Accessibility Tools

Skip to main content

React19 Research: The Spike Protein Problem

03 December 2022

Part 1: Spike Protein Lit Review   |   Part 2: Deeper Dive - The S1 Segment   |   Part 3: Persistence of Spike In the Body

 

 

Part 1: Spike Protein Lit Review

1st indicator that the spike protein was a problem is by looking at the vaccine adverse reaction literature. See attached, over 1300 articles. What you notice by reviewing these articles is that all the vaccines seem to have the same side effect profile. For example, myocarditis has been reported for both J&J and also AstraZeneca. Another example, the vaccine induced thrombotic thrombocytopenia that paused the J&J roll out in the US, which is unique in that it has positive PF4 antibodies without the patient being exposed to heparin, has also been described in the mRNA vaccines. These vaccines use different technology but the common theme among them is the spike protein.   There is also overlap between the symptoms of vaccine injured and COVID long haulers in that these patients look exactly the same. Check out these two articles, both co-authored by Dr.Nath of the NIH, regarding small fiber neuropathy and dysautonomia:

  And it’s not just long hauler syndrome, it’s any complication from COVID-19 infection reported in the medical literature, you can also find it for the vaccine injured. Examples:

  Again, the common factor here is the spike protein in order to explain how vaccine injured and covid actively infected/covid long haulers have the same illness.   Thus I started looking for autopsy and biopsy reports that would confirm the presence of spike protein. This is what I found:

  1. Endomyocardial biopsies of vaccine induced myocarditis shows presence of spike protein in 9 patients: https://www.mdpi.com/1422-0067/23/13/6940/htm?fbclid=IwAR2IVfyZICfANC42TsFE7qI73Ud1OhMsu3aOEASZEsnk4AEujxO9Zu9Vww4
  2. Spike protein in nonclassical monocytes in covid long haulers: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784688/
  3. Spike protein persistence in nonclassical monocytes on vaccine injured patients with similar/same symptoms of covid long hauler patients: https://www.researchsquare.com/article/rs-1844677/v1
  4. Spike protein found in skin biopsies of rashes in post-vaccine patients: https://www.sciencedirect.com/science/article/pii/S0738081X21001516
  5. Spike protein presence in a covid positive patient chilblain-like skin lesion biopsy: https://onlinelibrary.wiley.com/doi/10.1111/jdv.16970
  6. Spike protein presence in a skin biopsy of morphea rash following mRNA vaccine (found in sweat gland, inflammatory cells, and vessels): https://onlinelibrary.wiley.com/doi/10.1111/ijd.16062
  7. Spike protein detected in plasma in the plasma of MIS children who are weeks past their original infection: https://dm5migu4zj3pb.cloudfront.net/manuscripts/149000/149633/cache/149633.2-20210713124017-covered-e0fd13ba177f913fd3156f593ead4cfd.pdf

  An argument often made is that the vaccine spike was supposed to only stay in the deltoid. Here’s an article that demonstrated presence of whole spike protein and S1 subunit in the plasma of participants who received the Moderna vaccine: https://academic.oup.com/cid/article/74/4/715/6279075?login=false  

Part 2: Deeper Dive - The S1 Segment

So, the spike protein isn’t the whole virus; how can this particle itself be dangerous? Can this protein replicate itself outside of the whole virus?

The spike protein.

Here’s a picture of what this looks like on COVID.       The spike protein is how the virus attaches. Most often studied and proven is ACE-2 receptors as a site of its attachment.      

