https://doctors4covidethics.org/wp-cont ... xicity.pdfrisks that were clearly evident from these data, and the regulatory authorities failed
to enforce proper standards of oversight. This dual failure has caused the most
grievous harm to the public.
Before we discuss this study and its implications in detail, we will briefly review
how the Pfizer mRNA vaccine works. These explanations also apply to the Moderna
mRNA vaccine, whereas the AstraZeneca and the Johnson & Johnson vaccines differ
in some aspects.
1.1 How the mRNA COVID vaccines work
The Pfizer and Moderna mRNA vaccines consist of a synthetic messenger RNA
(mRNA) that encodes the SARS-CoV-2 “spike protein,” which normally is found
on the surface of the coronavirus particles. This mRNA is coated with a mixture of
synthetic lipids (fat-like molecules) that protect it during transport within the body,
and which also facilitate its uptake into the target cells through endocytosis.
After the vaccine has entered a cell, it initially finds itself enclosed by a mem-
brane vesicle—a little bubble that was pinched off from the cell membrane. The
subsequent accumulation of acid inside this bubble causes the lipids to be stripped
off, and the mRNA to be released into the cytosol (the intracellular fluid); this release
step is facilitated by the cationic lipid ALC-0315 (see later). The mRNA then binds
to ribosomes—the cell’s little protein factories—and induces the synthesis of the
actual spike protein molecules. Most of the spike protein molecules will then be
transported to the cell surface.
Once it appears there, it will be recognized by B-lymphocytes (B-cells), which will
then start making antibodies to it.2 Furthermore, some part of the spike protein can
also be cleaved off by proteases on the cell surface and released from the cell. If
this happens within the circulation, the released fragment—referred to as S1—can
bind to blood platelets (thrombocytes) and activate them. In this manner, the spike
protein directly promotes blood clotting.
2 B-cell activation involves additional steps and auxiliary cells that are here omitted for simplicity.
2
As with any protein that is synthesized within the cell, a small number of mole-
cules will undergo fragmentation, and the fragments will be presented on the cell
surface in association with specific (HLA-) carrier proteins. The purpose of this
mechanism is immune surveillance—as soon as fragments show up of some protein
which the immune system does not recognize as “self,” an immune response will be
mounted against that protein and against the cells that produce it. This response is
mediated by cytotoxic T-lymphocytes (CTLs, T-killer cells).
In mounting its cytotoxic response, the immune system will not distinguish
between a true virus infection and the expression of an mRNA vaccine—as long
as the spike protein fragments appear on the cell, the killer cells will be on the
march. If the vaccine is expressed in the cells that line the blood vessels—the
endothelial cells—the vascular lesion caused by the immune attack will again set off
blood clotting. Thus, we have at least two distinct paths toward blood clotting after
vaccination.
1.2 The lipid-coated mRNA vaccines acquire an apolipoprotein “corona”
Lipoprotein particles occur naturally in the bloodstream and within the tissues of
our body. They consist of a core of lipids that is surrounded with a shell of proteins
called apolipoproteins. Their purpose is to transport lipids such as cholesterol
and triacylglycerol (regular fat) between organs. For example, a specific type of
lipoprotein called chylomicrons transports dietary fats after they have been taken
up in the small intestine. Other lipoproteins called VLDL and LDL distribute fats
that have been synthesized in the liver to other organs and tissues.
The various apolipoproteins that encase the lipoproteins stabilize the particles,
and they also serve as “address tags” that bind to receptor molecules on cell surfaces.
This interaction will trigger the uptake of the lipoproteins into those cells. Artificial
lipid nanoparticles (LNPs) like those used in the COVID mRNA vaccines can acquire
a shell—a “corona”—of the body’s own apolipoprotein molecules [3]. This enables
these vaccines to be taken up into the cells of our body, too.
The liver has a central place in lipid and lipoprotein metabolic turnover. Ac-
cordingly, liver cells are rich in specific surface receptor molecules which mediate
3
lipoprotein uptake, suggesting that they will efficiently take up LNPs decorated with
apolipoproteins also. This is indeed the case. However, other organs have high rates
of lipoprotein uptake, too, and they must therefore be expected to accumulate the
apolipoprotein-decorated vaccine LNPs as well.
1.3 Receptor-mediated cellular uptake of lipoproteins and of vaccines
This slide illustrates the role of cellular receptors and the apolipoproteins in facili-
tating the uptake of vaccines into cells through endocytosis. They bind to the same
cellular receptors as the regular lipoprotein particles do, and they subsequently get
taken up in the same manner. The subsequent events—release of the mRNA and
protein synthesis—have already been discussed above.
