[ DONATE TO RMN ] [ View Thread ] [ Return to Main Page ] [ Read Prev Article ] [ Read Next Article ] [ CGI Media News Room ] [ SUBSCRIBE TO RMN ]

RMN is Reader Supported

Our Goal for
NOV 6 - DEC 5:
$2500

Powered by FundRazr

Click Widget
or Click Here to contribute.

Checks & Money Orders:

Raye Allan Smith
P.O. Box 95
Ashtabula, OH 44005


Users Online:
89

Who Founded RMNews?


Dewitt Jones' Video
"Celebrate What's Right
With The World"


"When the
Starships Fly!"

Listen at YouTube


The Theme for The Obergon Chronicles

Listen at YouTube


The Obergon Chronicles ebook


RUMOR MILL
NEWS RADIO


CGI ROOM
Common Ground
Independent Media


WHAT ARE
THE FACTIONS?


THE AMAZING
RAYELAN ALLAN


BIORHYTHMS

LOTTO PICKS

OTHER WAYS TO DONATE


CURRENT MOON




RUMOR MILL NEWS AGENTS WHO'VE BEEN INTERVIEWED ON RUMOR MILL NEWS RADIO

______________

NOVEMBER 2008

Kevin Courtois - Kcbjedi
______________

Dr Robin Falkov

______________

Melinda Pillsbury Hr1

Melinda Pillsbury Hr2

______________

Daneen Peterson

______________

Daneen Peterson

______________

Disclosure Hr1

Disclosure Hr2
______________

Scribe
______________

in_PHI_nitti
______________

Jasmine Hr1
Jasmine Hr2
______________

Tom Chittum Hr1
Tom Chittum Hr2
______________

Kevin Courtois
______________

Dr Syberlux
______________

Gary Larrabee Hr1
Gary Larrabee Hr2
______________

Kevin Courtois
______________

Pravdaseeker Hr1
Pravdaseeker Hr2
______________

DECEMBER 2008

Tom Chittum
______________

Crystal River
______________

Stewart Swerdlow Hr1
Stewart Swerdlow Hr2
______________

Janet Swerdlow Hr1
Janet Swerdlow Hr2
______________

Dr. Robin Falkov Hr1
Dr. Robin Falkov Hr2
Dr. Robin Falkov Hr3

JANUARY 2009 ______________

Patriotlad
______________

Patriotlad
______________

Crystal River
______________

Patriotlad
______________

Dr. Robin Falcov
______________

Patriotlad

FEBRUARY 2009

Find UFOs, The Apocalypse, New World Order, Political Analysis,
Alternative Health, Armageddon, Conspiracies, Prophecies, Spirituality,
Home Schooling, Home Mortgages and more, in:

The Rumor Mill News Reading Room

Ton of info on them here

Posted By: oldmaninthedesert
Date: Sunday, 15-Mar-2020 23:25:07
www.rumormill.news/142393

In Response To: Thanks To All Who Replied: Pirbrite Institute - Again (Lion)

Mr Lion
https://aim4truth.org/2020/03/14/cat-report-333/

: -------------------

: Can Anyone Look Up This Patent Information?

: The patent # is: U.S. patent: 10 130 701

: -------------------

: Thanks To All Who Replied: ------------------------

: ( 1 of 1 )
: United States Patent 10,130,701

: Bickerton , et al. November 20, 2018

: Coronavirus

: Abstract

: The present invention provides a live, attenuated coronavirus
: comprising a variant replicase gene encoding polyproteins
: comprising a mutation in one or more of non-structural
: protein(s) (nsp)-10, nsp-14, nsp-15 or nsp-16.

: The coronavirus may be used as a vaccine for treating and/or
: preventing a disease, such as infectious bronchitis, in a
: subject.

