AU2018380582B2 - Feline calicivirus vaccine - Google Patents

Abstract

The present invention provides new feline calicivirus vaccines, including multivalent vaccines. The present invention also provides methods of making and using the vaccines.

Description

FELINE CALICIVIRUS VACCINE

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119(e) of provisional application

U.S. Serial No. 62/596,508 filed December 8, 2017, U.S. Serial No. 62/582,050 filed

November 6, 2017, U.S. Serial No. 62/581,955 filed November 6, 2017, and

U.S. Serial No. 62/599,401 filed December 15, 2017, the contents of which are

hereby incorporated by reference in their entireties.

FIELD OF THE INVENTION The present invention relates to new vaccines for feline calicivirus. Methods

of making and using the vaccines alone or in combination with other protective

agents are also provided.

BACKGROUND Feline calicivirus (FCV) is usually associated with upper respiratory disease

in cats. FCV together with feline herpesvirus are thought to be responsible for

approximately 80% of all feline respiratory disease. The most common

characteristic and clinical signs of FCV infection is the development of vesicles

(ulcers) on the tongue and oral mucosa. These vesicles begin as small, individual

ulcers which may spread and affect a large part of the tongue. The vesicles usually

do not interfere with eating and drinking, and normally heal without incident. Fever

often also is observed in infected cats.

Certain strains of FCV also cause a disease in cats known as limping

syndrome. Limping syndrome is characterized by fever, joint and muscle soreness

(limping), and occasional lingual/oral ulceration. In addition, some strains of FCV

have been associated with chronic stomatitis in infected cats. Other, less common

clinical signs are conjunctivitis, rhinitis, and occasionally pneumonia. Cats infected

with FCV may become persistently infected, and may shed infectious virus for long

periods of time.

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FCV comprises a single-stranded, positive-sense RNA genome consisting of

three open reading frames (ORFs). The genome is polyadenylated at the 3' end

and bound by a virally-encoded protein at the 5'-end. The first open reading frame

encodes a viral protease and an RNA-dependent RNA polymerase, which are

expressed on a single polypeptide. This polypeptide then is post-translationally

cleaved by the viral protease. The second open reading frame encodes the major

capsid protein (i.e., the FCV capsid protein), which has six regions denoted as A-F

[Scott et al., 60 Am. J. Vet. Res.:652-658 (1999)]. Region A is cleaved to produce

the mature capsid protein. Whereas regions B, D, and F of ORF2 are relatively

conserved between FCV isolates, regions C and E are variable, with region E of

ORF2 ORF2 containing containingthe major the B-cell major epitopes B-cell [see, [see, epitopes Radford et al.,et Radford 38(2) al.,Vet Res.:319- 38(2) Vet :319-

335 (2007)]. ORF 3 encodes a minor structural protein [Sosnovtsev and Green, 277

Virology:193-203 (2000)]. Virology:193-203 (2000)].

A number of vector strategies have been employed through the years for

vaccines in an effort to protect against certain pathogens. One such vector strategy

includes the use of alphavirus-derived replicon RNA particles (RP) [Vander Veen, et

al. Anim Health Res Rev. 13(1):1-9. (2012) doi: 10.1017/S1466252312000011;

Kamrud Kamrud et etal., al.,J J GenGen Virol. . 91(Pt Virol. 7):1723-1727 91(Pt (2010)] 7):1723-1727 whichwhich (2010)] have been havedeveloped been developed

from several different alphaviruses, including Venezuelan equine encephalitis virus

(VEE) [Pushko et al., Virology 239:389-401 (1997)], Sindbis (SIN) [Bredenbeek et

al., Journal of Virology 67:6439-6446 (1993)], and Semliki Forest virus (SFV)

[Liljestrom and Garoff, Biotechnology (NY) 9:1356- 1361 (1991)]. RP vaccines

deliver propagation-defective alphavirus RNA replicons into host cells and result in

the expression of the desired antigenic transgene(s) in vivo [Pushko et al., Virology

239(2):389-401 (1997)]. RPs have an attractive safety and efficacy profile when

compared to some traditional vaccine formulations [Vander Veen, et al. Anim Health

Res Rev. 13(1):1-9. (2012) ] ]. RP The The RP platform platform has been has been used used to encode to encode pathogenic pathogenic

antigens and is the basis for several USDA-licensed vaccines for swine and poultry.

Although, long characterized as belonging to a single serotype, FCV isolates

are antigenically highly variable, and antibodies from cats vaccinated with older

vaccine strains of FCV, such as FCV F9, do not efficiently neutralize all current field

isolates. Moreover, new FCV strains associated with systemic disease and high mortality have been identified [see e.g., U.S. 7,449,323 B2]. These “virulent systemic” (VS-FCV) isolates are responsible for localized outbreaks, and current vaccines also do not appear to protect cats from disease caused by these strains. 5 This has led to concern that cats vaccinated with current vaccine strains are not fully protected from disease caused by such “antigenically heterologous” FCV strains, 2018380582

and that these heterologous strains may be responsible for outbreaks of disease, even in vaccinated cats. It is therefore desirable to develop new vaccines that stimulate more broadly reactive virus-neutralizing (VN) antibodies, and therefore 10 provide better protection against new field isolates.

The citation of any reference herein should not be construed as an admission that such reference is available as "prior art" to the instant application.

15 SUMMARY OF THE INVENTION Accordingly, in a first aspect, the present invention provides an immunogenic composition comprising a Venezuelan Equine Encephalitis (VEE) alphavirus RNA replicon particle that encodes a feline calicivirus (FCV) antigen; wherein the FCV antigen is a capsid protein. 20 In a second aspect, the present invention provides a vaccine comprising the immunogenic composition of the first aspect and a pharmaceutically acceptable carrier, wherein the vaccine elicits protective immunity to FCV infection upon administration to a feline subject. 25 In a third aspect, the present invention provides method of immunizing a feline against a pathogenic FCV comprising administering to the feline an immunologically effective amount of the vaccine of the second aspect.

