Ronavax Roulette: All About the Adjuvants

By Julie Beal

The coronavirus vaccines are supposed to be ‘intelligent’, because they use novel adjuvants that can target the immune system in specific ways. On the other hand, many of them could cause immediate ‘hypersensitive’ reactions such as anaphylaxis or seizures, or what seems like a delayed reaction, i.e. one that results in an autoimmune disorder. An adjuvant (say ‘add-joov-unt’) is a substance that’s deliberately added to a vaccine to make you have a certain kind of immune response.

Brainwashed kids Covid Vaccination

If a vaccine only contains an antigen (e.g. a virus), it doesn’t make people produce enough antibodies, so adjuvants are added to force a reaction and make it last longer:

Adjuvants may be molecules, compounds, or macromolecular complexes that boost the potency, quality, or longevity of specific immune responses to antigens, but which should cause minimal toxicity.

Note, in the quote above, they can only say ‘should’, because adjuvants are bad for the body – that’s the whole point of them. They’re supposed to make you react, but only a bit! So if you’re thinking, “Right, OK, so what are these things, and what will they do to me?”, the answer is, “It depends!”  This is partly because we’ve all got a different genetic make-up, and different kinds of microbes living inside us.  But it also depends on which vaccine we’re talking about, because every company makes its own special blend, so they’re all a bit different too.

https://pubmed.ncbi.nlm.nih.gov/25915555/

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“However!”  You can be empowered by learning about how genetic vaccines work, and how the reaction to a vaccine is linked to autoimmune disorders and the little bugs that live inside you! This means getting a bit ‘sciencey’, e.g. how the body works, and what microbes like bacteria and viruses actually do in the body, and maybe even what DNA is. All of this is well worth knowing, especially if you’re required to keep getting extra shots for the ‘health passports’ which are planned to be used globally.

An overview of the ronavax adjuvants

There are several new adjuvants being used in the coronavirus vaccines (or ‘ronavax’), sometimes in different combinations, and it’s all a bit bewildering, especially with all the strange names and stuff you’ve never heard of. So instead of trying to learn about the names of individual adjuvants, it’s easier and more helpful to think of them in terms of how they operate. To break it all down a bit, we’ll group them into these basic categories:

  1. Nanoparticles
  2. MRNA and DNA
  3. Genetic inserts
  4. Electroporation
  5. Aluminium
  6. Pretend bacteria (Lipid A and CPG ODNs)
  7. Xenobiotic chemical modifications
  8. Lipids

The old adjuvants, like mercury and thimerosol, have been largely abandoned, because the new ones are meant to be cleverer, and more desired for use in genetic vaccines. Some of the new adjuvants have been tested and some are already licensed in other vaccines, but on the whole, they’re still experimental, especially since they work in combination with the other parts of the vaccine, and yet few tests seem to be required or conducted for the ronavax. The Pfizer vaccine, for instance, contains mRNA, which in itself is an adjuvant, but so far they’ve only tested it on a few rats, because the World Health Organisation does not require further studies. This is in line with genetically engineered protein vaccines, for which tests of genotoxicity are not required.

Not only are the new adjuvants very powerful, they’ve also been largely omitted from the public conversation about coronavirus vaccines. They are now able to target specific parts of the immune system, so, like the genetic vaccines they’re added to, the adjuvants are considered smart or intelligent.

Modern genetic vaccines and the adjuvants they contain are trying to control every aspect of the body’s response. (For instance, the adjuvants in the coronavirus vaccines are trying to provoke a ‘Th1 response’ rather than a ‘Th2 response’.) Even the genetic code for mRNA can be tweaked “in order to fine tune or tailor physiologic responses and outcomes”. But the body isn’t that simple, the reaction is not under control, and skewing the immune response could have knock-on effects.

Human bodies are astonishingly complex and adaptive, each with its own genetic make-up, so we can all have a different reaction, because it’s genes that encode “immune receptors, transcription factors and signaling molecules involved in the immune responses”.  But it gets even more complicated, because there are trillions of microbes living inside us, and they’ve got ten times more genes than we have and they can all interact with the contents of a vaccine.

The ones with the contracts

In terms of which vaccine has which adjuvants in it, here’s a list of the ones that have the big contracts (but be warned, right now it’ll just look like gobbledygook! It takes a while to learn the new words so don’t expect to understand it all straight away!)

