Normally, Rand Paul is a clown with his war on Fauci, always for the wrong reasons mind you.  Generally, Rand Paul despite his medical background, sounds like a moron when talking science.

Rand Paul runs to Wuhan instead of a dozen American-funded labs, some here, some in Georgia, Romania, even Libya, that have Level 4 capabilities and USAID/CIA funding as we prove below.

Paul is now running the coverup.

Today he is reading something he didn’t write, something given him by his masters, and those masters, it now seems beyond any doubt, are using COVID 19 to destroy the US.



We believe Rand Paul is very much a part of it.

Watch Rand Paul lie, over and over and over, but carefully, reading a skillfully crafted disinformation packet that is filled with half-truths and utter lies.

The public is supposed to be too stupid to understand.

VT had the story right from the beginning.  Now that the CIA is officially tasked with finding a source for COVID, someone to blame, the vast American program of bio-warfare defense which creates every nasty virus we have seen, probably ebola, certainly swine flu, and then “accidentally” release them in such a way that the CIA’s agenda is served.

Since this article was written, we have begun to look at the worldwide operations of US nuclear/bio/chem contractor, Kushner-Trump’s favorite, Battelle, and their secret labs around the world.

When we began, our people started to be threatened.  That was a serious mistake.

Submit this paper to any physician or other qualified bio-sciences specialist.  See what they say.

TO SHARE THIS ARTICLE ON FACEBOOK, PLEASE COPY and PASTE this link:  https://www.veteranstodaynetwork.com/2020/04/29/documentary-proof-university-of-north-carolina-generated-covid-19/

Introduction

Documents below will show that research to create COVID 19 began in the United States in 2006 and culminated in a successful bio-weapon in 2015, with work done at the University of North Carolina and at Harvard, and at the Food and Drug Administration’s lab in Arkansas.

Their work was titled:

A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence

They did this and more, so much more as you will read below.

As Trump said, over and over and over, the Chinese were involved.

Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China supplied the Wuhan Bat Virus which was used in the American study.  Their name was included for that reason only.

COVID 19 was a US Army bio-weapons project to manufacture a pneumonia-causing disease that would be nearly impossible to vaccinate for in patients over 40 years old.

The proof is here, simply scroll down.  The study was run by the University of North Carolina and funded by USAID/CIA.  It chose a Chinese bat virus and chose to include a medical facility in Wuhan as well.

Now we know why, a smokescreen of the blame for a program China had little or nothing to do with, something satanically evil and purely American.

In November 2015, a study was published outlining the capability of producing the virus we are dealing with now.  Among the many involved was a lab in Wuhan, China.  It was listed from the beginning as one of the dozens, mostly American, working on this project.

However, one key participant was left out, USAID.  It is suspected, deeply so, that USAID is a front for American bio-warfare research such as that done in Tbilisi, Georgia, and elsewhere, much documented.  This is the citation that adds USAID to the research funding group.

Change history

20 November 2015

In the version of this article initially published online, the authors omitted to acknowledge a funding source, USAID-EPT-PREDICT funding from EcoHealth Alliance, to Z.-L.S. The error has been corrected for the print, PDF and HTML versions of this article.

[ Editor’s Note: We will now present Pravda’s biased article and, below that, the actual study proving the capability of producing COVID 19, proving it is not a naturally occurring virus once and for all.

As to who did what, this is not our job but we are proving, categorically, that when a Chinese lab is mentioned, it is a minor player in an American effort, as outlined exhaustively below.

This makes discussing the Wuhan lab possibly complicit in bio-warfare.

Similarly, when Forbes Magazine and others stated they could prove COVID 19 was made naturally, and of course, they had the same access we have, we suspect that they are part of a disinformation effort tied to USAID and bio-warfare.

Suspicion is not proof.  The proof is proof and there is proof enough to drown in.  Our thanks to the American medical professionals who pimped themselves out to the US Army and CIA and who helped bring us where we are now, a nation broken to pieces…VeteransToday ]

Pravda.Ru: Such material appeared in 2015 on the website of the scientific journal Natura 2015. Then the authors claimed that after the advent of the SARS virus (2002-2003) and the Middle East respiratory syndrome (MERS), scientists were aware of the risk of interspecific transmission that would lead to an epidemic among people.

Successful lab experiment

Among other things, the research team studied bats, which are the largest incubators of coronaviruses. Nevertheless, bats could not transmit the coronavirus to humans because they could not interact with human cells with ACE2 receptors.

The material also stated that horseshoe bats carry a strain of SARS coronavirus that can be transmitted to humans. It has been named the SHC014-CoV virus.

To better study this virus, scientists copied the coronavirus and infected it with laboratory mice. The results showed that the virus is really able to bind to human cells with ACE2 receptors and multiply in the cells of the respiratory system.

In the research work, it is noted that laboratory materials, samples and equipment that were used in the research were obtained from the Army Medical Research Institute of Infectious Diseases. Although it is not yet possible to say for sure that the virus that was tested in laboratory mice is the same as the SARS-Cove-2 coronavirus.

NATO policy

However, interesting things can be found in earlier documents.

For example:

  • The 2019 Alliance’s activity report says that in 2019, the Alliance’s first place in research and development was occupied by the topic of radiochemical and biological protection (29%), shifting the seemingly most pressing problem of Europe – counterterrorism (it turned out to be 4- m priority).
  • A year earlier, in 2018, the situation was exactly the opposite: terrorism, as it should be, was in the first place (28%), and radiochemical and biological protection in the fourth (13%).

As the Brussels snitch writes in the telegram channel, “given the absence of visible reasons for such a sharp change in scientific interests, there are two options and both are unpleasant:

  • or NATO now wags the fifth point, falsifying the data to show “and we always prepared for viruses, we are modern”,
  • or even in 2019 in the alliance, God forgive me, they knew where the trouble would come from.

Yes, the first option is much more real, but, you see, the facts are surprising.

Source:  Pravda

Original 2015 Research Unedited and Complete

Published: A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence by Vineet D Menachery, Boyd L Yount Jr, Kari Debbink, Sudhakar Agnihothram, Lisa E Gralinski, Jessica A Plante, Rachel L Graham, Trevor Scobey, Xing-Yi Ge, Eric F Donaldson, Scott H Randell, Antonio Lanzavecchia, Wayne A Marasco, Zhengli-Li Shi, Ralph S Baric

Nature Medicine volume 21, pages 1508–1513 (2015)

A Corrigendum to this article was published on 06 April 2016

This article has been updated

Abstract

The emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) and the Middle East respiratory syndrome (MERS)-CoV underscores the threat of cross-species transmission events leading to outbreaks in humans. Here we examine the disease potential of a SARS-like virus, SHC014-CoV, which is currently circulating in Chinese horseshoe bat populations1. Using the SARS-CoV reverse genetics system2, we generated and characterized a chimeric virus expressing the spike of bat coronavirus SHC014 in a mouse-adapted SARS-CoV backbone.

The results indicate that group 2b viruses encoding the SHC014 spike in a wild-type backbone can efficiently use multiple orthologs of the SARS receptor human angiotensin-converting enzyme II (ACE2), replicate efficiently in primary human airway cells and achieve in vitro titers equivalent to epidemic strains of SARS-CoV. Additionally, in vivo experiments demonstrate replication of the chimeric virus in mouse lung with notable pathogenesis.

Evaluation of available SARS-based immune-therapeutic and prophylactic modalities revealed poor efficacy; both monoclonal antibody and vaccine approach failed to neutralize and protect from infection with CoVs using the novel spike protein.

On the basis of these findings, we synthetically re-derived an infectious full-length SHC014 recombinant virus and demonstrate robust viral replication both in vitro and in vivo. Our work suggests a potential risk of SARS-CoV re-emergence from viruses currently circulating in bat populations.

Main

The emergence of SARS-CoV heralded a new era in the cross-species transmission of severe respiratory illness with globalization leading to rapid spread around the world and massive economic impact3,4. Since then, several strains—including influenza A strains H5N1, H1N1 and H7N9, and MERS-CoV—have emerged from animal populations, causing considerable disease, mortality and economic hardship for the afflicted regions5. Although public health measures were able to stop the SARS-CoV outbreak4, recent metagenomics studies have identified sequences of closely related SARS-like viruses circulating in Chinese bat populations that may pose a future threat1,6.