30 In a fourth aspect, the present invention provides use of the immunogenic composition of the first aspect, in the manufacture of a medicament for immunizing a feline against a pathogenic FCV.

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The present invention also provides vectors that encode one or more feline calicivirus (FCV) antigens. Such vectors can be used in immunogenic compositions comprising these vectors. The immunogenic compositions of the present invention may be used in vaccines. In one aspect of the present invention, a vaccine protects 5 the vaccinated subject (e.g., mammal) against FCV. In a particular embodiment of this type, the vaccinated subject is a feline. In a more particular embodiment, the 2018380582

vaccinated subject is a domestic cat. The present invention further provides combination vaccines for eliciting protective immunity against FCV and other diseases, e.g., other infectious diseases in cats. Methods of making and using the 10 immunogenic compositions and vaccines of the present invention are also provided.

In specific embodiments, the vector is an alphavirus RNA replicon particle that encodes one or more antigens that originate from a feline pathogen. In particular embodiments, the feline pathogen is a feline calicivirus (FCV). In specific 15 embodiments of this type, the alphavirus RNA replicon particle encodes an FCV capsid protein. In related embodiments, the alphavirus RNA replicon particle encodes an antigenic fragment of an FCV capsid protein. In certain embodiments,

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the FCV capsid protein is an FCV F9-Like capsid protein. In other embodiments,

the alphavirus RNA replicon particle encodes an antigenic fragment of an FCV F9-

Like capsid protein. In yet other embodiments, the FCV capsid protein is a virulent

systemic FCV (VS-FCV) capsid protein. In still other embodiments, the alphavirus

RNA replicon particle encodes an antigenic fragment of a VS-FCV capsid protein.

In yet other embodiments, the alphavirus RNA replicon particle encodes both the

FCV F9-Like capsid protein or antigenic fragment thereof and the VS-FCV capsid

protein or an antigenic fragment thereof.

In still more particular embodiments, the alphavirus RNA replicon particle is a

Venezuelan Equine Encephalitis (VEE) alphavirus RNA replicon particle. In yet

more specific embodiments the VEE alphavirus RNA replicon particle is a TC-83

VEE alphavirus RNA replicon particle. In other embodiments, the alphavirus RNA

replicon particle is a Sindbis (SIN) alphavirus RNA replicon particle. In still other

embodiments, the alphavirus RNA replicon particle is a Semliki Forest virus (SFV)

alphavirus RNA replicon particle. In an alternative embodiment a naked DNA vector

comprises a nucleic acid construct that encodes one or more antigens that originate

from a feline pathogen. In particular embodiments of this type, the naked DNA

vectors comprise a nucleic acid construct that encodes an FCV capsid protein, or

antigenic fragment thereof.

In certain embodiments, an alphavirus RNA replicon particle of the present

invention encodes one or more FCV antigens or antigenic fragments thereof. In

particular embodiments of this type, the alphavirus RNA replicon particles encode

two to four FCV antigens or antigenic fragments thereof. In related embodiments an

alphavirus RNA replicon particle of the present invention encodes one or more FCV

antigens or antigenic fragments thereof and one or more non-FCV antigens or

antigenic fragments thereof. In specific embodiments of this type, the alphavirus

RNA replicon particles encode one or more FCV capsid proteins or antigenic

fragments thereof and one to three non-FCV antigens or antigenic fragments

thereof. In more specific embodiments, the alphavirus RNA replicon particles

encode the VS-FCV capsid protein or an antigenic fragment thereof and the

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FCV-F9-like capsid protein or an antigenic fragment thereof and one to three

non-FCV antigens or antigenic fragments thereof.

In another aspect, the present invention provides immunogenic compositions

that comprise alphavirus RNA replicon particles that encode one or more FCV

antigens or antigenic fragments thereof. In related embodiments, the immunogenic

compositions comprise alphavirus RNA replicon particles that encode two to four

FCV antigens or antigenic fragments thereof. In particular embodiments of this

type, the alphavirus RNA replicon particle encodes an FCV capsid protein. In other

embodiments, the alphavirus RNA replicon particle encodes an antigenic fragment

of an FCV capsid protein. In certain embodiments, the immunogenic compositions

comprise an alphavirus RNA replicon particle that encodes an FCV F9-Like capsid

protein. In other embodiments, the immunogenic compositions comprise an

alphavirus RNA replicon particle that encodes an antigenic fragment of an FCV F9-

Like capsid protein. In yet other embodiments, the immunogenic compositions

comprise an alphavirus RNA replicon particle that encodes a virulent systemic FCV

(VS-FCV) capsid protein. In still other embodiments, the immunogenic

compositions comprise an alphavirus RNA replicon particle that encodes an

antigenic fragment of a VS-FCV capsid protein. In yet other embodiments, the

immunogenic compositions comprise an alphavirus RNA replicon particle that

encodes both the FCV F9-Like capsid protein or antigenic fragment thereof and the

VS-FCV capsid protein or an antigenic fragment thereof. In more particular

embodiments, the immunogenic composition comprises alphavirus RNA replicon

particles that are Venezuelan Equine Encephalitis (VEE) alphavirus RNA replicon

particles. In yet more specific embodiments the immunogenic compositions

comprise VEE alphavirus RNA replicon particles that are TC-83 VEE alphavirus

RNA replicon particles.

