For some of the COVID-19 vaccines, the drug substances released to market were manufactured differently than those used in clinical trials. Manufacturing nucleoside-modified mRNA (modRNA) for commercial COVID-19 vaccines relies on RNA polymerase transcription of a plasmid DNA template. Previous studies identified high levels of plasmid DNA in vials of modRNA vaccines, suggesting that the removal of residual DNA template is problematic. Therefore, we quantified the DNA load in a limited number of Pfizer-BioNTech and Moderna COVID-19 modRNA vaccine vials using two independent methods. Total DNA and specific DNA targets were quantified by Qubit fluorometry and quantitative polymerase chain reaction (qPCR), respectively on 32 vials representing 16 unique vaccine lots. RNase A treatment was used to assess the impact of RNA crosstalk in DNA fluorometry. A preliminary assessment of DNA fragment length and DNase I sensitivity were also performed. Total DNA ranged 371-1,548 ng/dose and 1,130-6,280 ng/dose in Pfizer and Moderna products, respectively. Specific DNA of multiple plasmid DNA targets ranged 0.22-7.28 ng/dose for Pfizer, and 0.01-0.78 ng/dose for Moderna. The SV40 promoter-enhancer-ori (0.25-23.72 ng/dose) was only detected in Pfizer vials. Oxford Nanopore sequencing of one vial found mean and maximum DNA fragment lengths of 214 bp and 3.5 kb, respectively. These data demonstrate the presence of 1.23 × 10⁸ to 1.60 × 10¹¹ plasmid DNA fragments per dose encapsulated in lipid nanoparticles. Using fluorometry, total DNA in all vials tested exceeded the regulatory limit for residual DNA set by the US Food & Drug Administration (FDA) and the World Health Authorization (WHO) by 36-153-fold for Pfizer and 112-627-fold for Moderna after accounting for nonspecific binding to modRNA. When tested by qPCR, all Moderna vials were within the regulatory limit, but 2/6 Pfizer lots (3 vials) exceeded the regulatory limit for the SV40 promoter-enhancer-ori by 2-fold. The presence of the SV40 promoter-enhancer element in Pfizer vials raises significant safety concerns. This study emphasizes the importance of methodological considerations when quantifying residual plasmid DNA in modRNA products, considering increased LNP transfection efficiency, and cumulative dosing presents significant and unquantified risks to human health.

Pharmacovigilance databases are showing evidence of injury in the context of the modified COVID-19 mRNA products. According to recent publications, adverse event reports linked to the mRNA COVID-19 injections largely point to the spike protein as an aetiological agent of adverse events, but we propose that the platform itself may be culpable. To assess the safety of current and future mRNA vaccines, further analysis is needed on the risks due to the platform itself, and not specifically the expressed antigen. If harm can be exclusively and conclusively attributed to the spike protein, then it is possible that future mRNA vaccines expressing other antigens will be safe. If harms are attributable to the platform itself, then regardless of the toxicity, or lack thereof, of the antigen to be expressed, the platform may be inherently unsafe, pending modification. In this work, we examine previous studies of RNA-based delivery by a lipid nanoparticle (LNP) and break down the possible aetiological elements of harm.

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Human cytomegalovirus (CMV) infection is a substantial cause of morbidity and mortality in immunocompromised hosts and globally is one of the most important congenital infections. The nucleoside analogue ganciclovir (GCV), which requires initial phosphorylation by the viral UL97 kinase, is the mainstay for treatment. To date, CMV decay kinetics during GCV therapy have not been extensively investigated and its clinical implications not fully appreciated. We measured CMV DNA levels in the blood of 92 solid organ transplant recipients with CMV disease over the initial 21 days of ganciclovir therapy and identified four distinct decay patterns, including a new pattern exhibiting a transient viral rebound (Hump) following initial decline. Since current viral dynamics models were unable to account for this Hump profile, we developed a novel multi-level model, which includes the intracellular role of UL97 in the continued activation of ganciclovir, that successfully described all the decline patterns observed. Fitting the data allowed us to estimate ganciclovir effectiveness in vivo (mean 92%), infected cell half-life (mean 0.7 days), and other viral dynamics parameters that determine which of the four kinetic patterns will ensue. An important clinical implication of our results is that the virological efficacy of GCV operates over a broad dose range. The model also raises the possibility that GCV can drive replication to a new lower steady state but ultimately cannot fully eradicate it. This model is likely to be generalizable to other anti-CMV nucleoside analogs that require activation by viral enzymes such as UL97 or its homologues. Cytomegalovirus (CMV) remains a cause of disease in individuals with weakened immune systems such as patients undergoing organ transplantation. The mainstay of treatment for CMV is ganciclovir: a CMV-specific drug that looks like a building block of the viral DNA and which requires activation by a protein contained within CMV. Understanding how patients respond to treatment can lead to a better understanding of the basic biology of CMV in the human and also help to optimize treatment regimens. In this study we observed that following treatment of CMV disease in organ transplant patients, CMV viral loads decay according to four distinct patterns. We have developed a novel mathematical model that is able to describe each of these decay patterns. The model could potentially be used to assess how different patients respond to treatment in clinical settings and also to design better drug regimens as per patient. These results have striking implications in terms of how long patients should remain on treatment and provide new insight into CMV infection in the human host.

