The Elephant and The Spike

“There is a debate tactic known as ‘elephant hurling’. This occurs when the critic throws summary arguments about complex issues to give the impression of weighty evidence, but with an unstated presumption that a large complex of underlying ideas is true, and failing to consider opposing data, usually because they have uncritically accepted the arguments from their own side. We should challenge elephant-hurlers to offer specifics and challenge the underlying assumptions.”

Excerpt from Refuting Evolution 2 by Jonathan Sarfati, Ph.D. with Michael Matthews

To those who have looked into virology both critically and logically, it is painfully obvious that there is no scientific evidence for the existence of any so-called “virus.” It simply does not exist. This holds equally true for the most recent fictitious boogeyman in “SARS-COV-2.” No “virus” has ever been properly purified nor isolated directly from the fluids taken from a sick host and then shown to be pathogenic in a natural way while adhering to the scientific method in order to demonstrate cause and effect. As no “virus” has ever been scientifically proven to exist, it should go without saying that no spike protein said to belong to one has ever met this same criteria either. I recently went through this lack of evidence for the “coronavirus” spike protein here. Thus, it is honestly a waste of time to even entertain the idea of a spike protein until such evidence can be produced for both the “virus” in question and its assumed proteins.

So why am I devoting another article to this 9-12 nm figment of the imagination? I was recently sent a post by Jeremy Hammond titled: Fact Check: COVID-19 Vaccine mRNA and Spike Protein Are Not Cleared ‘Within Days’ which was supposed to be fact-checking the “fact-checkers.” In this article, Mr. Hammond attempted to debunk a “fact-check” by Health Feedback which itself aimed to allay fears about the spike protein causing damage to the body after vaccination. According to “Covid” mythology, the spike protein is claimed to circulate inside the body after mRNA injection. After submitting oneself to the toxic experiment, an unobservable magical process is said to occur where the invisible mRNA puts on its teaching cap and instructs the body on how to create the invisible spike proteins in order to learn how to develop an immune response so that it can fight off the spike protein that it just discovered how to create. Seems like a “beLIEvable” story about the novel “virus,” right?

The Health Feedback “fact-check” argued that the spike protein is perfectly safe as it is produced in such small quantities in the body that it is harmless. Mr. Hammond, on the other hand, disagreed with their story and proceeded to provide his own version of events in order to claim that the spike protein by itself is harmful. While it is not my intent to make the case that the Health Feedback “fact-check” is correct (it most definitely is not) nor that the mRNA injections are safe by any means (I have previously shown that they are in fact very harmful with many unknown side effects), I must interject here as Mr. Hammond is attempting to debunk one fictional narrative with yet another, both of which are tied to the existence and supposed pathogenicity of a spike protein.

From what I’ve been told, Jeremy Hammond is for health freedom and he has a large following. Thus, it is very important that he gets his facts straight so as to not mislead people down a side road right back into the pharmaceutically-endorsed germ theory lie. While I do not know much about him, I have been told that, while he has consistently and rightfully called out the dangers of the vaccines, Mr. Hammond very much pushes the myth that “viruses” exist. That much is clear from his article insinuating that such a thing as a spike protein exists and is by itself pathogenic. Now, it is not my intent to attack Mr. Hammond. Rather, this is an attempt to set the record straight based on the evidence available. Hopefully, this will also stand as a good lesson in always vetting any study before submitting it as evidence to bolster your own argument.

While reading the article, it became clear that Mr. Hammond was using an unsavory tactic to try and debunk the Health Feedback “fact-check.” This tactic is known as elephant hurling, which is where one throws out numerous arguments and/or studies in order to create the illusion that the weight of the evidence is on their side. It is an intimidation tactic designed to overwhelm not only the “opponent” (Health Feedback in this case if it were an actual debate) but also those reading the article. It is an attempt to claim that a preponderance of evidence means that one’s argument is correct. However, if the preponderance of evidence is based upon a fraudulent foundation, such as the existence of a pathogenic spike protein said to belong to a “coronavirus,” the accumulated weight of the evidence is utterly meaningless. What we are left with is a gigantic pile of non-reproducible, non-replicable, and erroneous stories built around a fictional entity. This is very clear after looking at the list of studies supplied by Mr. Hammond.

I have gone through the main studies presented as evidence by Mr. Hammond that a spike protein exists and is pathogenic by itself. I have provided some (not all) of the many faults related to each of these papers. What you will see is that not a single study utilized purified and isolated particles said to be spike proteins. As is normally the case, the researchers engineered their own cell cultured creations claiming the existence of invisible spikes inside the petri dish. The experimental methods attempting to prove pathogenicity relate to indirect observations from 2D and 3D model systems as well as non-specific chemical reactions. In other words, the evidence is entirely in vitro, i.e. created in a lab, and has no bearing on what would or could occur naturally in vivo, i.e. within a living organism. The ensuing breakdown is rather long as Mr. Hammond hurled out quite a few unrelated sources so bear with me and we will get through them all.

The Spike Protein Studies

The Spike Protein Is Not Harmless

“Numerous studies have indicated that the spike protein of SARS‑CoV‑2 by itself, in the absence of whole viable virus, can have pathogenic and toxic effects. The following are some notable examples.”

“A study in Neurobiology of Disease in October 2020 found that the spike protein promotes loss of blood-brain barrier integrity and triggers an inflammatory response in brain endothelial cells.”

Before diving into this first study, it is extremely important to highlight something critical: not a single one of the proceeding studies submitted as proof of the pathogenicity of the spike protein by Jeremy Hammond use purified and isolated spike proteins from purified and isolated “SARS-COV-2.” What is used, instead, is known as a recombinant protein. What exactly is a recombinant protein?

“Recombinant protein is a manipulated form of protein, which is generated in various ways to produce large quantities of proteins, modify gene sequences and manufacture useful commercial products. The formation of recombinant protein is carried out in specialized vehicles known as vectors. Recombinant technology is the process involved in the formation of recombinant protein.

Recombinant Protein is a protein encoded by a gene — recombinant DNA — that has been cloned in a system that supports expression of the gene and translation of messenger RNA (see expression system). Modification of the gene by recombinant DNA technology can lead to expression of a mutant protein. Proteins coexpressed in bacteria will not possess post-translational modifications, e.g. phosphorylation or glycosylation; eukaryotic expression systems are needed for this.”

https://portlandpress.com/clinsci/article/136/6/431/231092/SARS-CoV-2-spike-protein-causes-cardiovascular

As can be seen, when discussing recombinant proteins, we are dealing with manipulated substances that are said to have been genetically engineered, modified, and cloned. In order to create the recombinant proteins, an appropriate expression system is needed in order to culture, maintain, and grow the desired amount of proteins for the intended use. These expression systems can come in the form of bacteria, yeast, insects, or mammalian cells.

“In such cases the system in which the protein is expressed must be easy to culture and maintain, grow rapidly, and produce large amounts of protein. Moreover, mammalian proteins also undergo various post-translational modifications. These requirements led to the discovery of protein expression systems. The various protein expression systems are bacteria, yeast, insect or mammalian systems.

The following factors determine the type of expression system used to produce recombinant proteins:

• time spent in expressing
• the proteinease of handling
• the expression system
• amount of protein needed
• mass of the protein
• type of post-translational modifications,
• number of disulfide bonds
• destination of the expressed protein

The process of expressing a recombinant protein in an expression system requires the following information/components.

• identification of the gene that encodes the protein of interest
• generation of cDNA from the respective mRNA
• selection of suitable expression vector to insert the gene sequence
• selection of suitable system that can express the vector
• appropriate screening and scaling up methods”

https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/protein-biology/protein-expression/protein-expression-systems

Without going into too much detail, it should be clear to see that what these studies are working with are nothing but lab-concocted creations said to contain the invisible spikes that have no relation to what may be theoretically free-floating inside a human body due to “natural infection” with a “virus.” There are no purified and isolated spike proteins that are ever observed nor utilized as a valid independent variable in order to determine cause and effect. Thus, any study using recombinant proteins (and/or pseudoviruses as will be seen later) are in fact pseudoscience, i.e. fake science, and should be immediately rejected as a form of valid evidence.

Now, on to Hammond’s first study. What you will find is that this study is entirely reliant upon, you guessed it, recombinant spike proteins.

The reagents used for the study included:

1. “SARS-CoV-2” subunit S1 (RayBiotech, Cat No 230–01101)
2. “SARS-CoV-2” RBD (RayBiotech, Cat No 230–01102)
3. “SARS-CoV-2” subunit S2 (RayBiotech, Cat No 230–01103)
   • All three are recombinant “spike proteins” expressed in E. Coli and maintained in a filtered solution in 50 mM Na2HPO4 (pH 7.4), 0.5 M NaCl, 450 mM imidazole, and 6 M urea.
4. “SARS-CoV-2 S1” (RayBiotech, Cat No 230–30161-10)
   • A recombinant “spike protein” grown in Human embryonic kidney 293 (HEK293) cells and supplied as a 0.2 um filtered solution in PBS (pH 7.4).
5. “SARS-CoV-2 S2” also derived from HEK293 cells were used in experiments where indicated

Beyond the use of recombinant spikes cultured and grown in bacteria and human kidneys cells, the systems used to study the blood-brain barrier (BBB) were 2D and 3D models (i.e. recreations). The brain cells were cultured in rat tail collagen coated flasks along with medium containing fetal calf serum. Three-dimensional models of the blood-brain barrier (3D BBB) were fabricated by polymerizing hydrogels. The models are considered the best recapitulations (closest resemblance) to the real BBB. In other words, we have fake spike proteins being examined in fake 2D and 3D models representing the blood-brain barrier. There is nothing real about the material used nor the experiments performed and thus no conclusions about what occurs inside a living organism can be gained from this paper, especially regarding the idea that a spike protein can cause the loss of blood-brain barrier integrity and can trigger an inflammatory response in brain endothelial cells:

The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier

“To test functional outcomes, two (a 2D and a 3D vessel-like) in-vitro models of the BBB using primary brain endothelial cells were used. In both models, the effects of the spike subunits of SARS-CoV-2 on barrier integrity were determined. Our results provide evidence of endothelial barrier permeability and pro-inflammatory responses upon exposure to these subunits. Moreover, our analysis points to barrier breach that may be independent of ACE2 since deleterious effects also occurred with the S2 subunit. To the authors knowledge, this is the first evaluation for the effects of SARS-CoV-2 spike protein on the BBB.”

2. Materials and methods

2.1. Reagents

“SARS-CoV-2 subunit S1 (RayBiotech, Cat No 230–01101), SARS-CoV-2 RBD (RayBiotech, Cat No 230–01102), and SARS-CoV-2 subunit S2 (RayBiotech, Cat No 230–01103) derived from E.coli and SARS-CoV-2 S1 (RayBiotech, Cat No 230–30161-10), SARS-CoV-2 S2 derived from HEK293 cells were used in experiments where indicated. We used SARS-CoV-2 spike protein subunits concentrations ranging from 0.1 nM to 50 nM based on a previous study that tested the effects of SARS-CoV-2 protein on stimulating human immune cells(Dosch et al., 2009). Other recombinant proteins (i.e TNFα) were purchased from R&D Systems (Minneapolis, MN, USA). Note, recombinant proteins diluted to the concentrations used for these studies were assayed for the presence of E.coli endotoxin using the ToxinSensor Chromogenic LAL Endotoxin Assay (GenScript) which found E.coli endotoxin levels to be only at negligible amounts.