In still other embodiments, the immunogenic composition comprises two or

more sets of alphavirus RNA replicon particles. In certain embodiments of this type,

one set of alphavirus RNA replicon particles encodes a first antigen, whereas the

other set of alphavirus RNA replicon particles encodes a second antigen. In

particular embodiments of this type, the first set of alphavirus RNA replicon particles

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encodes one or more FCV antigens or antigenic fragments thereof, and the second

set of alphavirus RNA replicon particles encode one or more FeLV antigens or

antigenic fragments thereof. In certain embodiments, the FCV antigen originates

from a virulent systemic feline calicivirus (VS-FCV) isolate. In other embodiments

the FCV antigen originates from a classical (F9-like) feline calicivirus isolate. In yet

other embodiments, the second set of alphavirus RNA replicon particles encode two

FCV antigens or antigens thereof, one of which originates from a virulent systemic

FCV isolate, whereas the other originates from a F9-like FCV. In still other

embodiments, an immunogenic composition comprises a first set of alphavirus RNA

replicon particles that encode an FCV F9-Like capsid protein or antigenic fragment

thereof and the second set of alphavirus RNA replicon particles encode a VS-FCV

capsid protein or an antigenic fragment thereof. In related embodiments, an

immunogenic composition comprises a first set of alphavirus RNA replicon particles

that encode a VS-FCV capsid protein or antigenic fragment thereof and the second

set of alphavirus RNA replicon particles encode a FeLV glycoprotein (e.g., gp85) or

an antigenic fragment thereof, (e.g., FeLV glycoprotein gp70 and/or gp45).

In yet other embodiments, the immunogenic composition comprises one set

of alphavirus RNA replicon particles that encode a first antigen, another set of

alphavirus RNA replicon particles that encode a second antigen, and a third set of

alphavirus RNA replicon particles that encode a third antigen. In a particular

embodiment of this type, the first set of alphavirus RNA replicon particles encode an

FCV antigen (e.g., the capsid protein) which originates from a classical (F9-like)

feline calicivirus or an antigenic fragment thereof, the second set of alphavirus RNA

replicon particles encode an FCV antigen (e.g., the capsid protein), which originates

from a virulent systemic feline calicivirus or an antigenic fragment thereof, and the

third set of alphavirus RNA replicon particles encode a FeLV antigen (e.g., the FeLV

gp85) or an antigenic fragment thereof.

Accordingly, in particular embodiments in which the immunogenic

compositions comprise multiple sets (e.g., 2-10) of alphavirus RNA replicon

particles, in which the first set of alphavirus RNA replicon particles encodes an FCV

F9-Like capsid protein or antigenic fragment thereof and/or a VS-FCV capsid protein

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or an antigenic fragment thereof, and the one or more other sets of alphavirus RNA

replicon particles encode one or more non-FCV antigens.

In certain embodiments of this type, the non-FCV antigen or antigenic

fragment thereof is a protein antigen that originates from feline herpesvirus (FHV).

In other embodiments, the non-FCV antigen is a protein antigen that originates from

feline leukemia virus (FeLV). In yet other embodiments, the non-FCV antigen is a

protein antigen that originates from feline pneumovirus (FPN). In still other

embodiments, the non-FCV antigen is a protein antigen that originates from feline

parvovirus (FPV). In yet other embodiments, the non-FCV antigen is a protein

antigen that originates from rabies virus. In still other embodiments, the non-FCV

antigen is a protein antigen that originates from feline infectious peritonitis virus

(FIPV). In yet other embodiments, the non-FCV antigen is a protein antigen that

originates from feline immunodeficiency virus. In still other embodiments, the non-

FCV antigen is a protein antigen that originates from borna disease virus (BDV). In

yet other embodiments, the non-FCV antigen is a protein antigen that originates

from feline influenza virus. In still other embodiments, the non-FCV antigen is a

protein antigen that originates from feline panleukopenia virus (FPLV). In yet other

embodiments the non-FCV antigen is a protein antigen that originates from feline

coronavirus (FCoV). In still other embodiments the non-FCV antigen is a protein

antigen that originates from feline rhinotracheitis virus (FVR). In yet other

embodiments the non-FCV antigen is a protein antigen that originates from

Chlamydophila felis.

The present invention also includes all of the alphavirus RNA replicon

particles of the present invention, the naked DNA vectors, the nucleic acid

constructs of the present invention including synthetic messenger RNA, and RNA

replicons, as well as all of the immunogenic compositions and/or vaccines that

comprise the nucleic acid constructs (e.g., synthetic messenger RNA, RNA

replicons), the alphavirus RNA replicon particles, and/or the naked DNA vectors of

the present invention.

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In particular embodiments, a nucleic acid construct of the present invention

encodes one or more FCV antigens or antigenic fragments thereof. In related

embodiments of this type, the nucleic acid construct encodes two to four FCV

antigens or antigenic fragments thereof. In other embodiments, alphavirus RNA

replicon particles comprise a nucleic acid construct that encodes one or more FCV

antigens or antigenic fragments thereof. In particular embodiments, alphavirus RNA

replicon particles comprise a nucleic acid construct that encodes two to four FCV

antigens or antigenic fragments thereof.

In still other embodiments, the immunogenic compositions comprise

alphavirus RNA replicon particles and/or naked DNA vectors that comprise a nucleic

acid construct that encodes two to four FCV antigens or antigenic fragments thereof.

In particular embodiments of this type, the alphavirus RNA replicon particles encode

an FCV F9-Like capsid protein or antigenic fragment thereof and/or a VS-FCV

capsid protein or an antigenic fragment thereof and an FeLV glycoprotein (gp85) or

an antigenic fragment thereof. In particular embodiments of this type, the antigenic

fragment of gp85 is the FeLV glycoprotein gp70. In other related embodiments, the

antigenic fragment of gp85 is the FeLV glycoprotein gp45. In more particular

embodiments, the immunogenic composition comprises alphavirus RNA replicon

particles that are Venezuelan Equine Encephalitis (VEE) alphavirus RNA replicon

particles. In yet more specific embodiments the VEE alphavirus RNA replicon

particles are TC-83 VEE alphavirus RNA replicon particles.

