Role In The Innate Immune Response.
It is a year since we first identified the significance of vitamin D in respect of SARS-CoV-2. Through the vitamin D pathways it can both prevent viral infection and attenuate disease progression and severity. Yet the role of vitamin D-modulated nitric oxide remains largely ignored and dismissed by the NHS as being of no value. Is it any wonder that so many have died unnecessarily in the UK when fundamental principles of the innate immune system have been disregarded?
A product of the reactive nitrogen species, nitric oxide (NO) is a potent part of the innate immune system. It is highly expressed in the upper respiratory tract and is heavily synthesised in the paranasal sinuses and nasaopharynx. As SARS-CoV-2 binds to apical ciliated epithelial cells in the upper respiratory tract, NO is ideally located, which is no coincidence since the upper respiratory tract represents one of the two main points of entry for invasive pathogens and therefore it is perhaps the most heavily protected area of the human body.
NO has a very short half-life of approx 5 milliseconds to 2 seconds within cellular tissue and is only effective within a very close range of 0.005 – 0.5 millimetres. Either independently or through cytokine-signalling pathways, NO recognises antigen, including coronaviruses, and lyses target cells.
SARS-CoV-2 Specific Response.
In addition to the broad innate response, NO also has an SARS-CoV-2 antigen-specific response, as it inhibits palmitoylation in the SARS-CoV-2 receptor binding domain, reducing binding affinity of the S2 subunit to the membrane of the target cell.
Following intracellular fusion, NO targets SARS-COV-2’s nsp3 & nsp4 [Xu et al., 2020], which disrupts mRNA transcription and assembly of the replication & transcription complex (RTC). In addition, NO triggers nitrosylation of the amino acid cysteine and is therefore effective in inhibiting cysteine proteases. The main protease of SARS-CoV-2, 3-chymotrypsin-like protease, is a cysteine protease. 3CLPro (also called MPro due to its main function), is necessary to unpack pp1a/ab, to cleave the nsps and – most importantly – to oversee the assembly of the RTC.
SARS-CoV-2 3CLPro showing cysteine residue
In other words, NO is effective at intefering with mRNA transcription and preventing the viral replication cycle. This is regardless of the variant as NO is unaffected by single nucleotide polymorphism or amino acid substitution in the virus’ genetic sequence. NO doesn’t care if it detects Alpha, Delta, Kappa or any other variant, it attacks it.
If that were not enough, NO also upregulates the production of mucus in the mucosal epithelium [Nagaki et al., 1995]. As well as creating a natural barrier to pathogen, mucus contains defensive mucins and sIgA istotype antibodies. IgA is a faster, stronger and more effective isotype than both IgM and IgG as we explained in January and expanded upon in March 2021.
In respect of attenuating COVID-19 disease progression and severity, our work from October 2020 on the benefits of the cholecalciferol→calcifediol→calcitriol pathway were validated in February 2021, with Nogués et al. showing that high dosage calcifediol when administered to those hospitalised with COVID-19 – thereby having at least moderate disease severity – produced a 74.3% reduction in disease progression and a 56.7% reduction in mortality rate.
Nitric Oxide’s Importance Has Been Known For Over 20 Years.
The relevance and importance of NO as a viral countermeasure for respiratory viruses in general and SARS-CoV-2 in particular have not just emerged since the the designation of SARS-CoV-2. Following SARS-CoV, studies were undertaken on the role of NO in preventing viral infection through inhibition/disruption of the replication process. Ask anyone involved in the relevant fields of study and you would know that NO (whether natural or through NO donor compounds, notably S-nitroso-N-acetylpenicillamine) is proven to reduce and help prevent viral infection in the upper respiratory tract.
For example, Saura et al., 1999; Keyaerts et al., 2004; Akerström et al., 2005; Klingström et al., 2006; Akerström et al. 2009; Regev-Shoshani et al., 2013; Akaberi et al., 2020; Lisi, Zelikin and Chandrawati 2021.
The authors of the last study make the prescient observation in respect of Regev-Shoshani et al. noting that “Remarkably, this study was all but neglected and all studies on antiviral effects of NO that appear in press continue to rely on NO donors.”
This comment is highly relevant as Regev-Shoshani is part of SaNOtize, the Canadian company that has been developing a nasal spray as a SARS-CoV-2 vaccine candidate. In January we first wrote about its Nitric Oxide Releasing System, which has now become Nitric Oxide Nasal Spray. It is a nasal spray that delivers focused NO in the upper respiratory tract. No reliance upon or wait for antibody seroconversion; no loss of effectiveness with neutralisation escape variants and no risk of adverse side effects through the use of phospholipids or polyethylene glycol.At the bare minimum, NONS is a highly effective antiviral – its clinical trials show >99% effectiveness – that offers passive immunity to those in high risk groups, principally those with immunosenescence. As early as October 2020, we stated that based upon average mortality age, comorbidities plus SARS-CoV-2’s pathogenesis and antigen shift (its adaptive camouflage or stealth mode), a prophylactic vaccine was not required and would be unlikely to be effective (given the continuous amino acid substitutions to gain fitness advantage through increased immunoevasion). We concluded;
“Perhaps the best outcome that can be hoped for is an antiviral that is 50% effective for those in high risk groups, where it can provide them with passive immunity. This would benefit those with immunosenescence and lacking immunological memory, with an antiviral in effect giving them a second line of defence.”
All of the ‘vaccines’ currently being injected are antivirals, with their reliance upon the structural spike protein being static meaning they show varying degrees of ineffectiveness against different variants. If a drug manufacturer is talking about third/fourth doses or annual ‘boosters’, they are describing a therapeutic antiviral not a prophylactic vaccine. Think oseltamivir or zanamivir, the annual flu antivirals which are not vaccines. The winter/seasonal flu antivirals cannot stop you catching or spreading the virus, they can only potentially reduce any disease progression and severity, thereby reducing recovery times and potential hospital admissions. Even if the data to support these benefits is inconclusive at best, with the NHS’ own conclusion of their effectiveness being:
“While it appears that these drugs have a modest benefit, there is no solid evidence that either drug can protect people from the more serious complications of influenza. Paracetamol or ibuprofen would seem to be a far more cost-effective method of relieving the symptoms of influenza.”
The effectiveness of Vaxzevria, BNT126b2, mRNA-1273 and Janssen will in the future most likely be considered in similar terms, before you factor in the known adverse side effects that have been ignored. We highlighted distinct risk factors in January, February, April and May 2021 and continue to research severe adverse side effects around antiphospholipid antibodies.
Conclusions.
One has to ask not just why the pivotal role of nitric oxide in preventing SARS-CoV-2 infection has been overlooked but why bodies such as Public Health England and the NHS have ridiculed those who have highlighted the medical benefits of vitamin D.
SARS-CoV-2’s Achilles Heel is vitamin D: it thrives in its deficiency and it is highly susceptible to its abundance.
Nitric oxide attacks the virion on multiple fronts, removing the reliance upon IgG seroconverion in the humoral system. As part of the innate immune response, it destroys SARS-CoV-2 virions and infected cells before viral replication, thereby removing the requirement for an adaptive immune response. Or a vaccine.