However, sequence data alone provides minimal insights to identify and prepare for future pre-pandemic viruses. Therefore, to examine the emergence potential (that is, the potential to infect humans) of circulating bat CoVs, we built a chimeric virus encoding a novel, zoonotic CoV spike protein—from the RsSHC014-CoV sequence that was isolated from Chinese horseshoe bats1—in the context of the SARS-CoV mouse-adapted backbone. The hybrid virus allowed us to evaluate the ability of the novel spike protein to cause disease independently of other necessary adaptive mutations in its natural backbone.

Using this approach, we characterized CoV infection mediated by the SHC014 spike protein in primary human airway cells and in vivo and tested the efficacy of available immune therapeutics against SHC014-CoV. Together, the strategy translates metagenomics data to help predict and prepare for future emergent viruses.

The sequences of SHC014 and the related RsWIV1-CoV show that these CoVs are the closest relatives to the epidemic SARS-CoV strains (Fig. 1a,b); however, there are important differences in the 14 residues that bind human ACE2, the receptor for SARS-CoV, including the five that are critical for host range: Y442, L472, N479, T487 and Y491 (ref. 7).

In WIV1, three of these residues vary from the epidemic SARS-CoV Urbani strain, but they were not expected to alter binding to ACE2 (Supplementary Fig. 1a,b and Supplementary Table 1). This fact is confirmed by both pseudotyping experiments that measured the ability of lentiviruses encoding WIV1 spike proteins to enter cells expressing human ACE2 (Supplementary Fig. 1) and by in vitro replication assays of WIV1-CoV (ref. 1). In contrast, 7 of 14 ACE2-interaction residues in SHC014 are different from those in SARS-CoV, including all five residues critical for the host range (Supplementary Fig. 1c and Supplementary Table 1).

These changes, coupled with the failure of pseudotyped lentiviruses expressing the SHC014 spike to enter cells (Supplementary Fig. 1d), suggested that the SHC014 spike is unable to bind human ACE2. However, similar changes in related SARS-CoV strains had been reported to allow ACE2 binding7,8, suggesting that additional functional testing was required for verification.

Therefore, we synthesized the SHC014 spike in the context of the replication-competent, mouse-adapted SARS-CoV backbone (we hereafter refer to the chimeric CoV as SHC014-MA15) to maximize the opportunity for pathogenesis and vaccine studies in mice (Supplementary Fig. 2a). Despite predictions from both structure-based modeling and pseudotyping experiments, SHC014-MA15 was viable and replicated to high titers in Vero cells (Supplementary Fig. 2b). Similar to SARS, SHC014-MA15 also required a functional ACE2 molecule for entry and could use human, civet and bat ACE2 orthologs (Supplementary Fig. 2c,d).

To test the ability of the SHC014 spike to mediate infection of the human airway, we examined the sensitivity of the human epithelial airway cell line Calu-3 2B4 (ref. 9) to infection and found robust SHC014-MA15 replication, comparable to that of SARS-CoV Urbani (Fig. 1c).

To extend these findings, primary human airway epithelial (HAE) cultures were infected and showed robust replication of both viruses (Fig. 1d). Together, the data confirm the ability of viruses with the SHC014 spike to infect human airway cells and underscore the potential threat of cross-species transmission of SHC014-CoV.

Figure 1: SARS-like viruses replicate in human airway cells and produce in vivo pathogenesis.

figure1

(a) The full-length genome sequences of representative CoVs were aligned and phylogenetically mapped as described in the Online Methods. The scale bar represents nucleotide substitutions, with only bootstrap support above 70% being labeled. The tree shows CoVs divided into three distinct phylogenetic groups, defined as α-CoVs, β-CoVs and γ-CoVs. Classical subgroup clusters are marked as 2a, 2b, 2c and 2d for the β-CoVs and as 1a and 1b for the α-CoVs. (b) Amino acid sequences of the S1 domains of the spikes of representative β-CoVs of the 2b group, including SARS-CoV, were aligned and phylogenetically mapped. The scale bar represents the amino acid substitutions. (c,d) Viral replication of SARS-CoV Urbani (black) and SHC014-MA15 (green) after infection of Calu-3 2B4 cells (c) or well-differentiated, primary air-liquid interface HAE cell cultures (d) at a multiplicity of infection (MOI) of 0.01 for both cell types. Samples were collected at individual time points with biological replicates (n = 3) for both Calu-3 and HAE experiments. (e,f) Weight loss (n = 9 for SARS-CoV MA15; n = 16 for SHC014-MA15) (e) and viral replication in the lungs (n = 3 for SARS-CoV MA15; n = 4 for SHC014-MA15) (f) of 10-week-old BALB/c mice infected with 1 × 104 p.f.u. of mouse-adapted SARS-CoV MA15 (black) or SHC014-MA15 (green) via the intranasal (i.n.) route. (g,h) Representative images of lung sections stained for SARS-CoV N antigen from mice infected with SARS-CoV MA15 (n = 3 mice) (g) or SHC014-MA15 (n = 4 mice) (h) are shown. For each graph, the center value represents the group mean, and the error bars define the s.e.m. Scale bars, 1 mm.

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To evaluate the role of the SHC014 spike in mediating infection in vivo, we infected 10-week-old BALB/c mice with 104 plaque-forming units (p.f.u.) of either SARS-MA15 or SHC014-MA15 (Fig. 1e–h). Animals infected with SARS-MA15 experienced rapid weight loss and lethality by 4 d post-infection (d.p.i.); in contrast, SHC014-MA15 infection produced substantial weight loss (10%) but no lethality in mice (Fig. 1e). Examination of viral replication revealed nearly equivalent viral titers from the lungs of mice infected with SARS-MA15 or SHC014-MA15 (Fig. 1f). Whereas lungs from the SARS-MA15–infected mice showed robust staining in both the terminal bronchioles and the lung parenchyma 2 d.p.i. (Fig. 1g), those of SHC014-MA15–infected mice showed reduced airway antigen staining (Fig. 1h); in contrast, no deficit in antigen staining was observed in the parenchyma or in the overall histology scoring, suggesting differential infection of lung tissue for SHC014-MA15 (Supplementary Table 2).

We next analyzed infection in more susceptible, aged (12-month-old) animals. SARS-MA15–infected animals rapidly lost weight and succumbed to infection (Supplementary Fig. 3a,b). SHC014-MA15 infection-induced robust and sustained weight loss, but had minimal lethality. Trends in the histology and antigen staining patterns that we observed in young mice were conserved in the older animals (Supplementary Table 3). We excluded the possibility that SHC014-MA15 was mediating infection through an alternative receptor on the basis of experiments using Ace2−/− mice, which did not show weight loss or antigen staining after SHC014-MA15 infection (Supplementary Fig. 4a,b and Supplementary Table 2). Together, the data indicate that viruses with the SHC014 spike are capable of inducing weight loss in mice in the context of a virulent CoV backbone.

Given the preclinical efficacy of Ebola monoclonal antibody therapies, such as ZMApp10, we next sought to determine the efficacy of SARS-CoV monoclonal antibodies against infection with SHC014-MA15. Four broadly neutralizing human monoclonal antibodies targeting SARS-CoV spike protein had been previously reported and are probable reagents for immunotherapy11,12,13. We examined the effect of these antibodies on viral replication (expressed as percentage inhibition of viral replication) and found that whereas wild-type SARS-CoV Urbani was strongly neutralized by all four antibodies at relatively low antibody concentrations (Fig. 2a–d), neutralization varied for SHC014-MA15. Fm6, an antibody generated by phage display and escape mutants11,12, achieved only background levels of inhibition of SHC014-MA15 replication (Fig. 2a). Similarly, antibodies 230.15 and 227.14, which were derived from memory B cells of SARS-CoV–infected patients13, also failed to block SHC014-MA15 replication (Fig. 2b,c). For all three antibodies, differences between the SARS and SHC014 spike amino acid sequences corresponded to direct or adjacent residue changes found in SARS-CoV escape mutants (fm6 N479R; 230.15 L443V; 227.14 K390Q/E), which probably explains the absence of the antibodies’ neutralizing activity against SHC014. Finally, monoclonal antibody 109.8 was able to achieve 50% neutralization of SHC014-MA15, but only at high concentrations (10 μg/ml) (Fig. 2d). Together, the results demonstrate that broadly neutralizing antibodies against SARS-CoV may only have marginal efficacy against emergent SARS-like CoV strains such as SHC014.