In yet other embodiments, the immunogenic composition comprises two or

more sets of alphavirus RNA replicon particles and/or naked DNA vectors. In

particular embodiments of this type, one set of alphavirus RNA replicon particles

and/or naked DNA vectors comprise a first nucleic acid construct, whereas the other

set of alphavirus RNA replicon particles and/or naked DNA vectors comprise a

second nucleic acid construct. In a specific embodiment of this type the first nucleic

acid construct encodes an FCV antigen or an antigenic fragment thereof, and the

second nucleic acid construct encodes a feline calicivirus (FCV) antigen or an

antigenic fragment thereof. In certain embodiments of this type, the FCV antigen

originates from a virulent systemic feline calicivirus (VS-FCV) isolate. In other

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embodiments the FCV antigen originates from a classical (F9-like) feline calicivirus

isolate. In yet other embodiments, the second nucleic acid construct encodes two

FCV antigens, one of which originates from a virulent systemic feline calicivirus

isolate, whereas the other originates from a classical (F9-like) feline calicivirus

isolate.

In still other embodiments, the immunogenic composition comprises one set

of alphavirus RNA replicon particles and/or naked DNA vectors that comprise a first

nucleic acid construct, another set of alphavirus RNA replicon particles and/or

naked DNA vectors that comprise a second nucleic acid construct, and a third set of

alphavirus RNA replicon particles and/or naked DNA vectors that comprise a third

nucleic acid construct. In a particular embodiment of this type, the first nucleic acid

construct encodes a FeLV antigen or an antigenic fragment thereof, the second

nucleic acid construct encodes a feline calicivirus (FCV) antigen which originates

from a virulent systemic feline calicivirus or an antigenic fragment thereof, and the

third nucleic acid construct encodes a feline calicivirus (FCV) antigen which

originates from a classical (F9-like) feline calicivirus or an antigenic fragment

thereof.

In yet other embodiments, the immunogenic composition comprises one set

of alphavirus RNA replicon particles and/or naked DNA vectors that comprise a first

nucleic acid construct, another set of alphavirus RNA replicon particles and/or

naked DNA vectors that comprise a second nucleic acid construct, a third set of

alphavirus RNA replicon particles and/or naked DNA vectors that comprise a third

nucleic acid construct, and a fourth set of alphavirus RNA replicon particles and/or

naked DNA vectors that comprise a fourth nucleic acid construct. In particular

embodiments of this type, the first nucleic acid construct encodes both a feline

calicivirus (FCV) antigen which originates from a virulent systemic feline calicivirus

or an antigenic fragment thereof, and a feline calicivirus (FCV) antigen which

originates from a classical (F9-like) feline calicivirus or an antigenic fragment

thereof.

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In still other embodiments, the immunogenic composition comprises a set of

alphavirus RNA replicon particles and/or naked DNA vectors that comprise a first

nucleic acid construct, another set of alphavirus RNA replicon particles and/or

naked DNA vectors that comprise a second nucleic acid construct, a third set of

alphavirus RNA replicon particles and/or naked DNA vectors that comprise a third

nucleic acid construct, a fourth set of alphavirus RNA replicon particles and/or

naked DNA vectors that comprise a fourth nucleic acid construct ,and a fifth set of

alphavirus RNA replicon particles and/or naked DNA vectors that comprise a fifth

nucleic acid construct. In such embodiments, the nucleotide sequences of the first

nucleic acid construct, the second nucleic acid construct, third nucleic acid

construct, the fourth nucleic acid construct, and the fifth nucleic acid construct are all

different. In particular embodiments of this type, the first nucleic acid construct

encodes both a feline calicivirus (FCV) antigen which originates from a virulent

systemic feline calicivirus or an antigenic fragment thereof, and a feline calicivirus

(FCV) antigen which originates from a classical (F9-like) feline calicivirus or an

antigenic fragment thereof.

Accordingly, an immunogenic composition of the present invention can

contain alphavirus RNA replicon particles and/or naked DNA vectors that comprise

a nucleic acid construct that encodes at least one non-FCV antigen for eliciting

protective immunity to a non-FCV pathogen. In particular embodiments of this type,

the non-FCV antigen is a protein antigen that originates from feline herpesvirus

(FHV). In other embodiments, the non-FCV antigen is a protein antigen that

originates from feline leukemia virus (FeLV). In yet other embodiments, the non-

FCV antigen is a protein antigen that originates from feline pneumovirus (FPN). In

still other embodiments, the non-FCV antigen is a protein antigen that originates

from feline parvovirus (FPV). In yet other embodiments, the non-FCV antigen is a a protein antigen that originates from feline infectious peritonitis virus (FIPV). In still

other embodiments, the non-FCV antigen is a protein antigen that originates from

feline immunodeficiency virus. In yet other embodiments, the non-FCV antigen is a

protein antigen that originates from rabies virus. In still other embodiments, the non-

FCV antigen is a protein antigen that originates from borna disease virus (BDV). In

yet other embodiments, the non-FCV antigen is a protein antigen that originates wo 2019/110213 WO PCT/EP2018/080096

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from feline influenza virus. In still other embodiments, the non-FCV antigen is a

protein antigen that originates from feline panleukopenia virus (FPLV). In yet other

embodiments the non-FCV antigen is a protein antigen that originates from feline

coronavirus (FCoV). In still other embodiments the non-FCV antigen is a protein

antigen that originates from feline rhinotracheitis virus (FVR). In still other

embodiments the non-FCV antigen is a protein antigen that originates from

Chlamydophila felis.

The present invention further provides combination immunogenic compositions

and/or vaccines (multivalent vaccines) that include alphavirus RNA replicon particles

that encode one or more antigens or antigenic fragments thereof originating from

FCV together (e.g., the FCV capsid protein) and further comprise one or more

modified live/attenuated or killed feline pathogens. In particular embodiments, the

immunogenic compositions further comprise a modified live or killed Chlamydophila

felis combined with alphavirus RNA replicon particles that encode an antigen or

antigenic fragment thereof originating from FeLV. In other embodiments, the

immunogenic compositions further comprise a modified live or killed feline

rhinotracheitis Virus (FVR) combined with alphavirus RNA replicon particles that

encode an antigen or antigenic fragment thereof originating from FeLV. In yet other

embodiments, the immunogenic compositions further comprise a modified live or

killed feline panleukopenia virus (FPL) combined with alphavirus RNA replicon

particles that encode an antigen or antigenic fragment thereof originating from

FeLV. In certain embodiments, a vaccine comprises an immunologically effective

amount of one or more of these immunogenic compositions.