 

Taking a deeper look at the spike protein composition:

  • “Spike protein consists of two subunits, S1 -- containing the receptor-binding domain (RBD) that engages ACE2, and S2 -- containing the fusion machinery required for virus-cell membrane fusion “
  • “Two cleavage sites, S1/S2 and S2’ separate S1 and S2 and must be cleaved by host proteases for Spike to mediate virus-cell fusion. Furin-like proteases, cathepsin L, and TMPRSS2 are able to cleave these sites, making them essential host factors for SARS-CoV-2 infection (10, 13–15). RBD engagement of ACE2 triggers conformational changes in S that result in S1 shedding and insertion of the fusion peptide into the host membrane”
  • “A unique element of the SARS-CoV-2 entry cascade is that the RBD-containing S1 portion of Spike can be shed from the surface of virions following engagement of the ACE2 receptor, suggesting that shed-S1 may also promote pathology via interactions with epithelial and endothelial cells independently of the virion”
  • “Our study revealed that full-length Spike and the RBD from SARS-CoV-2 were sufficient to mediate barrier dysfunction and vascular leak in an ACE2-independent manner. Further, transcriptional analyses showed that Spike modulates expression of transcripts involved in regulation of the extracellular matrix “
  • “Cell surface proteins on epithelial and endothelial surfaces are surrounded by a dense mesh of glycans termed the epithelial/endothelial glycocalyx layer (EGL). The EGL includes sialic acid and glycosaminoglycans (GAGs) and serves as a critical determinant of barrier function, protecting epithelial and endothelial cells from shear stress”
  • “We found that Spike treatment of both HPMEC and Calu-3 cells resulted in a significant decrease in EGL components compared to control conditions, and conversely, led to upregulation of EGL-degrading enzymes (Figures 2A–H). These data suggest that, like flavivirus NS1, SARS-CoV-2 Spike mediates disruption of the EGL.”
  • “Finally, we tested whether Spike from multiple SARS-CoV-2 variants of concern could trigger barrier dysfunction and found that, comparable to S protein from the ancestral Wuhan variant, Spike derived from alpha, beta, gamma, and delta variants could all facilitate EGL disruption of HPMEC, indicating that this phenomenon is not specific to one variant, but instead is a generalizable property of SARS-CoV-2 S.”
  • “we administered Spike intranasally and then measured accumulation of dextran-680 in various organs to evaluate both local (lungs) and distal (spleen, small intestine, liver, and brain) vascular leak. We found that SARS-CoV-2 Spike significantly induced vascular leak locally in the lungs as well as distally in the spleen and small intestine, with trending but non-significant leak measured in the liver and brain, as determined through accumulation of dextran-680
  • “Our study reveals the capacity and mechanism by which SARS-CoV-2 S mediates barrier dysfunction in epithelial and endothelial cells in vitroand vascular leak in vivo, thus suggesting that Spike alone can mediate barrier dysfunction independently from viral infection”
  • Source: Biering et al. bioRvix. Dec 2021 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8687463/

  In sum, when the covid virus binds to your cells, the S1 subunit is cleaved off. This S1 subunit independently causes cellular damage, separate from the virus as a whole, causing vascular leak and resulting immune response. All variant spikes can do this. Picture for reference, also from the paper above, which is a “Graphical abstract summarizing the ACE2-independent pathway by which SARS-CoV-2 S triggers barrier dysfunction.”: This idea of a viral protein independently being pathogenic, nor the idea of retain viral particles (viral RNA) causing ongoing cellular damage in absence of the infectious virus itself, is not new nor unique to COVID-19.

  • “ Little is known about the nature of the viral RNA that is detected in measles or other acute RNA viral infections or whether cells with viral RNA are the originally infected cells that survived acute infection and avoided immune elimination or newly infected cells through continued cell-to-cell transfer of viral RNA.
    • “the identification of measles virus as the cause of subacute sclerosing panencephalitis (SSPE), a progressive fatal central nervous system (CNS) disease that becomes manifest many years after apparent recovery from the original acute measles virus infection”
  • “After recovery of experimentally infected nonhuman primates from acute Ebola and Marburg filovirus infections, viral RNA is no longer detectable in primary sites of replication such as the liver but can often be found in the eyes and testes, where macrophages and Sertoli cells, respectively, remain RNA positive
    • “the late appearance of uveitis (Box 1) and recurrence of encephalomyelitis (Box 1) due to Ebola virus infection have emphasized the importance of RNA persistence in the eye, as well as the brain, and the potential for causing progressive disease”
  • “Sexual transmission of Zika, Marburg, and Ebola viruses months to years after recovery from acute disease has also highlighted the importance of virus persistence in the testes for triggering new chains of transmission and transfer to new geographic regions”
  • Source: https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3001687&fbclid=IwAR0IsdB8570vyFfG0hawIPGIOvL6ncwO-iGjvOKEC0CSIzMJ2wJA1SoTG2k%20long%20term%20persistent%20RNA%20is%20capable%20of%20reproduction;%20other%20viruses%20RNA%20have%20been%20found%20to%20be%20persistent%20and%20cause%20issues,%20such

