Pfizer mRNA Vexxine Reverse Transcribed To DNA in Human Liver

Remember those assurances that the mRNA could never ever be turned into DNA in the nucleus of our cells? Well…


https://www.mdpi.com/1467-3045/44/3/73/htm

Open Access Article

Intracellular Reverse Transcription of Pfizer BioNTech COVID-19 mRNA Vaccine BNT162b2 In Vitro in Human Liver Cell Line

by Markus Aldén 1ORCID,Francisko Olofsson Falla 1,Daowei Yang 1,Mohammad Barghouth 1,Cheng Luan 1,Magnus Rasmussen 2 andYang De Marinis 1

1 Department of Clinical Sciences, Lund University, 20502 Malmö, Sweden
2 Infection Medicine, Department of Clinical Sciences, Lund University, 22362 Lund, Sweden
[…]
Academic Editor: Stephen Malnick
Curr. Issues Mol. Biol. 2022, 44(3), 1115-1126;

https://doi.org/10.3390/cimb44030073

Received: 18 January 2022 / Revised: 19 February 2022 / Accepted: 23 February 2022 / Published: 25 February 2022

(This article belongs to the Topic Clinical, Translational and Basic Research on Liver Diseases)

Abstract

Preclinical studies of COVID-19 mRNA vaccine BNT162b2, developed by Pfizer and BioNTech, showed reversible hepatic effects in animals that received the BNT162b2 injection. Furthermore, a recent study showed that SARS-CoV-2 RNA can be reverse-transcribed and integrated into the genome of human cells. In this study, we investigated the effect of BNT162b2 on the human liver cell line Huh7 in vitro. Huh7 cells were exposed to BNT162b2, and quantitative PCR was performed on RNA extracted from the cells. We detected high levels of BNT162b2 in Huh7 cells and changes in gene expression of long interspersed nuclear element-1 (LINE-1), which is an endogenous reverse transcriptase. Immunohistochemistry using antibody binding to LINE-1 open reading frame-1 RNA-binding protein (ORFp1) on Huh7 cells treated with BNT162b2 indicated increased nucleus distribution of LINE-1. PCR on genomic DNA of Huh7 cells exposed to BNT162b2 amplified the DNA sequence unique to BNT162b2. Our results indicate a fast up-take of BNT162b2 into human liver cell line Huh7, leading to changes in LINE-1 expression and distribution. We also show that BNT162b2 mRNA is reverse transcribed intracellularly into DNA in as fast as 6 h upon BNT162b2 exposure.

Keywords: COVID-19 mRNA vaccine; BNT162b2; liver; reverse transcription; LINE-1; Huh7

So there you have it. This stuff gets reverse transcribed to DNA in the human liver cells. It looks like they first observed this from the actual virus (link below) so decided to look at the vaxxine in a cell line in the lab.

A recent study showed that SARS-CoV-2 RNAs can be reverse-transcribed and integrated into the genome of human cells [25]. This gives rise to the question of if this may also occur with BNT162b2, which encodes partial SARS-CoV-2 RNA. In pharmacokinetics data provided by Pfizer to European Medicines Agency (EMA), BNT162b2 biodistribution was studied in mice and rats by intra-muscular injection with radiolabeled LNP and luciferase modRNA. Radioactivity was detected in most tissues from the first time point (0.25 h), and results showed that the injection site and the liver were the major sites of distribution, with maximum concentrations observed at 8–48 h post-dose [26]. Furthermore, in animals that received the BNT162b2 injection, reversible hepatic effects were observed, including enlarged liver, vacuolation, increased gamma glutamyl transferase (γGT) levels, and increased levels of aspartate transaminase (AST) and alkaline phosphatase (ALP) [26]. Transient hepatic effects induced by LNP delivery systems have been reported previously [27,28,29,30], nevertheless, it has also been shown that the empty LNP without modRNA alone does not introduce any significant liver injury [27]. Therefore, in this study, we aim to examine the effect of BNT162b2 on a human liver cell line in vitro and investigate if BNT162b2 can be reverse transcribed into DNA through endogenous mechanisms.
[…]

4. Discussion
In this study we present evidence that COVID-19 mRNA vaccine BNT162b2 is able to enter the human liver cell line Huh7 in vitro. BNT162b2 mRNA is reverse transcribed intracellularly into DNA as fast as 6 h after BNT162b2 exposure. A possible mechanism for reverse transcription is through endogenous reverse transcriptase LINE-1, and the nucleus protein distribution of LINE-1 is elevated by BNT162b2.