In more specific embodiments, the immunogenic compositions comprise

alphavirus RNA replicon particles that encode a capsid protein or antigenic fragment

thereof originating from VS-FCV and further comprise a modified live or killed

F9-like FCV. In still other embodiments, the immunogenic compositions comprise

alphavirus RNA replicon particles that encode a capsid protein or antigenic fragment

thereof originating from VS-FCV and further comprise a modified live or killed

F9-like FCV, a modified live or killed Chlamydophila felis, a modified live or killed

FVR, and a modified live or killed FPL. In related embodiments, the immunogenic composition also comprises alphavirus RNA replicon particles that encode an antigen or antigenic fragment thereof originating from FeLV. In particular embodiments of this type, the feline antigen of the FeLV is the FeLV viral glycoprotein (gp85). In certain embodiments, the present invention provides vaccines that comprise an immunologically effective amount of one or more of these immunogenic compositions.

In particular embodiments, an alphavirus RNA replicon particle of the present

invention encodes a VS-FCV capsid protein or antigenic fragment thereof. In

specific embodiments of this type, the VS-FCV capsid protein comprises an amino

acid sequence comprising 95% identity or more with the amino acid sequence of

SEQ ID NO: 2. In more specific embodiments of this type, the VS-FCV capsid

protein comprises an amino acid sequence comprising 98% identity or more with the

amino acid sequence of SEQ ID NO: 2. In even more specific embodiments of this

type, the VS-FCV capsid protein comprises the amino acid sequence of SEQ ID

NO: 2. In specific embodiments of this type the VS-FCV capsid protein is encoded

by the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 12.

In related embodiments, an alphavirus RNA replicon particle of the present

invention encodes invention encodes a FCV a FCV F9-Like F9-Like capsid capsid protein protein or antigenic or antigenic fragmentfragment thereof. thereof. In In

specific embodiments of this type, the FCV F9-Like capsid protein comprises an

amino acid sequence comprising 95% identity or more with the amino acid

sequence of SEQ ID NO: 4. In more specific embodiments of this type, the FCV

F9-Like capsid protein comprises an amino acid sequence comprising 98% identity

or more with the amino acid sequence of SEQ ID NO: 4. In even more specific

embodiments of this type, the FCV F9-Like capsid protein comprises the amino acid

sequence of SEQ ID NO: 4. In specific embodiments of this type the FCV F9-Like

capsid protein is encoded by the nucleotide sequence of SEQ ID NO: 3 or SEQ ID

NO: 13.

In certain embodiments an alphavirus RNA replicon particle of the present

invention encodes a FeLV glycoprotein (gp85). In specific embodiments of this

type, the FeLV glycoprotein gp85 comprises an amino acid sequence comprising

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95% identity or more with the amino acid sequence of SEQ ID NO: 6. In more

specific embodiments of this type, the FeLV glycoprotein (gp85) comprises the

amino acid sequence of SEQ ID NO: 6. In even more specific embodiments of this

type the FeLV glycoprotein (gp85) is encoded by the nucleotide sequence of SEQ

ID NO: 5 or SEQ ID NO: 14.

In related embodiments, the FeLV glycoprotein gp70 comprises an amino

acid sequence comprising 95% identity or more with the amino acid sequence of

SEQ ID NO: 8. In more specific embodiments of this type, the FeLV glycoprotein

(gp85) comprises the amino acid sequence of SEQ ID NO: 8. In even more specific

embodiments of this type the FeLV glycoprotein (gp70) is encoded by the nucleotide

sequence of SEQ ID NO: 7 or SEQ ID NO: 15.

In yet other embodiments an alphavirus RNA replicon particle of the present

invention encodes a rabies virus glycoprotein (G). In specific embodiments of this

type, the rabies virus glycoprotein comprises an amino acid sequence comprising

95% identity or more with the amino acid sequence of SEQ ID NO: 10. In more

specific embodiments of this type, the rabies virus glycoprotein (G) comprises the

amino acid sequence of SEQ ID NO: 10. In even more specific embodiments of this

type the rabies virus glycoprotein (G) is encoded by the nucleotide sequence of

SEQ ID NO: 9 or SEQ ID NO: 16.

The present invention further comprises vaccines, including multivalent

vaccines, comprising the immunogenic compositions of the present invention. In

particular embodiments, the vaccines are nonadjuvanted vaccine. In certain

embodiments, the vaccine aids in the prevention of disease due to FCV. In related

embodiments, antibodies are induced in a feline subject when the feline is immunized

with the vaccine.

The present invention also provides methods of immunizing a feline against a

feline pathogen, e.g., FCV, comprising administering to the feline an immunologically

effective amount of a vaccine or multivalent of the present invention. In particular

embodiments the vaccine is administered via intramuscular injection. In alternative

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embodiments the vaccine is administered via subcutaneous injection. In other

embodiments the vaccine is administered via intravenous injection. In still other

embodiments the vaccine is administered via intradermal injection. In yet other

embodiments the vaccine is administered via oral administration. In still other

embodiments the vaccine is administered via intranasal administration. In specific

embodiments, the feline is a domestic cat.