: Inventors: Bickerton; Erica (Woking, GB), Keep; Sarah

: (Woking, GB), Britton; Paul (Woking, GB)

: Applicant: Name City State Country Type

: THE PIRBRIGHT INSTITUTE

: Pirbright, Woking
: N/A
: GB
: Assignee: THE PIRBRIGHT INSTITUTE (Woking, Pirbright, GB)

: Family ID: 51494985

: Appl. No.: 15/328,179

: Filed: July 23, 2015

: PCT Filed: July 23, 2015

: PCT No.: PCT/GB2015/052124

: 371(c)(1),(2),(4) Date: January 23, 2017

: PCT Pub. No.: WO2016/012793

: PCT Pub. Date: January 28, 2016

: Prior Publication Data

: Document Identifier Publication Date

: US 20170216427 A1 Aug 3, 2017

: Foreign Application Priority Data

: Jul 23, 2014 [GB] 1413020.7

: Current U.S. Class: 1/1
: Current CPC Class: C12N 7/00 (20130101); C12N 9/127
: (20130101); C07K 14/005 (20130101); A61K 39/215 (20130101);
: C12Y 207/07048 (20130101); C12N 2770/20062 (20130101); C12N
: 2770/20022 (20130101); C12N 2770/20034 (20130101); C12N
: 2770/20051 (20130101); A61K 2039/70 (20130101); C12N
: 2770/20021 (20130101); A61K 2039/5254 (20130101); A61K
: 2039/54 (20130101); C12N 2770/20071 (20130101)
: Current International Class: A61K 39/215 (20060101); C12N 9/12
: (20060101); C12N 7/00 (20060101); A61K 39/00 (20060101)
: References Cited [Referenced By]
: U.S. Patent Documents

: 7452542 November 2008 Denison
: Foreign Patent Documents

: WO-2004/092360 Oct 2004 WO

: WO-2005/049814 Jun 2005 WO

: WO-2007/078203 Jul 2007 WO

: WO-2011/004146 Jan 2011 WO

: Other References

: Sperry Journal of Virology, 2005, vol. 79, No. 6, pp.
: 3391-3400. cited by examiner .
: Altschul et al., Basic local alignment search tool. J. Mol.
: Biol. 215: 403-10 (1990). cited by applicant .
: Ammayapppan et al.,

: Identification of sequence changes responsible for the
: attenuation of avian infectious bronchitis virus strain
: Arkansas DPI, Arch. Virol., 154(3):495-9 (2009). cited by
: applicant .

: Anonymous: "EM_STD:KF377577", Oct. 30, 2013. cited
: by applicant .
: Armesto et al., A recombinant avian infectious bronchitis
: virus expressing a heterologous spike gene belonging to the
: 4/91 serotype, PLoS One, 6(8):e24352 (2011). cited by
: applicant .
: Armesto et al.,

: The replicase gene of avian coronavirus infectious bronchitis
: virus is a determinant of pathogenicity, PLoS One,
: 4(10):e7384 (2009). cited by applicant .
: Armesto et al.,

: Transient dominant selection for the modification and
: generation of recombinant infectious bronchitis
: coronaviruses, Methods Mol. Biol., 454:255-73 (2008). cited
: by applicant .
: Ausubel et al.,

: Short Protocols in Molecular Biology, 4th edition, Chapter 18
: (1999). cited by applicant .
: Britton et al.,

: Generation of a recombinant avian coronavirus infectious
: bronchitis virus using transient dominant selection, J.
: Virol. Methods, 123(2):203-11 (2005). cited by applicant .
: Britton et al.,

: Modification of the avian coronavirus infectious bronchitis
: virus for vaccine development, Bioeng. Bugs., 3(2):114-9
: (2012). cited by applicant .
: Casais et al.,

: Recombinant avian infectious bronchitis virus expressing a
: heterologous spike gene demonstrates that the spike protein
: is a determinant of cell tropism, J. Virol., 77(16):9084-9
: (2003). cited by applicant .
: Casais et al.,

: Reverse genetics system for the avian coronavirus infectious
: bronchitis virus, J. Virol., 75(24):12359-69 (2001). cited
: by applicant .
: Devereux et al.,

: A comprehensive set of sequence analysis programms for the
: VAX. Nucl. Acids Res.12: 387-95 (1984). cited by applicant
: .
: Cavanagh et al.,

: Manipulation of the infectious bronchitis coronavirus genome
: for vaccine development and analysis of the accessory
: proteins, Vaccine, 25(30):5558-62 (2007). cited by
: applicant .
: International Preliminary Report on Patentability,
: International Application No. PCT/GB2015/052124, dated Jan.
: 24, 2017. cited by applicant .

