPs, do you know off-hand if there is any research or theoretical argument regarding whether they can be "sequestered" somewhere in tissues or intracellularly without breaking up for a certain time? Emphasis on the "off-hand," I know you've been keeping the LNP question on the back-burner as have I so I'm not asking for any what-you-are-doings to be dropped.
It could be, you never know with these types of things but yeah I wouldn't be surprised if a bit of hubris in both designing the mRNA AND the LNP may have gotten themselves a bit over their own heads.
If you are referring to whether micellar LNP structures would exist inside the cell I don't think so. I believe fusion of the LNP with host membranes will pretty much cause them to become part of the membrane. It'd be pretty hard to find them alone because of their very high hydrophobic nature. Also the size of the LNPs also likely takes spacing of each molecule into account such that the typical sizes are the best size to provide coverage on all sides of the micelle that leaves no openings or gaps exposed to solution.
Let me know if there is anything else that might be helpful. Definitely scout out other information to get better information.
That paper is very interesting! I only looked at it superficially but the clotting in platelet poor plasma is very interesting (and concerning). A lot of the blood clots have been compared to heparin-induced thrombocytopenia especially in those given the adenovirus vaccines which would suggest clotting without platelets but caused by heparin and platelet factor aggregates. There's a lot to go through there. I had someone here post asking about fibrinolytic enzymes being helpful with COVID so this is very interesting!
Yes, a ton to go through. Pretorius has been doing spectacular work on coagulation and chronic inflammation with very little attention outside of South Africa as far as I can tell. So her team has already progressed very far with research on the spike protein.
The party line seems to be that fusion / disruption of the LNP can't occur until low pH of endosome, but I'm wondering if manufacturing quality would lead to ionizable lipids not always being present in enough quantity and if the result would be akin to a bomb that doesn't go off on landing, so I'm semi back to thinking intracellular sequestration is possible given how little we know about the quality control element. Overall it seems like an interaction with amyloid microclots might be excessive. Still, I'll probably run with it.
So this is very basic idea, but the endosomal route is likely not to be used by the vaccines and will rely solely on membrane fusion. The release of mRNA may be facilitated by endosomes but I think it may just be released freely into the cell.
Could you clarify on the ionizable nature? For the lipid tails many of them are typical hydrophobic chains and so they're hard to ionize and will likely be cleaved through similar fatty acid metabolism routes.
I was reading a year-old explainer that explained me that the heads are ionizable, and it showed a picture of the LNP being attracted to the endosome membrane once the ionizable lipids go positive in low pH, and at that point out shoots the magic transfection molecule. Prior, the endosome results from LDL binding. But the whole thing is very second-hand and could all be wrong. It has the famous "super orb" picture https://cen.acs.org/pharmaceuticals/drug-delivery/Without-lipid-shells-mRNA-vaccines/99/i8 - I would need another two week internet outage to sort out my understanding of the chemistry here.
Mar 5, 2022·edited Mar 5, 2022Liked by Modern Discontent
I understand redacting things to hide details of product construction, but it seems to me the things being redacted here are the effects. Is this necessary to prevent reverse engineering?
Because from my naive POV, it seems more they are hiding things that could prove inadequacy of clinical trial testing, rather than "we don't want competitor [X] to work out how we're doing what we're doing and copy our hard work".
Appreciate the follow up. Agreed too, that Mowrey guy is great value.
I would definitely agree it is to protect competitors from gaining any information. It's like anything that occurs downstream of the compound's design falls under the redaction as well.
And yes, his contribution helped provide so much more clarity to my post! Hence the correction. I also appreciate his perspective on OAS. I felt like there was a lot of ideas being misattributed or that the reading of the scientific evidence was off to me. I appreciate that he provides a counter argument and, I would argue, has a better grasp of the ideas of OAS and why it may not be an appropriate hypothesis to attribute everything to.
Ah yes, the OAS hypothesis presented elsewhere was the mirror of the "OMG NOVEL VIRUS" used to beat people into submission. And gave me similar pause.
Unglossed's take was a breath of rationality in amongst those competing storms. Fully agreed.
So nothing nefarious in the redactions? I found it interesting that despite this protection through redaction, they did not continue to redact with the Aussie report - or info slipped through.
Maybe they redact as thoroughly and consistently as they clinically trial these vaccines...
Well, I would say that for these LNP posts no, mainly because the formulation BioNTech decided to go with (LNP8) was already widely available and not proprietary and so the information for LNP was not redacted. However, the same can't be said for any other studies that may rely on mRNA using the spike protein. If any redactions obstruct any ability to assess the data then that would be something we should be really concerned about.