2.2. Endothelial cell culture

Primary human brain microvascular endothelial cells (hBMVECs) were isolated from fetal brain tissue as described (Andrews et al., 2018). Healthy tissue was provided (under informed consent) by the Laboratory of Developmental Biology (University of Washington, Seattle, WA) with approval granted by Temple University’s (Philadelphia, PA) Institutional Review Board and in full compliance by the National Institutes of Health’s (NIH) ethical guidelines. Cells were grown on rat tail collagen I coated flasks (BD Biosciences) in full growth medium (EBM-2 medium supplemented with EGM-2MV SingleQuots (Lonza, Cat No CC-3156 and CC-4147)) in an incubator set to 37 °C, 5% CO2, and 100% humidity. Experiments were performed in the basal medium (EBM2 supplemented with 10% FBS).

For certain studies (as indicated in the figures), the hCMEC/D3 (a gift from Dr. Pierre O Couraud, Institut Cochin, université Paris Descartes, Paris, France) cell line that are often used for modeling the BBB were used. The cell line is a telomerase-immortalized human brain endothelial cell line for which its barrier forming properties and cell culture conditions have been previously characterized (Weksler et al., 2005).

2.3. 3D BBB model

Three-dimensional models of the blood-brain barrier (3D BBB) were fabricated by polymerizing hydrogels composed of 5 mg/mL type I collagen, 1 mg/mL hyaluronan, and 1 mg/mL Matrigel within microfabricated devices. The full method for this approach is described in a previous study (Partyka et al., 2017). Briefly, hydrogels were injected into the reservoir of the device and 180-μm needles coated in 0.1% BSA were inserted prior to polymerization to create two parallel and cylindrical voids within the gel. hCMEC/D3 were injected into one channel at a density of 10 million per mL (15 μL per channel). Channels were incubated for 10 min to ensure cell attachment then injected with cells again and inverted for 10 min to coat the opposing side to ensure full coverage. Following cell seeding, channels were exposed to 0.7 dyn/cm2 of steady shear stress for four days using a linear syringe pump (Kent Scientific) to establish barrier function. Following the four-day perfusion, vessels were perfused for two hours with 50 nM of SARS-CoV-2 subunit S1. Following exposure to the viral protein, vessels were either placed in fixative or prepared for permeability testing. To quantify localization of ZO-1 to the cell-cell junction in these vessels, the fluorescence intensities along representative 100-μm sections of both the control and spike protein-treated condition were plotted as described in a previous study (DeOre et al., 2019). The variance of both these intensity plots were calculated, and the percent difference was calculated to quantify the reduction in localization of ZO-1 to the cell-cell junctions.”

“These systems (when also coupled with other cells) represent the most advanced recapitulation of the blood-brain barrier. Once the SARS-CoV-2 subunit S1 was introduced, the presence of barrier permeability (from lumen to parenchymal compartment) was clearly evident as early as 2 h. These results suggest that whether free viral spike proteins or those on the surface of the virus present during SARS-CoV-2 infection could induce barrier permeability (albeit once a certain threshold is reached) equivocal to the concentrations used here. As this is the first report on the topic, much work remains, particularly in regard to how permeability dynamics may change once these 3D microfluidic constructs are used with the whole SARS-CoV-2 virus.”

https://www.sciencedirect.com/science/article/pii/S096999612030406X?via%3Dihub

“A study in Vaccines in January 2021 noted that the pathogenicity of the spike protein is a relevant concern for COVID‑19 vaccines since the mRNA vaccines are designed to instruct human cells to produce the spike protein of the coronavirus.”

Hammond used this second study to claim that there is a concern that the spike protein, said to be created by the mRNA injection, can be pathogenic. What he didn’t tell his readers is that this study once again used recombinant spike proteins created from cell cultures to “infect” cultured primary human pulmonary artery smooth muscle cells (SMCs) or cultured human pulmonary artery endothelial cells. As in the previous study, there is nothing natural about the experimental set up and design. The theoretical spike proteins we are said to encounter are not lab-created concoctions nor do these invisible entities interact with human tissues that have been potentially subjected to fetal bovine serum and other additives (the exact ingredients were not disclosed in the paper). Thus, any experimental results can only relate to the lab-created materials and conditions and can not be extrapolated to what happens under natural conditions within a living organism.

However, due to the results from the culture experiments along with what they claim is recent suggestive evidence, the researchers stated that it was reasonable to assume that the lab-created concoction said to harbor the spike protein can act alone in order to be pathogenic. Even though they did not test this, the researchers felt it was important to consider the possibility that the “SARS-CoV-2” spike protein produced by the new “COVID-19” vaccines (in an unobservable process) triggers cell signaling events that promote PAH, other cardiovascular complications, and/or complications in other tissues/organs in certain individuals. In other words, they were speculating about how the fictional spike protein may affect a living organism based on results from fake cultured spike proteins further cultured with human tissues in a petri dish in a lab:

SARS-CoV-2 Spike Protein Elicits Cell Signaling in Human Host Cells: Implications for Possible Consequences of COVID-19 Vaccines

3. SARS-CoV-2 Spike Protein Elicits Cell Signaling in Human Cells

“It was found that the treatment of cultured primary human pulmonary artery smooth muscle cells (SMCs) or human pulmonary artery endothelial cells with the recombinant SARS-CoV-2 spike protein S1 subunit is sufficient to promote cell signaling without the rest of the viral components [21]. Furthermore, our analysis of the postmortem lung tissues of patients who died of COVID-19 has determined that these patients exhibited pulmonary vascular wall thickening, a hallmark of pulmonary arterial hypertension (PAH) [21]. Based on these results, we proposed that the SARS-CoV-2 spike protein (without the rest of the viral components) triggers cell signaling events that may promote pulmonary vascular remodeling and PAH as well as possibly other cardiovascular complications [21,22].

In our cell culture experiments, two recombinant SARS-CoV-2 spike proteins, both of which contain the RBD, were studied [21]. The full-length S1 subunit protein contains most of the S1 subunit (Val16–Gln690), while the RBD S1 subunit protein only contains the RBD region (Arg319–Phe541), as shown in Figure 1. Cultured primary human pulmonary artery SMCs and human pulmonary artery endothelial cells were treated with these proteins for 10 min. We found, using the phospho-specific MEK antibody, that the recombinant full-length S1 subunit of SARS-CoV-2 alone at a concentration as low as 130 pM activated MEK, the activator of extracellular signal-regulated kinase (ERK) and a well-known signal transduction mechanism for cell growth [23]. By contrast, such activation of cell signaling by the spike protein did not occur in rat pulmonary artery SMCs [21].”

6. Discussion

“It is generally thought that the sole function of viral membrane fusion proteins is to allow the viruses to bind to the host cells for the purpose of viral entry into the cells, so that the genetic materials can be released and the viral replication and amplification can take place. However, recent observations suggest that the SARS-CoV-2 spike protein can by itself trigger cell signaling that can lead to various biological processes. It is reasonable to assume that such events, in some cases, result in the pathogenesis of certain diseases.

Our laboratory only tested the effects of the SARS-CoV-2 spike protein in lung vascular cells and those implicated in the development of PAH. However, this protein may also affect the cells of systemic and coronary vasculatures, eliciting other cardiovascular diseases such as coronary artery disease, systemic hypertension, and stroke. In addition to cardiovascular cells, other cells that express ACE2 have the potential to be affected by the SARS-CoV-2 spike protein, which may cause adverse pathological events. Thus, it is important to consider the possibility that the SARS-CoV-2 spike protein produced by the new COVID-19 vaccines triggers cell signaling events that promote PAH, other cardiovascular complications, and/or complications in other tissues/organs in certain individuals (Figure 3). We will need to monitor carefully the long-term consequences of COVID-19 vaccines that introduce the spike protein into the human body. Furthermore, while human data on the possible long-term consequences of spike protein-based COVID-19 vaccines will not be available soon, it is imperative that appropriate experimental animal models are employed as soon as possible to ensure that the SARS-CoV-2 spike protein does not elicit any signs of the pathogenesis of PAH or any other chronic pathological conditions.”

https://www.mdpi.com/2076-393X/9/1/36/htm

“A study in Circulation Research in March 2021 showed that the spike protein alone can damage vascular endothelial cells.

“This study was cited by The Epoch Times to support its claim that the spike protein by itself is harmful, which Health Feedback attempted to rebut by noting that the research was done in hamsters using an engineered pseudovirus, which is incapable of intracellular replication, and was not a study of the effects of the spike protein in humans following vaccination. The study was “never designed to look at the toxicity of the spike protein after vaccination”, Health Feedback correctly notes.

Health Feedback also cites Dr. Peter Hotez, a vaccine developer with Baylor College of Medicine, correctly observing that the study “looks at cellular mechanisms of how viral spike protein works, not the immune response from a vaccine.”

Hammond used his third study to seemingly try and say that the spike protein can act alone to damage the vascular system. However, he immediately shares the Health Feedback critique stating that the study uses a pseudovirus, a limitation that even the researchers noted in their own paper.

Pseudoviruses are exactly what they sound like: fake (fake) “viruses.” These are said to be recombinant “viruses” (i.e. lab-created cultured concoctions from many sources) which mix and match genetic material from other “viruses” making them “less virulent.” This is how pseudoviruses are described according to the study Construction and applications of SARS-CoV-2 pseudoviruses: a mini review:

“Pseudoviruses are a kind of recombinant virus with their core or backbone and surface proteins derived from different viruses9. Genes inside pseudoviruses are usually altered or modified to abolish native surface protein expression. An additional plasmid is then used to express alternative surface proteins, producing a pseudovirus that can infect susceptible host cells but can only replicate intracellularly for a single round10, 11. As viral surface proteins play pivotal roles in gaining entry into host cells, the conformational structures of pseudoviral surface proteins have high similarity to that of the native viral proteins; however, pseudoviruses have attenuated virulence compared with wild-type (WT) viruses, allowing them to be safely handled in biosafety level 2 laboratories.”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8071765/figure/F1/?report=objectonly

Sadly, the researchers of the paper Hammond cited do not share how their pseudovirus was created, but from the above source we can obtain an idea of what was done through the cell culture process. The reseachers also claim that their findings using the fake (fake) “virus” need to be confirmed using the real (fake) “SARS-COV-2 virus.” Thus, once again, we can see that the study cited does not reflect reality in any way. Any and all conclusions about how the spike protein of “SARS-COV-2” would affect the vascular system can not be gained from this paper. All that can be taken away is that when mixing and matching different cultures, different indirect chemical results are generated outside of any living organism in a lab which can then be interpreted by the researchers into a story of how a fictional entity they never studied may act or behave while within the human body. Then the headlines and snippets from the conclusions can be used by bloggers and “fact-checkers” to bolster a fraudulent case claiming that part of the fictional entity is pathogenic by itself:

SARS-CoV-2 Spike Protein Impairs Endothelial Function via Downregulation of ACE 2

“We administered a pseudovirus expressing S protein (Pseu-Spike) to Syrian hamsters intratracheally. Lung damage was apparent in animals receiving Pseu-Spike, revealed by thickening of the alveolar septa and increased infiltration of mononuclear cells (Figure [A]).”