  • Lipid nanoparticles with mRNA (Pfizer/BioNTech, Moderna, Curevac)
  • Viral-vectored DNA (Johnson & Johnson, and AstraZeneca/Oxford University)
  • Bacterial plasmid, using electroporation (Inovio)
  • Protein vaccines containing novel lipid nanoparticle mixes, e.g. squalene and cholesterol (e.g. Novavax’s Matrix-M and GSK’s AS03)
  • Protein vaccine containing a CPG-ODN (Valenva is adding an adjuvant called CPG-1018 to its ronavax)

The first three vaccines in the list above contain genetic code that get you to manufacture the proteins in your body, whilst the last two contain genetically engineered proteins (parts of a virus) which have been produced in a bioreactor, e.g. in insect cells. Although protein vaccines are said to require strong adjuvants, the DNA/mRNA vaccines may not need anything added, because the method used by each one is in itself an adjuvant (foreign genetic material is an adjuvant, and so are nanoparticles). It is actually possible to chuck in some genetic adjuvants, though, so there’s a chance the DNA/mRNA vaccines have a little extra something added.

An Oxford University patent, for a MERS vaccine using the same ChAdOx viral vector as the AstraZeneca ronavax, says no adjuvant is added. Then again, there are other ingredients in the vaccine which could contribute to adverse effects on the body, i.e. excipients and contaminants.  The final recipe described in the patent includes histidine, sucrose, sodium chloride, magnesium chloride, Polysorbate 80, EDTA, ethanol and hydrochloric acid. But it might also contain bits of baby, because it’s grown in cells from an aborted foetus (same for the J&J vax).

There’s a wide range of disorders that could result from a coronavirus vaccine, most of which are classed as allergic, hypersensitive, or autoimmune reactions. Many of these disorders were summarised in an FDA meeting about possible adverse reactions to the ronavax. They include all the classics: anaphylaxis, arthritis, various types of myelitis, stroke, seizures, heart problems, pregnancy and birth problems, things that go wrong with the blood, Guillain-Barré syndrome, and, yes, even death. The CDC is advising ronavax administrators to make sure they’ve at least got injectable epinephrine and H1 antihistamines available, in case anyone suffers anaphylactic shock.

Anaphylaxis is an immediate and obvious reaction to a vaccine, but the other disorders would take a while to develop, and may eventually lead to a diagnosis of autoimmune disease, but, hey, not to worry! There is now a wide range of drugs marketed at people with these conditions, most of which aim to suppress the activity of the immune system. Apparently, “the immunosuppressive biologics used to treat autoimmune disease are the top-selling medications worldwide” so any collateral damage from the vaccines will net a tidy profit. Ker-ching!

Understanding the immune response

The immune system is a highly complex network which is constantly adapting to what’s happening. Vaccine makers have figured out which cells have specific responses to various substances, mostly based on how we respond to danger signals. Some cells do signalling, some activate other cells, and some attack the invader, for example.

Adjuvants operate by engaging innate immune cells, such as dendritic cells (DCs), and shape the subsequent adaptive immunity. Pathogen associated molecular patterns (PAMPs) and danger signals are potent adjuvants; they activate DCs by stimulating cell-surface or intracellular pathogen recognition receptors …

Adjuvants attempt to mimic the natural response to pathogens and other danger signals, and yet, any intervention can only mimic a small part of the whole reaction. We don’t all react in the same way as each other, and our responses depend on what our body needs overall, and what the threat is.

NANOPARTICLES

Adjuvants with tiny bits in (i.e. particulated adjuvants) are usually called nanoparticles (and if they’re a bit bigger, they’re called microparticles). Any foreign object that enters the body sets off alarm signals that begin an immune reaction, but modern nanoparticles are designed to trigger specific responses as a result of what they’re made from. This is possible because they can get into cells to release their payload. Nanoparticles can be formed from lipids, polysaccharides, virus-like particles, and/or alum; for instance, immune-stimulatory complexes (ISCOMs) are spherical cage-like nanoparticles formed via self-assembly of a mixture of Quil A (which is a saponin), cholesterol, phospholipids and antigens.