I think the fewer number of redactions with Australia may be a difference in bureaucratic structure, which may mean that the FDA chose to redact some information or that Australia may require more transparency with their data. What's weird is that the summary in the Australian report redacted the main findings which is quite strange considering the more elaborate explanations were left in.
As for the clinical trials, I guess I should add an addendum that clinical trials tend to be presented in a manner that benefits the pharmaceutical company the most. It's kind of a commonplace idea that, at least for SSRIs many companies may conduct several clinical trials and may tend to hide any trials that not show their SSRI in a good light.
Indeed it is difficult to parse through their redacted information! Kudos to Brian for providing the un-redacted information. I was looking briefly at another Pfizer document which does indicate that they studied a radioisotope LNP which also showed travel from injection site to mainly the liver (also spleen, adrenal gland, bone marrow, ovaries, as well as trace amounts pretty much everywhere they examined). Interestingly the LNP also contained the mRNA of luciferase (but not much else is mentioned about it - it was not measured). From the paper: Once intracellular, the [3H]-CHE does not recirculate and therefore allows assessment of distribution of the particle. So that would seem to indicate that they do not anticipate movement once an LNP has been taken up by a cell.
I am also quite intrigued by the modifications of the mRNA used in the vaccines. Seems they also added a poly-A tail as well as the pseudouridine swap to help evade immediate degradation by our immune system. So much we don't know ... yet!
Thank you for the document! I actually saw that this is the study right underneath the study I just examined within the Australian report. I think the Naked Emperor looked at this paper but focused mostly on that figure of dispersion so I think I will look at it a little deeper and see what else there is. It's ridiculous that the information is so blatant that there's a lot of circulation and dispersion on. Of course, the rebuttal now will be that "well it's not that bad!"
Total biology newb here, but my naive understanding was the lymph drains to the nodes from there into the blood stream. When the original "it stays at the injection site" claims were made, I was puzzled as that's not how I understand the lymph system to work from my studying of exercise physiology. I could be totally wrong, but my understanding was when you exercise you create lots of bits and pieces of detritus (damaged muscle cells, etc) and something like massage is great as it flushes all that "exterior to the damaged cells" gunk up to the lymph nodes and then out to the blood stream to be processed and expelled as necessary. (I remember an admonition to "massage towards the heart" for that reason).
So I recently started reading up on the immune system. In my reading on the immune system I see macrophages et al grab bits and pieces of pathogen (also vaccine), travel to the lymph nodes. Here they can present to T and B cells, get the adaptive response started, etc, etc.
If the lymph nodes drain into the blood, what's to stop the various bits and pieces of uneaten but successfully transported to the lymph node vaccine components (or generated spike) from being piped back into the blood system and thence around the body to wherever they can latch on?
Is it that only the cells with the right receptors (or what have you) can leave the lymph into the blood stream? Or is it more it's impossible to not get eaten by neutrophils, macrophages, etc on the way to the lymph node?
Apologies if this is really dumb. I am trying to learn :D
Never attribute your learning process to dumbness! Many of us are learning this as we go along. It's sometimes about the process of learning more than it is to always getting things down to a T.
But you're thinking is correct I believe. My reading of immunology is very basic it's one of the topics I really struggle with but I believe the pathway is to activate the immune system so that cells can travel to lymph nodes, ramp up immunity to then go out and target the pathogen.
I think one way to steelman the concerns over vaccine debris is to examine if similar issues may occur with viral infections. If viral debris can make their way into the bloodstream this may be a great concern with these spike proteins and really any prior vaccination regimen.
Now, the actual activities of the spike granting abilities to enter the bloodstream may be an issue. But one of the main factors here is the mRNA being found within the lymph node and that should be looked at in closer detail.
Otherwise there's so much here that I am still learning so much about.
The main issue with this study though is the presence of luciferase within the liver which certainly falls outside of the idea of "staying within the injection site".
It would be generous to call the official narrative of what happens after injection "coherent." Does the mRNA payload go to the lymph nodes, or just spike (after transfected muscle cells express it)? But we know that the official narrative would be wrong in either case since the LNPs go straight to the ovaries in the Pfizer study.
