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Discussion: Chinese Academy of Sciences (CAS) in Wuhan has been working with bats and coronavirus for many years - DNA manipulations, cloning....

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  • etudiant
    replied
    Originally posted by gsgs View Post
    all the genbank-known sequences are still very distant from covid , including RaTG13 .
    No signs for a lab-escape here

    They might have found covid in pangolins or bats and kept it secret, worked with it,
    maybe even manipulated the cleavage site and then it escaped.
    But that's not typical for their research, which requires publishing.

    They may have found the virus (in Yunnan ?), did some first testing, immediately recognised
    its military potential and kept everything secret while growing it, testing it further
    What possible military potential does this virus offer? It is highly infectious, but slow acting and only moderately lethal. I struggle to envision a practical military use for it.

    It seems more likely that this was just another research effort to try to understand these SARS related bat viruses. We know the scientists have at times been very blase about their study subjects, even carrying deadly germs in their airplane luggage, so the possibility of a slip up at a facility is quite real.
    Separately, I'd thought the virus was well over 90% identical to an earlier horseshoe bat carried specimen, but with some new sequences included which make it so dangerous. There does not yet appear to be a consensus as to where those new sequences come from.

    Leave a comment:


  • sharon sanders
    replied
    Originally posted by sharon sanders View Post
    hat tip Shiloh


    Apparently pulled from researchgate but preserved by Wayback Machine.

    https://web.archive.org/web/20200214...oV_coronavirus





    originsofcov.PNG

    Full paper here:

    Preprint Paper: The possible origins of 2019-nCov coronavirus

    Leave a comment:


  • gsgs
    replied
    all the genbank-known sequences are still very distant from covid , including RaTG13 .
    No signs for a lab-escape here

    They might have found covid in pangolins or bats and kept it secret, worked with it,
    maybe even manipulated the cleavage site and then it escaped.
    But that's not typical for their research, which requires publishing.

    They may have found the virus (in Yunnan ?), did some first testing, immediately recognised
    its military potential and kept everything secret while growing it, testing it further

    Leave a comment:


  • Shiloh
    replied
    Source: https://www.voachinese.com/a/5290767.html

    Hong Kong Fengyun (February 16, 2020) Wuhan P4 laboratory confirmed "poison"? Xi Jinping pushes "Biosecurity Law"
    Embed share


    Hong Kong Fengyun (February 16, 2020) Wuhan P4 laboratory confirmed "poison"? Xi Jinping pushes "Biosecurity Law"
    share it


    Washington —

    As Wuhan's pneumonia epidemic raged, Xi Jinping made a special mention of accelerating the promotion of biosafety legislation and incorporating biosafety into the national security system when he addressed a meeting of the Central Committee of Comprehensive and Deepening Reform on February 14, attracting the attention of all circles. New crown virus may come from the laboratory? Why did Xi Jiping push for the biosafety law at this time? Hong Kong Fengyun invited Su Yiren, former director of the Taiwan Bureau of Disease Management and lecturer of the Department of Technology of Nantai University of Science and Technology to analyze.

    P4 laboratory needs careful management

    Su Yiren: The P4 laboratory is a very important facility in the past in the production of vaccines or animal transmission models, or even in bioterrorism. So every country in this place is very careful to do management or legislation. According to the materials published by the professors of South China University of Technology, I am also very surprised. They have done a lot of bat experiments. The CDC, the Virus Research Institute, or the P4 laboratory in Wuhan are doing advanced work in several places. Experiment, if it is useful to the P4 laboratory, this place is the point we have been concerned about. In particular, some researchers were sprayed with blood or urine from bats. This part is actually the most important point for the P4 laboratory when doing some new viruses. It is the most important in the human protection or software supporting facilities of the P4 laboratory. of.

    Wuhan P4 laboratory's safety establishment does not meet regulations

    Su Yiren: The establishment of a P4 laboratory requires very high standards and requires national level management. It's not just hardware equipment. Generally speaking, there is a certain standard for the living environment. How many kilometers away can't be close to the people. Therefore, the distance between the South China Seafood Market and Wuhan P4 Lab is 280 meters. I was surprised when I heard this. I should not set up a P4 Lab at this distance. I don't know the background of the original establishment, but this is not in accordance with the regulations.

    Leave a comment:


  • sharon sanders
    replied
    hat tip Shiloh


    Apparently pulled from researchgate but preserved by Wayback Machine.

    https://web.archive.org/web/20200214...oV_coronavirus





    originsofcov.PNG

    Leave a comment:


  • JJackson
    replied
    I had a quick look at Pangolins a few weeks ago as an intermediary host. They are genetically on a branch with cats and dogs and they have ACE2 receptors which are a close match, but ACE2 is a common to many species. However they are critically endangered with very small population size, they are secretive by nature and like a wooded hill side habitat. The main population is in the south and well away from the epicenter. While possible this makes it poor potential host, something more common, even if seldom infected, would seem more likely.
    Last edited by JJackson; February 16, 2020, 06:39 PM.

    Leave a comment:


  • sharon sanders
    replied
    Also please see:

    nCoV-2019 Spike Protein Receptor Binding Domain Shares High Amino Acid Identity With a Coronavirus Recovered from a Pangolin Viral Metagenomic Dataset

    jan 30, 2020

    An outbreak of respiratory illness caused by a novel coronavirus (nCoV-2019, NC_045512.2 52) first identified in Wuhan China has resulted in over seven thousand confirmed cases.


    https://flutrackers.com/forum/forum/...emains-unclear


    Wondering if people can transmit SARS-like coronaviruses to pangolins? Supposedly numerous wild life was traded at the Wuhan Seafood market. Is pangolin the reservoir for COVID-19 or are humans?

    I am not a virologist but it seems that if humans can transmit disease to dogs then why not other mammals? We do not know how long COVID-19 was "in the making" in China. It may have been months - circulating inefficiently - stops and starts - until mutated through enough generations to become efficient human-to-human.

    Looking forward to the tribunal on this.


    Leave a comment:


  • sharon sanders
    replied
    From their own lab which they had not submitted before.

    Also please see this post. link


    Bat coronavirus isolate RaTG13, complete genome


    GenBank: MN996532.1

    FASTA Graphics


    Go to:
    LOCUS MN996532 29855 bp RNA linear VRL 10-FEB-2020

    DEFINITION Bat coronavirus isolate RaTG13, complete genome.

    ACCESSION MN996532 VERSION MN996532.1 KEYWORDS . SOURCE Bat coronavirus ORGANISM Bat coronavirus Viruses; Riboviria; Nidovirales; Cornidovirineae; Coronaviridae; Coronavirinae; unclassified Coronavirinae. REFERENCE 1 (bases 1 to 29855)

    AUTHORS Zhu,Y., Yu,P., Li,B., Hu,B., Si,H.R., Yang,X.L., Zhou,P. and Shi,Z.L.