Novavax make an adjuvant called Matrix-M and plan to use it in their coronavirus vaccine. It contains 40 nm nanoparticles, made with saponins, cholesterol, and phospholipids, so it’s similar to the GSK adjuvant, AS03. Novavax demonstrated how powerful their adjuvant is when they tested a vaccine containing “full-length spike nanoparticles” of MERS-CoV and SARS-CoV. They published a research paper in 2014 (‘Purified coronavirus spike protein nanoparticles induce coronavirus neutralizing antibodies in mice’) describing this experiment. The Novavax MERS vaccine was used to vaccinate mice, to test different combinations of adjuvants. Some mice got alum in the vax, others got Novavax’s own adjuvant called Matrix M, whilst a third group got only the antigen. The results clearly illustrate the intense effects of both alum and lipid nanoparticles; when they gave the mice 1 μg of nothing but the MERS protein, they produced GMTs (geometric mean titers) of 49 in terms of neutralizing antibodies; when alum was added, the GMT was 735; but with Matrix-M the GMT was 3,378! This means the neutralizing antibody levels were 15 times higher with alum, and a whopping 68 times higher with Matrix M. Adjuvants are supposed to ‘enhance’ the immune response, which you might imagine as being a gentle shove in the right direction, whereas these test results reveal an adjuvant that’s more like a hand grenade. It seems the success of a vaccine may more reliant on the adjuvant than the antigen.

Nanoparticles can be formed in oil in water emulsions by using a technique called microfluidization. The actual size of the particles affects the way the vaccine contents are taken up into cells, and how they interact with the antigen. “Alum microparticles remain localized at the site of injection”, whereas the nano-sized ones made from lipids move quickly into the draining lymph node. Lipid nanoparticles (LNPs) form different shapes depending on where they are in their phase transition state (as liquid crystals), and the shape of LNPs can influence their effect: for instance, if they’re rod‐like or needle‐shaped, they cause less of an inflammatory response compared to spherical ones. Other factors affecting LNPs are the type of lipids used, use of steric stabilizers, and their surface charge.

Nanoparticles are said to activate the inflammasome, which makes their use in vaccines a real concern, because the inflammasome has to be carefully regulated –if not, it can damage the body. The inflammasome senses danger signals from cells in distress, as well as threats from pathogens (including those in the gut microbiome). Once activated, it creates a cascade of inflammatory responses, but it can go too far and “cause uncontrolled tissue responses that may contribute to various diseases, including autoinflammatory disorders, cardiometabolic diseases, cancer and neurodegenerative diseases.”

Several vaccines containing nanoparticles have been licensed in the past decade, e.g. the Pandemrix vaccine that led to a number of young people getting narcolepsy. These vaccines use lipids to make the nanoparticles, which are basically types of oil. This could be an additional concern, since many autoimmune disorders are also linked to disruption of lipid metabolism in the body. All of our cells have membranes made from lipids, and the nanoparticles could affect the delicate balance of lipids in this membrane when they fuse with it.

The complement system is said to be activated by PEG lipids in liposomes, which could be an issue because complement is a part of the immune system that deals with infections, and inflammatory responses, and is also linked to autoimmune disorders such as lupus. However, this mechanism has not been properly studied, despite the fact that it can “dramatically alter the pharmacokinetics of particles on repeat administration, with significant consequences for their use in therapeutic applications.”

Lipid nanoparticles (LNPs) are being used to deliver mRNA in some of the coronavirus vaccines, and limited information about these has recently been provided by the manufacturers. For instance, the Pfizer vaccine uses LNPs composed of four lipids, two of which (cholesterol and DSPC) are described as “naturally occurring”, and it’s said that these lipids “will be metabolised and excreted” in the same way as ones that are found in the human body. The same is not said of the other two lipids (ALC-0315 and ALC-0159) which are proprietary blends, but the LNPs are said to hang around in the body for several days.

THE ADJUVANTING EFFECTS OF MRNA AND DNA

If injected into the body, both DNA and mRNA are attacked as foreign invaders before they can get inside a cell; they’re degraded by things called ribonucleases, or RNases (enzymes that digest mRNA). This made it difficult for researchers to use mRNA as a therapy, as well as other issues, such as trying to get mRNA through the cell membrane (both of them have a negative charge, and “two elements of a same charge repel each other”). Even if they managed to get it inside a cell, it was degraded by intracellular enzymes called exonucleases. These reactions demonstrate the power and flexibility of the immune response – all of which prevented development of mRNA treatments, until researchers came up with various fixes, which involve altering the genetic sequence and using non-natural chemical modifications (using versions of the real thing, e.g. pseudo-uridine) to trick the immune system so it doesn’t get destroyed, but is still understandable enough for the body to obey the instructions contained in the genetic code. It therefore retains its adjuvanting effect – it kind of triggers an alarm if it binds to special cell receptors. These receptors can recognize different kinds of RNA or DNA that are normally associated with pathogens like viruses and bacteria, which then sets off a chain-reaction in other immune cells.