“Although the use of a noninfectious pseudovirus is a limitation to this study, our data reveals that S protein alone can damage endothelium, manifested by impaired mitochondrial function and eNOS activity but increased glycolysis. It appears that S protein in ECs increases redox stress which may lead to AMPK deactivation, MDM2 upregulation, and ultimately ACE2 destabilization.4 Although these findings need to be confirmed with the SARS-CoV-2 virus in the future study, it seems paradoxical that ACE2 reduction by S protein would decrease the virus infectivity, thereby protecting endothelium. However, a dysregulated renin-angiotensin system due to ACE2 reduction may exacerbate endothelial dysfunction, leading to endotheliitis. Collectively, our results suggest that the S protein-exerted EC damage overrides the decreased virus infectivity. This conclusion suggests that vaccination-generated antibody and/or exogenous antibody against S protein not only protects the host from SARS-CoV-2 infectivity but also inhibits S protein-imposed endothelial injury.”

https://www.ahajournals.org/doi/10.1161/CIRCRESAHA.121.318902

“A study in The FASEB Journal in May 2021 found that the spike protein induces acute lung injury in mice genetically engineered to express the ACE2 receptor, which is the receptor on certain human cells that enables the coronavirus via the spike protein to fuse with and enter the cells where it then replicates.”

Sadly, this next study remains blocked to me as I am unable to access the full paper. However, Hammond did supply a link which discussed the findings and from that article we can find out some interesting details:

“Studying SARS-CoV-2 can be challenging because experiments involving the intact virus requires a Biosafety Level 3 laboratory. To overcome this hurdle, the researchers created a new model of acute lung injury that utilizes transgenic mice that express the human receptor for SARS-CoV-2 in their lungs.

“Our mouse model dramatically reduces the danger of doing this type of research by allowing COVID-19 lung injury to be studied without using the intact, live virus,” said Solopov. “This will greatly increase and diversify the ability to do COVID-19 research. Our model will also likely be useful for studying other coronaviruses.”

The researchers injected the genetically modified mice with a segment of the spike protein and analyzed their response 72 hours later. Another group of mice received only saline to serve as a control.

The researchers found that the genetically modified mice injected with the spike protein exhibited COVID-19-like symptoms that included severe inflammation, an influx of white blood cells into their lungs and evidence of a cytokine storm – an immune response in which the body starts to attack its own cells and tissues rather than just fighting off the virus. The mice that only received saline remained normal.”

What we can see is that transgenic mice were used in order to study the so-called spike protein. Transgenic mice are genetically-altered mice which are said to have been constructed by injecting cloned DNA into fertilized mouse eggs. The eggs that survive this process are then implanted into female mice in order to  develop them. These transgenic mice were then said to be injected with the spike protein into their throats. While I could not find any details about what spike protein was used in this study, it is reasonable to assume that this was yet another recombinant lab-created cultured protein as the researchers admitted to not using “live infectious virus” nor needing a level 3 biosafety laboratory.  The transgenic mice came down with “COVID-like” symptoms which consisted of severe inflammation, increased white blood cell counts in the lungs, and “evidence” of a cytokine storm (which was only mentioned in the article yet not in the abstract nor conclusion of the study).

Thus, we have a study utlizing genetically-altered mice that experienced inflammation when injected unnaturally in the throat with lab-created cultured recombinant goo. From these results, the researchers confidentally proposed that a single exposure of K18-hACE2 mice to “SARS-CoV-2” Spike Protein S subunit S1 may represent a valid model of “COVID-19” that may be useful for the preclinical investigation of new potential countermeasures against “COVID-19” and other “coronaviral” infections. Hardly a ringing endorsement, which is not surprising given the experimental set-up, yet this did not stop Hammond from including it as part of his litany of “favorable” evidence:

Single intratracheal exposure to SARS-CoV-2 S1 spike protein induces acute lung injury in K18-hACE2 transgenic mice

“The SARS-CoV-2 pandemic has infected more than 85,900,000 people and provoked the death of more than 1.9 million worldwide. Therapeutic options remain limited, and vaccines may exhibit narrow efficacy, due to short supplies, delays in distribution and the emergence of new resistant strains. It is mandatory to study new therapeutic approaches that modulate the strong inflammatory response observed in the lung, prevent respiratory failure and improve outcomes. The study of SARS-CoV-2 pathogenicity in vivo is challenging due to the necessary biosafety laboratory regulations. Thus, we developed an acute lung injury model by intratracheally instilling the S1 subunit of SARS-CoV-2 Spike S protein (400 µg/kg, 2 ml/kg body weight) in K18-hACE2 transgenic mice that overexpress the human receptor for SARS-CoV-2 Spike protein S, ACE2, and investigated outcomes 72 hours later. Mice exhibited an acute decline in body weight during the first 48 hours following instillation, compared to saline-instilled controls. At 72 hours, bronchoalveolar lavage fluid demonstrated a dramatic increase in white blood cell content, particularly neutrophils, and marked proteinosis compared to controls. Histologic examination of lung tissue revealed hyaline membranes, alveolar septal thickening, and a large number of neutrophils in the interstitial and alveolar spaces of Spike protein S exposed mice. We propose that a single exposure of K18-hACE2 mice to SARS-CoV-2 Spike Protein S subunit S1 may represent a valid model of COVID-19, allow the study of the molecular mechanisms of SARS-CoV-2 induced lung injury and be useful in the investigation of potential new therapeutic approaches to the management of COVID-19 as well as future coronavirus-dependent respiratory diseases.

Conclusions: We demonstrate for the first time that the intratracheal instillation of a single element of the SARS-CoV-2 virus, the subunit 1 of the Spike protein, in K18-hACE2 transgenic mice, is capable of eliciting strong pulmonary and systemic inflammation, including activation of the STAT3 and NFκB pathways in the lungs and biochemical and histological evidence of acute lung injury. We propose that this model may be useful for the preclinical investigation of new potential countermeasures against COVID-19 and other coronaviral infections.”

https://doi.org/10.1096/fasebj.2021.35.S1.04183

“A study in Clinical Infectious Diseases in May 2021 showed that the spike protein induced by Moderna’s mRNA COVID‑19 vaccine circulates throughout the body.”

The problem with Hammond’s next study relates once again to the use of recombinant lab-created spike proteins as well as the use of serological results to conclude anything meaningful from the experimental procedures. This study attempts to claim that antibody results show that the spike protein, said to be created during the unobservable process after vaccination, circulates throughout the body. The researchers used different assays during the study and commented on the fact that no one had ever shown these kinds of results before. They admitted that this may have to do with the limitations of the assays themselves, thus showcasing the potential inaccuracy of their own results due to technological limitations.

Beyond the technological limitations is the problem of claiming that theoretical antibodies can detect theoretical spike proteins. In order for this to be true, the spike protein and the antibodies themselves must both be purified and isolated first and then experimented with together in order to show that they are specific to each other. As neither the spike protein nor the antibodies have ever been scientifically proven to exist in purified/isolated form, these results are essentially meaningless. One can not use one fictional entity to determine the presence and/or absence of another fictional entity. This is why non-specific results regularly occur with antibodies and we end up seeing statements from researchers such as “possibly due to assay cross-reactivity with other human coronaviruses” as we see in this study.

In any case, only 3 of 13 participants saw spike protein antibody results for 15 days after the first injection and none saw any spike protein antibody results for upwards of 56 days after the second injection. If the spike protein is supposed to circulate in the body after injection, based on these results, it doesn’t appear to do so for very long if at all in the vast majority of the participants studied:

Circulating Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Vaccine Antigen Detected in the Plasma of mRNA-1273 Vaccine Recipients

“Here we provide evidence that circulating SARS-CoV-2 proteins are present in the plasma of participants vaccinated with the mRNA-1273 vaccine. We report antigen and serological data of the mRNA-1273 vaccine in 13 healthcare workers at the Brigham and Women’s Hospital. Ultrasensitive single-molecule array (Simoa) assays were used for the detection of SARS-CoV-2 antigens spike (S1–S2 unit), S1, and nucleocapsid and antibodies immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin M (IgM) against SARS-CoV-2 spike, S1, receptor binding domain (RBD), and nucleocapsid, as previously described [6, 7].”

“S1 antigen was detected as early as day 1 postvaccination, and peak levels were detected on average 5 days after the first injection (Figure 1A). The mean S1 peak level was 68 pg/mL ± 21 pg/mL. S1 in all participants declined and became undetectable by day 14. No antigen was detected at day zero for 12 of 13 participants, as expected. However, one individual presented detectable S1 on day zero, possibly due to assay cross-reactivity with other human coronaviruses or asymptomatic infection at the time of vaccination. Spike protein was detectable in 3 of 13 participants an average of 15 days after the first injection. The mean spike peak level was 62 pg/mL ± 13 pg/mL. After the second vaccine dose, no S1 or spike was detectable, and both antigens remained undetectable through day 56. For one individual (participant 8), spike was detected at day 29, 1 day after the second injection and was undetectable 2 days later.”

“We observe an increase in S1 over an initial period of 1–5 days, suggesting that mRNA translation begins immediately after vaccine inoculation. Interestingly, spike protein appears in 3 of 13 participants on average 8 days after S1 is produced. The Simoa antigen assays for the full spike protein are designed to require antibody binding to both the S1 and S2 subunits for detection, resulting in a cleaved spike protein to be undetectable. Additionally, spike protein concentrations in plasma of vaccinated participants may be below our assay limit of detection. We hypothesize that the cellular immune responses triggered by T-cell activation, which would occur days after the vaccination, lead to direct killing of cells presenting spike protein, and an additional release of spike into the blood stream [9]. The mechanisms underlying release of free S1 and the subsequent detection of the intact spike protein remain unclear and require further studies.”

“Limitations of the current study include the small sample size and potential biases that result from enrolling healthy, young adults, which may not be representative of the general population. Future studies should also examine the dynamics of antigen production with neutralization antibodies. Nonetheless, evidence of systemic detection of spike and S1 protein production from the mRNA-1273 vaccine is significant and has not yet been described in any vaccine study, likely due to limitations in assay sensitivity and timing assessment. The clinical relevance of this finding is unknown and should be further explored.”

From the supplemental material:

“SARS-CoV-2 Spike protein (produced in Bing Chen’s lab), Nucleocapsid recombinant protein (Ray Biotech 230-30164), S1 protein (Sino Biological 40591-V08H), and RBD (produced in Aaron Schmidt’s lab) were conjugated to 647 nm, 488 nm, 700 nm, and 750 nm dye-encoded carboxylated paramagnetic beads (Quanterix), respectively, using EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride) chemistry (ThermoFisher Scientific 77149).”

https://academic.oup.com/cid/article/74/4/715/6279075?login=false

“The authors of a paper published on the preprint website Authorea in May 2021 expressed concern that government authorities were minimizing or ignoring concerns about the potential toxicity and pathogenicity of the spike protein induced by vaccination. (A preprint study is one that has not yet undergone peer review.)”