Figure 2: SARS-CoV monoclonal antibodies have marginal efficacy against SARS-like CoVs.

figure2

(a–d) Neutralization assays evaluating efficacy (measured as a reduction in the number of plaques) of a panel of monoclonal antibodies, which were all originally generated against epidemic SARS-CoV, against infection of Vero cells with SARS-CoV Urbani (black) or SHC014-MA15 (green). The antibodies tested were fm6 (n = 3 for Urbani; n = 5 for SHC014-MA15)11,12 (a), 230.15 (n = 3 for Urbani; n = 2 for SHC014-MA15) (b), 227.15 (n = 3 for Urbani; n = 5 for SHC014-MA15) (c) and 109.8 (n = 3 for Urbani; n = 2 for SHC014-MA15)13 (d). Each data point represents the group mean and error bars define the s.e.m. Note that the error bars in SARS-CoV Urbani–infected Vero cells in b,c are overlapped by the symbols and are not visible.

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To evaluate the efficacy of existing vaccines against infection with SHC014-MA15, we vaccinated aged mice with double-inactivated whole SARS-CoV (DIV). Previous work showed that DIV could neutralize and protect young mice from challenge with a homologous virus14; however, the vaccine failed to protect aged animals in which augmented immune pathology was also observed, indicating the possibility of the animals being harmed because of the vaccination15. Here we found that DIV did not provide protection from challenge with SHC014-MA15 with regards to weight loss or viral titer (Supplementary Fig. 5a,b). Consistent with a previous report with other heterologous groups 2b CoVs15, serum from DIV-vaccinated, aged mice also failed to neutralize SHC014-MA15 (Supplementary Fig. 5c).

Notably, DIV vaccination resulted in robust immune pathology (Supplementary Table 4) and eosinophilia (Supplementary Fig. 5d–f). Together, these results confirm that the DIV vaccine would not be protective against infection with SHC014 and could possibly augment disease in the aged vaccinated group.

In contrast to the vaccination of mice with DIV, the use of SHC014-MA15 as a live, attenuated vaccine showed potential cross-protection against challenge with SARS-CoV, but the results have important caveats. We infected young mice with 104 p.f.u. of SHC014-MA15 and observed them for 28 d. We then challenged the mice with SARS-MA15 at day 29 (Supplementary Fig. 6a). The prior infection of the mice with the high dose of SHC014-MA15 conferred protection against challenge with a lethal dose of SARS-MA15, although there was only a minimal SARS-CoV neutralization response from the antisera elicited 28 d after SHC014-MA15 infection (Supplementary Fig. 6b, 1:200). In the absence of a secondary antigen boost, 28 d.p.i. represents the expected peak of antibody titers and implies that there will be diminished protection against SARS-CoV over time16,17. Similar results showing protection against challenge with a lethal dose of SARS-CoV were observed in aged BALB/c mice with respect to weight loss and viral replication (Supplementary Fig. 6c,d). However, the SHC014-MA15 infection dose of 104 p.f.u. induced >10% weight loss and lethality in some aged animals (Fig. 1 and Supplementary Fig. 3). We found that vaccination with a lower dose of SHC014-MA15 (100 p.f.u.), did not induce weight loss, but it also failed to protect aged animals from a SARS-MA15 lethal dose challenge (Supplementary Fig. 6e,f). Together, the data suggest that the SHC014-MA15 challenge may confer cross-protection against SARS-CoV through conserved epitopes, but the required dose induces pathogenesis and precludes use as an attenuated vaccine.

Having established that the SHC014 spike has the ability to mediate infection of human cells and cause disease in mice, we next synthesized a full-length SHC014-CoV infectious clone based on the approach used for SARS-CoV (Fig. 3a)2. Replication in Vero cells revealed no deficit for SHC014-CoV relative to that for SARS-CoV (Fig. 3b); however, SHC014-CoV was significantly (P < 0.01) attenuated in primary HAE cultures at both 24 and 48 h after infection (Fig. 3c). In vivo infection of mice demonstrated no significant weight loss but showed reduced viral replication in lungs of full-length SHC014-CoV infection, as compared to SARS-CoV Urbani (Fig. 3d,e). Together, the results establish the viability of full-length SHC014-CoV but suggest that further adaptation is required for its replication to be equivalent to that of epidemic SARS-CoV in human respiratory cells and in mice.

Figure 3: Full-length SHC014-CoV replicates in human airways but lacks the virulence of epidemic SARS-CoV.

figure3

(a) Schematic of the SHC014-CoV molecular clone, which was synthesized as six contiguous cDNAs (designated SHC014A, SHC014B, SHC014C, SHC014D, SHC014E and SHC014F) flanked by unique BglI sites that allowed for directed assembly of the full-length cDNA expressing open reading frames (for 1a, 1b, spike, 3, envelope, matrix, 6–8 and nucleocapsid). Underlined nucleotides represent the overhang sequences formed after restriction enzyme cleavage. (b,c) Viral replication of SARS-CoV Urbani (black) or SHC014-CoV (green) after infection of Vero cells (b) or well-differentiated, primary air-liquid interface HAE cell cultures (c) at an MOI of 0.01. Samples were collected at individual time points with biological replicates (n = 3) for each group. Data represent one experiment for both Vero and HAE cells. (d,e) Weight loss (n = 3 for SARS-CoV MA15, n = 7 for SHC014-CoV; n = 6 for SARS-Urbani) (d) and viral replication in the lungs (n = 3 for SARS-Urbani and SHC014-CoV) (e) of 10-week-old BALB/c mice infected with 1 × 105 p.f.u. of SARS-CoV MA15 (gray), SHC014-CoV (green) or SARS-CoV Urbani (black) via the i.n. route. Each data point represents the group mean, and error bars define the s.e.m. **P < 0.01 and ***P < 0.001 using two-tailed Student’s t-test of individual time points.

During the SARS-CoV epidemic, links were quickly established between palm civets and the CoV strains that were detected in humans4.

Building on this finding, the common emergence paradigm argues that epidemic SARS-CoV originated as a bat virus, jumped to civets, and incorporated changes within the receptor-binding domain (RBD) to improve binding to civet Ace2 (ref. 18).

Subsequent exposure to people in live-animal markets permitted human infection with the civet strain, which, in turn, adapted to become the epidemic strain (Fig. 4a).

However, phylogenetic analysis suggests that early human SARS strains appear more closely related to bat strains than to civet strains18.

Therefore, a second paradigm argues that direct bat-human transmission initiated SARS-CoV emergence and that palm civets served as a secondary host and reservoir for continued infection (Fig. 4b)19. For both paradigms, spike adaptation in a secondary host is seen as a necessity, with most mutations expected to occur within the RBD, thereby facilitating improved infection. Both theories imply that pools of bat CoVs are limited and that host-range mutations are both random and rare, reducing the likelihood of future emergence events in humans.

Figure 4: Emergence paradigms for coronaviruses.

figure4

Coronavirus strains are maintained in quasi-species pools circulating in bat populations. (a,b) Traditional SARS-CoV emergence theories posit that host-range mutants (red circle) represent random and rare occurrences that permit infection of alternative hosts. The secondary-host paradigm (a) argues that a nonhuman host is infected by a bat progenitor virus and, through adaptation, facilitates transmission to humans; subsequent replication in humans leads to the epidemic viral strain. The direct paradigm (b) suggests that transmission occurs between bats and humans without the requirement of an intermediate host; selection then occurs in the human population with closely related viruses replicating in a secondary host, permitting continued viral persistence and adaptation in both. (c) The data from chimeric SARS-like viruses argue that the quasi-species pools maintain multiple viruses capable of infecting human cells without the need for mutations (red circles). Although adaptations in secondary or human hosts may be required for epidemic emergence, if SHC014 spike–containing viruses recombined with virulent CoV backbones (circles with green outlines), then epidemic disease may be the result in humans. Existing data support elements of all three paradigms.

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Although our study does not invalidate the other emergence routes, it does argue for a third paradigm in which circulating bat CoV pools maintain ‘poised’ spike proteins that are capable of infecting humans without mutation or adaptation (Fig. 4c). This hypothesis is illustrated by the ability of a chimeric virus containing the SHC014 spike in a SARS-CoV backbone to cause robust infection in both human airway cultures and in mice without RBD adaptation.