  So the first paper suggests that the spike protein, specifically the S1 subunit, causes endothelial dysfunction. But this is not the only paper published that demonstrates this.

  • https://pubmed.ncbi.nlm.nih.gov/34156871/ Mice model injecting isolated S1 protein
    • “we show that the activated (cleaved) form of the SP, S1SP, elicits strong pulmonary and systemic inflammatory responses in K18-hACE2 mice and milder responses in WT mice, whereas the intact SP provokes minimal or no responses.”
    • “we report that S1SP, when instilled intratracheally, produces biochemically, immunologically, and histologically evident COVID-19-like ALI, including “cytokine storm.” In agreement with these findings, we also report a direct effect of S1SP on human lung microvascular endothelial cell barrier integrity, in culture. This mouse model is easily reproducible and can be used for the study of new therapeutic approaches to COVID-19 outside BSL3 environments.”
  • https://pubmed.ncbi.nlm.nih.gov/34943787/ This paper looks at spike protein and evaluate dendritic cell response to it
    • “Here we evaluated human DCs in response to SARS-CoV-2 Spike protein, or to a fragment encompassing the receptor binding domain (RBD) challenge. Both proteins increased the expression of maturation markers, including MHC molecules and costimulatory receptors. DCs interaction with the SARS-CoV-2 Spike protein promotes activation of key signaling molecules involved in inflammation, including MAPK, AKT, STAT1, and NFκB, which correlates with the expression and secretion of distinctive proinflammatory cytokines. Differences in the expression of ACE2 along the differentiation of human monocytes to mature DCs and inter-donor were found. Our results show that SARS-CoV-2 Spike protein promotes inflammatory response and provides molecular links between individual variations and the degree of response against this virus.
  • https://www.jbc.org/article/S0021-9258(22)00135-1/fulltext This paper looks at spike protein effects using human umbilical vein endothelial cells and also in rats
    • “These results show that spike alone activates the proinflammatory program in endothelial cells and suggest that the RGD sequence located in the spike receptor–binding domain is responsible for this effect.”
    • “spike interfered with the peripheral distribution of cluster of differentiation 31 (CD31), an adhesion protein that maintains the junctional integrity of Endothelial Cells (27) (5B). These observations provided direct evidence of spike-induced increase in Endothelial cell permeability.”
    • “To investigate whether the spike protein alone could promote EC hyperpermeabilityin vivo, 2 μl of vehicle (PBS) alone or containing 0.5 μg of spike was injected into the vitreous of rats, and the retinal vasculature was evaluated in flat-mounted retinas 24 h after intravitreal injection. Spike treatment caused multiple retinal hemorrhagic areas that were absent in control retinas injected with PBS ( 6A). In other experiments, the accumulation of Evans blue–linked albumin was measured as an index of enhanced retinal vasopermeability. Spike (0.25 μg) induced a significant (greater than two-fold) increase in tracer accumulation in the retina 24 h after intravitreal injection (Fig. 6B). These findings show that spike disrupts the barrier function of retinal capillaries resulting in vascular leakage.”