1.4 Transcytosis of lipoproteins from the bloodstream into the tissues
All substrate exchange between the tissues and the bloodstream occurs in the capil-
laries. In these finest of all blood vessels, the blood is separated from the extracel-
lular matrix of the tissues by only one cellular layer—namely, the endothelial cells.
The capillary wall permits free passage only to small molecules such as for example
4
blood sugar (glucose) or amino acids. The lipoproteins, which are far larger, must
be transported across the capillary wall by transcytosis. In this two-stage process,
endocytosis on one side of the cell is followed by exocytosis, that is, by release of
the particles, which occurs on the other side.
While this figure shows transcytosis from the bloodstream to the tissue, the
process actually works in both directions. In this manner, cells in the tissues can
avail themselves of cholesterol carried by circulating LDL, but they can also return
surplus cholesterol through the bloodstream to the liver via other lipoproteins
(HDL).
Transcytosis will also apply to the “corona”-decorated vaccine LNPs and enable
them to reach the tissues in various organs. Reverse transcytosis of vaccine might
contribute to its uptake from the muscle tissue into the circulation after injection
(see below).
2 The Pfizer vaccine pharmacokinetics study on rats
•A “model vaccine” was used—same LNPs, different mRNA (coding for luciferase)
•Cholesterol contained in the LNPs was labeled with radioactivity (3H) for tracing
•The distribution of the lipid between different organs was measured at various
time points following intramuscular injection
This is the key experiment in Pfizer’s animal study [1]. The technical approach used
here is quite common, since radioactivity can be very sensitively and accurately
measured. The radioactively labeled vaccine preparation was injected into rats. The
animals were “sacrificed” (cut up) at various time points after the injection, and the
amount of radioactivity in different organs was measured.
The model protein used in this study was a firefly protein called luciferase. This
is the very protein that permits fireflies to glow in the dark. When the rats’ body
cells take up the mRNA that encodes luciferase and then synthesize the protein,
they, too, will begin to glow in the dark. Light, like radioactivity, is convenient to
measure; the more light that emanates from a given tissue, the more mRNA uptake
and protein synthesis have occurred.3 Therefore, between the radiolabel on the lipid
and the luminescence elicited by luciferase, it was possible to determine both the
distribution of the model vaccine within the body and its biological activity.
2.1 Key data from the lipid distribution study
The first thing to note is that the labeled lipid shows up in the blood plasma after a
very short time. The highest plasma level is reached at two hours after the injection;
however, even after only 15 minutes (0.25 hours) the level already reaches almost
half of that maximal value. Reverse transcytosis might in part account for this
rapid uptake process. A more important factor may be drainage of tissue fluid
3 To generate light, luciferase also requires a specific small-molecule substrate named luciferin
and adenosine triphosphate (ATP). The luminescence assay is therefore more complex and less quan-
titatively accurate than measurements of radioactivity.
5
through the lymphatic vessels into the bloodstream. Lymphatic drainage will likely
be accelerated by the acute release of inflammatory mediators within the muscle
tissue. 0
5
10
15
20
25
30
0.25 1 2 4 8 24 48
Lipid equivalent (μg/g)
Time after injection (h)
blood plasma
liver
spleen
ovaries
testes
adrenal glands
As the blood plasma level drops off, the activity rises in several other organs. The
fastest and highest rise is observed in the liver and the spleen. Both of these organs
are rich in macrophages, a cell type that is in charge of clearing particles such as
microbes or the fragments of decayed cells from the bloodstream. Macrophages are
also numerous in the bone marrow, where the vaccine reaches somewhat lower but
still substantial levels (not shown).
While the macrophages are likely responsible for most of the uptake in the spleen,
this may not be the case in the liver. Here, the vaccine likely ends up mostly
in the organ-specific epithelial cells, which are very rich in lipoprotein receptors.
Uptake into the ovaries and into the adrenal glands is most likely also mediated
by lipoprotein receptors. Both organs take up lipoproteins to obtain cholesterol,
which they use as a precursor for producing steroid hormones—corticosteroids in
the adrenal glands, and female sexual hormones (estrogens and progestins) in the
ovaries.
The testes, too, produce sexual hormones (in particular testosterone) from chol-
esterol, but here the accumulation of vaccine lipid is remarkably much lower. The
scientific literature does not offer a full, straightforward explanation for the re-
stricted uptake into the testes, but it may be related to the so-called blood-testes-
barrier. In most other organs examined the levels were similarly low as in the testes.
We note, however, that at least the blood vessels will be affected in every organ and
in every tissue.