    TITLE Direct Submission JOURNAL Submitted (27-JAN-2020) CAS Key Laboratory of Special Pathogens, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, No. 44 Xiao Hong Shan, Wuhan, Hubei 430071, China

    COMMENT ##Assembly-Data-START## Assembly Method :: CLC Genomics Workbench v. 12.0 Sequencing Technology :: Illumina ##Assembly-Data-END## https://www.ncbi.nlm.nih.gov/nuccore/1802633852

    Leave a comment:


  • sharon sanders
    replied
    Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses


    Lei-Ping Zeng1 , Xing-Yi Ge 1 , Cheng Peng1 , Wanbo Tai 2 , Shibo Jiang2,3 , Lanying Du2* & Zheng-Li Shi 1* 1 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China; 2 Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY 10021, USA; 3 Key Laboratory of Medical Molecular Virology of MOE/MOH, School of Basic Medical Sciences, Fudan University, Shanghai 200433, China

    Received July 9, 2017; accepted September 16, 2017; published online November 10, 2017 Citation: Zeng, L.P., Ge, X.Y., Peng, C., Tai, W., Jiang, S., Du, L., and Shi, Z.L. (2017).

    Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses. Sci China Life Sci 60, 1399–1402. https://doi.org/10.1007/s11427-017-9189-3


    Dear Editor, The 2002–2003 global pandemic caused by severe acute respiratory syndrome coronavirus (SARS-CoV) infected around 8,000 people with 10% mortality (http://www.who.int/csr/sars/en/). The virus has a positive-stranded RNA genome that encodes a large polyprotein (1a and 1ab), four structural proteins, including spike (S), small envelop (E), membrane (M), and nucleocapsid (N), as well as several accessory proteins (Hu et al., 2015). The S protein plays a key role in cellular entry and is functionally divided into two subunits: S1 at the N-terminal end responsible for cell attachment and S2 at the C-terminal end responsible for membrane fusion (Jiang et al., 2005). Angiotensin I converting enzyme 2 (ACE2) was identified as a cellular receptor of SARS-CoV (Li et al., 2003). A fragment (residues 318-520) of the SARS-CoV S1 subunit was identified as the receptor-binding domain (RBD) that plays a key role in binding ACE2 and, hence, mediating virus entry (Wong et al., 2004). Therefore, RBD in the S1 subunit of S protein contains the major neutralizing epitopes for inducing neutralizing antibodies, thus serving as an *Corresponding authors (Lanying Du, email: ldu@nybc.org; Zheng-Li Shi, email: zlshi@wh.iov.cn) important target for developing immunotherapeutics and vaccines (Jiang et al., 2005).

    SARS-CoV is considered to be an emerging zoonotic pathogen crossing species barriers to infect humans (Hu et al., 2015). Since the first outbreak, genetically diverse SARS-like coronaviruses (SL-CoVs) have been discovered in horseshoe bats worldwide (Hu et al., 2015). These bat viruses are highly similar to SARS-CoV in most gene encoding regions, but they display wide diversity in S proteins, particularly at their RBDs. Based on the alignment of the RBD sequences of SL-CoVs with those of SARS-CoV, SL-CoVs can be classified into two clades (Figure S1 in Supporting Information). Clade one includes WIV1 and SHC014, which are identical in size and share the same receptor as SARS-CoV, and clade two, as represented by Rp3, has deletions in the RBD and does not use ACE2 (Hu et al., 2015; Zhou et al., 2013). WIV1 and SHC014 both use the ACE2 receptor, but their respective RBDs are highly variable in that the RBD of W1V1 is very similar to that of SARS-CoV, while the SHC014 RBD is much more variable, suggesting that their neutralizing epitopes are different. Therefore, in this study, we tested the cross-neutralization activity of SARS-CoV RBD-specific antibodies after in vitro infection by these two bat SL-CoV strains.

    The SL-CoV strains were propagated in Vero E6 cells ? Science China Press and Springer-Verlag Berlin Heidelberg 2017 life.scichina.com link.springer.com Figure 1 Cross-neutralization activity of anti-SARS-CoV RBD antibodies against infection of bat SL-CoV strain WIV1 and SHC014. The mAbs (A) and mouse sera (pAbs) (B) were diluted as indicated in the figure. The neutralization activities were measured by plaque reduction neutralization assay. NC, negative control. (ATCC). Four mouse monoclonal antibodies (mAbs) (ID number: 19B2, 10E7, 13B6, and 121D7) and four mouse polyclonal antibodies (pAbs) (ID number: 433, 435, 691, and 692) against SARS-CoV RBD were produced in the Viral Immunology Laboratory, Lindsley F. Kimball Research Institute, New York Blood Center. The mAbs were generated by immunization of BALB/c mice with recombinant SARS-CoV RBD protein containing a C-terminal Fc (RBD-Fc), and purified from culture supernatants of stable hybridoma cell lines using protein A-Sepharose 4 Fast Flow (He et al., 2005). The pAbs were generated by immunization of BALB/c mice with SARS-CoV RBD-Fc (for 433 and 435) or RBD protein containing a C-terminal His6 (RBD-His) (for 691 and 692), and collection of respective sera at 10 days after the 3rd dose. These mouse mAbs and pAbs demonstrated potent neutralization activity against infection of pseudotyped SARS-CoV that expresses the S protein of SARS-CoV (Figure S2A and B in Supporting Information). A human mAb, m396, previously isolated from a large antibody Fab library and shown to potently 1400 Zeng, L.P., et al. Sci China Life Sci December (2017) Vol. 60 No. 12 cross-neutralize SARS-CoV isolates (Zhu et al., 2007), was kindly provided by Dr. Dimiter S. Dimitrov (Cancer and Inflammation Program, NCI, NIH, USA).

    Since SHC014 could not be successfully isolated, a recombinant virus (rWIV1-SHC014S) was constructed based on the WIV1 backbone with the replacement of SHC014S gene, as described previously (Zeng et al., 2016). The S sequence of SHC014 was amplified with primer pair (F-SHC014-Bsa I, 5′-AGTGGTCTCAACGAACATGAAATTGTTAGTTTTAGTTTTTGCTAC-3′ and R-SHC014-Bsa I, 5′-TCAGGTCTCAGTTCGTTTATGTGTAATGTAATTTGACACCCTTG-3′), digested with Bsa I, and inserted into an artificial bacterial chromosome along with the other viral cDNA fragments.

    Vero E6 cells were plated at 1.5?105  cells well −1 in 24-well tissue culture plates and grown overnight. The aforementioned mAbs and mouse pAbs (antisera) were diluted in DMEM starting at a ratio of 1:20. Serial 2-fold dilutions were mixed with an equal volume of 50 plaque-forming units (PFU) of SL-CoV WIV1 or rWIV1-SHC014S, and incubated at 37?C for 1 h. Serum from a C57BL/6 na?ve mouse was used as negative control. The mixture was added to monolayers of Vero E6 cells, incubated at 37?C for 1 h, and then changed to a semi-solid medium (DMEM containing 1% methylcellulose and 2% FBS). Plaques were counted at 4–5 days post-infection. The neutralization titers of the antibodies were calculated.