 

  • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8757925/ This paper summaries research regarding the spike protein as being pathogenic
    • “One paper reported perivascular inflammation in brains of deceased patients with COVID-19, while others showed that the spike protein could damage the endothelium in an animal model, that it could disrupt an in vitro model of the blood-brain barrier (BBB), and that it can cross the BBB resulting in perivascular inflammation. “
    • “the spike protein appears to share antigenic epitopes with human molecular chaperons resulting in autoimmunity and can activate toll-like receptors (TLRs), leading to release of inflammatory cytokines. Moreover, some antibodies produced against the spike protein may not be neutralizing, but may change its conformation rendering it more likely to bind to its receptor. As a result, one wonders whether the spike protein entering the brain or being expressed by brain cells could activate microglia, alone or together with inflammatory cytokines, since protective antibodies could not cross the BBB, leading to neuro-inflammation and contributing to long-COVID
    • “Free spike protein could have a number of direct pathologic actions on different cell types (Fig.​(Fig.1A).1A). These include direct stimulation of peripheral nerves [105] and stimulation of release of pro-inflammatory and vasoactive mediators [106, 107], especially platelet-activating factor (PAF)”
    • “One paper reported that the spike protein could damage the endothelium in an animal model [110], while another paper showed that recombinant S1 RBD can damage mouse brain endothelial cells in vitro by inducing degradation of endothelial junction proteins, thus affecting endothelial barrier function [111].”
    • “recent paper reported rapid internalization of S1 RBD and of the spike RBD active trimer by cultured human brain microvascular endothelial cells, followed by increased permeability of transferrin and dextran, as well as mitochondrial damage [112].”
    • “Another recent paper using a 3D-BBB microfluidic model showed that S1 upregulated ACE2 expression and triggered RhoA activation, a key molecule regulating endothelial cytoskeleton [113].”
    • “Yet, another paper reported that spike-transfected human epithelial cells showed increased senescence-associated secretory and inflammatory proteins [114].”
    • “Two other papers reported that the spike protein could disrupt the barrier function in an in vitro model of the blood-brain barrier (BBB) [115] and that the S1 protein can actually cross the BBB and enter the brain in mice [116] (Fig.​(Fig.1A).1A)”
    • “Using transgenic mice expressing the human sigma protein, it was shown that intranasal infection with SARS-CoV-2 rapidly induced ischemic-like reactivity in brain pericytes and the S protein reached the brain of the mice [117].”
    • “In addition to direct damage, the spike protein appears to share antigenic epitopes with human molecular chaperons resulting in autoimmunity against endothelial cells [118]. Moreover, a recent paper showed that spike epitopes could form heterodimeric complexes with selected human glial proteins [119].“
    • “The S1 subunit can also bind to the surface glycoprotein neuropilin-1 (NRP-1), thus increasing infectivity, but also dysregulating angiogenesis, immune responses, and neuronal development [138,139].
    • ”New evidence indicates that the spike protein also binds to heparan sulfate (HS) molecules expressed on the surface of target cells, with mutant variants having higher binding affinity to HS [135]. This binding may be due to the fact that the SARS-CoV-2 spike protein contains four more positively charged and five fewer negatively charged residues than SARS-CoV, thus increasing the binding affinity of SARS-CoV-2 for HS [136].”
    • “Lastly, a recent paper analyzed human fetal expression of six different S protein “interactors” and showed weak expression of ACE2 and TMPRSS2, but high expression of furin with peak expression 12–26 weeks post conception; moreover, using publically available single-cell RNA sequencing datasets, it was shown that these interactors showed higher co-expression with neurons [155]. This finding indicates that the spike protein can adversely affect the developing brain and potentially lead to neurologic complications in neonates of infected mothers [156], including autism spectrum disorder [157].”

In conclusion, there is a preponderance of evidence that the spike protein both as a whole and also as its S1 subunit are biologically active and cause harm. Keep note of the cleavage of the S1 subunit will be important later on with the vaccine safety. Still the question remains, why is the spike persistent in long haulers and in vaccine injured? Is it the same original spike or is there a replication process of the protein? Why do most people do well with COVID and clear the spike protein where as others become devastated from it?  

Summary so far:

-Spike protein is the link that explains why covid long haulers/covid complications and covid vaccine adverse reactions are medically the same - Here's a new autopsy report of a Pfizer vaccine death proven on autopsy by finding spike protein in vessels, brain, and cardiac tissue:  https://www.mdpi.com/2076-393X/10/10/1651 -Spike protein, especially the S1 subunit, is shed after full spike binding and causes damage via vascular endothelial damage and once it makes holes in the vascular lining, it then enters organs; this is a generalizable property of all COVID spike proteins in both virus variants and also the S1 subunit shedding has been proven to occur from vaccine spike protein too Lingering next question from the last email is why is the spike protein persistent? Can it replicate without the virus?