Coupled with the observation of previously identified pathogenic CoV backbones3,20, our results suggest that the starting materials required for SARS-like emergent strains are currently circulating in animal reservoirs. Notably, although full-length SHC014-CoV probably requires additional backbone adaption to mediate human disease, the documented high-frequency recombination events in CoV families underscores the possibility of future emergence and the need for further preparation.

To date, genomics screens of animal populations have primarily been used to identify novel viruses in outbreak settings21. The approach here extends these data sets to examine questions of viral emergence and therapeutic efficacy. We consider viruses with the SHC014 spike a potential threat owing to their ability to replicate in primary human airway cultures, the best available model for human disease. In addition, the observed pathogenesis in mice indicates a capacity for SHC014-containing viruses to cause disease in mammalian models, without RBD adaptation.

Notably, differential tropism in the lung as compared to that with SARS-MA15 and attenuation of full-length SHC014-CoV in HAE cultures relative to SARS-CoV Urbani suggest that factors beyond ACE2 binding—including spike processivity, receptor bio-availability or antagonism of the host immune responses—may contribute to the emergence. However, further testing in nonhuman primates is required to translate these findings into the pathogenic potential in humans.

Importantly, the failure of available therapeutics defines a critical need for further study and for the development of treatments. With this knowledge, surveillance programs, diagnostic reagents, and effective treatments can be produced that are protective against the emergence of group 2b–specific CoVs, such as SHC014, and these can be applied to other CoV branches that maintain similarly heterogeneous pools.

In addition to offering preparation against future emerging viruses, this approach must be considered in the context of the US government-mandated pause on gain-of-function (GOF) studies22.

On the basis of previous models of emergence (Fig. 4a,b), the creation of chimeric viruses such as SHC014-MA15 was not expected to increase pathogenicity. Although SHC014-MA15 is attenuated relative to its parental mouse-adapted SARS-CoV, similar studies examining the pathogenicity of CoVs with the wild-type Urbani spike within the MA15 backbone showed no weight loss in mice and reduced viral replication23. Thus, relative to the Urbani spike–MA15 CoV, SHC014-MA15 shows again in pathogenesis (Fig. 1).

On the basis of these findings, scientific review panels may deem similar studies building chimeric viruses based on circulating strains too risky to pursue, as increased pathogenicity in mammalian models cannot be excluded.

Coupled with restrictions on mouse-adapted strains and the development of monoclonal antibodies using escape mutants, research into CoV emergence and therapeutic efficacy may be severely limited moving forward. Together, these data and restrictions represent a crossroads of GOF research concerns; the potential to prepare for and mitigate future outbreaks must be weighed against the risk of creating more dangerous pathogens. In developing policies moving forward, it is important to consider the value of the data generated by these studies and whether these types of chimeric virus studies warrant further investigation versus the inherent risks involved.

Overall, our approach has used metagenomics data to identify a potential threat posed by the circulating bat SARS-like CoV SHC014. Because of the ability of chimeric SHC014 viruses to replicate in human airway cultures, cause pathogenesis in vivo and escape current therapeutics, there is a need for both surveillance and improved therapeutics against circulating SARS-like viruses. Our approach also unlocks the use of metagenomics data to predict viral emergence and to apply this knowledge in preparing to treat future emerging virus infections.

Methods

Viruses, cells, in vitro infection and plaque assays

Wild-type SARS-CoV (Urbani), mouse-adapted SARS-CoV (MA15) and chimeric SARS-like CoVs were cultured on Vero E6 cells (obtained from United States Army Medical Research Institute of Infectious Diseases), grown in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, CA) and 5% fetal clone serum (FCS) (Hyclone, South Logan, UT) along with antibiotic/antimycotic (Gibco, Carlsbad, CA). DBT cells (Baric laboratory, source unknown) expressing ACE2 orthologs have been previously described for both human and civet; bat Ace2 sequence was based on that from Rhinolophus leschenaulti, and DBT cells expressing bat Ace2 were established as described previously8.

Pseudotyping experiments were similar to those using an HIV-based pseudovirus, prepared as previously described10, and examined on HeLa cells (Wuhan Institute of Virology) that expressed ACE2 orthologs. HeLa cells were grown in minimal essential medium (MEM) (Gibco, CA) supplemented with 10% FCS (Gibco, CA) as previously described24.

Growth curves in Vero E6, DBT, Calu-3 2B4, and primary human airway epithelial cells were performed as previously described8,25. None of the working cell line stocks were authenticated or tested for mycoplasma recently, although the original seed stocks used to create the working stocks are free from contamination. Human lungs for HAE cultures were procured under the University of North Carolina at Chapel Hill Institutional Review Board–approved protocols. HAE cultures represent highly differentiated human airway epithelium containing ciliated and non-ciliated epithelial cells as well as goblet cells. The cultures are also grown on an air-liquid interface for several weeks before use, as previously described26.

Briefly, cells were washed with PBS and inoculated with the virus or mock-diluted in PBS for 40 min at 37 °C. After inoculation, cells were washed three times and a fresh medium was added to signify time ‘0’. Three or more biological replicates were harvested at each described time point. No blinding was used in any sample collections nor was samples randomized. All virus cultivation was performed in a biosafety level (BSL) 3 laboratory with redundant fans in the biosafety cabinets, as described previously by our group2. All personnel wore powered air-purifying respirators (Breathe Easy, 3M) with Tyvek suits, aprons, and booties and were double-gloved.

Sequence clustering and structural modeling.

The full-length genomic sequences and the amino acid sequences of the S1 domains of the spike of representative CoVs were downloaded from Genbank or Pathosystems Resource Integration Center (PATRIC), aligned with ClustalX and phylogenetically compared by using maximum likelihood estimation using 100 bootstraps or by using the PhyML package, respectively. The tree was generated using maximum likelihood with the PhyML package. The scale bar represents nucleotide substitutions. Only nodes with bootstrap support above 70% are labeled.

The tree shows that CoVs are divided into three distinct phylogenetic groups defined as α-CoVs, β-CoVs, and γ-CoVs. Classical subgroup clusters are marked as 2a, 2b, 2c, and 2d for β-CoVs, and 1a and 1b for the α-CoVs. Structural models were generated using Modeller (Max Planck Institute Bioinformatics Toolkit) to generate homology models for SHC014 and Rs3367 of the SARS RBD in complex with ACE2 based on crystal structure 2AJF (Protein Data Bank). Homology models were visualized and manipulated in MacPyMol (version 1.3).

Construction of SARS-like chimeric viruses.

Both wild-type and chimeric viruses were derived from either SARS-CoV Urbani or the corresponding mouse-adapted (SARS-CoV MA15) infectious clone (ic) as previously described27.

Plasmids containing spike sequences for SHC014 were extracted by restriction digest and ligated into the E and F plasmid of the MA15 infectious clone. The clone was designed and purchased from Bio Basic as six contiguous cDNAs using published sequences flanked by unique class II restriction endonuclease sites (BglI). Thereafter, plasmids containing wild-type, chimeric SARS-CoV, and SHC014-CoV genome fragments were amplified, excised, ligated, and purified.

In vitro transcription reactions were then performed to synthesize full-length genomic RNA, which was transfected into Vero E6 cells as previously described2. The medium from transfected cells was harvested and served as seed stocks for subsequent experiments. Chimeric and full-length viruses were confirmed by sequence analysis before use in these studies. Synthetic construction of chimeric mutant and full-length SHC014-CoV was approved by the University of North Carolina Institutional Biosafety Committee and the Dual Use Research of Concern committee.

Ethics statement

This study was carried out in accordance with the recommendations for the care and use of animals by the Office of Laboratory Animal Welfare (OLAW), NIH. The Institutional Animal Care and Use Committee (IACUC) of The University of North Carolina at Chapel Hill (UNC, Permit Number A-3410-01) approved the animal study protocol (IACUC #13-033) used in these studies.

Mice and in vivo infection

Female, 10-week-old, and 12-month-old BALB/cAnNHsD mice were ordered from Harlan Laboratories. Mouse infections were done as previously described20. Briefly, animals were brought into a BSL3 laboratory and allowed to acclimate for 1 week before infection. For infection and live-attenuated virus vaccination, mice were anesthetized with a mixture of ketamine and xylazine and infected intranasally, when challenged, with 50 μl of phosphate-buffered saline (PBS) or diluted virus with three or four mice per time point, per infection group per dose as described in the figure legends.