Lymph-transported spike would likely dump into the bloodstream, though pre-existing antibodies may form a complex with loose spike which results in better interception. The LNPs are not clear-cut, but I think they would be intercepted in the Subcapsular Sinus floor which normally screens bacteria and viruses (though with the caveat that what we call an "infection" usually implies that the SCS has been breached, and so innate immunity has been overcome) - https://www.frontiersin.org/articles/10.3389/fimmu.2019.00347/full - Figure 2 is a good illustration. Excitingly, the authors tease the idea that the SCS can "grow" pathogens to generate more attention from the immune cells underneath. This could account for how memory immunity doesn't always have to wait for innate immunity to fail before developing tools against a specific bug. (SCS uniformity may be much more relevant to immunodeficiency/AIDS than lymphocytes but I still haven't looked into the whole AIDS thing at all.) However, once again the LNP dispersion suggests that this stuff isn't being intercepted at all, so I think that suggests it is just leaking into the blood from the injection site rather than evading the SCS. In this quote the last sentence answers your question about the bits and pieces.
"The “flypaper” function of SCS macrophages is also applicable to lymph-borne bacteria. Fluorescently labeled Pseudomonas aeruginosa, an extracellular bacterium, was found in the LN parenchyma and blood 8 h post-injection when the macrophages were depleted, while bacteria were limited to the SCS when the macrophage layer was intact (36). More specifically, lipid antigens, such as lipopolysaccharide found on bacteria, has also been shown to localize with the SCS macrophages (37). This further defines the “flypaper” function of the SCS macrophages as it is not only preventing a systemic spread, but specifically limits pathogens to the SCS in the LN. Restricting pathogens to the SCS is at least partially achieved by the expression of CD169 on the macrophages, as CD169 interacts with α2,3-linked sialic acids expressed on the surface of cells or microbes. Biotinylated exosomes specifically bound to SCS macrophages on tissue sections while biotinylated bovine serum did not, suggesting the CD169+ macrophages retain extracellular vesicles and microbes rather than free flow proteins at the sinus "
Your explanation of the % encapsulation part helps me think up a rationale why an "inferior" Acuitas LNP5 was included to begin with - perhaps as a benchmark for the in-house LNP12 formulation. Presumably BioNTech was hopeful but not confident that their own version was ready for prime-time.
Thank you for the kind remarks and links.
Speaking of LNPs, do you know off-hand if there is any research or theoretical argument regarding whether they can be "sequestered" somewhere in tissues or intracellularly without breaking up for a certain time? Emphasis on the "off-hand," I know you've been keeping the LNP question on the back-burner as have I so I'm not asking for any what-you-are-doings to be dropped.
It could be, you never know with these types of things but yeah I wouldn't be surprised if a bit of hubris in both designing the mRNA AND the LNP may have gotten themselves a bit over their own heads.
If you are referring to whether micellar LNP structures would exist inside the cell I don't think so. I believe fusion of the LNP with host membranes will pretty much cause them to become part of the membrane. It'd be pretty hard to find them alone because of their very high hydrophobic nature. Also the size of the LNPs also likely takes spacing of each molecule into account such that the typical sizes are the best size to provide coverage on all sides of the micelle that leaves no openings or gaps exposed to solution.
Thanks - that's actually very helpful as I was hoping to just focus on the extracellular potential.
If you want a preview of where my thinking is going, see figure 6H in https://www.ncbi.nlm.nih.gov/labs/pmc/articles/PMC8380922/
Let me know if there is anything else that might be helpful. Definitely scout out other information to get better information.
That paper is very interesting! I only looked at it superficially but the clotting in platelet poor plasma is very interesting (and concerning). A lot of the blood clots have been compared to heparin-induced thrombocytopenia especially in those given the adenovirus vaccines which would suggest clotting without platelets but caused by heparin and platelet factor aggregates. There's a lot to go through there. I had someone here post asking about fibrinolytic enzymes being helpful with COVID so this is very interesting!
Yes, a ton to go through. Pretorius has been doing spectacular work on coagulation and chronic inflammation with very little attention outside of South Africa as far as I can tell. So her team has already progressed very far with research on the spike protein.
The party line seems to be that fusion / disruption of the LNP can't occur until low pH of endosome, but I'm wondering if manufacturing quality would lead to ionizable lipids not always being present in enough quantity and if the result would be akin to a bomb that doesn't go off on landing, so I'm semi back to thinking intracellular sequestration is possible given how little we know about the quality control element. Overall it seems like an interaction with amyloid microclots might be excessive. Still, I'll probably run with it.
So this is very basic idea, but the endosomal route is likely not to be used by the vaccines and will rely solely on membrane fusion. The release of mRNA may be facilitated by endosomes but I think it may just be released freely into the cell.