: International Search Report and Written Opinion, International
: Application No. PCT/GB2015/052124, dated Oct. 9, 2015.
: cited by applicant .
: Larkin et al.,

: Clustal W and Clustal X version 2.0, Bioinformatics,
: 23(21):2947-8 (2007). cited by applicant .
: Menachery et al.,

: Attenuation and restoration of severe acute respiratory
: syndrome coronavirus mutant lacking 2'-o-methyltransferase
: activity, J. Virol., 88(8):4251-64 (2014). cited by
: applicant .
: Tatusova et al., BLAST 2 Sequences, a new tool for comparing
: protein and nucleotide sequences, FEMS Microbiol. Lett.,
: 174(2):247-50 (1999). cited by applicant .
: Wang et al.,

: Attenuation of porcine reproductive and respiratory syndrome
: virus strain MN184 using chimeric construction with vaccine
: sequence, Virology, 371(2):418-29 (2008). cited by
: applicant .
: Wei et al.,

: Development and characterization of a recombinant infectious
: bronchitis virus expressing the ectodomain region of S1
: gene of H120 strain, Appl. Microbiol. Biotechnol.,
: 98(4):1727-35 (2014). cited by applicant.

: Primary Examiner: Li; Bao Q
: Attorney, Agent or Firm: Marshall, Gerstein & Borun LLP
: Claims

: The invention claimed is: 1. A live, attenuated coronavirus
: comprising a variant replicase gene encoding polyproteins
: comprising a mutation in one or both of non-structural
: protein(s) nsp-10 and nsp-14, wherein the variant replicase
: gene encodes a protein comprising an amino acid mutation of
: Pro to Leu at the position corresponding to position 85 of
: SEQ ID NO: 6, and/or wherein the variant replicase gene
: encodes a protein comprising an amino acid mutation of Val
: to Leu at the position corresponding to position 393 of SEQ
: ID NO: 7.

: 2. The coronavirus according to claim 1 wherein the variant
: replicase gene encodes a protein comprising one or more
: amino acid mutations selected from: an amino acid mutation
: of Leu to Ile at the position corresponding to position 183
: of SEQ ID NO: 8; and an amino acid mutation of Val to Ile
: at the position corresponding to position 209 of SEQ ID NO:
: 9.

: 3. The coronavirus according to claim 1 wherein the replicase
: gene encodes a protein comprising the amino acid mutations
: Val to Leu at the position corresponding to position 393 of
: SEQ ID NO: 7; Leu to Ile at the position corresponding to
: position 183 of SEQ ID NO: 8; and Val to Ile at the
: position corresponding to position 209 of SEQ ID NO: 9.

: 4. The coronavirus according to claim 1 wherein the replicase
: gene encodes a protein comprising the amino acid mutations
: Pro to Leu at the position corresponding to position 85 of
: SEQ ID NO: 6; Val to Leu at the position corresponding to
: position 393 of SEQ ID NO: 7; Leu to Ile at the position
: corresponding to position 183 of SEQ ID NO: 8; and Val to
: Ile at the position corresponding to position 209 of SEQ ID
: NO: 9.

: 5. The coronavirus according to claim 1 wherein the replicase
: gene comprises at least one nucleotide substitutions
: selected from: C to Tat nucleotide position 12137; and G to
: C at nucleotide position 18114; compared to the sequence
: shown as SEQ ID NO: 1; and optionally, comprises one or
: more nucleotide substitutions selected from T to A at
: nucleotide position 19047; and G to A at nucleotide
: position 20139; compared to the sequence shown as SEQ ID
: NO: 1.

: 6. The coronavirus according to claim 1 which is an infectious
: bronchitis virus (IBV).

: 7. The coronavirus according to claim 1 which is IBV M41.

: 8. The coronavirus according to claim 7, which comprises an S
: protein at least, part of which is from an IBV serotype
: other than M41.