For individual mice, notations for infection including failure to inhale the entire dose, bubbling of inoculum from the nose, or infection through the mouth may have led to exclusion of mouse data at the discretion of the researcher; post-infection, no other pre-established exclusion or inclusion criteria are defined. No blinding was used in any animal experiments, and animals were not randomized. For vaccination, young and aged mice were vaccinated by footpad injection with a 20-μl volume of either 0.2 μg of double-inactivated SARS-CoV vaccine with alum or mock PBS; mice were then boosted with the same regimen 22 d later and challenged 21 d thereafter.

For all groups, as per protocol, animals were monitored daily for clinical signs of disease (hunching, ruffled fur, and reduced activity) for the duration of the experiment. Weight loss was monitored daily for the first 7 d, after which weight monitoring continued until the animals recovered to their initial starting weight or displayed weight gain continuously for 3 d.

All mice that lost greater than 20% of their starting body weight were ground-fed and further monitored multiple times per day as long as they were under the 20% cutoff. Mice that lost greater than 30% of their starting body weight were immediately sacrificed as per protocol. Any mouse deemed to be moribund or unlikely to recover was also humanely sacrificed at the discretion of the researcher. Euthanasia was performed using an isoflurane overdose and death was confirmed by cervical dislocation. All mouse studies were performed at the University of North Carolina (Animal Welfare Assurance #A3410-01) using protocols approved by the UNC Institutional Animal Care and Use Committee (IACUC).

Histological analysis.

The left lung was removed and submerged in 10% buffered formalin (Fisher) without inflation for 1 week. Tissues were embedded in paraffin and 5-μm sections were prepared by the UNC Lineberger Comprehensive Cancer Center histopathology core facility. To determine the extent of antigen staining, sections were stained for viral antigen using a commercially available polyclonal SARS-CoV anti-nucleocapsid antibody (Imgenex) and scored in a blinded manner by for staining of the airway and parenchyma as previously described20. Images were captured using an Olympus BX41 microscope with an Olympus DP71 camera.

Virus neutralization assays.

Plaque reduction neutralization titer assays were performed with previously characterized antibodies against SARS-CoV, as previously described11,

12,13

. Briefly, neutralizing antibodies or serum was serially diluted twofold and incubated with 100 p.f.u. of the different infectious clone, SARS-CoV strains for 1 h at 37 °C. The virus and antibodies were then added to a 6-well plate with 5 × 105 Vero E6 cells/well with multiple replicates (n ≥ 2). After a 1-h incubation at 37 °C, cells were overlaid with 3 ml of 0.8% agarose in a medium. Plates were incubated for 2 d at 37 °C, stained with neutral red for 3 h, and plaques were counted. The percentage of plaque reduction was calculated as (1 − (no. of plaques with an antibody/no. of plaques without antibody)) × 100.

Statistical analysis

All experiments were conducted contrasting two experimental groups (either two viruses, or vaccinated and unvaccinated cohorts). Therefore, significant differences in viral titer and histology scoring were determined by a two-tailed Student’s t-test at individual time points. Data were normally distributed in each group being compared and had similar variance.

Biosafety and biosecurity

Reported studies were initiated after the University of North Carolina Institutional Biosafety Committee approved the experimental protocol (Project Title: Generating infectious clones of bat SARS-like CoVs; Lab Safety Plan ID: 20145741; Schedule G ID: 12279).

These studies were initiated before the US Government Deliberative Process Research Funding Pause on Selected Gain-of-Function Research Involving Influenza, MERS, and SARS Viruses.  This paper has been reviewed by the funding agency, the NIH. Continuation of these studies was requested, and this has been approved by the NIH.

SARS-CoV is a select agent. All work for these studies was performed with approved standard operating procedures (SOPs) and safety conditions for SARS-CoV, MERs-CoV, and other related CoVs. Our institutional CoV BSL3 facilities have been designed to conform to the safety requirements that are recommended in the Biosafety in Microbiological and Biomedical Laboratories (BMBL), the US Department of Health and Human Services, the Public Health Service, the Centers for Disease Control (CDC) and the NIH. Laboratory safety plans were submitted to, and the facility has been approved for use by, the UNC Department of Environmental Health and Safety (EHS) and the CDC. Electronic card access is required for entry into the facility.

All workers have been trained by EHS to safely use powered air-purifying respirators (PAPRs), and appropriate work habits in a BSL3 facility and active medical surveillance plans are in place. Our CoV BSL3 facilities contain redundant fans, emergency power to fans and biological safety cabinets and freezers, and our facilities can accommodate SealSafe mouse racks. Materials classified as BSL3 agents consist of SARS-CoV, bat CoV precursor strains, MERS-CoV and mutants derived from these pathogens. Within the BSL3 facilities, experimentation with an infectious virus is performed in a certified Class II Biosafety Cabinet (BSC).

All members of the staff wear scrubs, Tyvek suits and aprons, PAPRs and shoe covers, and their hands are double-gloved. BSL3 users are subject to a medical surveillance plan monitored by the University Employee Occupational Health Clinic (UEOHC), which includes a yearly physical, annual influenza vaccination and mandatory reporting of any symptoms associated with CoV infection during periods when working in the BSL3. All BSL3 users are trained in exposure management and reporting protocols, are prepared to self-quarantine and have been trained for safe delivery to a local infectious disease management department in an emergency situation. All potential exposure events are reported and investigated by EHS and UEOHC, with reports filed to both the CDC and the NIH.

Accession codes

Accessions

Protein Data Bank

  • 2AJF

Change history

20 November 2015

In the version of this article initially published online, the authors omitted to acknowledge a funding source, USAID-EPT-PREDICT funding from EcoHealth Alliance, to Z.-L.S. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgments

Research in this manuscript was supported by grants from the National Institute of Allergy & Infectious Disease and the National Institute of Aging of the US National Institutes of Health (NIH) under awards U19AI109761 (R.S.B.), U19AI107810 (R.S.B.), AI085524 (W.A.M.), F32AI102561 (V.D.M.) and K99AG049092 (V.D.M.), and by the National Natural Science Foundation of China awards 81290341 (Z.-L.S.) and 31470260 (X.-Y.G.), and by USAID-EPT-PREDICT funding from EcoHealth Alliance (Z.-L.S.). Human airway epithelial cultures were supported by the National Institute of Diabetes and Digestive and Kidney Disease of the NIH under award NIH DK065988 (S.H.R.). We also thank M.T. Ferris (Dept. of Genetics, University of North Carolina) for the reviewing of statistical approaches and C.T. Tseng (Dept. of Microbiology and Immunology, University of Texas Medical Branch) for providing Calu-3 cells. Experiments with the full-length and chimeric SHC014 recombinant viruses were initiated and performed before the GOF research funding pause and have since been reviewed and approved for continued study by the NIH. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Author information

Affiliations

  1. Department of Epidemiology, the University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    • Vineet D Menachery
    • , Boyd L Yount Jr
    • , Kari Debbink
    • , Lisa E Gralinski
    • , Jessica A Plante
    • , Rachel L Graham
    • , Trevor Scobey
    • , Eric F Donaldson
    •  & Ralph S Baric
  2. Department of Microbiology and Immunology, the University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    • Kari Debbink
    •  & Ralph S Baric
  3. National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arkansas, USA
    • Sudhakar Agnihothram
  4. Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China
    • Xing-Yi Ge
    •  & Zhengli-Li Shi
  5. Department of Cell Biology and Physiology, the University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    • Scott H Randell
  6. Cystic Fibrosis Center, Marsico Lung Institute, the University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
    • Scott H Randell
  7. Institute for Research in Biomedicine, Bellinzona Institute of Microbiology, Zurich, Switzerland
    • Antonio Lanzavecchia
  8. Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA
    • Wayne A Marasco
  9. Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA
    • Wayne A Marasco

Contributions

V.D.M. designed, coordinated and performed experiments, completed the analysis and wrote the manuscript. B.L.Y. designed the infectious clone and recovered chimeric viruses; S.A. completed neutralization assays; L.E.G. helped perform mouse experiments; T.S. and J.A.P. completed mouse experiments and plaque assays; X.-Y.G. performed pseudotyping experiments; K.D. generated structural figures and predictions; E.F.D. generated phylogenetic analysis; R.L.G. completed RNA analysis; S.H.R. provided primary HAE cultures; A.L. and W.A.M. provided critical monoclonal antibody reagents; and Z.-L.S. provided SHC014 spike sequences and plasmids. R.S.B. designed experiments and wrote the manuscript.