 

 

Part 3: Persistence of Spike In the Body - It's all in the delivery system

The landmark paper for mRNA research was in 1990 which revealed that injecting viral particles in a cell, with no adjustments to the viral particles, causes the cell to start reproducing the viral particle independent of the virus. https://pubmed.ncbi.nlm.nih.gov/1690918/

    • "RNA and DNA expression vectors containing genes for chloramphenicol acetyltransferase, luciferase, and beta-galactosidase were separately injected into mouse skeletal muscle in vivo. Protein expression was readily detected in all cases, and no special delivery system was required for these effects. The extent of expression from both the RNA and DNA constructs was comparable to that obtained from fibroblasts transfected in vitro under optimal conditions. In situ cytochemical staining for beta-galactosidase activity was localized to muscle cells following injection of the beta-galactosidase DNA vector. After injection of the DNA luciferase expression vector, luciferase activity was present in the muscle for at least 2 months."

At this point, the focus became how do we optimize this process, as the longer a cell can be made to reproduce the viral protein, the strong the immune response will be and thus the longer lasting immunity generated from the vaccine. One of the ways to enhance this process was by using lipid nanoparticles, which allows for better evasion of the body’s defenses to allow delivery of the viral protein into the cells. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8940982/

  • "The fundamental mechanism underlying the mRNA vaccine technology is based on a vehicle that enables the delivery of a nucleic acid molecule encoding the antigen of interest into the target cell in the human host, thus allowing the host cell to fabricate the target protein and express the antigen to elicit the immune response"
  • "mRNA delivered by vaccine vehicles based on lipid nanoparticles (LNPs) enters cells exclusively by endocytosis, forming an endosome without destroying the cell membrane. After entering the cytoplasm, the endosome is directed immediately to lysosomes for degradation. Therefore, in order to ensure structural integrity and thus translation of injected mRNA, endosomal fusion with lysosomes and disruption must be evaded. Studies have shown that the ionizable lipids in LNPs play a role in mRNA release and endosomal escape."
  • "mRNA is then translated into proteins by ribosomes"
  • "translated proteins based on the information contained in the mRNA can be secreted into the extracellular environment"

Another way that the process was enhanced was by adjusting the genetics of the spike protein itself to modify robust expression of the protein. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6453507/

  • "Unlike proteins, mRNA vaccines need to be expressed in situ to induce an antigen-specific immune response; therefore, understanding the magnitude and durability of antigen expression after mRNA injection is important for vaccine optimization. Results by Vogel et al.55 showed that naked, unmodified mRNA induced protein expression in vivo at 12–24 h after intramuscular (i.m.) injection and lasted for at least 6 days. Kinetics and magnitude of translation can be influenced by mRNA formulation, route of administration, nucleoside modifications, and sequence optimization.45, 56, 57"
  • "In addition to modified nucleosides, sequence optimization is also used to ensure robust protein expression and immunogenicity. Thes et al.45 have shown that an mRNA sequence enriched in G:C content and carrying optimized UTRs is superior to a nucleoside-modified counterpart in vitro and in vivo. Sequence-optimized mRNA delivered with lipid nanoparticles (LNPs) also elicited robust functional antibody titers "

The Pfizer vaccine utilizes both the lipid nanoparticle technology and modified nucleosides to introduce the spike protein of COVID into the body for your body to make the protein, which is released into circulation, to generate an immune response. Emailing detailing this using the released Pfizer documents to follow.   However, interestingly, the COVID virus appears to at baseline use the virus particle spike protein to have it be reproduced separately from the virus itself to cause damage. This in my reading and understanding of virology may be the most biologically active protein we have ever encountered. Those patients stuck in the ICU for months with ARDS may have persistent replication of spike protein causing their persistent symptoms and immune response, even though the virus as a whole is not present. Thus the immune response is appropriately activated in response to spike protein damage to tissue but the immune system is failing to completely clear the spike protein where as other patients easily do this.  