    With the exception of mAb 121D7, which had only marginal neutralization activity against WIV1 infection, all tested mAbs, including 19B2, 10E7, 13B6, and m396, had NT50 (50% neutralization titer) less than 1.25 μg mL−1 against WIV1 infection. However, none of these mAbs showed sufficiently high neutralization activity against rWIV1-SHC014S infection, with only ~20% of rWIV1-SHC014S virus neutralized at most antibody concentrations (Figure 1A). While all four mouse antisera (pAbs 433, 435, 691, and 692) had only low neutralization activity against rWIV1-SHC014 infection, they all demonstrated neutralization >90% of WIV1 virus at a dilution of 1:320 (Figure 1B and Figure S3 in Supporting Information).


    In summary, our results have demonstrated that most SARS-CoV RBD-specific antibodies tested in this study could cross-neutralize SL-CoV strain WIV1, but not SHC014. While SARS-CoV and WIV1 have comparable RBD, the RBD of SHC014 is much more variable, as previously noted (Figure S1 in Supporting Information). More specifically, the RBD of SHC014 has a difference of 24 amino acids (aa) compared to that of SARS-CoV, while the RBD of WIV1 only has a difference of 8 aa. This may explain why SHC014 could not be cross-neutralized effectively by most antibodies (mAbs or pAbs) targeting SARS-CoV RBD. The fact that SHC014 retains its ability to infect human cells implies that the available antibodies and vaccines based on SARS-CoV RBD will not protect the next SARS-like disease caused by bat SL-CoV SHC014 strain.

    Thus, further development of effective vaccines and treatments for potential infection by the SL-CoV strains, as represented by SHC014, is urgently needed.


    Compliance and ethics The author(s) declare that they have no conflict of interest. Acknowledgements This work was supported by the National Natural Science Foundation of China (81290341, 31621061 to Zheng-Li Shi), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDPB0301 to Zheng-Li Shi), National Institute of Allergy and Infectious Diseases of National Institutes of Health grant (R01AI110964 to Zheng-Li Shi), the National Key Research and Development Program of China (2016YFC1201000 to Shibo Jiang) and NIH grant (R01AI098775 to Shibo Jiang and Lanying Du). He, Y., Lu, H., Siddiqui, P., Zhou, Y., and Jiang, S. (2005). Receptor-binding domain of severe acute respiratory syndrome coronavirus spike protein contains multiple conformation-dependent epitopes that induce highly potent neutralizing antibodies. J Immunol 174, 4908–4915. Hu, B., Ge, X., Wang, L.F., and Shi, Z. (2015). Bat origin of human coronaviruses. Virol J 12, 221. Jiang, S., He, Y., and Liu, S. (2005). SARS vaccine development. Emerg Infect Dis 11, 1016–1020. Li, W., Moore, M.J., Vasilieva, N., Sui, J., Wong, S.K., Berne, M.A., Somasundaran, M., Sullivan, J.L., Luzuriaga, K., Greenough, T.C., Choe, H., and Farzan, M. (2003). Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 426, 450–454. Wong, S.K., Li, W., Moore, M.J., Choe, H., and Farzan, M. (2004). A 193- amino acid fragment of the SARS coronavirus S protein efficiently binds angiotensin-converting enzyme 2. J Biol Chem 279, 3197–3201. Zeng, L.P., Gao, Y.T., Ge, X.Y., Zhang, Q., Peng, C., Yang, X.L., Tan, B., Chen, J., Chmura, A.A., Daszak, P., and Shi, Z.L. (2016). Bat severe acute respiratory syndrome-like coronavirus WIV1 encodes an extra accessory protein, ORFX, involved in modulation of the host immune response. J Virol 90, 6573–6582. Zhou, P., Han, Z., Wang, L.F., and Shi, Z. (2013). Identification of immunogenic determinants of the spike protein of SARS-like coronavirus. Virol Sin 28, 92–96. Zhu, Z., Chakraborti, S., He, Y., Roberts, A., Sheahan, T., Xiao, X., Hensley, L.E., Prabakaran, P., Rockx, B., Sidorov, I.A., Corti, D., Vogel, L., Feng, Y., Kim, J.O., Wang, L.F., Baric, R., Lanzavecchia, A., Curtis, K.M., Nabel, G.J., Subbarao, K., Jiang, S., and Dimitrov, D.S. (2007). Potent cross-reactive neutralization of SARS coronavirus isolates by human monoclonal antibodies. Proc Natl Acad Sci USA 104, 12123–12128. Zeng, L.P., et al. Sci China Life Sci December (2017) Vol. 60 No. 12 1401 SUPPORTING INFORMATION Figure S1 Alignment of the RBDs of SARS-CoV Tor2 strain and bat SL-CoV strains WIV1, SHC014, and Rp3. Figure S2 Neutralization activity of anti-SARS-CoV RBD antibodies against infection of pseudotyped SARS-CoV. Figure S3 Plaque neutralization of SL-CoV WIV1 and SHC014 strains by anti-SARS-CoV RBD sera. The supporting information is available online at http://life.scichina.com and https://link.springer.com. The supporting materials are published as submitted, without typesetting or editing. The responsibility for scientific accuracy and content remains entirely with the authors.


    https://link.springer.com/content/pd...017-9189-3.pdf

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  • Pathfinder
    replied
    This article was written in 2017:

    Translation Google



    13 years of unremitting pursuit of Chinese scientists to find the source of SARS virus

    CCTV News Client CCTV News Client December 29, 2017 14:06A-A +

    In 2002, the SARS epidemic was first discovered in Guangdong, China, and gradually spread to Southeast Asia and the world, triggering a global epidemic of infectious diseases. Fifteen years have passed since the first reported case of SARS in December 2002. Where is the source of the SARS virus? Recently, a scientific research result attracted the attention of the scientific community: experts from the Wuhan Institute of Virology, Chinese Academy of Sciences, found a natural gene bank of bat SARS coronavirus in Yunnan, China, and research revealed the possible recombination origin of SARS virus.

    The SARS epidemic broke out in 2002. Two years later, that is, from 2004, the research team began to search for the source of the epidemic. This finding was 13 years. How was the host of the virus source locked on the bat? Finding a possible recombination origin of the SARS virus does it mean that the source of the SARS virus has been found? Today's story, we approached this scientific research team to reveal the secret of the source of the SARS virus and explore the scientific process behind the search for the "SARS" virus source.

    Atypical pneumonia SARS, or Severe Acute Respiratory Syndrome, has been published by the WHO. From the end of 2002 to August 2003, SARS affected 32 countries and regions. There were 8422 infections and 916 deaths worldwide, with an average mortality rate of 10.8%. When the deadly epidemic finally subsided in 2004, research into the origin of the unknown virus began immediately.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences, and her new virus research team, also started the research on the source of SARS virus at this time.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology of the Chinese Academy of Sciences, will ask if there is a relative, saying that SARS is gone, what's the point of doing this thing? It ’s ridiculous, it ’s possible (disease) that it will never come, but I think we do a lot of work, and if one of your jobs can prevent the outbreak of disease, I think we are very meaningful.