The vaccines and multivalent vaccines of the present invention can be

administered as a primer vaccine and/or as a booster vaccine. In specific

embodiments, a vaccine of the present invention is administered as a one shot vaccine

(one dose), without requiring subsequent administrations. In certain embodiments, in

the case of the administration of both a primer vaccine and a booster vaccine, the

primer vaccine and the booster vaccine can be administered by the identical route. In

certain embodiments of this type, the primer vaccine and the booster vaccine are both

administered by subcutaneous injection. In alternative embodiments, in the case of the

administration of both a primer vaccine and a booster vaccine, the administration of the

primer vaccine can be performed by one route and the booster vaccine by another

route. In certain embodiments of this type, the primer vaccine can be administered by

subcutaneous injection and the booster vaccine can be administered orally.

The invention further provides for a method of immunizing a feline against

FCV comprising injecting the feline with an immunologically effective amount of the

above described vaccines. In particular embodiments the vaccines can include from about about 11X X104 10 to toabout about1 1 X 10 10 RPs X 10¹ RPsororhigher, forfor higher, example. In more example. particular In more particular

embodiments the vaccines can include from about 1 X 105 toabout 10 to about11XX10 109 RPs. RPs. InIn

even more particular embodiments the vaccines can include from about 1 X 106 to 10 to

about 1 X 108 RPs. In 10 RPs. In particular particular embodiments embodiments the the feline feline is is aa domestic domestic cat. cat.

In particular embodiments the vaccines of the present invention are

administered in 0.05 mL to 3 mL doses. In more particular embodiments the dose

administered is 0.1 mL to 2 mLs. In still more particular embodiments the dose

administered is 0.2 mL to 1.5 mLs. In yet other embodiments the dose administered

is 0.5 mL to 2.0 mLs. In still other embodiments the dose administered is 0.3 to 1.0

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mLs. In yet more particular embodiments the dose administered is 0.4 ml mL to 0.8

mLs. mLs.

These and other aspects of the present invention will be better appreciated

by reference to the following Detailed Description.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides efficacious, safe FCV vaccines. In particular

embodiments the vaccine is nonadjuvanted. In one aspect, the vaccines of the

present invention do not induce feline injection-site sarcomas, yet still aid in the

protection of the vaccinates from the upper respiratory disease and/or limping

syndrome caused by FCV infection. In a particular embodiment, the FCV capsid

protein originates from a virulent systemic FCV (VS-FCV). In related embodiments,

the FCV capsid protein originates from an older strain, such as an FCV F9 strain

(F9-Like FCV).

Accordingly, the vaccine compositions of the present invention include an

immunologically effective amount of a vector encoding an antigen from one or more

strains of feline calicivirus that aids in eliciting protective immunity in the recipient

vaccinated animal. Furthermore, the present invention provides new immunogenic

compositions to improve the reliability of the vaccination to aid in the reduction of

upper respiratory disease in cats in a feline infected by FCV and to thereby yield

more transient or mild disease and/or lead to the reduction of the infection. In a

particular aspect of the present invention, the vaccines comprise an alphavirus RNA

replicon particle (RP) encoding an FCV capsid, e.g., originating from a VS-FCV or

alternatively, a classical strain, such as an FCV F9-Like strain.

In more specific embodiments, the vaccines comprise alphavirus RNA

replicon particles (RPs) that comprise the capsid protein and glycoproteins of

Venezuelan Equine Encephalitis Virus (VEE) and encode the FCV capsid protein

and/or an antigenic fragment thereof. In even more specific embodiments, the

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vaccines comprise alphavirus RNA replicon particles (RPs) that comprise the capsid

protein and glycoproteins of the avirulent TC-83 strain of VEE and encode one or

more FCV capsid proteins and/or one or more antigenic fragments thereof.

In another aspect of the present invention, the vaccines comprise naked DNA

vectors that encode one or more FCV capsid proteins and/or one or more antigenic

fragments thereof. The vaccines of the present invention can be administered to a

feline in the absence of an adjuvant and still effectively aid in the protection of the

vaccinated feline against FCV.

In order to more fully appreciate the invention, the following definitions are

provided. provided.

The use of singular terms for convenience in description is in no way

intended to be so limiting. Thus, for example, reference to a composition

comprising "a polypeptide" includes reference to one or more of such polypeptides.

In addition, reference to an "alphavirus RNA replicon particle" includes reference to

a plurality of such alphavirus RNA replicon particles, unless otherwise indicated.

As used herein the term "approximately" is used interchangeably with the

term "about" and signifies that a value is within fifty percent of the indicated value

i.e., a composition containing "approximately" 1 X 108 alphavirus RNA 10 alphavirus RNA replicon replicon

particles per milliliter contains from 0.5 X 108 to 1.5 10 to 1.5 XX 10 108 alphavirus alphavirus RNA RNA replicon replicon

particles per milliliter.

As used herein, the term "feline" refers to any member of the Felidae family.

Domestic cats, pure-bred and/or mongrel companion cats, and wild or feral cats are

all felines.

As used herein, the term "replicon" refers to a modified RNA viral genome

that lacks one or more elements (e.g., coding sequences for structural proteins) that

if they were present, would enable the successful propagation of the parental virus

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in cell cultures or animal hosts. In suitable cellular contexts, the replicon will amplify

itself and may produce one or more sub-genomic RNA species.

As used herein, the term "alphavirus RNA replicon particle", abbreviated

"RP", is an alphavirus-derived RNA replicon packaged in structural proteins, e.g.,

the capsid and glycoproteins, which also are derived from an alphavirus, e.g., as

described by Pushko et al., [Virology 239(2):389-401 (1997)]. An RP cannot

propagate in cell cultures or animal hosts (without a helper plasmid or analogous

component), because the replicon does not encode the alphavirus structural

components (e.g., capsid and glycoproteins).

The terms "FCV F9-Like" and "F9-Like FCV" are used interchangeably with

each other and with the term "classical FCV" and as used herein is an FCV that can

be characterized as an older and formerly, universal vaccine strain of FCV, for

which the FCV F9 strain is considered a typical representative. In direct contrast,

the FCV termed virulent systemic "VS-FCV" or as used herein interchangeably "(VS)

FCV", is a newer class of FCV, which is unusually virulent, and cannot be

neutralized by antibodies raised against the FCV F9-Like strains [see,

U.S. 7,449,323; Radford et al., 38(2) Vet res. 319-335 (2007)].