Intracellular accumulation of LNP in hepatocytes has been demonstrated in vivo [36]. A preclinical study on BNT162b2 showed that BNT162b2 enters the human cell line HEK293T cells and leads to robust expression of BNT162b2 antigen [37]. Therefore, in this study, we first investigated the entry of BNT162b2 in the human liver cell line Huh7 cells. The choice of BNT162b2 concentrations used in this study warrants explanation. BNT162b2 is administered as a series of two doses three weeks apart, and each dose contains 30 µg of BNT162b2 in a volume of 0.3 mL, which makes the local concentration at the injection site at the highest 100 µg/mL [31]. A previous study on mRNA vaccines against H10N8 and H7N9 influenza viruses using a similar LNP delivery system showed that the mRNA vaccine can distribute rather nonspecifically to several organs such as liver, spleen, heart, kidney, lung, and brain, and the concentration in the liver is roughly 100 times lower than that of the intra-muscular injection site [38]. In the assessment report on BNT162b2 provided to EMA by Pfizer, the pharmacokinetic distribution studies in rats demonstrated that a relatively large proportion (up to 18%) of the total dose distributes to the liver [26]. We therefore chose to use 0.5, 1, and 2 μg/mL of vaccine in our experiments on the liver cells. However, the effect of a broader range of lower and higher concentrations of BNT162b2 should also be verified in future studies.
In the current study, we employed a human liver cell line for in vitro investigation. It is worth investigating if the liver cells also present the vaccine-derived SARS-CoV-2 spike protein, which could potentially make the liver cells targets for previously primed spike protein reactive cytotoxic T cells. There has been case reports on individuals who developed autoimmune hepatitis [39] after BNT162b2 vaccination. To obtain better understanding of the potential effects of BNT162b2 on liver function, in vivo models are desired for future studies.

In the BNT162b2 toxicity report, no genotoxicity nor carcinogenicity studies have been provided [26]. Our study shows that BNT162b2 can be reverse transcribed to DNA in liver cell line Huh7, and this may give rise to the concern if BNT162b2-derived DNA may be integrated into the host genome and affect the integrity of genomic DNA, which may potentially mediate genotoxic side effects. At this stage, we do not know if DNA reverse transcribed from BNT162b2 is integrated into the cell genome. Further studies are needed to demonstrate the effect of BNT162b2 on genomic integrity, including whole genome sequencing of cells exposed to BNT162b2, as well as tissues from human subjects who received BNT162b2 vaccination.

Human autonomous retrotransposon LINE-1 is a cellular endogenous reverse transcriptase and the only remaining active transposon in humans, able to retrotranspose itself and other nonautonomous elements [40,41], and ~17% of the human genome are comprised of LINE-1 sequences [42]. The nonautonomous Alu elements, short, interspersed nucleotide elements (SINEs), variable-number-of-tandem-repeats (VNTR), as well as cellular mRNA-processed pseudogenes, are retrotransposed by the LINE-1 retrotransposition proteins working in trans [43,44]. A recent study showed that endogenous LINE-1 mediates reverse transcription and integration of SARS-CoV-2 sequences in the genomes of infected human cells [25]. Furthermore, expression of endogenous LINE-1 is often increased upon viral infection, including SARS-CoV-2 infection [45,46,47]. Previous studies showed that LINE-1 retrotransposition activity is regulated by RNA metabolism [48,49], DNA damage response [50], and autophagy [51]. Efficient retrotransposition of LINE-1 is often associated with cell cycle and nuclear envelope breakdown during mitosis [52,53], as well as exogenous retroviruses [54,55], which promotes entrance of LINE-1 into the nucleus. In our study, we observed increased LINE-1 ORF1p distribution as determined by immunohistochemistry in the nucleus by BNT162b2 at all concentrations tested (0.5, 1, and 2 μg/mL), while elevated LINE-1 gene expression was detected at the highest BNT162b2 concentration (2 μg/mL). It is worth noting that gene transcription is regulated by chromatin modifications, transcription factor regulation, and the rate of RNA degradation, while translational regulation of protein involves ribosome recruitment on the initiation codon, modulation of peptide elongation, termination of protein synthesis, or ribosome biogenesis. These two processes are controlled by different mechanisms, and therefore they may not always show the same change patterns in response to external challenges. The exact regulation of LINE-1 activity in response to BNT162b2 merits further study.