This next paper is more of a commentary than an actual study. While the authors are right to suggest that the adverse vaccine reactions that occur can and will be counted as cases of “SARS-COV-2,” the evidence used to claim that the spike protein is pathogenic and the potential cause is based on Hammond’s earlier fraudulent study which used pseudoviruses and hamsters. Again, I am not stating that the vaccines are not dangerous. They most definitely are dangerous and toxic. However, the story that there is a pathogenic spike protein created from the mRNA injection is nothing but pure science fiction based upon fraudulent evidence that is circulated by those aiming to keep the confused public believing in the germ theory lie. Ironically, the authors end their commentary by saying that “relying on a careful evaluation of the relevant scientific research, is urgent” and that it “is imperative to follow the science.” Sadly, it appears that they did no such thing:

SARS-CoV-2 mass vaccination: Urgent questions on vaccine safety that demand answers from international health agencies, regulatory authorities, governments and vaccine developers   

“Furthermore, even in the absence of SARS-CoV-2 virus, Spike glycoprotein alone causes endothelial damage and hypertension in vitro and in vivo in Syrian hamsters by down-regulating angiotensin-converting enzyme 2 (ACE2) and impairing mitochondrial function [26]. Although these findings need to be confirmed in humans, the implications of this finding are staggering, as all vaccines authorized for emergency use are based on the delivery or induction of Spike glycoprotein synthesis. In the case of mRNA vaccines and adenovirus-vectorized vaccines, not a single study has examined the duration of Spike production in humans following vaccination. Under the cautionary principle, it is parsimonious to consider vaccine-induced Spike synthesis could cause clinical signs of severe COVID-19, and erroneously be counted as new cases of SARS-CoV-2 infections. If so, the true adverse effects of the current global vaccination strategy may never be recognized unless studies specifically examine this question. There is already non-causal evidence of temporary or sustained increases in COVID-19 deaths following vaccination in some countries (Fig. 1) and in light of Spike’s pathogenicity, these deaths must be studied in depth to determine whether they are related to vaccination.”

“An open scientific dialogue is urgent and indispensable to avoid erosion of public confidence in science and public health and to ensure that the WHO and national health authorities protect the interests of humanity during the current pandemic. Returning public health policy to evidence-based medicine, relying on a careful evaluation of the relevant scientific research, is urgent. It is imperative to follow the science.”

https://www.authorea.com/users/414448/articles/522499-sars-cov-2-mass-vaccination-urgent-questions-on-vaccine-safety-that-demand-answers-from-international-health-agencies-regulatory-authorities-governments-and-vaccine-developers?commit=123b84611353b243b6d09320ac98cb07db022771

“Ironically, Dr. Peter Hotez, whom Health Feedback cites to support its argument that the spike protein induced by vaccination is harmless, was among the authors of a study published in the journal Circulation in July 2021 noting that mRNA COVID‑19 vaccines can cause myocarditis, or inflammation of the heart, and hypothesizing that this might be due to some individuals’ immune response to either vaccine mRNA or the vaccine-induced spike protein.”

Hammond next supplied a review article by Dr. Peter Hotez which he used as evidence that the mRNA vaccine can cause myocarditis. In the review, Dr. Hotez and the other authors hypothesized (an educated guess) that myocarditis could be due to the spike protein. However, in the paper it was admitted that the mechanisms for the development of myocarditis were unknown and several possible proposals were listed. The study that was cited in the review which was used as evidence for the hypothesized role that the spike protein plays in myocarditis, used recombinant spike protein and commercially available antibodies in order to obtain the results. Once again, we have hypothetical conclusions crafted around results from lab-created concoctions which have no bearing on reality:

“Although the mechanisms for development of myocarditis are not clear, molecular mimicry between the spike protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and self-antigens, trigger of preexisting dysregulated immune pathways in certain individuals, immune response to mRNA, and activation of immunologic pathways, and dysregulated cytokine expression have been proposed.”

“Another important potential mechanism for myocarditis is molecular mimicry between the spike protein of SARS-CoV-2 and self-antigens.50 Antibodies against SARS-CoV-2 spike glycoproteins have been experimentally shown to cross-react with structurally similar human peptide protein sequences, including α-myosin.50 However, severe adverse events or autoimmune reactions have been very rare.46,47 Although COVID-19 vaccination does not appear to provoke de novo immune-mediated adverse events, it is possible that it may trigger preexisting dysregulated pathways in certain individuals with predisposition, resulting in a polyclonal B-cell expansion, immune complex formation, and inflammation.48″

https://www.ahajournals.org/doi/10.1161/CIRCULATIONAHA.121.056135

Link 50 takes us to this study:

Potential antigenic cross-reactivity between SARS-CoV-2 and human tissue with a possible link to an increase in autoimmune diseases

“Commercially available mouse monoclonal antibody made against recombinant SARS coronavirus spike protein and rabbit monoclonal antibody made against SARS coronavirus nucleoprotein were applied at optimal dilution to the SARS-CoV-2 proteins and to 50 different tissue antigens using enzyme-linked immunosorbent assay (ELISA). Recombinant SARS-CoV-2 spike protein S1 and recombinant SARS-CoV-2 nucleocapsid protein were purchased from RayBiotech. ELISA wells were coated with nuclear antigens, dsDNA, F-actin, and mitochondria (M2) antigen purchased from different companies. An additional 45 tissue antigens used in this study have been previously described [9].”

https://www.sciencedirect.com/science/article/pii/S1521661620304253?via%3Dihub

“A study published at the preprint server bioRxiv in July 2021 found an association between “Long Covid” and persistence of the spike protein in the absence of persistence of whole viable virus, once again indicating that the spike protein alone is pathogenic.”

This next study provided by Hammond tried to make the case that detecting the spike protein RNA without finding whole “SARS-COV-2” RNA/genome was sufficient to claim that the spike protein itself was pathogenic and persisting within the body. To deternine this, the researchers first used digital droplet PCR to find “SARS-COV-2” RNA in the blood of a few patients, thus relying on a fraudulent test and genome to determine the initial findings. This step resulted in 36% (4 of 11) of severe “COVID-19” patients’ PBMCs contained “SARS-CoV-2” RNA compared to 4% (1/26) of PASC patients’ PBMCs. The researchers then decided to use flow cytometry with antibodies said to define B cell, T-cell, and monocytic subsets in addition to simultaneous staining of these cells with an antibody claimed to be specific to the “SARS-CoV-2” S1 protein to try and determine the reservoir for the detected RNA. To confirm the presence of “SARS-CoV-2” S1 protein, they then sorted CD14lo, CD16+ monocytes and performed Ultra High-Performance Liquid Chromatography. After various preparation processes, the researchers measured the peptides coming from peripheral blood monolayers (stored in 90% fetal bovine serum) and compared them to a peptide database from a commercially made recombinant spike protein and found up to 44% (i.e. up to = not always that high) of the S1 subunit peptides could be identified in patient samples. In other words, the researchers used indirect measurements of peptides matched between patient samples and recombinant spike proteins which were mapped to a peptide database created from the commercially made recombinant protein in order to claim INDIRECTLY that the spike protein was in the patients samples. Can you spot the circular methods there?

Worse yet, full length sequencing of the five cases submitted for genomic analysis failed to identify any full-length sequence in the spike protein gene, or any other gene, that could account for the observed spike protein detected by proteomic analysis. All they could find were fragments of “SARS-COV-2” genomes. Thus, they concluded that it must not be replication competent “virus” that was found but non-infectious leftover spike proteins which were not cleared from the body. As can be seen, this smattering of indirect evidence is based upon a “virus” and spike protein that was never properly purified and isolated first in order to accurately determine any specific antibody responses nor determine an accurate genome and peptide database. Thus, all measurements used to indirectly claim the presence of the spike protein are based on results coming from fraudulent lab-generated cultured creations:

Persistence of SARS CoV-2 S1 Protein in CD16+ Monocytes in Post-Acute Sequelae of COVID-19 (PASC) Up to 15 Months Post-Infection

“Since the reports by our group and others found that monocyte subsets can be infected by HIV, HCV, Zika virus and Dengue fever virus (10–12), we screened peripheral blood mononuclear cells (PBMCs) from PASC individuals, as well as acute severe COVID-19 as controls, for SARS-CoV-2 RNA (Table 1). Using the highly sensitive, quantitative digital droplet PCR (ddPCR), we found that 36% (4 of 11) of severe COVID-19 patients’ PBMCs contained SARS-CoV-2 RNA compared to 4% (1/26) of PASC patients’ PBMCs. The one PASC patient that was RNA positive was 15 months post infection.

To further establish the exact reservoir contributing to the positive signal detected using ddPCR, we performed high parameter flow cytometry with antibodies that define B cell, T-cell, and monocytic subsets in addition to simultaneous staining of these cells with an antibody for the SARS-CoV-2 S1 protein. As demonstrated in Figure 2, we found distinct subpopulations of SARS-CoV-2 containing cells in the CD14lo, CD16+ monocytic subset for 73% (19 out of 26) of PASC patients and 91% (10 out of 11) of severe COVID-19 patients. As demonstrated in Figure 3, the quantity of SARS-CoV-2 S1 containing cells were statistically significant in both the severe patients (P=0.004) and in the PASC patients (P=0.02). Neither classical monocytes nor intermediate monocytes expressed the SARS-CoV-2 S1 protein.

To confirm the presence of SARS-CoV-2 S1 protein, we sorted CD14lo, CD16+ monocytes and performed Ultra High-Performance Liquid Chromatography (UHPLC). Following immunoprecipitation, the elution fractions were dried down in vacuo, resuspended in ddH₂O and purified by to remove any non-crosslinked SARS-CoV-2 S1 antibody as well as any detergents from the commercial immunoprecipitation buffers. The UHPLC collected fractions were dried in vacuo, resuspended in 100 mM HEPES (pH 8.0, 20% Acetonitrile), and subjected to cistern: reduction and alkylation with chloroacetamide. The samples were then digested with AspN and LysC endopeptidases for 16h at 37°C. The digested peptides were analyzed on an Agilent 6550 IonFunnel QTOF and 1290 UHPLC by comparing patient samples to identical digests performed on commercially available SARS-CoV-2 S1 subunit. S1 subunit peptides from patient samples were mapped to a peptide database generated using commercial S1 subunit digests. Peptide identification consisted of matches in exact mass, isotope distribution, peptide charge state, and UHPLC retention time. As shown in Figure 4, the retention time of the representative peptide NLREFVFK in the digested commercial S1 subunit and Sample LH1-6 matched. Additionally, the Mass Spectra in Figure 4 show identical mass, isotope distribution, and charge states for the representative peptide NLREFVFK in the representative LH1 sample and commercial S1 subunit (also observed in LH 2-6, not shown). Using these metrics, up to 44% of the S1 subunit peptides could be identified in patient samples LH1-LH6 (Supplementary Table 1), providing complementary evidence to flow cytometry experiments that demonstrate the presence of S1 subunit protein in these patient cells.”

“Cells from 4 out of 11 severe COVID-19 patients and 1 out of 26 PASC patients contained ddPCR+ peripheral blood mononuclear cells, however, only fragmented SARS-CoV-2 RNA was found in PASC patients. No full length sequences were identified, and no sequences that could account for the observed S1 protein were identified in any patient.”

“It is important to note that the S1 protein detected in these patients appears to be retained from prior infection or phagocytosis of infected cells undergoing apoptosis and is not the result of persistent viral replication. Full length sequencing of the five cases submitted for genomic analysis failed to identify any full-length sequence in the spike protein gene, or any other gene, that could account for the observed spike protein detected by proteomic analysis.”