The term "non-FCV", is used to modify terms such as pathogen, and/or

antigen (or immunogen) to signify that the respective pathogen, and/or antigen (or

immunogen) is neither an FCV pathogen nor an FCV antigen (or immunogen) and

that a non-FCV protein antigen (or immunogen) does not originate from an FCV.

The terms "originate from", "originates from" and "originating from" are used

interchangeably with respect to a given protein antigen and the pathogen or strain of

that pathogen that naturally encodes it, and as used herein signify that the

unmodified and/or truncated amino acid sequence of that given protein antigen is

encoded by that pathogen or strain of that pathogen. The coding sequence, within a

nucleic acid construct of the present invention for a protein antigen originating from

a pathogen may have been genetically manipulated so as to result in a modification

and/or truncation of the amino acid sequence of the expressed protein antigen

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relative to the corresponding sequence of that protein antigen in the pathogen or

strain of pathogen (including naturally attenuated strains) it originates from.

As used herein, the terms "protecting", or "providing protection to", or

"eliciting protective immunity to", "aids in prevention of disease", and "aids in the

protection" do not require complete protection from any indication of infection. For

example, "aids in the protection" can mean that the protection is sufficient such that,

after challenge, symptoms of the underlying infection are at least reduced, and/or

that one or more of the underlying cellular, physiological, or biochemical causes or

mechanisms causing the symptoms are reduced and/or eliminated. It is understood

that "reduced," as used in this context, means relative to the state of the infection,

including the molecular state of the infection, not just the physiological state of the

infection.

As used herein, a "vaccine" is a composition that is suitable for application to

an animal, e.g., feline (including, in certain embodiments, humans, while in other

embodiments being specifically not for humans) comprising one or more antigens

typically combined with a pharmaceutically acceptable carrier such as a liquid

containing water, which upon administration to the animal induces an immune

response strong enough to minimally aid in the protection from a disease arising

from an infection with a wild-type micro-organism, i.e., strong enough for aiding in

the prevention of the disease, and/or preventing, ameliorating or curing the disease.

As used herein, a multivalent vaccine is a vaccine that comprises two or

more different antigens. In a particular embodiment of this type, the multivalent

vaccine stimulates the immune system of the recipient against two or more different

pathogens.

The terms "adjuvant" and "immune stimulant" are used interchangeably

herein, and are defined as one or more substances that cause stimulation of the

immune system. In this context, an adjuvant is used to enhance an immune

response responsetotoone or or one more vaccine more antigens/isolates. vaccine Accordingly, antigens/isolates. "adjuvants" Accordingly, are "adjuvants" are

agents that nonspecifically increase an immune response to a particular antigen,

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thus reducing the quantity of antigen necessary in any given vaccine, and/or the

frequency of injection necessary in order to generate an adequate immune

response to the antigen of interest. In this context, an adjuvant is used to enhance

an immune response to one or more vaccine antigens/isolates. The American

Association of Feline Practitioners Feline Vaccination Guidelines, for example,

suggest the use of nonadjuvanted FeLV vaccines [AAFP Feline Advisory Panel, 15:

785-808 (2013)].

As used herein, a "nonadjuvanted vaccine" is a vaccine or a multivalent

vaccine that does not contain an adjuvant.

As used herein, the term "pharmaceutically acceptable" is used adjectivally to

mean that the modified noun is appropriate for use in a pharmaceutical product.

When it is used, for example, to describe an excipient in a pharmaceutical vaccine,

it characterizes the excipient as being compatible with the other ingredients of the

composition and not disadvantageously deleterious to the intended recipient animal,

e.g., feline.

Parenteral administration" includes subcutaneous injections, submucosal

injections, intravenous injections, intramuscular injections, intradermal injections,

and infusion.

As used herein the term "antigenic fragment" in regard to a particular protein

(e.g., a protein antigen) is a fragment of that protein that is antigenic, i.e., capable of

specifically interacting with an antigen recognition molecule of the immune system,

such as an immunoglobulin (antibody) or T cell antigen receptor. For example, an

antigenic fragment of an FCV capsid protein is a fragment of the capsid protein that

is antigenic. In specific embodiments, the antigenic fragment of an FCV capsid

protein comprises region E of the ORF2, which contains the major B-cell epitopes.

Preferably, an antigenic fragment of the present invention is immunodominant for

antibody and/or T cell receptor recognition. In particular embodiments, an antigenic

fragment with respect to a given protein antigen is a fragment of that protein that

retains at least 25% of the antigenicity of the full length protein. In preferred

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embodiments an antigenic fragment retains at least 50% of the antigenicity of the

full length protein. In more preferred embodiments, an antigenic fragment retains at

least 75% of the antigenicity of the full length protein. Antigenic fragments can be

as small as 20 amino acids or at the other extreme, be large fragments that are

missing as little as a single amino acid from the full-length protein. In particular

embodiments the antigenic fragment comprises 25 to 150 amino acid residues. In

other embodiments, the antigenic fragment comprises 50 to 250 amino acid

residues. For FeLV, for example, the FeLV gp45 glycoprotein and the FeLV gp70

glycoprotein are antigenic fragments of the FeLV gp85 glycoprotein.

As used herein one amino acid sequence is 100% "identical" or has 100%

"identity" to a second amino acid sequence when the amino acid residues of both

sequences are identical. Accordingly, an amino acid sequence is 50% "identical" to to

a second amino acid sequence when 50% of the amino acid residues of the two

amino acid sequences are identical. The sequence comparison is performed over a

contiguous block of amino acid residues comprised by a given protein, e.g., a

protein, or a portion of the polypeptide being compared. In a particular embodiment,

selected deletions or insertions that could otherwise alter the correspondence

between the two amino acid sequences are taken into account.