    Soon, researchers detected SARS coronavirus from civets on the wildlife market. However, further experiments found that although the civet was a direct source of infection, it did not seem to be the "originator".

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: Is the most direct source of infection the most fundamental source? This is not necessarily the case. In order to find the source of the virus, we need to find its natural host. Only by finding its source can we fundamentally prevent this disease.

    Assistant researcher Hu Ben is responsible for the genetic evolution of the virus. The natural host he mentioned must carry the virus for a long period of time without causing disease. In addition, under natural conditions, these natural host animals must have a certain group infection rate. However, investigations of farms and wild civets turned out to be surprising.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: Both wild civet and farm civet are not infected with SARS virus, and if you artificially infect the civet with this SARS virus in the laboratory, it will also get sick It also shows symptoms. This shows that the civet cat does not meet such a characteristic of the natural host of the SARS virus. (Civet) is more of just playing the role of an intermediate host, and the true root cause is something else.

    So where did these deadly viruses originate? Who is its natural host? Some experts put forward a bold idea.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: Which animal should I look for if I look for the next animal? Some experts have suggested that we should look for bats.

    For virus researchers, bats have a special status. Among mammals, it is the second largest group after rodents. Its species account for 20% of mammal species, and it is widely distributed throughout the world. Bats are also natural hosts for many viruses, including Ebola virus, Marburg virus, rabies virus, Hendra virus, and Nipah virus. Due to the bat's special immune system, it is rare for it to carry the virus. In the long evolutionary process, bats have become natural hosts for hundreds of viruses.

    In the 1990s, there were two serious zoonotic virus infectious diseases, which broke out in Australia and Southeast Asia, respectively. The root cause came from the same animal-bats. This raises the possibility for the research team to track the source of SARS virus, and whether the source of SARS is also bats.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: The first sample was taken from Guangxi. At that time, our experts were advised that the work they did was another virus. We called Nipah and Hendra, a virus that was also transmitted by bats. At that time, the virus was mainly found in fruit bats, so We basically picked fruit bats. After we took the samples back, we did a genetic test. After 8 months of testing, nothing was found.

    Stumbled across the team's hopes

    Eight months of hard work has yielded nothing. After all, there are more than 1,300 bats in the world, and more than 130 in China alone. Fruit bats, are there other bats? How to find it? It's a haystack.

    At that time, the entire team was very frustrated, and Shi Zhengli was almost giving up, but just at this juncture, an accidental discovery made them hope again.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: The neighbor ’s research group was doing SARS virus detection in the early stage. They have some kits that can detect antibodies to SARS virus. We also took sera from bats at the time and found that there were Several (bat) species do have antibodies to SARS virus in their sera.

    By detecting 408 bats from different families and monitoring antibodies and nucleic acids, the targets became clearer. Finally, a coronavirus similar to the SARS virus was found in bats. Daisy-headed bats are named for their complex horseshoe-shaped nasal leaves and are important hosts for many animal-derived viruses such as rabies. The discovery was published in the scientific journal of the international authoritative academic journal of the year, which aroused many concerns. But soon, doubts came again.

    Shi Zhengli, Wuhan Institute of Virology, Chinese Academy of Sciences: Many experts have questioned that you still detected similar to it (SARS virus), but did not find a direct source,

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: Although it is similar to SARS virus and belongs to a family, it has a large difference in many genes in the genome, such as the S gene.

    The "S gene" is also called spike protein gene, which is responsible for the binding of the virus to the receptor. When the virus infects the cell, it must first bind to the receptor on the cell surface to invade the cell and cause the infection.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: It ’s like the relationship between the key and the lock. What key unlocks what lock? If the S genes are different, the virus may use different receptors. We cannot infect humans and civet cats like the SARS virus, so we have to keep looking.

    Shi Zhengli, Researcher, Wuhan Institute of Virology, Chinese Academy of Sciences: Guizhou, Guangxi, Guangdong, Hubei, Hunan, Henan, Henan, running around, wherever there is a bat cave , we go

    Because SARS is a highly pathogenic and potent virus, after 2005, as the epidemic went away, there were fewer domestic researchers. But Shi Zhengli and her team did not stop tracking the source of the virus. They went southwest, south, and middle of China to find bat virus samples across the country. The farthest people have been to Motu, Tibet, Xishuangbanna, Yunnan, old forests in deep mountains, and bat caves in the wilderness.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: where there are bats, one is not easy to find. Second, it has a relatively long road and a steep terrain. Sometimes there is no road.

    Luo Dongsheng, Ph.D. student, Wuhan Institute of Virology, Chinese Academy of Sciences: when catching bats in Guangdong, the water was deep in the cave. When I went in, my shoes didn't take off, and the water was almost at the waist. Net, you can hit the net, there is a hole that needs to be pushed in, you have to go in one or two hundred meters.

    Bats are mostly in dark, humid, inaccessible caves, walking through deep mountains and dense forests, and often encounter some unknown dangers.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: We still made sufficient preparations. We put on socks and tied the socks to our knees, but that did n’t work. We walked along the road, and we found that there was blood in the bathtub Only then did I know that the moth has entered the body.

    Luo Dongsheng, Ph.D. student, Wuhan Institute of Virology, Chinese Academy of Sciences: the most thrilling trip I took was to slide down the top of a cave next to the Yarlung Zangbo River Grand Canyon. Below is the waterfall. What I thought at the time was that I would not swim. It's over in the water.

    The bats appear day and night, and in the evening, researchers will enter the bat cave fully or capture the bats at the mouth of the cave. After returning, they will take a virus sample on the workbench temporarily set up in the wild. Despite wearing gloves, the risk of being bitten by a bat remains.

    Cui Jie, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: You can bite your hand through gloves, just like a needle.

    Can be bitten by bats before being vaccinated

    Bats carry a variety of severe viruses, including rabies. For this reason, before each field sampling, the team members will be vaccinated against rabies in advance. Wherever they go, they insist on non-invasive sampling. Minimize damage to bats and habitats.

    Yunnan Xiaoshan Cave finds "key" for SARS virus

    The samples were taken back to the laboratory and stored in a freezer at minus 80 ? C. Until 2011, a SARS-like coronavirus S gene closer to the SARS virus was first detected in a bat cave in Yunnan. In 2013, the Laboratory of Wuhan Institute of Virology, Chinese Academy of Sciences, isolated the first live virus of bat SARS-like coronavirus from the sample, and a more similar S gene, allowing this virus to use the same receptor as SARS virus and to Infect human cells. It was named "WIV1" by the English abbreviation of Wuhan Virus Research Institute to highlight the important value of this discovery. The results were published in the November 2013 issue of Nature. The "key" was finally found. But the secret uncovered by this small cave is far more than researchers expected.