COVID-19 can have short- and long-term health consequences, including various cardiovascular, respiratory, hematologic, autoimmune, and neurological conditions. Although it is often claimed that COVID-19 mRNA vaccinations reduce COVID-19 severity and post-acute sequelae, these assertions are refuted by evidence of extensive mRNA immunization-related harms that appear to be amplified by SARS-CoV-2 infection, resulting in considerable overlap in reported adverse outcomes. Spike proteins from both sources persist in the human body over the long-term, leading to immune dysfunction, inflammation, autoimmunity, organ dysfunction, and overlapping toxicities. We hypothesize that the mRNA vaccinations create a persistent toxic milieu of spike protein, inflammatory lipid nanoparticles, and DNA impurities, amplifying morbidity and mortality risks commonly ascribed to SARS-CoV-2 infection. Many 2021-2024 morbidity/mortality events in highly vaccinated populations, though often attributed solely to COVID-19 illness (due to close temporal associations with laboratory-confirmed infection), were more likely to result from these interactions or “hybrid harms”. Evidence supporting our hypothesis includes studies of negative efficacy, overlapping pathologies (e.g., myocarditis and thrombosis), redundant mechanisms, and epidemiological surges in excess mortality during the Omicron era (since December 2021) in extensively vaccinated countries. Case report data indicate that spike protein production along with associated “spikeopathies” may persist for at least three years, during which a coronavirus infection could trigger a new disease syndrome that would logically be attributed to the infection based on the timing. In contrast there is a relatively mild course for Omicron infections in the unvaccinated. Ongoing spike production from prior mRNA vaccinations is likely to predispose Omicron-infected individuals to cumulative adverse effects over time. The amplified toxicities and immunopathologic effects may help account for near-synchronous waves of COVID-19 and all-cause mortality in the Omicron era. This novel framework calls for re-examining the unique immunopathological consequences of SARS-CoV-2 breakthrough infection in COVID-19 mRNA-vaccinated individuals and consideration of the implications for future public health strategies.

The coronavirus disease of 2019 (COVID-19) has the potential to cause or promote a number of health problems in both the short and long terms. It is often claimed that the COVID-19 messenger RNA (mRNA) vaccinations can reduce the severity of COVID-19 and its sequelae. We counter this claim with evidence and moreover hypothesize that the mRNA vaccinations are triggering mechanisms that result in an amplification of COVID-19 morbidity along with the “post-acute sequelae of COVID-19” or PASC. Since 2021, the coronavirus infections were often superimposed on a preexisting mRNA vaccine-induced milieu of toxic spike protein, inflammatory lipid nanoparticles, and residual process-related DNA impurities. Spike proteins resulting from both the mRNA vaccination and the natural SARS-CoV-2 infection have been shown to persist for extended periods, raising parallel concerns regarding potential implications for long-term safety with this vaccination. Many, if not most, of the morbidity and mortality events attributed to COVID-19 in extensively-vaccinated populations in 2022-2023 were due to the long-term background persistence of spike protein and other vaccine-associated components resulting from previous COVID-19 mRNA vaccinations. Although close temporal associations with laboratory-confirmed SARS-CoV-2 often engendered the assumption of viral causation, the morbidity and mortality events are plausibly attributed to a combination of mass vaccination and coronavirus infections. The overlapping spike-related toxicities and immunological effects of mRNA vaccinations and coronavirus infections have resulted in pronounced immune dysregulation and inflammatory cascades that likely account for near-synchronous waves of COVID-19 and all-cause mortality. We present research suggesting a potential for coronavirus infection-induced amplification of adverse events (AEs) associated with the COVID-19 mRNA vaccinations, including a diverse range of cardiovascular, hematological, autoimmune, and neurological conditions that frequently overlap with PASC. In this paper, we coin the phrase “Hybrid Harms Hypothesis”, which calls for re-examining the unique immunopathological dynamics of coronavirus infection in COVID-19 mRNA-vaccinated individuals and their implications for public health strategies.