The vaccines made by Inovio have two types of DNA, because they contain DNA from the bacterial plasmid that acts as a vector, as well as the DNA that encodes the coronavirus antigen. Bacterial DNA is recognized by the body because of its CpG pattern; this triggers the immune system, e.g. production of TH1 cytokines.

Electroporation is a technique used to get DNA plasmids into cells, and Inovio have their own device, called CELLECTRA, for this purpose. DNA has to get all the way to the nucleus, to be transcribed into mRNA, which is do-able with viral vectors because they are able to get through cell membranes, but DNA plasmids need more help. Electroporation involves applying a series of electrical pulses to the skin, straight after being vaccinated (in the same place). This makes cell membranes more permeable by temporarily creating little holes in it so the DNA can get in. It also “increases DNA distribution throughout the tissue and causes a local inflammatory reaction, both of which contribute to a stronger immune response”. This is because the body senses danger, either from cell damage, or cell death.



Genetic adjuvants

These can be added to vaccines that use viral vectors, bacterial plasmids, or mRNA in nanoparticles, since it just involves chucking in some extra genes, from somewhere or other. The use of additional adjuvant molecules, such as GM-CSF, has been shown to shift a Th2-type response to a Th1-type response. Although there’s no sign of genetic adjuvants being used in the coronavirus vaccines, they’re worth knowing about because . They are sometimes called molecular or endogenous adjuvants and they’re based on human genes, so that specific proteins are produced which are able to target “tumors, autoimmunity, viral infections, and transplants”. They can include heat shock proteins, cytokines (e.g. IL-12), chemokines, antibodies, small peptides and proteins.

Moderna’s patent for a betacoronavirus vaccine describes the possibility of adding a flagellin adjuvant. Flagellin is a type of bacteria, which has little tentacle-like things called ‘flagella’ to help it move. When it’s added to nanoparticles, it can be more ‘potent’ than the real thing. Curevac make RNAdjuvant, which is said to avoid inducing splenomegaly (enlarged spleen) in mice “as described for standard adjuvants such as CpG-DNA”. Curevac’s RNActive® vaccines have two components – one is a natural form of mRNA, encoding the antigen, and the other is protamine which boosts the immune response by activating the TLR7 receptor. Protamine is a nuclear protein that stabilizes DNA during the creation of sperm, said to help stop mRNA being degraded by cells in the blood.

(For a full run-down of adjuvants being used in the ronavax, see ‘Potential adjuvants for the development of a SARS-CoV-2 vaccine based on experimental results from similar coronaviruses’, by Guptaa and Guptab.)

Alum

Alum has … been used as a carrier/vehicle for a number of new age adjuvants including IL-12, GM-CSF, CpG, and MPL. Most importantly, the recent approval of MPL-Alum® for two different vaccines, Fendrix and Cervarix, has set precedence for the use of TLR4 agonists within human vaccines.

Aluminium can be absorbed through the skin and can collect in tissues, e.g. it can collect in breast tissue from using antiperspirants. It’s been used in vaccines for years because it makes the antigen hang around for longer, thus prolonging the immune response. Alum also kills off your cells, and the effect of this is said to be “an endogenous adjuvant” (one you produce yourself); basically, when cells die, they release DNA and uric acid, both of which are perceived as danger signals, leading to even more immune reactions. Alum-based vaccines initiate Th2-type antibody responses which are associated with allergic reactions and autoimmune disorders.

Yet more to consider

Blimey, did you actually read all that? It’s proper hard-going, I know, so I’ve written other articles which explain more about these things, but in a more general way. There’s one about nothing but lipids, and another about the use of ‘alien’ genetics in the coronavirus vaccines. These are also a bit heavy, so I’ve tried to make them more readable! If you want to know more about how autoimmune disorders are linked to the human microbiome and its interaction with vaccine contents, check out my article on molecular mimicry. Last but not least, there’s a ‘ronavax primer’ that explains some of the new words and concepts a bit more.

See Julie Beal’s entire archive HERE.

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