Corresponding authors

Correspondence to Vineet D Menachery or Ralph S Baric.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary Text and Figures

Supplementary Figures 1–6 and Supplementary Tables 1–4 (PDF 4747 kb)

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Cite this article

Menachery, V., Yount, B., Debbink, K. et al. A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence. Nat Med 21, 1508–1513 (2015). https://doi.org/10.1038/nm.3985

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81 COMMENTS

  1. So many comments!

    My brainless take on Rand Paul:

    He is one in Congress who was never informed that the US used genetic biological warfare first on China in Wuhan, then again on Iran’ government.
    Trump Pompeo, Graham, etc. all were in the know and all lying about it to cover it all up, but RP by being an outsider, goes with the manufactured :explanations” coming from another angle and that’s just what they want ..a useful idiot in other words to serve master

  2. The world’s first lab-certified Covid death was America’s Lovell “Cookie” Brown. She died in Kansas on Jan. 9, 2020, according to her CDC-certified death certificate. The first Chinese Covid death occurred two days later.

    Also, Covid was endemic throughout the USA by December, 2019, by which time 4-6 million Americans had been infected: “Serologic testing of U.S. blood donations to identify SARS-CoV-2-reactive antibodies: December 2019-January 2020”. Sridhar V Basavaraju, MD, et al. https://academic.oup.com/cid/advance-article/doi/10.1093/cid/ciaa1785/6012472

    Francis Collins, M.D., Ph.D., director of the US National Institutes of Health, said that “the Coronavirus might have been spreading quietly in humans for years, or even decades, without causing a detectable outbreak”.

  3. Yup, nobody except Head Mammon Worshipper Rand caught it…

    Till DramaQueenDon’s debate meltdown.
    Then suddenly everyone was catching it.

    Jesus:
    “You can’t serve Mammon”.

    Trump’s Bible Belt Base:
    “Hold my beer!”

    They deserve to be trolled mercilessly that they brought this pestilence on themselves. (To the extent they are… Pro-life about the 651,000 deaths; which is almost nil.)

  4. The powerful evils are still ruling and ruining the world, including China and USA.

    In China, besides forcing every adult to be vaccinated, the powerful evils are requiring babies 2 months and above and children 3 years and above to be vaccinated. The reason they gave is that vaccination is a major way to fight COVID-19 virus and Delta variant though they know very well that their vaccination resulted in almost 100% infection of the vaccinated by Delta about a month ago. It seems their only reason is to throw the whole China into great danger for any variant that can break through vaccination. So evil that they only have match of their evil US “husband”!

    Yes, Delta variant is also a bioweapon bio-engineered by US devils to mainly infect the unvaccinated in USA, the opposite of China. Please see this short article for full details:

    Delta Variant, a bioweapon with sharper modifications targeting the vaccinated in China and the unvaccinated in USA?
    https://stateofthenation.co/?p=82351

    • China did not and does not require anyone to be vaccinated, despite having the world’s best Covid stats. Please don’t make up stuff and post it here.

  5. I received the third poke of the Pfizer vaccine last week because I’m being treated for a perineurial cancer behind my left eye, which was NOT caused by the first two pokes of the vaccine, for those pseudo-scientists out there who have so much to say about the bioweapon COVID-19.

  6. That COVID-19 was a bioweapon was certainly known to the Israelis in November of 2019. That’s when the Times of Israel reported that US intelligence first notified them and the Trump administration, which reportedly was “not interested.” This means that everything Trump has said about COVID-19 since then is a lie.

    Trumpsters, you’ve been duped again. The fiends who first deployed COVID-19 in China are now deploying the new Delta variant of this bioweapon in the red states where vaccination rates are low and they refuse to wear masks. That makes it a much more infectious and virulent killer!

    • We at VT are totally aware of what Fauci is withholding and how he has messed up, by trying to play political doctor and hiding very unpleasant facts.

      What Fauci is doing is treating us all like children and playing god

      What rand paul is doing is endlessly worse….

      We think he may have been involved in the planning of COVID from day one.

  9. ,,, Only if you agree that US-Doctors are stupid, in excess of 12,000 per annum are covid-deaths😨❗
    . US-CDC INFLUENZA DEATHS DATA:
    . YEAR HOSPITALIZATION DEATHS
    .
    . 2018 – 2019 490,600 34,200 ——:
    . 2017 – 2018 810,000 61,000 :
    . 2016 – 2017 500,000 38,000 :
    . 2015 – 2016 280,000 23,000 :— FULL COVID-19 RELEASE RATIO TEST – PER VIAL EFFECTIVITY
    . 2014 – 2015 591,000 51,000 :
    . 2013 – 2014 347,000 38,000 :
    . 2012 – 2013 571,000 43,000 ——:
    . 2011 – 2012 140,000 12,000 ————- NORMAL FLU DEATHS
    . 2010 – 2011 290,000 37,000 ————- CONTROLLED COVID-19 PRIMARY/PILOT TEST
    . _______
    . TOTAL 337,200 DEATHS
    . 2009 – 2010 274,304 12,469 ————- NORMAL FLU DEATHS
    . _______
    . NEW TOTAL 349,669 DEATHS
    ,,, actually it should be approx 500,000 American elderly “unexplained cause of deaths” during influenza symptoms hospitalization considering you already have COVID-19 since 2003 and possibly much earlier.🥵 🥶 😱
    .
    ,,, I’m still looking for US-CDC Burden records/data 2000-2009 at present.

    • ,,, it will be much better if VT install an edit and photo-upload function in “comment box” so I can send the US-CDC documents too🤪❗

  10. . US Patent no.: US 7,220,852 BI on May 22, 2007 – “CORONAVIRUS ISOLATED FROM HUMANS”
    Application US10/822,904 events
    2003-04-25
    Priority to US46592703P
    2004-04-12
    Application filed by Centers of Disease Control and Prevention CDC
    2004-04-12
    Priority to US10/822,904
    2007-05-22
    Application granted
    2007-05-22
    Publication of US7220852B1
    2007-06-28
    Assigned to THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY OF THE DEPARTMENT OF HEALTH AND HUMAN SERVICES, CENTERS FOR DISEASE CONTROL AND PREVENTION
    Status
    Expired – Fee Related
    2024-04-12
    Anticipated expiration
    ,,, see @ https://patents.google.com/patent/US7220852B1/en

  11. . US Patent no.: US 7,220,852 BI on May 22, 2007 – “CORONAVIRUS ISOLATED FROM HUMANS”.
    ,, time spent to create the virus, test it on “HUMANS” then applied for US patent on Apr. 25, 2003.
    ,, means US-Government got the COVID-19 even much earlier !
    . “HUMANS” with an “S” means a lot of people.
    ,,, how many American elderly did US-Government infect and killed so it can amplify the COVID-19 ?
    ,,, history will show that American govt has no qualm in killing American civilians or American War Heroes in testing it’s “Bio-weapons”.

  12. I still think Trump was neck deep in the release, received an antidote shortly after the World Military Games in Wuhan, and thus could publicly eschew wearing a mask for months and create plausible deniability with his over-the-top display of ignorance during daily COVID briefings. At this point, the highest priority should be for Trump facing federal charges, be it for corruption, racketeering, obstruction of justice, attempted election fraud, or inciting a riot on the US Capitol.

    • he and manalia officially changed their address to Florida end of sept 2019.
      When Nixon resigned, he went to florida and rehabbed Mara Lago.
      After the revolution, many loyalists fled to florida.