Below are some papers that talk more about the spike protein production in infected patients:

  1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC434101/
    • "Coronavirus S proteins undergo extensive co- and posttranslational modifications and conformational maturation (for a review, see reference 3). They are extensively glycosylated, become acylated, and undergo formation of multiple intrachain disulfide bonds (20, 21). Most of these events occur during and immediately after synthesis in the endoplasmic reticulum and are critical for the subsequent oligomerization, assembly, and transport processes. In infected cells, the spike complexes are incorporated into viral particles and released with virions from the cell, but a fraction of the complexes is also transported to the plasma membrane where it causes fusion with neighboring cells. Likewise, fusion occurs when the S proteins are expressed individually in the proper cells."
  2. The spike protein is made separately in the cell from the rest of the virus proteins https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7575906/
    • "The SARS-CoV-2 Spike glycoprotein is synthesized as a 1273-amino acid polyprotein precursor on the rough endoplasmic reticulum (RER) (Figure 1) (13). The unprocessed precursor harbors an endoplasmic reticulum (ER) signal sequence located at the N terminus, which targets the Spike glycoprotein to the RER membrane and is removed by cellular signal peptidases in the lumen of the ER (14, 15). A single stop-transfer, membrane-spanning sequence located at the C terminus of the Spike protein prevents it from being fully released into the lumen of the ER and subsequent secretion from the infected cell (16, 17). Co-translationally, N-linked, high-mannose oligosaccharide side chains are added during synthesis (18, 19). Shortly after synthesis, the Spike glycoprotein monomers trimerize, which might be thought to facilitate the transport from the ER to the Golgi complex. Once in the Golgi complex, most of the high-mannose oligosaccharide side chains are modified to more complex forms (20, 21), and O-linked oligosaccharide side chains are also added (22, 23).
    • "a fraction of mature SARS-CoV-2 S proteins travel through the secretory pathway to the plasma membrane, where they can mediate fusion of infected with uninfected cells to form multinucleated giant cells (syncytia) (24, 31). "
  3. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC111495/
    • "Coronavirus assembly is not dependent on the Spike protein. Studies in which the glycosylation and thus the proper folding of the protein were inhibited by treatment of mouse hepatitis virus strain A59 (MHV-A59)-infected cells with tunicamycin revealed that spikeless, noninfectious particles can be formed"
    • "Coronavirus Spike proteins undergo extensive co- and posttranslational modifications and conformational maturation (for a review, see reference 3). They are extensively glycosylated, become acylated, and undergo formation of multiple intrachain disulfide bonds (20, 21). Most of these events occur during and immediately after synthesis in the endoplasmic reticulum and are critical for the subsequent oligomerization, assembly, and transport processes. In infected cells, the spike complexes are incorporated into viral particles and released with virions from the cell, but a fraction of the complexes is also transported to the plasma membrane where it causes fusion with neighboring cells. Likewise, fusion occurs when the S proteins are expressed individually in the proper cells."

Thus in sum, the capability of a viral protein to self-replicate is what the mRNA vaccines in particular were based on. Vaccine manufacturers have added coating and changes to the proteins nucleosides to help the viral protein better evade your immune system, invade cells and begin replicating. However, for COVID, the spike protein already at baseline is made independently from the rest of the virus and excreted from cells in an infected patient. The many papers on vaccine injuries and Covid long-haul proves the persistence of spike protein, its ability to harm, all of which point to this persistence and its resulting harm on the body as the cause of symptoms. Its ability to invade any tissue is why there is such a large body of literature of different organs impacted. Now that we have identified the problem, the next step is what part of the immune system fails to clear the spike compared to the majority of people that can successfully clear it, and how to fix this.  

This will be the next summary to follow, as well as a summary with the Pfizer specific info about their vaccine.

  Author: Anonymous Covid-19 vaccine-injured MD