2.2 Direct vs. indirect transport of radiolabel to the ovaries
It is noteworthy that the level of radioactivity in the liver rises very fast within
the first eight hours but then stagnates, whereas in the ovaries and the adrenal
glands the rise continues even two full days after the injection. This suggests
that the radioactivity may be redistributed from the liver to these glands. In this
context, we must remember that the LNP component which carried the label was
cholesterol. The labeled cholesterol would behave just like endogenous (unlabeled)
6
cholesterol, and after uptake into the liver we would expect it to be recycled and
redistributed to other organs. Cholesterol redistributed from the liver would likely
be unaccompanied by the mRNA. Therefore, the question whether the cholesterol
found in the ovaries is acquired in this indirect manner or through direct uptake of
the vaccine is of considerable importance.
In addition to cholesterol, the vaccine LNPs contain another naturally occuring lipid
(distearoyl-phosphatidylcholine) and two non-natural ones (see below). Thus, we
must ask to what extent these other lipids would undergo redistribution from the
liver and then also accumulate e.g. in the ovaries.
Finally, it must also be noted that the distribution of the vaccine might be affected
by the protein encoded by its mRNA component. If instead of the presumably inert
luciferase enzyme the spike protein had been expressed, this might have affected
vascular integrity, particularly also at the blood brain barrier. This might translate
into increased uptake into other organs, including the central nervous system.
Each of the posed questions could readily have been answered using experiments
similar to those reported by Pfizer—in particular, each of the relevant lipids should
have been radioactively labeled in turn, and the proper mRNA encoding the actual
spike protein should have been used instead of the one encoding luciferase. It
should go without saying that the FDA, the EMA and other regulators should never
have authorized the use of the vaccine without mandating and reviewing thorough
studies of this kind.
2.3 Very slow elimination of the cationic lipid ALC-0315 from rat liver
Of the two non-natural lipids contained in the vaccine LNPs, one (ALC-0315) is
weakly basic, whereas the other (ALC-0159) carries a polyethyleneglycol (PEG) moi-
ety. As just discussed, no comprehensive distribution studies on these lipids were
carried out. However, Pfizer did report the change over time of their concentrations
within the liver. The level of the PEG-modified lipid dropped slowly but regularly
7
with time. The other one, however—the cationic lipid ALC-0315—remained at very
high levels at two weeks (336 hours) after the injection. Even after 6 weeks some of
the compound was still detected in liver. As discussed in the preceding section, we
cannot rule out that these synthetic lipids, too, are redistributed from the liver to
other organs, where they might then be stored for even longer periods of time. 0.01
0.1
1
10
100
1000
0.1 1 10 100
Concentration (μg/ml)
Time after injection (h)
Cationic lipid
PEG−modified lipid
You may have heard that some pesticides such as DDT can persist in the human
body for months and even years. This typically occurs with compounds which are
very lipophilic, meaning that they partition into fat droplets within fat tissue and
other organs. As long as the fat within these droplets is not utilized, the chemicals
dissolved within them will be safe from metabolic turnover and degradation. The
cationic lipid ALC-0315 is likely able to accumulate in the same manner. If so, we
can expect persistence for even longer periods of time than evident from this graph
in tissues that have lower metabolic activity than the liver.
2.4 Slow degradation is built into the structure of ALC-0315HO N
O
O
O
O
HO N
OH
OH HO
O
HO
O
2 H2 O
8
This topic is rather technical, and it is not necessary for the big picture. If you can’t
make out what this diagram is showing, feel free to skip it.
The structure at the top shows the intact cationic lipid referred to as ALC-0315.
Hydrolysis of the two ester (C(=O)O) bonds produces the three fragments at the
bottom; according to Pfizer’s document, this is the initial step in the degradation
and elimination pathway of this lipid. The following features suggest that inside the
body this step will occur rather slowly:
1. The entire molecule contains no permanent charge and only one ionizable atom
(the nitrogen, N), which is linked to three alkyl chains. Aside from the one polar
hydroxy (OH) group, the entire remainder of the molecule is hydrophobic. This
means that the molecule will partition very strongly not only into lipid bilayers
(membranes) but also into lipid droplets, where it will be effectively hidden from
any degradative enzymes.
2. When this molecule is part of a lipid bilayer, as is the case within the vaccine
LNPs, the two ester bonds will still be buried deep within the hydrophobic por-
tion of that bilayer, which will protect them from hydrolytic cleavage.
3. Hydrolysis of the ester bonds will to some degree be sterically hindered by the
the adjacent branches in the fatty acyl residues.
With the possible exception of the lack of a permanent charge, none of these
features is essential for the desired function of the molecule, namely to release
the mRNA from the vaccine particles after the latter have been taken up into our
body cells. There are many ways in which this molecule could have been modified
for faster degradation in vivo. It is therefore noteworthy that this was not done—
the vaccine was deliberately formulated with a compound that is degraded and
eliminated from the body very slowly. Given that this lipid will most likely stay in our
tissues for months, we must expect cumulative toxicity with repeated vaccination
more at the site, with lovely diagrams .... chilling