  16. VPN’s

    unix based applications require U to change the Box’s Clock, or time to get the VPN redirecting properly..

    https://www.doileak.com/

    Clock change is cmd or cmnd line – somethng on the order of Mk File and Arc…

    the ip check may sho germany/amsterdam.. but the pointer will B bak to USA w the time stamp from your bot.. i.e. plain jane ‘ hurrricane electric bakbone” >> tunnels require a new stopwach

    as to that quacka jibberish – looks like anybody who had a petri dish of this snot – cut it loose.. the virus appears to B in the Hep C Famile’ ( never isolated intatc) and chiral like HIV w a forward and reverse on the same strand.. i.e. reverse transcriptase…

  18. https://www.dailymail.co.uk/news/article-9240321/WHO-backs-Beijings-claim-theres-no-evidence-Covid-China-December.html
    This may be an olive branch being extended to the Biden Administration; China knowing that the SARS-CoV-2 virus did not originate in a Wuhan lab, that it was brought by US operatives participating in military games, but offering the “frozen meat” explanation, in a gesture posited as a way to repair relations now that Trump has been removed.

  22. Has anyone heard of studies on Ivermectin? Dr Pierre Kory seems to see this as a very effective covid19 total prevention and treatment for the virus. The drug is also widely used in animals. DR Pierre Kory seems to have some convincing information on Fox NewsNow and YouTube.

  23. I watched this video a couple of days ago with Dr. Pierre Kory, president of the FLCCC Alliance
    (Front Line Covid-19 Critical Care Alliance) speaking about Ivermectin and was pretty blown away by what he was saying. He is begging the authorities to look at their data and not just solely focus on new drugs. His speech raises extremely serious questions about what’s currently going on, and the motivations behind them. Thanks VT for another great article.

    https://www.youtube.com/watch?v=Tq8SXOBy-4w
    https://covid19criticalcare.com/

    • The two mRNA vaccines, by Pfizer and Moderna, apparently only target the spike of bat coronavirus SHC014, as mentioned above in the Abstract, being the spike protein in the chimeric virus created at UNC, which is the part of the SARS-CoV-2 virus that binds with the ACE2 cell receptor. If the virus can’t bind with the receptor, it can’t enter the cell and replicate. Only an antibody for the spike protein is created by the immune response.

    • Yes, All those in the study, all those who have been involved in the certification both in the companies and the several layers of govenmental approval, all of whom have had a lot of experience in doing this. When we have vaccines in the 90 to 95% effectiveness range it is late in the day to be circling back to day one like none of this is real. Will there be some allergic reactions by some? Sure. But that does not matter because allergic reactions are commone with a huge variety of triggers, but they effect a very small number of people, and hence the vaccine baby is not thrown out with the bath water.

  25. Not much has changed since last August. We still know that Trump knew about COVID-19 back in November of last year when he and his inner circle were vaccinated for it. We still know that everything he’s said about it since is total BS. He lied to us about it so his Likudnik pals could shake down the USA for trillions and trillions. This we knew back in August.

    We now know that the Trumpsters, those running around without masks and having COVID-19 parties, are part of the plan to spread this bioweapon around as much as possible. We now know that they voted for him anyway, which is the text-book definition of True-believer syndrome. He’s turned much of the Republican Party into a personality cult that will do whatever he says. This we now know.

  26. The connection between the response to COVID19, and the 2010 Rockefeller Foundation’s LOCKSTEP scenario can’t be ignored. The same with Fauci’s 2017 statement: “‘There will be a challenge to the coming administration in the arena of infectious diseases.’ Both reek of foreknowledge, and planning. If one cares to look, the trail goes all the way back to 2005.
    Here’s a link to the Rockefeller paper, to save time scroll to page 18 titled LOCKSTEP. https://archive.org/details/pdfy-tNG7MjZUicS-wiJb/page/n51/mode/2up
    And, here’s a link to a video of Fauci making his prediction, choice of words is interesting “there will be”
    https://www.msn.com/en-us/health/medical/2017-dr-fauci-predicted-a-pandemic-under-trump/vi-BB14h4x7
    And, here’s Pompeo, surrounded by Trump and Fauci telling us that we are in a live exercise:
    https://www.youtube.com/watch?v=kCmHlp97YNk

    • Joe, connecting too many dots

      the dots that do connect, the kosher nostra/deep state steals an election, trump gets in, a manufactured virus is unleashed on the US and half a million die while the nation is destroyed by donald trump and the deep state. not much dot connecting needed here, they connect themselves

  27. A very good review. You do however not rememnber tha research in Finnish West Careiia (Karelia/Karelen) by US researchers <bout jow te local "jumping sheerp might be uses to caryy pathogens intio the Soviet Union. Both Mannerheim an d Kekkonen vigourouslu prohibete such attempta, But Norwegiaian US – commandióes still did attempt. Fotunatly, hey too did recist towards the final ends.

    • Corrrectiona:
      “A very good review. You do however not rememnber the research in Finnish West Careiia (Karelia/Karelen) by US researchers <about how the local "jumping sheep might be uses to carry pathogens into the Soviet Union. Both marshal Mannerheim and PMs Svinuhuvud end (later) Kekkonen vigprously prohibeted such attempts, But Norwegiaian/ US – commandioes still did attempt it. Fortunatly, hey too did start torecist towards the final ends after theyunderstood their mission was to infest Leningrad (now Seint Petersburg ´) .. then even some ov the US henchmen revolted in disgust. This happened around 1950 plus or minus a few years. .

  28. A similar fire mishap seems to me to be main reason whyUS marines now do evecuate all their aberams tanknks from centa¨raø (or mittle) Norway veryh fast — several hundreds of them, possibly?

  29. At scool in Finalnd, Missoury and Chib´na, I leaeber that Alcocole burned at 72 dergees Celalcius, Would anyone say no to thet+

    so,: wery low heat and undetected <<<<8undetrctrd by fitr.wrrants)fire might have been the cause also.

    This is not meant o cast into suspicion nor negste what hs been posted on Vt! Just a thought.

  30. Much of what Harold says is false and was the first time he posted this. 5.59 percent death rate. The CDC doesn’t track flu deaths, check. There is no proof of any flu deaths at all. Look.

  31. The Great Wuhan Well Poisoning. Courtesy of the Baal and Malikah ‘Gators’ Lost Nation. The Death Roll. Starring Doctor “Fausty” Fauci.

  32. Harold, you know how much we love you but you are using shit science. False negatives run 30 to one over false positives based on real world experience by those who give tests and treat patients. Your pseudo facts are totally upside down and agenda driven. Current fatality/death rates in the US for those you say test positive though not really sick at all is 5.59 percent.

    What we must surmise is that they are being murdered by the nurses that test them.

    Ok, I can believe that.

  33. Well, here’s a different take on things…

    On the Occult Meaning of the Term COVID
    ‘(T)ranscribed into Hebrew and read accordingly from right to left,…
    COVID becomes DIVOC transcribed as ‫ק‬ ‫ו‬ ‫ב‬ ‫י‬ ‫ד‬ in Hebrew and it actually means something – it means possession by an evil spirit. (The word is transcribed into English as dybbuk, b and v being represented through the same Hebrew character, Bet – ב‬’

    And as for the Atom Bomb…
    ‘The date chosen for the experiment played also a major role…
    ‘The first atomic blast was programmed to coincide with Tisha B’Av, the Jewish holiday commemorating the destruction of the Temple of Solomon as well as the one of the second Jewish temple destroyed by the Romans on 70 A.D. First programmed between July 18 and 21 (in 1945 Tisha B’Av fell on July 19 th )’

    https://astutenews.com/2020/06/on-the-occult-meaning-of-the-term-covid/

    • ….someone parked the info, that divoc means : SEPARATION… which fits, the Grandparents suffered from the Separation by the ASHKENAZIS-Corporation NSDAP, with the JEWISH-STAR at the coat and at the entrance door to be picked up for the CAMPS,(today FEMA/DHS) just Moderna ” A*Beit macht frei” Work makes free” , in which they all were to rape them there as human experiments for the IG-FARBEN Cooperation of the Pharma and Chemistry& WEAPON Ind. of some big Family-Clans. Whether infants to old people. The warehouses, were biologic organic aligned, so that the test results of the TESTlabs 99% success promising products were ready for the mass-production, Good one, as bad one. What caused the mass death was not poison gas and the gassing plants, which is drilled into us, but miserable hunger, they starved to death, because at the time everything was broken down. RHEINWIESEN is the 6 MIO KEY!! They tried to destroy EVIDENCES, called HOLOCAUST, holocaustus: lat. To burn. So it happens today again, the fast pirvate commercial small business holocaustus, so that no forensic proof of the genocide with the associated products in the body can be proven, that just NOT CoVID or Corona, or Delta in all spellings for confusion and disinformation, the cause of the sudden and unexpected death was the one and only reason to keep the SCAMDEMIC-PLANDEMIC BUSINESS in the PROFIT-Zone. All nasa : hebrew: deceive, but thick unscrupulous business, at the expense of the population.”]