    Significance of SARS virus traceability and disease control

    The successful isolation of WIV1 has allowed global scientists to diverge and debate the origin of the SARS virus. The target of this multi-year hunt became clearer-the SARS coronavirus originated from the chrysanthemum.

    After years of hard work, the source of SARS has finally been traced, but scientists have not stopped, because in the bat cave where the S gene was first discovered, there are still more secrets hidden.

    According to Hu Ben, there was an information gap left in the entire evidence chain at that time.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: In fact, the question about the bat origin of SARS virus has been answered almost, but because it still has some genes that are different from SARS virus, so it is still alive. Some questions are exactly how the SARS virus appeared in this bat.

    The sampling of this bat cave in Yunnan continued for another 5 years. From these samples, the research team isolated 3 live viruses in succession, and obtained the full-length genomic sequences of a total of 15 bat SARS-like coronaviruses. Surprisingly, the 15 strains contained all the genome components of the SARS virus.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: We found a gene library of SARS virus in this place, that is, the different genes that make up these parts of the SARS genome can be carried by these bats in this hole. Found on the genome.

    Hu Ben imported the viral genome sequence found in the bat cave of Yunnan and the genome sequence of SARFS virus into the gene recombination analysis software. As a result, it was found that the genome sequences of several strains of viruses after recombination had caused atypical pneumonia. The gene sequence of SARS virus is highly consistent.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: The fuchsia line and the dark blue line represent the two bat SARS-like coronaviruses that we found here, so the section in front of the SARS virus genome It is highly similar to the fuchsia bat virus gene. In the middle, it appeared a cross, and it became highly homologous with the dark blue bat SARS-like virus. We further speculate that the SARS virus might have been formed by recombination between these bat-like SARS-like coronaviruses that year.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: All SARS-like coronaviruses and various types of SARS-like coronaviruses were found here. And there are some viruses inside, and some viruses have potential for cross-species infection.

    By comparison, the highest similarity between SARS-like coronaviruses found in bat caves and their respective genes is above 97%, which is highly homologous. Genetically, this means that the most direct ancestor of the SARS virus came from these bat viruses.

    Experts said that although no virus exactly the same as SARS virus was found, the discovery fully confirmed that SARS virus originated in bats and revealed its possible way of production-genetic recombination.

    Strengthen prevention and control to reduce invasion of wild animals

    After 13 years of virus tracing, the origin of the SARS virus was finally found. But researchers also warned that new SARS-like viruses could emerge in the future.

    In addition to increasing vigilance, reducing the invasion of wild animals and their living environment, and preventing market transactions of wild animals, we can effectively prevent the spread of new infectious viruses.

    Shi Zhengli told us that there is a risk of SARS-like virus outbreaks in the future, but the study of virus traceability has found a direction for the prevention of new infectious diseases.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, the Chinese Academy of Sciences: In fact, you don't need to panic. There are actually many pathogens that wild animals carry in nature, but the chance of infecting people is very, very small, and it will not come to actively infect people. We now often see a new infectious disease in one or two years. This is a result of human activities. We humans are going to invade wild animal territories, we are going to travel, we are going to develop, we are going to cultivate, and there are many such (human activities). ), Is actually an infection caused by our human activities.

    Hu Ben, an assistant researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: Minimize exposure to these infectious animals and minimize the risk of this SARS-like outbreak.

    In the eyes of researchers, staying away from wildlife and protecting their habitat is the best source of prevention and control. Virus traceability can also lay the foundation for downstream prevention, diagnosis, vaccine development, and other basic research.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: The results of the work you are doing can be used for disease prevention or for future agricultural prevention. This is also something we are very happy about.

    It took 13 years of research to finally answer people's questions. However, researchers tell us that for this topic, they still have to answer many unsolved mysteries related to it, such as: How did the virus on Yunnan bats spread to animals and people in distant Guangdong? Why do bats carry so many powerful viruses without getting sick? Wait, the road ahead will be even longer.

    Shi Zhengli, a researcher at the Wuhan Institute of Virology, Chinese Academy of Sciences: What you see is a small step, actually we have obtained many, many smaller steps. In fact, doing this basic science is inherently lonely and you need to persist, you will always be a pioneer.

    Almost everyone had a scientist dream when they were young, and they felt that scientists knew more, especially powerful. I do n’t know until I grow up. Why do scientists know so much? Because when everyone starts to forget, only they are persistently searching for the answer. Their greatest strength is not the instrument in their hands, but the spirit in their hearts. For the truth of science and the well-being of mankind, they are willing to endure any loneliness.

    Scientists are the people who are advancing the world. They take truth as their mission and use curiosity and the spirit of exploration to continuously expand the upper limit of human cognition. Every discovery is a new beginning, and every answer will lead to new questions . Rather than trace the source of SARS, it is better to say that it has just begun, and scientists have traveled a long way and have never stopped. For their persistence and determination, we sincerely offer our most sincere respect!

    http://m.news.cctv.com/2017/12/29/AR...dO171229.shtml

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  • Shiloh
    replied
    Source: https://www.sinchew.com.my/content/content_2207254.html
    Website refers to "P4" laboratory virus leak in Wuhan
    1/27 2020 Monday

    2020-01-27 19:30:44 90

    Website refers to "P4" laboratory virus leak in Wuhan
    National Comprehensive

    (Petaling Jaya, 27th) According to an article published on the international news site Gnews, this new coronavirus pneumonia was leaked by the "P4" laboratory set up by the Chinese government in Wuhan, and the Chinese Embassy spokesman Tang Yan today In response, this is a vicious rumor that is baseless and speculative.

    "I saw this report. I believe that people with rational thinking ability and basic judgments of right and wrong know that this is a vicious rumor that is baseless and speculative."


    He said that China is in a critical period of prevention and control of the epidemic, but part of it has made use of the current epidemic fever to throw out so-called explosive news and make "hot" and "deceive traffic" behaviors, without the basic ethics of the news media.

    He said that since the outbreak of pneumonia, the Chinese government has attached great importance to it, and the people have become a city. A large number of medical personnel and equipment are being collected from other provinces in China and even overseas to the epidemic area. At the same time, in order to effectively block the spread of the virus, the Chinese government has extended the Spring Festival. Holidays, restrictions on travel, closure of the city and other extraordinary measures.

    Virus comes from wild animals

    "The Chinese government adheres to the principle of timely, accurate, transparent and open disclosure, and has repeatedly held news conferences on the epidemic, detailing the development of the epidemic, the source of the virus, and control measures, including pointing out that the virus originated from wild animals in the seafood market."

    He also said that the Chinese government has established a daily epidemic release system, and also notified the WHO and relevant countries of the epidemic in a timely manner and maintained close communication. The relevant measures reflect China's high attention to epidemic prevention and control and a highly responsible attitude towards global health security.

