1/🧵 🚨🚨🚨 💉
ds DNA Causes RAPID CLOT FORMATION in MICE in the
LINING of BLOOD VESSELS (arteries/veins/capillaries=vascular endothelium),
but ONLY in the presence of a
POSITIVELY (+) CHARGED LIPID TRANSFECTION AGENT.
RNA Vaccines contain ds DNA Contamination
A Study:
ds DNA Causes RAPID CLOT FORMATION in MICE in the
LINING of BLOOD VESSELS (arteries/veins/capillaries=vascular endothelium),
but ONLY in the presence of a
POSITIVELY (+) CHARGED LIPID TRANSFECTION AGENT.
RNA Vaccines contain ds DNA Contamination
A Study:
2/ THE STUDY:
"Double-stranded DNA induces a prothrombotic phenotype in the vascular endothelium"
Gaitzsch, E., Czermak, T., Ribeiro, A. et al. Double-stranded DNA induces a prothrombotic phenotype in the vascular endothelium. Sci Rep 7, 1112 (2017). doi.org/10.1038/s41598…
"Double-stranded DNA induces a prothrombotic phenotype in the vascular endothelium"
Gaitzsch, E., Czermak, T., Ribeiro, A. et al. Double-stranded DNA induces a prothrombotic phenotype in the vascular endothelium. Sci Rep 7, 1112 (2017). doi.org/10.1038/s41598…
3/
The study investigates the prothrombotic (CLOT) effects of double-stranded DNA (dsDNA) in vascular endothelial cells.
Little is known (at the time of the study) about the pathophysiological relevance of dsDNA for the vascular endothelium. nature.com/articles/s4159…
The study investigates the prothrombotic (CLOT) effects of double-stranded DNA (dsDNA) in vascular endothelial cells.
Little is known (at the time of the study) about the pathophysiological relevance of dsDNA for the vascular endothelium. nature.com/articles/s4159…
4/ dsDNA is a potent activator of the innate immune system.
Viral infections involving dsDNA can lead to systemic diseases with vascular complications and thrombotic events (hepatitis is one).
vascular endothelium plays a major role in defense, hemostasis, and inflammation.
Viral infections involving dsDNA can lead to systemic diseases with vascular complications and thrombotic events (hepatitis is one).
vascular endothelium plays a major role in defense, hemostasis, and inflammation.
5/ Exogenous dsDNA (double stranded DNA), ONLY in the presence of a positively charged (cationic lipid agent) disrupts the "haemostatic balance" and drives a prothrombotic state (clot formation.
The study uses poly(dA:dT), a synthetic dsDNA analog,
The study uses poly(dA:dT), a synthetic dsDNA analog,
6/ to mimic the effects of viral DNA. Similar prothrombotic effects were observed with hepatitis B virus DNA-containing immunoprecipitates.
7/ dsDNA induces upregulation of procoagulant proteins (tissue factor and plasminogen activator inhibitor-1 (PAI-1)) in endothelial cells.
Accelerated blood clotting in vitro and increased thrombus formation in vivo (IN MICE) were observed after exposure to dsDNA, but only
Accelerated blood clotting in vitro and increased thrombus formation in vivo (IN MICE) were observed after exposure to dsDNA, but only
8/ in the presence of positively charged (cationic) lipids!
The mechanisms and the molecular signaling pathways are:
dsDNA induces nuclear translocation of transcription factors IRF3 and NF-κB, engaging proinflammatory and prothrombotic gene regulation.
Multiple receptors,
The mechanisms and the molecular signaling pathways are:
dsDNA induces nuclear translocation of transcription factors IRF3 and NF-κB, engaging proinflammatory and prothrombotic gene regulation.
Multiple receptors,
9/ including TLR9 and RIG-I, are involved in dsDNA signaling in endothelial cells.
dsDNA triggers surface expression of von Willebrand factor (vWF), leading to platelet-endothelium interactions.
(multiple threads on dsDNA and TLR 9/cGAS STING: )
dsDNA triggers surface expression of von Willebrand factor (vWF), leading to platelet-endothelium interactions.
(multiple threads on dsDNA and TLR 9/cGAS STING: )
https://twitter.com/_HeartofGrace_/status/1724373484602544226
10/ *Von Willebrand Factor (vWF) is a glycoprotein that is critical in blood clotting and hemostasis. It is primarily produced and released by endothelial cells and megakaryocytes. The main functions of vWF: activates platelet adhesion to the lining of vessels, aggregation
11/ of platelets, and vWF binds and stabilizes factor VIII in the blood, preventing its degradation.
dsDNA is implicated in viral infections like hepatitis B, which often result in systemic diseases affecting various organs, including the vascular system
dsDNA is implicated in viral infections like hepatitis B, which often result in systemic diseases affecting various organs, including the vascular system
12/ endothelium (inner layer of blood vessels) plays a crucial role in host defense by acting as a barrier against pathogens. dsDNA is a pathogen and is recognized by our innate immune system.