: 9. The coronavirus according to claim 8, wherein the S1
: subunit is from an IBV serotype other than M41.

: 10. The coronavirus according to claim 8, wherein the S
: protein is from an IBV serotype other than M41.

: 11. The coronavirus according to claim 1 which has reduced
: pathogenicity compared to a coronavirus expressing a
: corresponding wild-type replicase, wherein the virus is
: capable of replicating without being pathogenic to the
: embryo when administered to an embryonated egg.

: 12. A variant replicase gene as defined in claim 1.

: 13. A protein encoded by a variant coronavirus replicase gene
: according to claim 12.

: 14. A plasmid comprising a replicase gene according to claim
: 12.

: 15. A method for making the coronavirus according to claim 1
: which comprises the following steps: (i) transfecting a
: plasmid according to claim 14 into a host cell; (ii)
: infecting the host cell with a recombining virus comprising
: the genome of a coronavirus strain with a replicase gene;
: (iii) allowing homologous recombination to occur between
: the replicase gene sequences in the plasmid and the
: corresponding sequences in the recombining virus genome to
: produce a modified replicase gene; and (iv) selecting for
: recombining virus comprising the modified replicase gene.

: 16. The method according to claim 15, wherein the recombining
: virus is a vaccinia virus.

: 17. The method according to claim 15 which also includes the
: step: (v) recovering recombinant coronavirus comprising the
: modified replicase gene from the DNA from the recombining
: virus from step (iv).

: 18. A cell capable of producing a coronavirus according to
: claim 1.

: 19. A vaccine comprising a coronavirus according to claim 1
: and a pharmaceutically acceptable carrier.

: 20. A method for treating and/or preventing a disease in a
: subject which comprises the step of administering a vaccine
: according to claim 19 to the subject.

: 21. The method of claim 20, wherein the disease is infectious
: bronchitis (IB).

: 22. The method according to claim 20 wherein the method of
: administration is selected from the group consisting of;
: eye drop administration, intranasal administration,
: drinking water administration, post-hatch injection and in
: ovo injection.

: 23. The method according to claim 21 wherein the
: administration is in ovo vaccination.

: 24. A method for producing a vaccine according to claim 19,
: which comprises the step of infecting a cell according to
: claim 18 with a coronavirus according to claim 1.

: 25. The coronavirus according to claim 1, further comprising a
: mutation in one or both of nsp-15 and nsp-16.
: Description

: FIELD OF THE INVENTION

: The present invention relates to an attenuated coronavirus
: comprising a variant replicase gene, which causes the virus
: to have reduced pathogenicity. The present invention also
: relates to the use of such a coronavirus in a vaccine to
: prevent and/or treat a disease.

: BACKGROUND TO THE INVENTION

: Avian infectious bronchitis virus (IBV), the aetiological
: agent of infectious bronchitis (IB), is a highly infectious
: and contagious pathogen of domestic fowl that replicates
: primarily in the respiratory tract but also in epithelial
: cells of the gut, kidney and oviduct.

: IBV is a member of the Order Nidovirales, Family
: Coronaviridae, Subfamily Corona virinae and Genus
: Gammacoronavirus;

: genetically very similar coronaviruses cause disease in
: turkeys, guinea fowl and pheasants.

: Clinical signs of IB include sneezing, tracheal rales, nasal
: discharge and wheezing.

: Meat-type birds have reduced weight gain, whilst egg-laying
: birds lay fewer eggs and produce poor quality eggs.

: The respiratory infection predisposes chickens to secondary
: bacterial infections which can be fatal in chicks.

: The virus can also cause permanent damage to the oviduct,
: especially in chicks, leading to reduced egg production and
: quality; and kidney, sometimes leading to kidney disease
: which can be fatal.

: IBV has been reported to be responsible for more economic loss
: to the poultry industry than any other infectious disease.

: Although live attenuated vaccines and inactivated vaccines are
: universally used in the control of IBV, the protection
: gained by use of vaccination can be lost either due to
: vaccine breakdown or the introduction of a new IBV serotype
: that is not related to the vaccine used, posing a risk to
: the poultry industry.