    He pointed out that the international community and the media have generally affirmed the Chinese government's rapid and decisive response, including the Director-General of the World Health Organization, Tan Tesai said that China's current measures "help to stop the spread of the epidemic", and Prime Minister of Malaysia Tun Maha Di pointed out that the move to isolate Wuhan shows the Chinese government's determination to prevent and control the epidemic.

    "I hope friends from all walks of life in Malaysia will pay attention to the progress of the epidemic through the authoritative news officially released by China. Do not listen to the letter, spread false or even vicious rumors. With the hard work of the Chinese government and people and the support and help of countries around the world, we are confident, determined, It also has the ability to achieve the ultimate victory in epidemic prevention and control. "
    Author: Hu Yaosen
    Article source: Xingzhou Daily 2020-01-27


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  • Pathfinder
    replied
    Virus-hit Wuhan has two laboratories linked to Chinese bio-warfare program
    ...

    By Bill Gertz - The Washington Times - Friday, January 24, 2020
    ...
    Mr. Shoham holds a doctorate in medical microbiology. From 1970 to 1991 he was a senior analyst with Israeli military intelligence for biological and chemical warfare in the Middle East and worldwide, holding the rank of lieutenant colonel.
    ...
    “Coronaviruses (particularly SARS) have been studied in the institute and are probably held therein,” he said. “SARS is included within the Chinese BW program, at large, and is dealt with in several pertinent facilities.”

    It is not known if the institute’s array of coronaviruses are specifically included in biological weapons program but it is possible, he said.

    Asked if the new coronavirus may have leaked, Mr. Shoham said: “In principle, outward virus infiltration might take place either as leakage or as an indoor unnoticed infection of a person that normally went out of the concerned facility. This could have been the case with the Wuhan Institute of Virology, but so far there isn’t evidence or indication for such incident.”

    After researchers sequence of the genome of the new coronavirus it might be possible to determine or suggest its origin or source.

    Mr. Shoham, now with the Begin-Sadat Center for Strategic Studies at Bar Ilan University in Israel, said the virology institute is the only declared site in China known as P4 for Pathogen Level 4, a status indicating it uses the strictest safety standards to prevent the spread of the most dangerous and exotic microbes being studied.

    The former Israeli military intelligence doctor also said suspicions were raised about the Wuhan Institute of Virology when a group of Chinese virologists working in Canada improperly sent samples to China of what he said were some of the deadliest viruses on earth, including the Ebola virus.
    ...
    The biosafety lab is located about 20 miles from the Hunan Seaford Market that reports from China say may have been origin point of the virus.

    Rutgers University microbiologist Dr. Richard Ebright told London’s Daily Mail that “at this point there’s no reason to harbor suspicions” the lab may be linked to the virus outbreak.

    https://www.washingtontimes.com/news...linked-chines/

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  • sharon sanders
    replied
    The China scientific community has the genetic material. In this 2018 study they used mammals, and proposed further such studies.

    Key word: infectivity



    Emerg Microbes Infect. 2018; 7: 154.
    Published online 2018 Sep 12. doi: 10.1038/s41426-018-0155-5
    PMCID: PMC6135831
    PMID: 30209269


    Genomic characterization and infectivity of a novel SARS-like coronavirus in Chinese bats


    Dan Hu,1,2Changqiang Zhu,2Lele Ai,2Ting He,2Yi Wang,3Fuqiang Ye,2Lu Yang,2Chenxi Ding,2Xuhui Zhu,2Ruicheng Lv,2Jin Zhu,2Bachar Hassan,4Youjun Feng,5Weilong Tan,2 and Changjun Wang1,2
    Author information Article notes Copyright and License information Disclaimer
    1Department of Epidemiology, College of Preventive Medicine, Third Military Medical University, Chongqing, 400038 China
    2Department of Epidemiology, Research Institute for Medicine of Nanjing Command, Nanjing, 210002 China
    3Jiangsu Institute of Parasitic Diseases, Wuxi, Jiangsu Province 214064 P.R. China
    4Stony Brook University, Stony Brook, 11794 USA
    5Department of Pathogen Biology & Microbiology and Department of General Intensive Care Unit of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058 China
    Youjun Feng, Email: nc.ude.ujz@jygnef.
    Contributor Information.
    Corresponding author.
    This article has been cited by other articles in PMC.

    Associated Data

    Supplementary Materials
    Go to: Abstract


    SARS coronavirus (SARS-CoV), the causative agent of the large SARS outbreak in 2003, originated in bats. Many SARS-like coronaviruses (SL-CoVs) have been detected in bats, particularly those that reside in China, Europe, and Africa. To further understand the evolutionary relationship between SARS-CoV and its reservoirs, 334 bats were collected from Zhoushan city, Zhejiang province, China, between 2015 and 2017. PCR amplification of the conserved coronaviral protein RdRp detected coronaviruses in 26.65% of bats belonging to this region, and this number was influenced by seasonal changes. Full genomic analyses of the two new SL-CoVs from Zhoushan (ZXC21 and ZC45) showed that their genomes were 29,732 nucleotides (nt) and 29,802 nt in length, respectively, with 13 open reading frames (ORFs). These results revealed 81% shared nucleotide identity with human/civet SARS CoVs, which was more distant than that observed previously for bat SL-CoVs in China. Importantly, using pathogenic tests, we found that the virus can reproduce and cause disease in suckling rats, and further studies showed that the virus-like particles can be observed in the brains of suckling rats by electron microscopy. Thus, this study increased our understanding of the genetic diversity of the SL-CoVs carried by bats and also provided a new perspective to study the possibility of cross-species transmission of SL-CoVs using suckling rats as an animal model.


    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6135831/

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  • Shiloh
    replied
    Source: http://www.rfi.fr/cn/%E4%B8%AD%E5%9B...A1%B9%E7%9B%AE

    Translated:
    French media explodes Sino-French Wuhan virus laboratory P4 cooperation project causes controversy

    Post time: 25/01/2020-16:40 Change time: 25/01/2020-16:42
    Author:
    Fa Guang

    The new coronavirus that originated in Wuhan is raging all over China and is spreading rapidly overseas. The French challenge website publishes the book "Dangerous Relations between France and China" on Sino-French cooperation to build a P4 virus laboratory. The project is to help China build according to the template of the “box-in-box” of Merrill ’s P4 laboratory in Lyon. It is called a model of the “Belt and Road” by Chinese media. Today, the Wuhan P4 Laboratory is the scientific research base for the prevention and control of acute infectious diseases in China, and it is also the deposit center for severe pathogens and the WHO Reference Laboratory for severe infectious diseases.

    The challenge website article pointed out that France is a leading country in the field of global virus research. In 1999, France set up the largest virus research center in Europe in Lyon. In 2003, the Chinese Academy of Sciences proposed to the French government to assist China to open a similar virus research center in China. Requirements. The Chinese request has caused differences between the French government and virus experts in France, because although the Chinese Virus Center can fight outbreaks of infectious diseases, some French experts worry that China will use technology provided by France to develop chemical weapons. French intelligence agencies A serious warning was presented to the government.