High Parameter Immune Profiling/Flow Cytometry

Peripheral blood mononuclear cells were isolated from peripheral blood using Lymphoprep density gradient (STEMCELL Technologies, Vancouver, Canada). Aliquots 200 of cells were frozen in media that contained 90% fetal bovine serum (HyClone, Logan, UT) and 10% dimethyl sulfoxide (Sigma-Aldrich, St. Louis, MO) and stored at -70°C. Cells were stained and analyzed using a 17-color antibody cocktail including a PE-labeled SARS-CoV-2 S1 antibody (BioTechne, Minneapolis MN).

LC-MS analysis

Digested recombinant SARS-CoV-2 Spike S1 protein was analyzed by a high mass accuracy mass spectrometer to generate a list of detectable peptides with retention time and accurate masses. An Agilent 1290 Infinity II high pressure liquid chromatography (HPLC) system and an AdvanceBio Peptide Mapping column (2.1 × 150 mm, 2.7 μm) were used for peptide separation prior to mass analysis.”

https://www.biorxiv.org/content/10.1101/2021.06.25.449905v3.article-info

“A study in Viruses in October 2021 found that the spike protein can enter the nucleus of cells and inhibit DNA damage repair. The authors noted that this finding was relevant for mRNA COVID‑19 vaccines since they are designed to elicit human cells to produce the spike protein.”

Interestingly, this next source supplied by Hammond to state that the spike protein can enter the nucleus of cells and inhibit DNA damage repair was actually retracted May 10th, 2022, well over a month before his article was published on June 28th, 2022.

Retraction published on 10 May 2022, see Viruses 2022, 14(5), 1011.

This is a perfect example as to why one should never just read the headline and throw any study out there in support of an argument without actuallly having read the study first. All claims from this paper which were used by Hammond in support of his spike protein argument are therefore false and invalid:

SARS–CoV–2 Spike Impairs DNA Damage Repair and Inhibits V(D)J Recombination In Vitro

“The published article [1] has been retracted. Following publication, the first author contacted the editorial office regarding an improper experimental design with the potential to significantly affect the integrity of the resultant experimental data.

Adhering to our complaint procedure, an investigation was conducted. Both the chosen construct of the spike plasmid that contained a C-terminal fused with 6xHis tag and use of a GFP reporter system under overexpression conditions in the protocol were identified as having the potential to introduce significant ambiguity regarding the nature of the reported observations. The reliability of the results and conclusions presented have therefore been undermined. Furthermore, statements regarding the effect of the spike protein on the adaptive immunity are misleading as in this article no experiments related to the adaptive immunity were performed, and the full-length spike-based vaccine was not studied. Therefore, conclusions related to vaccine safety are not validated and lacked experimental support. This article [1] is retracted and shall be marked accordingly. This retraction was approved by the Editor-in-Chief of the journal Viruses.”

https://www.mdpi.com/1999-4915/13/10/2056/htm

“A study in the Journal of Immunology in November 2021 found that the spike proteins induced by the Pfizer-BioNTech COVID‑19 vaccine are carried by extracellular vesicles called exosomes and circulate throughout the body, which helps to explain the blood antibody response elicited by vaccination.”

In Hammond’s next study, it is claimed that the vaccines create exosomes which carry spike proteins on their surface that circulate throughout the body. This is yet another serological study attempting to claim that the antibodies used are specific to a spike protein which was used in order to stain the exosome so that the spike proteins can be visualized. However, once again it must be stated that in order to deternine specific antibody responses, the “virus,” it’s spike protein, the antibody, and the exosomes would have needed to have been properly purified and isolated directly from the fluids of humans first. This has never been done and thus these all remain unproven fictitious entities which can not be used in order to verify the existence of the other. This is yet another study using recombinant cultured goo said to contain spike proteins and not purified/isolated particles proven to be spike proteins. It all amounts to nothing more than staining fluids and pointing and declaring at the dots which stick to a blob in EM and claiming this as proof that exosomes with spike proteins were created from vaccination:

Cutting Edge: Circulating Exosomes with COVID Spike Protein Are Induced by BNT162b2 (Pfizer–BioNTech) Vaccination prior to Development of Antibodies: A Novel Mechanism for Immune Activation by mRNA Vaccines

“In the current study, we analyzed eight healthy adults who received both doses of the SARS-CoV-2 vaccine (Pfizer–BioNTech). Our results demonstrated the induction of circulating exosomes carrying the SARS-CoV-2 spike protein by day 14, when Abs to the spike protein were not detectable in the sera using an ELISA method developed in our laboratory. Circulating Abs were detectable only after the second booster dose of vaccine (days 14), and the amount of exosomes containing spike protein was increased up to ∼12-fold maximum.”

Materials and Methods

Patient cohort and demographics

We analyzed eight healthy adult volunteers vaccinated with the mRNA-based SARS-CoV-2 vaccine (Pfizer–BioNTech). Blood was collected before vaccination, days 7 and 14 after the first dose, day 14 after the second dose, and 4 mo after both the doses. This study was approved by the Institutional Review Boards (IRB) at St. Joseph’s Hospital (IRB number PHXB16-0027-10-18).

Exosome isolation and nanoparticle tracking analysis

Exosomes were isolated from 500 µl of plasma using Invitrogen Exosome Isolation Kit followed by 0.22-micron filtration (7). All exosomes were analyzed for size by NanoSight NS300 (Malvern Panalytical, Great Malvern, U.K.), and the mean size of the particles used in our experiments was <200 nm (8).

Detection of Abs to SARS-CoV-2 spike protein and nucleocapsid protein from human plasma samples

Development of Abs to SARS-CoV-2 spike Ag was determined using an ELISA developed in our laboratory. In brief, 1 μg/ml SARS-CoV-2 spike protein (Sino Biological) suspended in PBS was coated on an ELISA plate and incubated overnight at 4°C. Human plasma was added to these plates at 1:750 dilution. Detection was performed using secondary anti-human IgG-HRP (1:10,000) and developed using tetramethylbenzidine substrate and read at 450 nm.”

Transmission electron microscopy of isolated exosomes for SARS-CoV-2 spike protein

“Exosomes were labeled with immunogold and mouse anti–SARS-CoV-2 spike Ab, and coronavirus FIPV3-70 Ab (1:100) was added to the grids. Grids were washed and stained with uranyl acetate and viewed by transmission electron microscopy (JEOL USA, Peabody, MA) (10).”

“We performed transmission electron microscopy using Abs specific for SARS-CoV-2 spike to demonstrate the presence of SARS-CoV-2 Ags on the surface of exosomes from controls and healthy vaccinated individuals. Exosomes from vaccinated individuals are positive for SARS-CoV-2 Ag (Fig. 1B). We have also stained both the exosome samples with coronavirus FIPV3-70 Ab as negative control and did not observe any positive reaction in exosomes (Fig. 1B).”

https://www.jimmunol.org/content/207/10/2405#ref-7

“A study in Clinical Science in December 2021 tested the hypothesis that the spike protein “may act as a ligand to induce non-infective cellular stress”. They showed that exposure to the spike protein alone “elicited signalling and functional alterations”, including “secretion of pro-inflammatory molecules typically involved in the cytokine storm” and “production of pro-apoptotic factors” causing endothelial cell death.

Health Feedback briefly notes this study by saying that it “showed that the spike protein can affect heart cells in the lab, but they used higher amounts of the protein than those found in COVID‑19 patients.”

Nevertheless, the study did demonstrate a biologically plausible mechanism by which blood clots could occur with SARS‑CoV‑2 infection or vaccination.”

This was another study used by Hammond to try and claim that the spike protein alone is pathogenic. He believed that the study showed a plausible way blood clots may occur due to “SARS-COV-2” and/or injection from the mRNA vaccine. However, once again, this study did not use purified and isolated particles but instead experimented with lab-created recombinant S proteins made from insect cells. The primary cell cultures used for the study were grown in dedicated medium supplemented with human recombinant growth factors and 2% fetal calf serum which hardly sounds like something the cells would encounter within a living organism. These cells were passaged between 4 to 7 times which can have detrimental effects on the culture as the passage number increases. The cell line cultures consisted of human gut epithelial cell line, Caco2, expressing hACE2 as well as African green monkey kidney cell line VeroE6 engineered to overexpress the human ACE2 and TMPRSS2. All cells were cultured in Dulbecco’s modified Eagle’s medium plus GlutaMAX supplemented with 10% FBS, 1% sodium pyruvate, and 0.1 mM non-essential amino acids. The human lung epithelial cell line Calu3 (ATCC HTB-55) was cultured in Eagle’s minimum essential medium plus GlutaMAX with 10% FBS, 0.1 mM non-essential amino acids, and 1% sodium pyruvate. In other words, there is absolutely nothing natural about the materials nor the chemical additives that they were kept in and experimented with.

The researchers stated that their study provided novel (as in fictional) proof-of-concept evidence for S protein capacity to cause molecular and functional changes in human vascular PCs. However, they admitted that their small sample size was inadequate and that further investigation in a larger population of patients was warranted to determine the cause for the inter-individual variability in PC infection. They also could not exclude that different scenarios may happen in vivo, i.e. within a living organism, as compared to that seen in vitro, i.e. inside a petri dish in a lab, thus essentially admitting that their results can not be applied to what occurs within a human body. Interestingly, the researchers also admitted that low amounts of the S protein could be detected in pre-pandemic control sera. They stated that this could be explained by the sequence homology between some regions of the S protein and other human proteins/peptides. In other words, the S protein contains similar sequences to normal human proteins/peptides and thus the tests that the researchers were using may have been picking up nothing more than normal human proteins/peptides rather than the theoretical S protein. Sadly, the immunogen sequence for the ELISA kit they used was locked away behind proprietary information (as is always the case), and therefore they could not determine if it recognised the S protein residues that have homology with unrelated peptides. Thus, the results from this study truly were worthless:

The SARS-CoV-2 Spike protein disrupts human cardiac pericytes function through CD147 receptor-mediated signalling: a potential non-infective mechanism of COVID-19 microvascular disease 

“Exposure to the recombinant S protein alone elicited signalling and functional alterations, including: (1) increased migration, (2) reduced ability to support endothelial cell (EC) network formation on Matrigel, (3) secretion of pro-inflammatory molecules typically involved in the cytokine storm, and (4) production of pro-apoptotic factors causing EC death.”

Primary cell cultures

“Cardiac PCs were immunosorted as CD31neg/CD34pos cells from human myocardial samples, and expanded in a dedicated medium supplemented with human recombinant growth factors and 2% v/v foetal calf serum (FCS) (ECGM2 complete kit, C-22111, PromoCell) as previously described [11,28]. Briefly, samples were finely minced using scissors and scalpel until nearly homogenous and digested with Liberase (Roche) for up to 1 h at 37 C, with gentle rotation. The digest was passed through 70-, 40-, and 30-μm strainers. Finally, the cells were recovered and sorted using anti-CD31 and -CD34 microbeads (Miltenyi) to deplete the population of CD31pos ECs and select CD31neg/CD34pos cells, which distinguish a population of perivascular cells in situ [11,28]. After expansion to passage 3, the purity of the cell population was verified using immunocytochemistry (ICC) or flow cytometry [11,28].

Human coronary artery ECs (CAECs) were purchased from PromoCell and expanded in the same medium used for PCs. All cells used in the present study tested negative for mycoplasma contamination (assessed using the PCR Mycoplasma Test Kit I/C, PromoCell, cat# PK-CA91-1096). Cells were used between passages 4 and 7.