Could you clarify on the ionizable nature? For the lipid tails many of them are typical hydrophobic chains and so they're hard to ionize and will likely be cleaved through similar fatty acid metabolism routes.
I was reading a year-old explainer that explained me that the heads are ionizable, and it showed a picture of the LNP being attracted to the endosome membrane once the ionizable lipids go positive in low pH, and at that point out shoots the magic transfection molecule. Prior, the endosome results from LDL binding. But the whole thing is very second-hand and could all be wrong. It has the famous "super orb" picture https://cen.acs.org/pharmaceuticals/drug-delivery/Without-lipid-shells-mRNA-vaccines/99/i8 - I would need another two week internet outage to sort out my understanding of the chemistry here.
I understand redacting things to hide details of product construction, but it seems to me the things being redacted here are the effects. Is this necessary to prevent reverse engineering?
Because from my naive POV, it seems more they are hiding things that could prove inadequacy of clinical trial testing, rather than "we don't want competitor [X] to work out how we're doing what we're doing and copy our hard work".
Appreciate the follow up. Agreed too, that Mowrey guy is great value.
I would definitely agree it is to protect competitors from gaining any information. It's like anything that occurs downstream of the compound's design falls under the redaction as well.
And yes, his contribution helped provide so much more clarity to my post! Hence the correction. I also appreciate his perspective on OAS. I felt like there was a lot of ideas being misattributed or that the reading of the scientific evidence was off to me. I appreciate that he provides a counter argument and, I would argue, has a better grasp of the ideas of OAS and why it may not be an appropriate hypothesis to attribute everything to.
Ah yes, the OAS hypothesis presented elsewhere was the mirror of the "OMG NOVEL VIRUS" used to beat people into submission. And gave me similar pause.
Unglossed's take was a breath of rationality in amongst those competing storms. Fully agreed.
So nothing nefarious in the redactions? I found it interesting that despite this protection through redaction, they did not continue to redact with the Aussie report - or info slipped through.
Maybe they redact as thoroughly and consistently as they clinically trial these vaccines...
Well, I would say that for these LNP posts no, mainly because the formulation BioNTech decided to go with (LNP8) was already widely available and not proprietary and so the information for LNP was not redacted. However, the same can't be said for any other studies that may rely on mRNA using the spike protein. If any redactions obstruct any ability to assess the data then that would be something we should be really concerned about.
I think the fewer number of redactions with Australia may be a difference in bureaucratic structure, which may mean that the FDA chose to redact some information or that Australia may require more transparency with their data. What's weird is that the summary in the Australian report redacted the main findings which is quite strange considering the more elaborate explanations were left in.
As for the clinical trials, I guess I should add an addendum that clinical trials tend to be presented in a manner that benefits the pharmaceutical company the most. It's kind of a commonplace idea that, at least for SSRIs many companies may conduct several clinical trials and may tend to hide any trials that not show their SSRI in a good light.
Indeed it is difficult to parse through their redacted information! Kudos to Brian for providing the un-redacted information. I was looking briefly at another Pfizer document which does indicate that they studied a radioisotope LNP which also showed travel from injection site to mainly the liver (also spleen, adrenal gland, bone marrow, ovaries, as well as trace amounts pretty much everywhere they examined). Interestingly the LNP also contained the mRNA of luciferase (but not much else is mentioned about it - it was not measured). From the paper: Once intracellular, the [3H]-CHE does not recirculate and therefore allows assessment of distribution of the particle. So that would seem to indicate that they do not anticipate movement once an LNP has been taken up by a cell.
I am also quite intrigued by the modifications of the mRNA used in the vaccines. Seems they also added a poly-A tail as well as the pseudouridine swap to help evade immediate degradation by our immune system. So much we don't know ... yet!
I certainly appreciate your attempt to sift through these papers - surely it seems they were aware that "it never leaves the injection site" was a blatant lie. Here is the paper I mentioned: https://phmpt.org/wp-content/uploads/2022/03/125742_S1_M4_4223_185350.pdf
Thank you for the document! I actually saw that this is the study right underneath the study I just examined within the Australian report. I think the Naked Emperor looked at this paper but focused mostly on that figure of dispersion so I think I will look at it a little deeper and see what else there is. It's ridiculous that the information is so blatant that there's a lot of circulation and dispersion on. Of course, the rebuttal now will be that "well it's not that bad!"
"Of course, the rebuttal now will be that "well it's not that bad!""
Or a self-serving / obfuscaring / obfuscating (I like obfuscaring now haha),
"It proves the (lymph system / immune response / vaccine) is working!"