  34. All of these comments are from a time when most people didn’t know “the awful secret”, that Trump knew about COVID-19 back in November of last year when he and his inner circle were vaccinated for it.

    https://veteranstoday.com/2020/06/03/disclosure-the-pandemic-the-awful-secret-theyve-been-hiding-an-intel-drop

    Everything he’s said about it since is total BS. I hope Trumpsters, those who’re running around without masks and having COVID-19 parties know that you’ve been lied to by your fearless leader, a consummate con man.

  35. NOW IS THE TIME FOR CUTTING Cockroaches In Action COMPLETELY FROM USA along with HOMELAND SECURITY !!!

  36. Superb well documents article that is a must-read by members of Congress, mainstream media, investigative reporters, lawyers, and aggrieved family members who lost loved ones, indeed the medical staff who are on the front line. What we need is an independent coroner investigation into the whole thing, separate from the US government, or its agencies, independent of the UN and its agencies. Still, a professional coroner inquest by independent professionals to get the bottom of this and investigate the hidden role the CDC played in funding the continued research when the US government outlawed the continued work on the project. There must be civil and criminal liability spread around the globe representing families who lost their loved one millions of people who lost their jobs and lost their business. This must not be a white wash investigation like the JFK and the 9/11 but open professional investigations like never before.

    • Ooh, now that does look like a useful find, cheers Edward, it shall be studied with interest.

  41. Thanks very much.
    Why coronovirus is more deadly with elderly people ? Answer is in this paper… Please read with much attention around these 2 sentences, close to notes 14 and 15 :

    “however, the vaccine failed to protect aged animals in which augmented immune pathology was also observed, indicating the possibility of the animals being harmed because of the vaccination15.”
    …
    “Together, these results confirm that the DIV vaccine would not be protective against infection with SHC014 and could possibly augment disease in the aged vaccinated group.”

  42. There is some anectodal evidence coming out of the New Orleans “hot spot” that a particularly virulent and lethal strain is at work there. Although there may be a possibility that the area was “salted” with an additional strain (or perhaps sarin?) to make the effects lethal within a few days. Thank god I moved out of NOLA and went far out into a more rural environment after Katrina.

  43. Excellent work. The detail requires study. Geroge Webb/Brassballs.com mentions Dr. Sani Bavari.
    Former Head of the Army’s Medical Research Institute of infectious diseases at Ft. Detrick, Maryland and a certain spraying technology as critical elements in play here.

  45. Quite frankly, what I don’t get is this, once they created the new coronavirus that be couldn’t be stopped by any of the current Sar vaccines, why on earth didn’t they develop an antidote or a vaccine ?

  46. Nature.com which published a copy of the paper recently added this:
    Editors’ note, March 2020: We are aware that this article is being used as the basis for unverified theories that the novel coronavirus causing COVID-19 was engineered. There is no evidence that this is true; scientists believe that an animal is the most likely source of the coronavirus.
    You have to laugh as the paper itself is the proof!

  48. Highly informative article with undeniable proof. Thanks to Duff and his team. All biolabs operated by military and intel agencies should be shot down and outlawed for a better world.

  49. Now, would it be possible or plausible to find out who, if anyone, from the US Military
    Sporting team attending the World’s military sporting “olympiad” & “competing” (unsuccessfully)
    in Wuhan, during October 2019, had any connection, contact, or work to/with any US bio lab, be
    it in North Carolina or Tbilisi? They had been reportedly even housed close to the famous
    “wild animals” (selling horse shoe bats too) market. Is there any substance to a short Chinese
    announcement that the 0 patient for CV contamination was one of, or even leader of the US
    Military. “sports” team? Also, is there any lead to the “floating” suggestion that the mortality of
    the present CV, was/is in some cases, enhanced by “diluted” sarin – any testing?
    Finally, pls. look at Madona’s glorious production at Eurovision 2019, believe in May – 20 monks
    on her each side =2020? – why the female dancers were sporting facial masks? – why the blow
    of air from Madona’s mouth made dancers fall as dead? – Wake up – the final words? How
    come – who knew then?

  50. ‘Plates were incubated for 2 d at 37 °C’ Human body temp. nice of them. And ‘Human lungs were provided…’
    ‘Viruses, cells, in vitro infection and plaque assays.’ Basically they were investigating a ‘plague’ of course ..then it went live in 2019. The rest is history now.

  51. Thanks a lot, i would stringly suggest to reread this sentence :
    “Together, these results confirm that the DIV vaccine would not be protective against infection with SHC014 and **could possibly augment disease in the aged vaccinated group.**”

  52. ETHICS STATE

    Hardly any animals were harmed in the experiment, all we really wanted to harm was HUMANS.

  53. correct, it was weaponized elsewhere, Tbilisi, Georgia by Ba##### for USAID and the Department of Defense

    • we know for sure by the investigations of Dyliana Galantsieva (excuse-me for error and for little english tongue) http://www.dyliana.bg which there are a lot of evidences of development of many many many drugs and chemic biological weapons of the same pharmaceutical industry (proxi developers for pentagon and american government of the same chemic weapons Usa promised to stop to develop many years ago) that now are selling billions of doses of a gmo eugenetical products developed as the perfect weapon to make money and help the evil zionist criminal goal of a new world order and government. Evidences are many, exists a lot of documents and so many smoking guns to take in charge and putting them on trial, obviously condamning , the poena can’t be other than death in public places in many western countries (they are many, they are culprits and with all the accomplices, which are many too, included all the governments official of ALL WESTERN GOVERNMENTS which now we know are better involved and are gaining the fruits of the conspiration as well). At same level are the social medias, good censors of all the discordant voices with their narration. The trial will be public , media after having been one of the weapons used to kill millions of people. When they will be prosecuted? It’s late, for many people dead ……… and many that will die in next future

  54. The North Carolina specifically reports that using the Wuhan bat virus they created a chimera, meaning one that would infect humans, and tested its leathlity. In the Intel business we call that bio-weapons testing, done in the lab of course. And you all know what the next step is. Second, the funding by USAID was left out of the first online publication as that was a direct CIA funding tie in, but that was later remedied. And the US Army bio division provided equipment needed to conduct the study. And third, even our story has gotten zero coverage, not even one official call wanting to know more, (where typically we would point them in a particular direction, which is usually a place they don’t want to go.

  55. Wow, I don’t even know where to start with this one. But, just as our Federal and State elected officials cannot seem to refrain from commenting as if they were public health officials or researchers, neither can Gordon or VT for that matter.

    If anyone dares to read the abstract of the 2015 article, they obviously do not claim to have “created” “covid-19.” Both “created” and “covid-19” are in quotation marks to highlight the ignorant use of both terms. The virus was not “created” in North Carolina. Furthermore, the virus itself is SARS-CoV-2. Covid-19 is the disease (sickness) that results from the viral infection.

    Please, if you don’t know what you’re talking about, please don’t do armchair scientific analysis and post that as if it were actually news. I’ve been a VT reader for the better part of a decade, but this type of content is just plain ignorant.

    • “generated” is “created”….and you don’t seem to have a science background. They generated a chimera, why they got the samples from China. Your reply is time wasting and senseless.

    • Mr. Duff, I am none of those other people. Maybe some others are also using Opera with a VPN – which puts my IP address typically in Ukraine, Sweden, or the Netherlands (Ukrainian Whiskas cat food ads are often funny, but I use Royal Canin). I would have thought VT would have better capabilities of identifying people than this. But I will give you a hint – my actual Internet provider is a cooperative located in a suburb of a Big 10 university town in the Upper Midwest.

      If you want to waste a bit of time, once the travel and social distancing restrictions are lifter, I will ride my F 800 GT over to Michigan or Ohio and meet you for a beer somewhere (warning – I am quite the dull conversationalist). But where Kevin Barrett lives would be closer for me (DC Comics has the pronunciation wrong).

  57. Would it be reasonable to assume that a vaccine was also created(therefore, already exists) before the decision to release this virus?

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