Cell line cultures

The human gut epithelial cell line, Caco2, expressing hACE2 (Caco-2-ACE2) was a kind gift from Dr Yohei Yamauchi, University of Bristol. The African green monkey kidney cell line VeroE6 engineered to overexpress the human ACE2 and TMPRSS2 (VeroE6/ACE2/TMPRSS2) [29] was a kind gift from Dr Suzannah Rihn, MRC-University of Glasgow Centre for Virus Research. All cells were cultured in Dulbecco’s modified Eagle’s medium plus GlutaMAX (DMEM, Gibco, Thermo Fisher, cat# 10567014) supplemented with 10% v/v FBS (Gibco, Thermo Fisher, A3840001), 1% v/v sodium pyruvate, and 0.1 mM non-essential amino acids. The human lung epithelial cell line Calu3 (ATCC HTB-55) was cultured in Eagle’s minimum essential medium plus GlutaMAX (MEM, Gibco, Thermo Fisher, cat# 41090036) with 10% v/v FBS, 0.1 mM non-essential amino acids, and 1% v/v sodium pyruvate.”

Measurement of S protein in patients’ sera

“The presence of S protein in COVID-19 patients’ serum was evaluated using the COVID-19 Spike Protein ELISA Kit from Abcam (ab274342), according to manufacturer’s instructions. Pre-pandemic sera were employed as controls. All test sera were diluted 1:2. The S protein concentration was expressed as nanogram per millilitre serum. The antibody supplied in the kit recognised the S2 domain.”

Production and purification of the recombinant SARS-CoV-2 S protein

“SARS-CoV-2 S protein was expressed in insect cells and purified as described previously [33,35]. Briefly, the S construct encoded amino acids 1–1213 (extracellular domain – ECD) fused with a thrombin cleavage site, followed by a T4-foldon trimerisation domain and a hexahistidine (HIS) affinity purification tag at the C-terminus. The polybasic furin cleavage site was mutated (RRAR to A) to increase the stability of the protein for in vitro studies [33,35]. S protein was expressed in Hi5 cells using the MultiBac system [36]. Secreted S protein was harvested 3 days after infection by centrifuging the cell culture at 1000×g for 10 min followed by another centrifugation of supernatant at 5000×g for 30 min. S protein-containing medium was incubated with HisPur Ni-NTA Superflow Agarose (Thermo Fisher Scientific) for 1 h at 4°C. Resin bound with S protein was separated from unbound proteins and medium using a gravity flow column, followed by 30 column volume wash with wash buffer (65 mM NaH₂PO₄, 300 mM NaCl, 20 mM imidazole, pH 7.5). Finally, the protein was eluted with a step-gradient of elution buffer (65 mM NaH₂PO₄, 300 mM NaCl, 235 mM imidazole, pH 7.5). Eluted fractions were analysed by reducing SDS/PAGE. Fractions containing the S protein were pooled and concentrated using 50-kDa MWCO Amicon centrifugal filter units (EMD Millipore). During concentration, proteins were buffer-exchanged in PBS, pH 7.5. Concentrated protein was aliquoted, flash-frozen in liquid nitrogen, and stored at −80°C until use. In all the in vitro experiments of the manuscript, we will refer to the S-ECD protein simply as S protein.

Recombinant Spike S1 (#10522-CV) and S2 (#10584-CV) were purchased from R&D, resuspended in PBS according to manufacturer’s instructions, aliquoted and stored at −80°C until use. Similarly to the S-ECD, the S1 and S2 proteins were produced in insect cells.”

Discussion

“Our study provides novel proof-of-concept evidence for S protein capacity to cause molecular and functional changes in human vascular PCs, either dependently or independently of the CD147 receptor (summarised in Figure 11).”

“Here, we report that two-third of patients tested did not have their PCs infected by SARS-CoV-2, while the rate of infection was below 8% in the remaining subjects, suggesting a very low permissiveness of these cells to the coronavirus, at least in vitro.”

“Further investigation in a larger population of patients is warranted to determine the cause for the inter-individual variability in PC infection. Moreover, we cannot exclude different scenarios may happen in vivo.”

“In our study, low amounts of the S protein could be detected in pre-pandemic control sera. This could be explained by the sequence homology between some regions of the S protein and other human proteins/peptides. A previous report identified pathogenic regions of SARS-CoV-1 S protein, which share sequence homology with Angrgm-52 (GenBank accession number AAL62340), a novel gene up-regulated in human mesangial cells stimulated by angiotensin II and bradykinin [53]. Unfortunately, the immunogen sequence for this particular ELISA kit ab274342 is proprietary information, therefore we could not determine if it can recognise the S protein residues that have homology with unrelated peptides.”

Study limitations

“The study was conducted on isolated cells and therefore the evidence must be confirmed in vivo.

The amount of S protein used for in vitro studies was higher than the average S protein concentration detected in COVID-19 patients’ serum. However, circulating S protein represents the spill-over from infected organs, where concentration may be higher due to retention at the receptor level. Because we do not have access to post-mortem myocardial samples, we could not verify this hypothesis.”

https://portlandpress.com/clinsci/article/135/24/2667/230273/The-SARS-CoV-2-Spike-protein-disrupts-human

“A study at bioRxiv on December 2021 showed that the SARS‑CoV‑2 spike protein by itself promoted platelet activation, which the authors suggested was “a pathogenic mechanism to explain thrombosis associated to COVID‑19 lung disease, by which Spike present in SARS‑CoV‑2 virions or exposed on the surface of infected cells, leads to platelet stimulation and subsequent activation of the coagulation cascade.” (Emphasis added.)

That study was also cited by The Epoch Times, to which Health Feedback responded by noting that the researchers “studied the mechanisms for formation of blood clots, showing that spike protein can activate platelets in the lab but they did not establish if this happened at concentrations found in patients or vaccinated people.”

In this second to last study, Hammond relied on a preprint non-peer-reviewed paper to share the author’s suggestion that the spike protein can act alone in order to cause thrombosis in “Covid” patients. The paper comes with the below warning that the study should not be reported as conclusive:

A reminder: they have not been formally peer-reviewed and should not guide health-related behavior or be reported in the press as conclusive.

Interestingly, in a moment of shooting his own argument in the foot, Hammond pointed out that the Health Feedback article rightly stated that the results were only relevant in the lab and that the researchers did not establish if the results with the recombinant spike protein happened at concentrations found in patients or vaccinated people. This may have to do with the fact that, once again, the study relied on cell-cultured creations and “pseudoviruses” to generate their conclusions.

For some reason, I was unable to copy/paste the relevant sections from the study but I am providing the methods section from the abstract as well as a few images from the PDF showing that the researchers utilized cell cultured creations and lab-created “pseudoviruses” for their experiments. You can see the various processes used to create their fictional entity. Thus, once again, the experimental results do not reflect reality in any way:

SARS-CoV-2 Spike protein activates TMEM16F-mediated platelet pro-coagulant activity

“Methods We produced SARS-CoV-2 Spike or VSV-G protein-pseudotyped virions, or generated cells expressing Spike on their plasma membrane, and tested their effects on platelet adhesion (fluorescence), aggregation (absorbance), exposure of phosphatidylserine (flow cytometry for annexin V binding), calcium flux (flow cytometry for fluo-4 AM), and clot formation and retraction. These experiments were also conducted in the presence of the TMEM16F activity inhibitors Niclosamide and Clofazimine.”

https://www.biorxiv.org/content/10.1101/2021.12.14.472668v2

“A commentary in Clinical Science on March 29 of this year summarized the state of knowledge about the pathogenicity of the spike protein alone:”

Finally, we have made it to the last of Hammond’s exhausting list of pseudoscientific sources he seemingly failed to read. This one is a commentary on the state of “knowledge” on the lab-created recombinant and/or pseudovirus spike protein acting pathogenically alone. In other words, it added nothing new and actually used the same Avolio et al. study Hammond used which I outlined two sections above. If you can recall, this was the study which utilized lab-created recombinant S proteins made from insect cells for their experiments. The reseachers could not exclude that different scenarios may happen in vivo, i.e. within a living organism, as compared to that seen in vitro, i.e. inside a petri dish in a lab, thus essentially admitting that their results can not be applied to what occurs within a human body. They also admitted that the S protein contains similar sequences to normal human proteins/peptides and that the tests that the researchers were using may have been picking up nothing more than normal human proteins/peptides rather than the theoretical S protein. In other words, the evidence that the spike protein acts alone pathogenically from this commentary is as worthless as the Avolio et al. study from which it came:

“The concept that the S protein can cause detrimental effects in COVID-19 patients independent of infection could partially explain the long-term health issues.”

“There has been extensive evaluation of SARS-CoV-2 infection and COVID-19 on cardiovascular health [1,2]; however, there is emerging evidence that the S protein shed from SARS-CoV-2 can circulate in the blood of patients and have detrimental consequences [3,10,11]. (Figure 1) The study by Avolio et al. in this issue of Clinical Science provides strong evidence that the circulating S protein could be more detrimental to cardiac health than infection of SARS-CoV-2 to the heart [3]. More specifically, the S protein was found to act through the CD147 receptor on human cardiac pericytes to cause microvascular dysfunction [3]. The authors also determined that the S protein caused human cardiac pericyte inflammation via yet to be determined mechanisms [3].”

“The current study by Avolio et al. found similar results with the Alpha and Delta variants of the S protein on human cardiac pericytes [3]. Although the findings of the current study are provocative, future investigations need to expand the doses of S proteins to lower levels and other S protein variants for in vitro studies. Lastly, as we investigate SARS-CoV-2 and the circulating S protein and the impact on human health, we will be better prepared to treat patients as COVID-19 moves from a pandemic to an endemic status.”

https://portlandpress.com/clinsci/article/136/6/431/231092/SARS-CoV-2-spike-protein-causes-cardiovascular

In Summary:

• According to Jeremy Hammond, numerous studies have indicated that the spike protein of “SARS‑CoV‑2” by itself, in the absence of whole viable “virus,” can have pathogenic and toxic effects
• However, the studies supplied do not use purified/isolated spike proteins but recombinant proteins which are a manipulated form of protein, which is generated in various ways to produce large quantities of proteins, modify gene sequences and manufacture useful commercial products
• Recombinant protein is encoded by a gene — recombinant DNA — that has been cloned in a system that supports expression of the gene and translation of messenger RNA
• In such cases the system in which the protein is expressed must be easy to culture and maintain, grow rapidly, and produce large amounts of protein
• The various protein expression systems are:
  1. Bacteria
  2. Yeast
  3. Insect
  4. Mammalian
• A few of Hammond’s studies also relied on “pseudoviruses” which are a kind of recombinant “virus” with their core or backbone and surface proteins derived from different “viruses”
• Genes inside “pseudoviruses” are usually altered or modified to abolish native surface protein expression
• An additional plasmid is then used to express alternative surface proteins, producing a “pseudovirus” that can infect susceptible host cells but can only replicate intracellularly for a single round
• As “viral” surface proteins are said to play pivotal roles in gaining entry into host cells, the conformational structures of “pseudoviral” surface proteins have high similarity to that of the native “viral” proteins; however, “pseudoviruses” have attenuated virulence compared with wild-type (WT) “viruses,” allowing them to be safely handled in biosafety level 2 laboratories