The cell model that we used in this study is a carcinoma cell line, with active DNA replication which differs from non-dividing somatic cells. It has also been shown that Huh7 cells display significant different gene and protein expression including upregulated proteins involved in RNA metabolism [56]. However, cell proliferation is also active in several human tissues such as the bone marrow or basal layers of epithelia as well as during embryogenesis, and it is therefore necessary to examine the effect of BNT162b2 on genomic integrity under such conditions. Furthermore, effective retrotransposition of LINE-1 has also been reported in non-dividing and terminally differentiated cells, such as human neurons [57,58].

The Pfizer EMA assessment report also showed that BNT162b2 distributes in the spleen (<1.1%), adrenal glands (<0.1%), as well as low and measurable radioactivity in the ovaries and testes (<0.1%) [26]. Furthermore, no data on placental transfer of BNT162b2 is available from Pfizer EMA assessment report. Our results showed that BNT162b2 mRNA readily enters Huh7 cells at a concentration (0.5 µg/mL) corresponding to 0.5% of the local injection site concentration,M.b< induce changes in LINE-1 gene and protein expression, and within 6 h, reverse transcription of BNT162b2 can be detected. It is therefore important to investigate further the effect of BNT162b2 on other cell types and tissues both in vitro and in vivo.

5. Conclusions
Our study is the first in vitro study on the effect of COVID-19 mRNA vaccine BNT162b2 on human liver cell line. We present evidence on fast entry of BNT162b2 into the cells and subsequent intracellular reverse transcription of BNT162b2 mRNA into DNA.

https://pubmed.ncbi.nlm.nih.gov/33958444/

Reverse-transcribed SARS-CoV-2 RNA can integrate into the genome of cultured human cells and can be expressed in patient-derived tissues

Liguo Zhang 1, Alexsia Richards 1, M Inmaculada Barrasa 1, Stephen H Hughes 2, Richard A Young 1 3, Rudolf Jaenisch 4 3
Affiliations expand
PMID: 33958444 PMCID: PMC8166107 DOI: 10.1073/pnas.2105968118

Abstract

Prolonged detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA and recurrence of PCR-positive tests have been widely reported in patients after recovery from COVID-19, but some of these patients do not appear to shed infectious virus. We investigated the possibility that SARS-CoV-2 RNAs can be reverse-transcribed and integrated into the DNA of human cells in culture and that transcription of the integrated sequences might account for some of the positive PCR tests seen in patients. In support of this hypothesis, we found that DNA copies of SARS-CoV-2 sequences can be integrated into the genome of infected human cells. We found target site duplications flanking the viral sequences and consensus LINE1 endonuclease recognition sequences at the integration sites, consistent with a LINE1 retrotransposon-mediated, target-primed reverse transcription and retroposition mechanism. We also found, in some patient-derived tissues, evidence suggesting that a large fraction of the viral sequences is transcribed from integrated DNA copies of viral sequences, generating viral-host chimeric transcripts. The integration and transcription of viral sequences may thus contribute to the detection of viral RNA by PCR in patients after infection and clinical recovery. Because we have detected only subgenomic sequences derived mainly from the 3′ end of the viral genome integrated into the DNA of the host cell, infectious virus cannot be produced from the integrated subgenomic SARS-CoV-2 sequences.

Keywords: LINE1; SARS-CoV-2; chimeric RNAs; genomic integration; reverse transcription.

Copyright © 2021 the Author(s). Published by PNAS.

One wonders how many PCR Detected “cases” are in recovered folks or in folks who had the jab and do NOT have active infection, just have had their DNA modified to make virus parts…

I think this shows the “unsymptomatic spreader” and the “PCR Diagnostic” both have really big doubts verging on “just broken idea” status?

Clearly the claim that the mRNA Never Ever gets into your cells is shown to be bogus.

So I’m now very much never ever going to accept ANY mRNA “therapy” or “vaccine” unless I’m terminally ill with cancer and it’s the only possible hope.

“Not ready for prime time” comes to mind.

Also it looks like the CCP / Fauxi Wuhan Covid Virus is messing up the global genome too. Thanks for that “gain of function” R&D Fauxi…/sarc;

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About E.M.Smith

A technical managerial sort interested in things from Stonehenge to computer science. My present "hot buttons' are the mythology of Climate Change and ancient metrology; but things change...
This entry was posted in Covid, Political Current Events. Bookmark the permalink.