    However, with the support of then Prime Minister Raffarin, China and France finally signed a cooperation agreement during Chirac's visit to China in 2004. France will assist China in building a P4 virus center, but the agreement stipulates that Beijing cannot use this technology for offensive activities. The agreement was controversial when it was signed, and Raffarin once said: "The heads of government of the two countries signed a cooperation agreement, but the administration has blocked it after all.

    The French General Administration of Foreign Security pointed out that RTV, an architectural design firm in Lyon, France was originally responsible for the laboratory's engineering, but in 2005 China officially chose Wuhan's local design agency IPPR (Zhongyuan International Engineering Co., Ltd.) for the project. According to the department's investigation, the IPPR design institute is closely related to the subordinate departments of the Chinese military. These departments have long been the target of supervision by the US Central Intelligence Agency. Due to the aforementioned security concerns and repeated delays in the implementation of the agreement, as well as the diplomatic crisis between China and France in 2008, this led to the Wuhan P4 virus center's official operation until 2017. The then French Prime Minister Bernard Cazeneuve attended the launching ceremony of the laboratory.

    Wuhan Virus Laboratory is accredited to conduct research on three types of viruses: Ebola, Congo-Crimea hemorrhagic fever and Nipah virus.

    Laboratories must also be certified by WHO in order to be officially included in partner laboratories to help identify internationally spread viruses. The laboratory was originally planned to be certified by WHO in 2020, making the laboratory fully functional.

    Should France help China build a P4 laboratory? Has China kept its original commitment? If the head of the Wuhan P4 laboratory is certain that the project will help Asia and the world to fight the epidemic more effectively, the French government does not agree with the above views. A government official told reporters that China has repeatedly violated its commitments in the course of more than a decade of cooperation. For example, China promised to build only one laboratory in Wuhan. Today, it was discovered that China has built multiple laboratories. And some labs are very suspicious.


    Leave a comment:


  • sharon sanders
    replied

    but wait....there is more....


    J Gen Virol. 2010 Apr;91(Pt 4):1058-62. doi: 10.1099/vir.0.016378-0. Epub 2009 Dec 16.


    Intraspecies diversity of SARS-like coronaviruses in Rhinolophus sinicus and its implications for the origin of SARS coronaviruses in humans.

    Yuan J1, Hon CC, Li Y, Wang D, Xu G, Zhang H, Zhou P, Poon LL, Lam TT, Leung FC, Shi Z. Author information

    1 State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences (CAS), Wuhan, PR China.


    Abstract


    The Chinese rufous horseshoe bat (Rhinolophus sinicus) has been suggested to carry the direct ancestor of severe acute respiratory syndrome (SARS) coronavirus (SCoV), and the diversity of SARS-like CoVs (SLCoV) within this Rhinolophus species is therefore worth investigating. Here, we demonstrate the remarkable diversity of SLCoVs in R. sinicus and identify a strain with the same pattern of phylogenetic incongruence (i.e. an indication of recombination) as reported previously in another SLCoV strain. Moreover, this strain possesses a distinctive 579 nt deletion in the nsp3 region that was also found in a human SCoV from the late-phase epidemic. Phylogenetic analysis of the Orf1 region suggested that the human SCoVs are phylogenetically closer to SLCoVs in R. sinicus than to SLCoVs in other Rhinolophus species. These findings reveal a closer evolutionary linkage between SCoV in humans and SLCoVs in R. sinicus, defining the scope of surveillance to search for the direct ancestor of human SCoVs.


    https://www.ncbi.nlm.nih.gov/pubmed/20016037


    --------------------------------------------


    Arch Virol. 2010 Oct;155(10):1563-9. doi: 10.1007/s00705-010-0729-6. Epub 2010 Jun 22.


    Angiotensin-converting enzyme 2 (ACE2) proteins of different bat species confer variable susceptibility to SARS-CoV entry.

    Hou Y1, Peng C, Yu M, Li Y, Han Z, Li F, Wang LF, Shi Z. Author information

    1 State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences (CAS), Wuhan, Hubei, China.


    Abstract


    The discovery of SARS-like coronavirus in bats suggests that bats could be the natural reservoir of SARS-CoV. However, previous studies indicated the angiotensin-converting enzyme 2 (ACE2) protein, a known SARS-CoV receptor, from a horseshoe bat was unable to act as a functional receptor for SARS-CoV. Here, we extended our previous study to ACE2 molecules from seven additional bat species and tested their interactions with human SARS-CoV spike protein using both HIV-based pseudotype and live SARS-CoV infection assays. The results show that ACE2s of Myotis daubentoni and Rhinolophus sinicus support viral entry mediated by the SARS-CoV S protein, albeit with different efficiency in comparison to that of the human ACE2. Further, the alteration of several key residues either decreased or enhanced bat ACE2 receptor efficiency, as predicted from a structural modeling study of the different bat ACE2 molecules. These data suggest that M. daubentoni and R. sinicus are likely to be susceptible to SARS-CoV and may be candidates as the natural host of the SARS-CoV progenitor viruses. Furthermore, our current study also demonstrates that the genetic diversity of ACE2 among bats is greater than that observed among known SARS-CoV susceptible mammals, highlighting the possibility that there are many more uncharacterized bat species that can act as a reservoir of SARS-CoV or its progenitor viruses. This calls for continuation and expansion of field surveillance studies among different bat populations to eventually identify the true natural reservoir of SARS-CoV.


    https://www.ncbi.nlm.nih.gov/pubmed/20567988


    -------------------------------------------


    Virol Sin. 2010 Feb;25(1):36-44. doi: 10.1007/s12250-010-3096-2. Epub 2010 Feb 12.


    Immunogenicity of the spike glycoprotein of bat SARS-like coronavirus.

    Hou YX1, Peng C, Han ZG, Zhou P, Chen JG, Shi ZL. Author information

    Abstract


    A group of SARS-like coronaviruses (SL-CoV) have been identified in horseshoe bats. Despite SL-CoVs and SARS-CoV share identical genome structure and high-level sequence similarity, SL-CoV does not bind to the same cellular receptor as for SARS-CoV and the N-terminus of the S proteins only share 64% amino acid identity, suggesting there are fundamental differences between these two groups of coronaviruses. To gain insight into the basis of this difference, we established a recombinant adenovirus system expressing the S protein from SL-CoV (rAd-Rp3-S) to investigate its immune characterization. Our results showed that immunized mice generated strong humoral immune responses against the SL-CoV S protein. Moreover, a strong cellular immune response demonstrated by elevated IFN-γ and IL-6 levels was also observed in these mice. However, the induced antibody from these mice had weaker cross-reaction with the SARS-CoV S protein, and did not neutralize HIV pseudotyped with SARS-CoV S protein. These results demonstrated that the immunogenicity of the SL-CoV S protein is distinct from that of SARS-CoV, which may cause the immunological differences between human SARS-CoV and bat SL-CoV. Furthermore, the recombinant virus could serve as a potential vaccine candidate against bat SL-CoV infection.


    https://www.ncbi.nlm.nih.gov/pubmed/20960282


    --------------------------------------------



    J Gen Virol. 2012 Feb;93(Pt 2):275-81. doi: 10.1099/vir.0.033589-0. Epub 2011 Oct 19.