: Further, there is a need in the industry to develop vaccines
: which are suitable for use in ovo, in order to improve the
: efficiency and cost-effectiveness of vaccination
: programmes.

: A major challenge associated with in ovo vaccination is that
: the virus must be capable of replicating in the presence of
: maternally-derived antibodies against the virus, without
: being pathogenic to the embryo.

: Current IBV vaccines are derived following multiple passage in
: embryonated eggs, this results in viruses with reduced
: pathogenicity for chickens, so that they can be used as
: live attenuated vaccines.

: However such viruses almost always show an increased virulence
: to embryos and therefore cannot be used for in ova
: vaccination as they cause reduced hatchability. A 70%
: reduction in hatchability is seen in some cases.

: Attenuation following multiple passage in embryonated eggs
: also suffers from other disadvantages.

: It is an empirical method, as attenuation of the viruses is
: random and will differ every time the virus is passaged, so
: passage of the same virus through a different series of
: eggs for attenuation purposes will lead to a different set
: of mutations leading to attenuation.

: There are also efficacy problems associated with the process:
: some mutations will affect the replication of the virus and
: some of the mutations may make the virus too attenuated.
: Mutations can also occur in the S gene which may also
: affect immunogenicity so that the desired immune response
: is affected and the potential vaccine may not protect
: against the required serotype.

: In addition there are problems associated with reversion to
: virulence and stability of vaccines.

: It is important that new and safer vaccines are developed for
: the control of IBV. Thus there is a need for IBV vaccines
: which are not associated with these issues, in particular
: vaccines which may be used for in ovo vaccination.

: SUMMARY OF ASPECTS OF THE INVENTION

: The present inventors have used a reverse genetics approach in
: order to rationally attenuate IBV. This approach is much
: more controllable than random attenuation following
: multiple passages in embryonated eggs because the position
: of each mutation is known and its effect on the virus, i.e.
: the reason for attenuation, can be derived.

: Using their reverse genetics approach, the present inventors
: have identified various mutations which cause the virus to
: have reduced levels of pathogenicity.

: The levels of pathogenicity may be reduced such that when the
: virus is administered to an embryonated egg, it is capable
: of replicating without being pathogenic to the embryo.

: Such viruses may be suitable for in ovo vaccination, which is
: a significant advantage and has improvement over attenuated
: IBV vaccines produced following multiple passage in
: embryonated eggs.

: Thus in a first aspect, the present invention provides a live,
: attenuated coronavirus comprising a variant replicase gene
: encoding polyproteins comprising a mutation in one or more
: of non-structural protein(s) (nsp)-10, nsp-14, nsp-15 or
: nsp-16.

: The variant replicase gene may encode a protein comprising one
: or more amino acid mutations selected from the list of: Pro
: to Leu at position 85 of SEQ ID NO: 6, Val to Leu at
: position 393 of SEQ ID NO: 7; Leu to Ile at position 183 of
: SEQ ID NO: 8; Val to Ile at position 209 of SEQ ID NO: 9.

: The replicase gene may encode a protein comprising the amino
: acid mutation Pro to Leu at position 85 of SEQ ID NO: 6.

: The replicase gene may encode a protein comprising the amino
: acid mutations Val to Leu at position 393 of SEQ ID NO: 7;
: Leu to Ile at position 183 of SEQ ID NO: 8; and Val to Ile
: at position 209 of SEQ ID NO: 9.

: The replicase gene may encodes a protein comprising the amino
: acid mutations Pro to Leu at position 85 of SEQ ID NO: 6;
: Val to Leu at position 393 of SEQ ID NO:7; Leu to Ile at
: position 183 of SEQ ID NO:8; and Val to Ile at position 209
: of SEQ ID NO: 9.

: The replicase gene may comprise one or more nucleotide
: substitutions selected from the list of: C to T at
: nucleotide position 12137;

: G to C at nucleotide position 18114;

: T to A at nucleotide position 19047; and

: G to A at nucleotide position 20139;

: compared to the sequence shown as SEQ ID NO: 1.