• “SARS-CoV-2” subunit S1 (RayBiotech, Cat No 230–01101)
• “SARS-CoV-2” RBD (RayBiotech, Cat No 230–01102)
• “SARS-CoV-2” subunit S2 (RayBiotech, Cat No 230–01103)
  • All three are recombinant “spike proteins” expressed in E. Coli and maintained in a filtered solution in 50 mM Na2HPO4 (pH 7.4), 0.5 M NaCl, 450 mM imidazole, and 6 M urea
• “SARS-CoV-2 S1” (RayBiotech, Cat No 230–30161-10)
  • A recombinant “spike protein” grown in Human embryonic kidney 293 (HEK293) cells and supplied as a 0.2 um filtered solution in PBS (pH 7.4)
• “SARS-CoV-2 S2” also derived from HEK293 cells were used in experiments where indicated
• Other recombinant proteins (i.e TNFα) were purchased from R&D Systems (Minneapolis, MN, USA)
• To test functional outcomes, two (a 2D and a 3D vessel-like) in-vitro models of the blood brain barrier (BBB) using primary brain endothelial cells were used
• Primary human brain microvascular endothelial cells (hBMVECs) were isolated from fetal brain tissue
• Cells were grown on rat tail collagen I coated flasks (BD Biosciences) in full growth medium (EBM-2 medium supplemented with EGM-2MV SingleQuots (Lonza, Cat No CC-3156 and CC-4147)) in an incubator set to 37 °C, 5% CO2, and 100% humidity and experiments were performed in the basal medium (EBM2 supplemented with 10% FBS)
• These systems (when also coupled with other cells) represent the most advanced recapitulation (to reproduce or closely resemble) of the blood-brain barrier
• These results suggest that whether free “viral” spike proteins or those on the surface of the “virus” present during “SARS-CoV-2” infection could induce barrier permeability (albeit once a certain threshold is reached) equivocal to the concentrations used here (i.e. these results only relate to the lab-created conditions and concentrations used for the study)
• As this is the first report on the topic, they admit much work remains, particularly in regard to how permeability dynamics may change once these 3D microfluidic constructs are used with the whole “SARS-CoV-2 virus”

• It was found that the treatment of cultured primary human pulmonary artery smooth muscle cells (SMCs) or human pulmonary artery endothelial cells with the recombinant “SARS-CoV-2” spike protein S1 subunit is sufficient to promote cell signaling without the rest of the “viral” components
• In the cell culture experiments, two recombinant “SARS-CoV-2” spike proteins were studied
• Cultured primary human pulmonary artery SMCs and human pulmonary artery endothelial cells were treated with these proteins for 10 min
• They used the “phospho-specific” MEK antibody to determine that the recombinant full-length S1 subunit of “SARS-CoV-2” alone at a concentration as low as 130 pM, activated MEK
• The authors claim that recent observations suggest that the “SARS-CoV-2” spike protein can by itself trigger cell signaling that can lead to various biological processes
• Thus, they determined it is reasonable to assume that such events, in some cases, result in the pathogenesis of certain diseases
• According to the authors, this protein may also affect the cells of systemic and coronary vasculatures, eliciting other cardiovascular diseases such as coronary artery disease, systemic hypertension, and stroke (i.e. it is pure speculation as there is no evidence of this)
• In addition to cardiovascular cells, other cells that express ACE2 have the potential to be affected by the “SARS-CoV-2” spike protein, which may cause adverse pathological events (i.e. it is pure speculation as there is no evidence of this)
• The authors state that it is important to consider the possibility that the “SARS-CoV-2” spike protein produced by the new “COVID-19” vaccines triggers cell signaling events that promote PAH, other cardiovascular complications, and/or complications in other tissues/organs in certain individuals (i.e. it is pure speculation as there is no evidence of this)

• The researchers administered a pseudovirus expressing S protein (Pseu-Spike) to Syrian hamsters intratracheally (through the throat)
• Lung damage was apparent in animals receiving Pseu-Spike, (i.e. fake-spike…granted that’s an ironic admission as they all are) revealed by thickening of the alveolar septa and increased infiltration of mononuclear cells
• The researchers admitted that the use of a noninfectious pseudovirus is a limitation to this study
• However, they claim that their data revealed that (fake) S protein alone can damage endothelium, manifested by impaired mitochondrial function and eNOS activity but increased glycolysis
• It appeared that (fake) S protein in ECs increases redox stress which may lead to AMPK deactivation, MDM upregulation, and ultimately ACE2 destabilization
• They then admitted that their findings needed to be confirmed with the “SARS-CoV-2 virus” in the future study thus showing that their paper is utterly meaningless and irrelevant

• Studying “SARS-CoV-2” can be challenging because experiments involving the intact “virus” requires a Biosafety Level 3 laboratory
• To overcome this hurdle, the researchers created a new model of acute lung injury that utilizes transgenic mice (genetically-altered mice) that express the human receptor for “SARS-CoV-2” in their lungs
• “Our mouse model dramatically reduces the danger of doing this type of research by allowing COVID-19 lung injury to be studied without using the intact, live virus”
• The researchers injected the genetically modified mice with a segment of the spike protein and analyzed their response 72 hours later
• The genetically modified mice injected with the spike protein exhibited “COVID-19-like” symptoms that included:
  1. Severe inflammation
  2. An influx of white blood cells into their lungs
  3. Evidence of a cytokine storm
• Is injecting cultured goo down the throats of genetically altered mice not supposed to result in severe inflammation and increased WBC’s in the blood?
• Thus, they claim to have developed an acute lung injury model by intratracheally instilling the S1 subunit of “SARS-CoV-2” Spike S protein (400 µg/kg, 2 ml/kg body weight) in K18-hACE2 transgenic mice that overexpress the human receptor for “SARS-CoV-2” Spike protein S, ACE2, and investigated outcomes 72 hours later
• They proposed that a single exposure of K18-hACE2 mice to “SARS-CoV-2” Spike Protein S subunit S1 may represent a valid model of “COVID-19,” allow the study of the molecular mechanisms of “SARS-CoV-2” induced lung injury and be useful in the investigation of potential new therapeutic approaches to the management of “COVID-19” as well as future “coronavirus-dependent respiratory diseases”
• No mention of how the spike protein was created/aquired but it was most likely yet again a recombinant protein due to the fact that the researchers were not using “intact live viruses” nor in need of a biosafety level 3 facility

• Ultrasensitive single-molecule array (Simoa) assays were used for the detection of “SARS-CoV-2” antigens spike (S1–S2 unit), S1, and nucleocapsid and antibodies immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin M (IgM) against SARS-CoV-2 spike, S1, receptor binding domain (RBD), and nucleocapsid
• One individual presented detectable S1 on day zero, possibly due to assay cross-reactivity with other human “coronaviruses” or asymptomatic infection at the time of vaccination
• Spike protein was detectable in 3 of 13 participants an average of 15 days after the first injection
• After the second vaccine dose, no S1 or spike was detectable, and both antigens remained undetectable through day 56
• They observed an increase in S1 over an initial period of 1–5 days, suggesting that mRNA translation begins immediately after vaccine inoculation
• Spike protein concentrations in plasma of vaccinated participants may be below the assay limit of detection
• The researchers hypothesized that the cellular immune responses triggered by T-cell activation, which would occur days after the vaccination, lead to direct killing of cells presenting spike protein, and an additional release of spike into the blood stream
• The mechanisms underlying release of free S1 and the subsequent detection of the intact spike protein remain unclear and require further studies
• Limitations of the current study include the small sample size and potential biases that result from enrolling healthy, young adults, which may not be representative of the general population
• Nonetheless, they concluded that the evidence of systemic detection of spike and S1 protein production from the mRNA-1273 vaccine is significant and has not yet been described in any vaccine study, likely due to limitations in assay sensitivity and timing assessment
• In other words, results such as the ones presented here were likely never reported before due to the admitted limitations of the technology used
• The clinical relevance of this finding is unknown and should be further explored
• The spike proteins used were all lab-generated recombinant creations:
  1. “SARS-CoV-2” Spike protein (produced in Bing Chen’s lab)
  2. Nucleocapsid recombinant protein (Ray Biotech 230-30164)
  3. S1 protein (Sino Biological 40591-V08H)
  4. RBD (produced in Aaron Schmidt’s lab)

• This commentary states that even in the absence of “SARS-CoV-2 virus,” Spike glycoprotein alone causes endothelial damage and hypertension in vitro and in vivo in Syrian hamsters by down-regulating angiotensin-converting enzyme 2 (ACE2) and impairing mitochondrial function
• The study cited by the authors as evidence for the spike protein being pathogenic alone was Hammond’s previous study admitting to limitations of using a pseudovirus
• They admitted that although these findings need to be confirmed in humans, the implications of this finding are staggering, as all vaccines authorized for emergency use are based on the delivery or induction of Spike glycoprotein synthesis
• In the case of mRNA vaccines and adenovirus-vectorized vaccines, not a single study has examined the duration of Spike production in humans following vaccination
• The authors state that under the cautionary principle, it is parsimonious to consider vaccine-induced Spike synthesis could cause clinical signs of severe “COVID-19,” and erroneously be counted as new cases of “SARS-CoV-2” infections
• They state that relying on a careful evaluation of the relevant scientific research, is urgent and that it is imperative to follow the science (both of which they obviously did not do)
• While the authors are right in that the severe reactions from vaccines are attributed to “SARS-COV-2,” they are promoting a fraudulent theory that a spike protein was created by the injection and is the cause

• Although the mechanisms for development of myocarditis are not clear, several hypotheses have been proposed:
  1. Molecular mimicry between the spike protein of “severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)” and self-antigens
  2. Trigger of preexisting dysregulated immune pathways in certain individuals
  3. Immune response to mRNA, and activation of immunologic pathways
  4. Dysregulated cytokine expression
• Antibodies against “SARS-CoV-2” spike glycoproteins have been experimentally shown to cross-react with structurally similar human peptide protein sequences, including α-myosin
• The study cited within the review as evidence for the role the spike protein may have in causing myocarditis used recombinant “SARS-CoV-2” spike protein S1 and recombinant “SARS-CoV-2” nucleocapsid protein purchased from RayBiotech along with commercially made antibodies