    Bat severe acute respiratory syndrome-like coronavirus ORF3b homologues display different interferon antagonist activities.

    Zhou P1, Li H, Wang H, Wang LF, Shi Z. Author information

    1 State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Republic of China.


    Abstract


    The ORF3b protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has a nuclear localization signal (NLS) at its C terminus and antagonizes interferon (IFN) function by modulating the activity of IFN regulatory factor 3 (IRF3). SARS-like coronaviruses (SL-CoVs) found in bats share an identical genome organization and high sequence identity for most of their gene products. In this study, ORF3b homologues were identified from three bat SL-CoV strains. These ORF3b homologues were C-terminally truncated and lacked the C-terminal NLS of SARS-CoV. IFN antagonist activities analysis demonstrated that one SL-CoV ORF3b still possessed IFN antagonist and IRF3-modulating activities. These results indicate that different ORF3b proteins display different IFN antagonist activities and this function is independent of the protein's nuclear localization, suggesting a potential link between bat SL-CoV ORF3b function and viral pathogenesis.


    https://www.ncbi.nlm.nih.gov/pubmed/22012463


    --------------------------------------------------

    J Virol. 2016 Jun 24;90(14):6573-6582. doi: 10.1128/JVI.03079-15. Print 2016 Jul 15.


    Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIV1 Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response.

    Zeng LP1, Gao YT1, Ge XY1, Zhang Q1, Peng C1, Yang XL1, Tan B1, Chen J1, Chmura AA2, Daszak P2, Shi ZL3. Author information

    1 Key Laboratory of Special Pathogens, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China. 2 EcoHealth Alliance, New York, New York, USA. 3 Key Laboratory of Special Pathogens, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China zlshi@wh.iov.cn.


    Abstract


    Bats harbor severe acute respiratory syndrome (SARS)-like coronaviruses (SL-CoVs) from which the causative agent of the 2002-2003 SARS pandemic is thought to have originated. However, despite the fact that a large number of genetically diverse SL-CoV sequences have been detected in bats, only two strains (named WIV1 and WIV16) have been successfully cultured in vitro These two strains differ from SARS-CoV only in containing an extra open reading frame (ORF) (named ORFX), between ORF6 and ORF7, which has no homology to any known protein sequences. In this study, we constructed a full-length cDNA clone of SL-CoV WIV1 (rWIV1), an ORFX deletion mutant (rWIV1-ΔX), and a green fluorescent protein (GFP)-expressing mutant (rWIV1-GFP-ΔX). Northern blotting and fluorescence microscopy indicate that ORFX was expressed during WIV1 infection. A virus infection assay showed that rWIV1-ΔX replicated as efficiently as rWIV1 in Vero E6, Calu-3, and HeLa-hACE2 cells. Further study showed that ORFX could inhibit interferon production and activate NF-κB. Our results demonstrate for the first time that the unique ORFX in the WIV1 strain is a functional gene involving modulation of the host immune response but is not essential for in vitro viral replication.


    IMPORTANCE:


    Bats harbor genetically diverse SARS-like coronaviruses (SL-CoVs), and some of them have the potential for interspecies transmission. A unique open reading frame (ORFX) was identified in the genomes of two recently isolated bat SL-CoV strains (WIV1 and -16). It will therefore be critical to clarify whether and how this protein contributes to virulence during viral infection. Here we revealed that the unique ORFX is a functional gene that is involved in the modulation of the host immune response but is not essential for in vitro viral replication. Our results provide important information for further exploration of the ORFX function in the future. Moreover, the reverse genetics system we constructed will be helpful for study of the pathogenesis of this group of viruses and to develop therapeutics for future control of emerging SARS-like infections.


    https://www.ncbi.nlm.nih.gov/pubmed/27170748


    ---------------------------------------------------------------


    Viruses. 2019 Apr 24;11(4). pii: E379. doi: 10.3390/v11040379.


    Characterization of a New Member of Alphacoronavirus with Unique Genomic Features in Rhinolophus Bats.

    Wang N1,2, Luo C3,4, Liu H5, Yang X6, Hu B7, Zhang W8, Li B9, Zhu Y10, Zhu G11, Shen X12,13, Peng C14, Shi Z15. Author information

    1 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. wuzhongxiaoliu@163.com. 2 College of Life Science, University of Chinese Academy of Sciences, Beijing 100864, China. wuzhongxiaoliu@163.com. 3 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. chine.lcm@163.com. 4 College of Life Science, University of Chinese Academy of Sciences, Beijing 100864, China. chine.lcm@163.com. 5 Center for Emerging Infectious Disease, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. liuhz@wh.iov.cn. 6 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. yangxl@wh.iov.cn. 7 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. ouihaagendazs@hotmail.com. 8 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. happyzhang_2007@163.com. 9 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. shenqi0622@126.com. 10 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. zhuyan@wh.iov.cn. 11 EcoHealth Alliance, New York, NY 10001, USA. zhugj609@126.com. 12 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. pukulovesu@163.com. 13 College of Life Science, University of Chinese Academy of Sciences, Beijing 100864, China. pukulovesu@163.com. 14 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. pengcheng@wh.iov.cn. 15 CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. zlshi@wh.iov.cn.


    Abstract


    Bats have been identified as a natural reservoir of a variety of coronaviruses (CoVs). Several of them have caused diseases in humans and domestic animals by interspecies transmission. Considering the diversity of bat coronaviruses, bat species and populations, we expect to discover more bat CoVs through virus surveillance. In this study, we described a new member of alphaCoV (BtCoV/Rh/YN2012) in bats with unique genome features. Unique accessory genes, ORF4a and ORF4b were found between the spike gene and the envelope gene, while ORF8 gene was found downstream of the nucleocapsid gene. All the putative genes were further confirmed by reverse-transcription analyses. One unique gene at the 3' end of the BtCoV/Rh/YN2012 genome, ORF9, exhibits ~30% amino acid identity to ORF7a of the SARS-related coronavirus. Functional analysis showed ORF4a protein can activate IFN-β production, whereas ORF3a can regulate NF-κB production. We also screened the spike-mediated virus entry using the spike-pseudotyped retroviruses system, although failed to find any fully permissive cells. Our results expand the knowledge on the genetic diversity of bat coronaviruses. Continuous screening of bat viruses will help us further understand the important role played by bats in coronavirus evolution and transmission.


    https://www.ncbi.nlm.nih.gov/pubmed/31022925




    .
    Last edited by sharon sanders; January 25, 2020, 12:21 PM.

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