12 Responses to Pfizer mRNA Vexxine Reverse Transcribed To DNA in Human Liver

  1. andysaurus says:

    I’m glad I only had the Astra Zeneca shot, which as I understand it is more like traditional vaccines. I have refused the Pfizer booster, even though my age, and health puts me in a vulnerable category (overweight, 72YO, sleep apnoea, cardiac problems etc.) I take loads of supplements (Vitamins C, D, methylated B12, K2, minerals zinc, magnesium, enzymes Co-Q10, NAC) and avoid masks wherever possible. I eat meat, fish, cheese, chicken, and eggs, butter and I cook in beef fat (which I call dripping). I am secretary of my local Men’s Shed which brings me into contact with many people. I practice normal hygiene, such as washing my hands regularly. I am located in a semi-rural satellite community and have about 4 acres of land (1.34 Ha) I have not had Wuflu.
    I know that n=1 studies are of limited value, but that’s mine.

  2. E.M.Smith says:

    @AndySaurus:

    You are a 100% Sample of YOU. Just as I am a 100% sample of me. I’m 100% success at dodging Covid via “Cattle Drench Ivrmectin” or perhaps just some incredibly magical personal resistance. Whatever…

    I’m increasingly of the opinion that the “Covid Cure” is way way worse than the disease, and that is the goal. To herd us into something else. So I’m not going to be herded.

  3. andysaurus says:

    Thanks @E.M.Smith. If my coercive Australian Government had not banned the use of Ivermectin for anything other than Lupus, I would be on it too. I now see the value of the second amendment, I don’t think they would go quite that far in the U.S., although they seem to be trying everything else. Somehow I don’t think the big pharma profits are disconnected from the lockdown/mask/jab at all costs dogma. There are some politicians making a fortune from it.
    If, as I think probable, the value of prophylactic medicines has been suppressed at the cost of countless lives, then they (Government, media,big tech) should swing for it. IMHO.

  4. AC Osborn says:

    EM, those studies have been around a while now and ignored by the MSM of course.

  5. E.M.Smith says:

    @AC Osborn:

    About 3 months per the date stamp. The older ones were in mice or rats IIRC. But yeah, it all points one way and the MSM is looking anywhere but that direction.

  6. H.R. says:

    No one is going to ‘look there’ unless they are told to look there.

    The YSM is just the propaganda department for the GEBs. They receive the narrative from the bosses who own the YSM.

    I’ve seen quotes from editors and jurinalists that indicate that they believe they are ‘in charge’ and calling the shots. They are too dumb to realize they are merely useful idiots. They’d know that if they ever realized that the narratives are handed down to them and are not originated by them. Too dumb to even figure that out.

    I find it hard to believe that at least a few in the YSM haven’t figured this out. It’s plain to thinking, observant outsiders what is going on. I guess that small percentage are just a bunch of cynical sellouts.

  7. Simon Derricutt says:

    H.R – a few people get it at GB News. See https://www.gbnews.uk/gb-views/the-state-is-no-longer-working-to-serve-us-and-to-protect-our-shared-heritage-says-neil-oliver/310649 for Neil Oliver’s latest. I’m surprised he’s allowed to publish that, though.

  8. H.R. says:

    @Simon – Yah but… then they are no longer employed by the YSM.

    All the counternarrative information comes from alternative sources. All, every last one, of the legacy MSM is in the tanks for the GEBs.

  9. David A says:

    More direct evidence of vaccine harms…
    https://childrenshealthdefense.org/defender/covid-vaccines-incurable-fatal-degenerative-brain-disorder/
    and this one is rapidly fatal.

  10. another ian says:

    “Sudden Adult Death Syndrome: because young people die, just like that”

    https://joannenova.com.au/2022/06/sudden-adult-death-syndrome-because-young-people-die-just-like-that/#comment-2556036

    “Eva Vlaardingerbroek discusses the rise in sudden adult death syndrome with Mark Steyn – YouTube”

    And

    “There was an unexpected 40% increase in ‘all cause deaths’ in 2021”

  11. Trad says:

    MDPI, acronym of Multidisciplinary Digital Publishing Institute, is not a very well respected scientific publisher. In fact, it has had to withdraw many open access research papers due to gross errors and mistakes.Take whatever you read there with a large grain of salt.

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