13/ Previous studies show vascular endothelium can sense intracellular dsDNA, eliciting a strong inflammatory response.
This study shows the prothrombotic (clotting) effects of dsDNA in the vascular endothelium, in the presence of cationic (positively) charged lipid transfection
This study shows the prothrombotic (clotting) effects of dsDNA in the vascular endothelium, in the presence of cationic (positively) charged lipid transfection
14/ Methods: Human microvascular endothelial cells (HMEC) were treated with poly(dA:dT) at a concentration of 5 µg/mL.
Two conditions were tested: poly(dA:dT) alone and poly(dA:dT) complexed with cationic lipids (Lipofectamine 2000).
Only the dsDNA in the cationic lipids
Two conditions were tested: poly(dA:dT) alone and poly(dA:dT) complexed with cationic lipids (Lipofectamine 2000).
Only the dsDNA in the cationic lipids
15/ interacted inside the cells [this could occur transiently, but for this study, they only looked at "transfection". ]
This engaged the nuclear translocation of transcription factors IRF3.Nuclear translocation of NF-κB was also observed (Immunofluorescent staining)
This engaged the nuclear translocation of transcription factors IRF3.Nuclear translocation of NF-κB was also observed (Immunofluorescent staining)
16/ Bright field was also used to detect the uptake and interactions in the cells.
*********************************************
🐭Then, mice were used. 🐭
A transfection agent with and without dsDNA was injected into mice.
dsDNA significantly accelerated thrombus formation
*********************************************
🐭Then, mice were used. 🐭
A transfection agent with and without dsDNA was injected into mice.
dsDNA significantly accelerated thrombus formation
17/ the injection of dsDNA complexed with Lipofectamine (cationic lipid reagent) into mice accelerates clot formation-- a prothrombotic effect of double-stranded DNA in the microvasculature, leading to quicker onset and complete vessel occlusion, especially with vessel injury.
18/ the study conducted the in vivo experiments using C57Bl/6 mice, a common strain used in biomedical research
P < 0.05 vs. control
The dsDNA injection with cationic lipids resulted in a more rapid onset of thrombus formation in arterioles
P < 0.05 vs. control
The dsDNA injection with cationic lipids resulted in a more rapid onset of thrombus formation in arterioles
19/ CLOT FORMATION:
dsDNA induces upregulation of procoagulant proteins: tissue factor and plasminogen activator inhibitor-1 (PAI-1) in endothelial cells.
Accelerated blood clotting in vitro and increased thrombus formation in vivo were observed after exposure to dsDNA.
dsDNA induces upregulation of procoagulant proteins: tissue factor and plasminogen activator inhibitor-1 (PAI-1) in endothelial cells.
Accelerated blood clotting in vitro and increased thrombus formation in vivo were observed after exposure to dsDNA.
20/ And again. dsDNA triggers surface expression of von Willebrand factor (vWF), leading to platelet-endothelium interactions, with the upregulation of tissue factor and PAI-1-- a prothrombotic phenotype with increased tissue plasminogen activator (tPA)
21/ Layman's: double stranded DNA was used in this study both in cells and in mice, with positively charged lipids in a transfection reagent and without. dsDNA was found to cause faster blood clotting both in cells, and in mice, in the presence of positively charged lipids.
22/ one might make the connection, that because this study shows both in cells and in mice that double stranded DNA, which is also the main backbone of the DNA plasmid contamination discovered in the rna vaccines, that this is also a mechanism for clotting (on top of the
23/ spike protein, positive net charged LNP going to the lungs and causing clots, and the negatively charged lipids causing clots. This is yet another mechanism to cause clots.
Also, the anti phospholipid syndrome would cause clots.
Also, the anti phospholipid syndrome would cause clots.
https://twitter.com/_HeartofGrace_/status/1722031142851817706
25/
https://twitter.com/_HeartofGrace_/status/1722985047651663903
https://twitter.com/_HeartofGrace_/status/1721959669231419416
https://twitter.com/_HeartofGrace_/status/1725484645985251683
https://twitter.com/_HeartofGrace_/status/1714311719961477455
@P_J_Buckhaults
@drdrew
@DrJBhattacharya
(is it OK that I typically tag you two together?)
;)
@DrJBhattacharya
(is it OK that I typically tag you two together?)
;)
26/ amendment: poly dA:dT IS the dsDNA
(endothelium--my bad)
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