• Using digital droplet PCR, the researchers found that 36% (4 of 11) of severe “COVID-19” patients’ PBMCs contained “SARS-CoV-2” RNA compared to 4% (1/26) of PASC patients’ PBMCs
• The one PASC patient that was RNA positive was 15 months post infection
• They performed high parameter flow cytometry with antibodies that define B cell, T-cell, and monocytic subsets in addition to simultaneous staining of these cells with an antibody for the “SARS-CoV-2 S1” protein to find the reservoir of the RNA
• To confirm the presence of “SARS-CoV-2” S1 protein (as they could not see it), they sorted CD14lo, CD16+ monocytes and performed Ultra High-Performance Liquid Chromatography (UHPLC)
• After numerous preparation processes, the samples were then digested with AspN and LysC endopeptidases for 16h at 37°C
• The digested peptides were analyzed on an Agilent 6550 IonFunnel QTOF and 1290 UHPLC by comparing patient samples to identical digests performed on commercially available “SARS-CoV-2” S1 subunit (a commercially made recombinant spike protein)
• S1 subunit peptides from patient samples were mapped to a peptide database generated using commercial S1 subunit digests
• The retention time of the representative peptide NLREFVFK in the digested commercial S1 subunit and Sample LH1-6 matched
• Additionally, the Mass Spectra show identical mass, isotope distribution, and charge states for the representative peptide NLREFVFK in the representative LH1 sample and commercial S1 subunit
• Using these metrics, up to 44% of the S1 subunit peptides could be identified in patient samples LH1-LH6 providing complementary evidence to flow cytometry experiments that demonstrate the presence of S1 subunit protein in these patient cells
• As can be seen, they used indirect measurements of peptides matched between patient samples and recombinant spike proteins which were mapped to a peptide database created from the lab-created recombinant protein in order to claim INDIRECTLY that the spike protein was in the patients samples
• Only fragmented “SARS-CoV-2” RNA was found in PASC patients
• No full length sequences were identified, and no sequences that could account for the observed S1 protein were identified in any patient
• They felt it was important to note that the S1 protein detected in these patients appeared to be retained from prior infection or phagocytosis of infected cells undergoing apoptosis and is not the result of persistent “viral” replication
• Full length sequencing of the five cases submitted for genomic analysis failed to identify any full-length sequence in the spike protein gene, or any other gene, that could account for the observed spike protein detected by proteomic analysis
• Aliquots 200 of peripheral blood monolayer cells were frozen in media that contained 90% fetal bovine serum and 10% dimethyl sulfoxide and stored at -70°C.
• Cells were stained and analyzed using a 17-color antibody cocktail including a PE-labeled “SARS-CoV-2” S1 antibody

• Retraction published on 10 May 2022, see Viruses 2022, 14(5), 1011
• After publication, the first author contacted the editorial office regarding an improper experimental design with the potential to significantly affect the integrity of the resultant experimental data
• Both the chosen construct of the spike plasmid that contained a C-terminal fused with 6xHis tag and use of a GFP reporter system under overexpression conditions in the protocol were identified as having the potential to introduce significant ambiguity regarding the nature of the reported observations
• The reliability of the results and conclusions presented have therefore been undermined
• Furthermore, statements regarding the effect of the spike protein on the adaptive immunity are misleading as in this article no experiments related to the adaptive immunity were performed, and the full-length spike-based vaccine was not studied
• Therefore, conclusions related to vaccine safety are not validated and lacked experimental support

• The authors claim that the results demonstrated the induction of circulating exosomes carrying the “SARS-CoV-2” spike protein by day 14, when Abs to the spike protein were not detectable in the sera using an ELISA method developed in their laboratory
• Blood was collected before vaccination, days 7 and 14 after the first dose, day 14 after the second dose, and 4 mo after both the doses
• They analyzed eight healthy adult volunteers (small sample size) vaccinated with the mRNA-based “SARS-CoV-2” vaccine (Pfizer–BioNTech)
• Exosomes were “isolated” from 500 µl of plasma using Invitrogen Exosome Isolation Kit followed by 0.22-micron filtration
• Development of Abs to “SARS-CoV-2” spike Ag was determined using an ELISA developed in their laboratory
• In brief, 1 μg/ml “SARS-CoV-2” spike protein (a recombinant protein from Sino Biological) suspended in PBS was coated on an ELISA plate and incubated overnight at 4°C
• Exosomes were labeled with immunogold and mouse “anti–SARS-CoV-2” spike Ab, and “coronavirus” FIPV3-70 Ab was added to the grids
• They performed transmission electron microscopy using Abs specific for “SARS-CoV-2” spike to demonstrate the presence of “SARS-CoV-2” Ags on the surface of exosomes from controls and healthy vaccinated individuals

• Exposure to the recombinant S protein alone elicited signalling and functional alterations
• Cardiac PCs were immunosorted as CD31neg/CD34pos cells from human myocardial samples, and expanded in a dedicated medium supplemented with human recombinant growth factors and 2% v/v foetal calf serum (FCS)
• Samples were finely minced using scissors and scalpel until nearly homogenous and digested with Liberase (Roche) for up to 1 h at 37 C, with gentle rotation
• Human coronary artery ECs (CAECs) were purchased from PromoCell and expanded in the same medium used for PCs
• Cells were used between passages 4 and 7
• The human gut epithelial cell line, Caco2, expressing hACE2 and the African green monkey kidney cell line VeroE6 engineered to overexpress the human ACE2 and TMPRSS2 were also used
• The presence of S protein in “COVID-19” patients’ serum was evaluated using the “COVID-19” Spike Protein ELISA Kit from Abcam (ab274342), according to manufacturer’s instructions (i.e. they used antibodies to indirectly claim the presence of the spike protein)
• The antibody supplied in the kit (i.e. commercially made lab created antibodies) recognised the S2 domain
• “SARS-CoV-2” S protein was expressed in insect cells
• Secreted S protein was harvested 3 days after infection by centrifuging the cell culture at 1000×g for 10 min followed by another centrifugation of supernatant at 5000×g for 30 min
• S protein-containing medium was incubated with HisPur Ni-NTA Superflow Agarose (Thermo Fisher Scientific) for 1 h at 4°C
• Fractions containing the S protein were pooled (i.e. they were mixed together) and concentrated using 50-kDa MWCO Amicon centrifugal filter units (EMD Millipore)
• Recombinant Spike S1 (#10522-CV) and S2 (#10584-CV) were purchased from R&D, resuspended in PBS according to manufacturer’s instructions, aliquoted and stored at −80°C until use
• Similarly to the S-ECD, the S1 and S2 proteins were produced in insect cells
• The authors reported that two-third of patients tested did not have their PCs infected by “SARS-CoV-2,” while the rate of infection was below 8% in the remaining subjects, suggesting a very low permissiveness of these cells to the “coronavirus,” at least in vitro (in the lab)
• Further investigation in a larger population of patients is warranted to determine the cause for the inter-individual variability in PC infection
• They could not exclude that different scenarios may happen in vivo (in a living organism)
• Low amounts of the S protein could be detected in pre-pandemic control sera
• This could be explained by the sequence homology between some regions of the S protein and other human proteins/peptides (i.e. the S protein has the same sequences as human proteins/peptides)
• Unfortunately, the immunogen sequence for this particular ELISA kit ab274342 is proprietary information, therefore they could not determine if it can recognise the S protein residues that have homology with unrelated peptides
• The study was conducted on isolated cells and therefore the evidence must be confirmed in vivo (i.e. within a living organism)
• The amount of S protein used for in vitro studies was higher than the average S protein concentration detected in “COVID-19” patients’ serum (in other words, the researchers needed to use more S protein material than what is “seen” in regular patients in order to get the results they wanted)
• The authors claim that circulating S protein represents the spill-over from infected organs, where concentration may be higher due to retention at the receptor level, however, as they did not have access to post-mortem myocardial samples, they could not verify this hypothesis

• The researchers produced “SARS-CoV-2” Spike or VSV-G protein-pseudotyped “virions,” or generated cells expressing Spike on their plasma membrane, and tested their effects on:
  1. Platelet adhesion (fluorescence)
  2. Aggregation (absorbance)
  3. Exposure of phosphatidylserine (flow cytometry for annexin V binding)
  4. Calcium flux (flow cytometry for fluo-4 AM)
  5. Clot formation and retraction
• I was unable to highlight more from this study as it would not allow me to copy/paste but seeing as to how the experiments involved “pseudoviruses” and cell cultured creations, it’s pretty safe to conclude that the results hold no relation to anything that would naturally occur within a living organism

• The concept that the S protein can cause detrimental effects in “COVID-19” patients independent of infection could partially explain the long-term health issues
• The author sites the study by Avolio et al. (discussed two sections above) and state that it provides strong evidence that the circulating S protein could be more detrimental to cardiac health than infection of “SARS-CoV-2” to the heart
• More specifically, the S protein was found to act through the CD147 receptor on human cardiac pericytes to cause microvascular dysfunction
• Avolio et al. also determined that the S protein caused human cardiac pericyte inflammation via yet to be determined mechanisms
• The author admits that although the findings of the current study are provocative, future investigations need to expand the doses of S proteins to lower levels and other S protein variants for in vitro studies

Back in June 2020, CNN’s very own Dr. Sanjay Gupta, of all people, wrote an article which warned the public to be wary of the flood of scientific studies coming out during the early days of the pandemic. The article was titled Science by press release: When the story gets ahead of the science and Dr. Gupta wrote about his observations after deciding to look at the available data pouring in from the scientific community. In his own assessment, he was “surprised at how thin the available data actually is in peer-reviewed medical journals.” He rightfully pointed out that most of what was seen came in the form of press releases or pre-print reports that had not undergone the scientific scrutiny of independent review. In fact, Gupta stated that the studies being offered up and showcased were not ready for “prime time.” He observed that many were not studies at all, but rather subjective conclusions based on data, and methods which remained hidden and difficult to validate. 

It seems that Jeremy Hammond must have missed this article by Dr. Gupta as his own “fact-check” of the “fact-checkers” is full of commentaries, preprints, unverified studies with hidden and faulty methods, and even one paper that was fully retracted more than a month before Hammond’s own article was published. None of the studies supplied ever utilized any so-called spike proteins purified and isolated directly from human fluids. Instead, lab-created genetically engineered cloned and cultured recombinant concoctions grown in bacteria and insect cells and/or “pseudoviruses” created from similar cultured manipulations were used. The studies were done in artificial cultured conditions in vitro, i.e. in a petri dish in a lab, and the results could not be extrapolated to what occurs in vivo, i.e. within a living organism. Thus, any conclusions, hypotheses, and theories based off of the fraudulent data supplied from these studies amounts to nothing more than science fiction novels written by your favorite virologist, blogger, “fact-check,” and/or media personality.

It does not matter how many studies someone like Hammond chooses to throw out in support of their argument if the studies themselves are built upon fraudulent foundations and have not been properly reviewed, reproduced, replicated, and verified. What ensues is an article spewing forth a preponderance of faulty evidence. This is a problem as, to the layperson, the numerous links and conclusions from the studies looks impressive upon first glance. This false narrative generated from deceitful practices helps to sell the idea that a spike protein exists and can be created from the mRNA injections in some fantastical unobservable process. It helps to keep the “virus” lie alive in that these researchers are able to work with, manipulate, and generate pieces of the invisible “virus” in order to study. It helps to keep the pharmaceutical lies alive in that there are “safer” therapeutics and drugs available that can specifically target the spike protein of the fictional entity. It helps to keep confidence and belief in pseudosciences like genomics said to identify and recreate the protein through recombination as well as immunology in order to tag these invisible entities with fictional antibodies and determine indirectly how the body responds to them. It helps to keep the confused, who are looking for a way out of the germ theory and pharmaceutical lies, entrenched into this deceptive web of disinformation and misrepresentation.

There is no need to create a hypothetical narrative around a fictional spike protein in order to claim how the mRNA vaccines harm a person. The very act of injecting anything into the body is a potential cause of harm as this is an unnatural way for the body to encounter any substance. Even injecting something as simple as water can cause disruption of red blood cells as well potential kidney damage. The “vaccines” are full of contaminants both known and unknown and people who are allowing the injection of these substances into their bodies are playing Russian Roulette with their own health. No fictional spike protein is necessary to explain the harm these injections can cause. People like Jeremy Hammond are doing nothing more than perpetuating fear founded upon unscientific fraud.