: The coronavirus may be an infectious bronchitis virus (IBV).

: The coronavirus may be IBV M41.

: The coronavirus may comprise an S protein at least part of
: which is from an IBV serotype other than M41.

: For example, the S1 subunit or the entire S protein may be
: from an IBV serotype other than M41.

: The coronavirus according to the first aspect of the invention
: has reduced pathogenicity compared to a coronavirus
: expressing a corresponding wild-type replicase, such that
: when the virus is administered to an embryonated egg, it is
: capable of replicating without being pathogenic to the
: embryo.

: In a second aspect, the present invention provides a variant
: replicase gene as defined in connection with the first
: aspect of the invention.

: In a third aspect, the present invention provides a protein
: encoded by a variant coronavirus replicase gene according
: to the second aspect of the invention.

: In a fourth aspect, the present invention provides a plasmid
: comprising a replicase gene according to the second aspect
: of the invention.

: In a fifth aspect, the present invention provides a method for
: making the coronavirus according to the first aspect of the
: invention which comprises the following steps: (i)
: transfecting a plasmid according to the fourth aspect of
: the invention into a host cell; (ii) infecting the host
: cell with a recombining virus comprising the genome of a
: coronavirus strain with a replicase gene; (iii) allowing
: homologous recombination to occur between the replicase
: gene sequences in the plasmid and the corresponding
: sequences in the recombining virus genome to produce a
: modified replicase gene; and (iv) selecting for recombining
: virus comprising the modified replicase gene.

: The recombining virus may be a vaccinia virus.

: The method may also include the step: (v) recovering
: recombinant coronavirus comprising the modified replicase
: gene from the DNA from the recombining virus from step
: (iv).

: In a sixth aspect, the present invention provides a cell
: capable of producing a coronavirus according to the first
: aspect of the invention.

: In a seventh aspect, the present invention provides a vaccine
: comprising a coronavirus according to the first aspect of
: the invention and a pharmaceutically acceptable carrier.

: In an eighth aspect, the present invention provides a method
: for treating and/or preventing a disease in a subject which
: comprises the step of administering a vaccine according to
: the seventh aspect of the invention to the subject.

: Further aspects of the invention provide: the vaccine
: according to the seventh aspect of the invention for use in
: treating and/or preventing a disease in a subject. use of a
: coronavirus according to the first aspect of the invention
: in the manufacture of a vaccine for treating and/or
: preventing a disease in a subject.

: The disease may be infectious bronchitis (IB).

: The method of administration of the vaccine may be selected
: from the group consisting of; eye drop administration,
: intranasal administration, drinking water administration,
: post-hatch injection and in ovo injection.

: Vaccination may be by in ova vaccination.

: The present invention also provides a method for producing a
: vaccine according to the seventh aspect of the invention,
: which comprises the step of infecting a cell according to
: the sixth aspect of the invention with a coronavirus
: according to the first aspect of the invention.

: More data here:
: http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&p=1&u=%2Fnetahtml%2FPTO%2Fsearch-bool.html&r=1&f=G&l=50&co1=AND&d=PTXT&s1=10130701.PN.&OS=PN/10130701&RS=PN/10130701

: --------------------





If you enjoyed this article,
Please consider a monthly subscription to Rumor Mill News!!


RMN is an RA production.

Articles In This Thread

The only pay your RMN moderators receive
comes from ads.
Please consider putting RMN in
your ad blocker's whitelist.

Serving Truth and Freedom
Worldwide since 1996
 
Politically Incorrect News
Stranger than Fiction
Usually True!


Powered
by FundRazr
Click Widget
or Click Here to contribute.


Organic Sulfur 4 Health

^


AGENTS WEBPAGES

Provided free to RMN Agents

Organic Sulfur 4 Health

^


AGENTS WEBPAGES

Provided free to RMN Agents



[ DONATE TO RMN ] [ View Thread ] [ Return to Main Page ] [ Read Prev Article ] [ Read Next Article ] [ CGI Media News Room ] [ SUBSCRIBE TO RMN ]

The Rumor Mill News Reading Room is maintained by Forum Admin with WebBBS 5.12.