As used herein, nucleotide and amino acid sequence percent identity can be

determined using C, MacVector (MacVector, Inc. Cary, NC 27519), Vector NTI

(Informax, Inc. MD), Oxford Molecular Group PLC (1996) and the Clustal W

algorithm with the alignment default parameters, and default parameters for identity.

These commercially available programs can also be used to determine sequence

similarity using the same or analogous default parameters. Alternatively, an

Advanced Blast search under the default filter conditions can be used, e.g., using

the GCG (Genetics Computer Group, Program Manual for the GCG Package,

Version 7, Madison, Wisconsin) pileup program using the default parameters.

As used herein, the term "inactivated" microorganism is used interchangeably

with the term "killed" microorganism. For the purposes of this invention, an

"inactivated" microorganism is an organism which is capable of eliciting an immune

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response in an animal, but is not capable of infecting the animal. An antigen of the

present invention (e.g., an inactivated feline calicivirus) may be inactivated by an

agent selected from the group consisting of binary ethyleneimine, formalin, beta-

propiolactone, thimerosal, or heat. In a particular embodiment, inactivated feline

calicivirus isolates combined with an RP of the present invention are inactivated by

binary ethyleneimine.

The alphavirus RNA replicon particles of the present invention may be

lyophilized and rehydrated with a sterile water diluent. On the other hand, when the

alphavirus RNA replicon particles are stored separately, but intended to be mixed

with other vaccine components prior to administration, the alphavirus RNA replicon

particles can be stored in the stabilizing solution of those components, e.g., a high

sucrose solution.

A vaccine of the present invention can be readily administered by any

standard route including intravenous, intramuscular, subcutaneous, oral, intranasal,

intradermal, and/or intraperitoneal vaccination. The skilled artisan will appreciate

that the vaccine composition is preferably formulated appropriately for each type of

recipient animal and route of administration.

Thus, the present invention also provides methods of immunizing a feline

against FCV and/or other feline pathogens. One such method comprises injecting a

feline with an immunologically effective amount of a vaccine of the present

invention, so that the feline produces appropriate FCV antibodies.

Multivalent Vaccines:

The present invention also provides multivalent vaccines. For example, the

coding sequence of a protein antigen or antigenic fragment thereof, or combination

of such coding sequences of protein antigens useful in a feline vaccine can be

added to an alphavirus RNA replicon particle (RP) or combined in the same RP as

one that encodes a feline antigen of the FCV [e.g., the FCV capsid protein] in the

vaccine. In specific embodiments, the alphavirus RNA replicon particle encodes

both the FCV F9-Like capsid protein or an antigenic fragment thereof and the

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VS-FCV capsid protein or an antigenic fragment thereof and encodes a non-FCV

antigen. Accordingly, such multivalent vaccines are included in the present

invention.

Examples of pathogens that one or more of such protein antigens can

originate from include feline rhinotracheitis Virus (FVR), feline leukemia virus

(FeLV), feline panleukopenia Virus (FPL) feline herpesvirus (FHV), other FCV

strains, feline parvovirus (FPV), feline infectious peritonitis virus (FIPV), feline

immunodeficiency virus, borna disease virus (BDV), rabies virus, feline influenza

virus, canine influenza virus, avian influenza, canine pneumovirus, feline

pneumovirus, Chlamydophila felis (FKA Chlamydia psittaci), Bordetella

bronchiseptica, and Bartonella spp. (e.g., B. henselae). In particular embodiments,

a coding sequence for a capsid protein or analogous protein from one or more of

these feline or canine pathogens can be inserted into the same RP as the FCV

antigen. Alternatively, or in combination therewith, a coding sequence for a capsid

protein or analogous protein from one or more of these feline or canine pathogens

can be inserted into one or more other RPs, which can be combined in a vaccine

with an RP that encodes the FCV F9-Like capsid protein or an antigenic fragment

thereof and/or the VS-FCV capsid protein or an antigenic fragment thereof.

In addition, an alphavirus RNA replicon particle(RP) that encodes one or more

antigens of the FCV [e.g., the FCV F9-Like capsid protein or an antigenic fragment

thereof and/or the VS-FCV capsid protein or an antigenic fragment thereof] can be

added together with one or more other live, attenuated virus isolates, e.g., a live

attenuated FCV F9-Like virus (e.g., modified live FCV F9) and/or a live attenuated

feline herpesvirus and/or a live attenuated feline parvovirus and/or a live, attenuated

feline leukemia virus, and/or a live, attenuated feline infectious peritonitis virus

and/or a live, attenuated feline immunodeficiency virus and/or a live, attenuated

borna disease virus and/or a live, attenuated rabies virus, and/or a live, attenuated

feline influenza virus and/or a live, attenuated canine influenza virus, and/or a live,

attenuated avian influenza, and/or a live, attenuated canine pneumovirus, and/or a

live, attenuated feline pneumovirus. In addition, a live, attenuated Chlamydophila

felis, and/or a live, attenuated Bordetella bronchiseptica and/or a live, attenuated

Bartonella spp. (e.g., B. henselae) can also be included in such multivalent

vaccines.

Furthermore, an alphavirus RNA replicon particle (RP) that encodes one or

more antigens of the FCV [e.g., the FCV F9-Like capsid protein or an antigenic

fragment thereof and/or the VS-FCV capsid protein or an antigenic fragment thereof]

can be added together with one or more other killed virus isolates such as a killed

FCV strain, and/or a killed feline herpesvirus and/or a killed feline parvovirus and/or

a killed feline leukemia virus, and/or a killed feline infectious peritonitis virus and/or a

killed feline immunodeficiency virus and/or a killed borna disease virus and/or a

killed rabies virus, and/or a killed feline influenza virus and/or a killed canine

influenza virus, and/or a killed avian influenza virus, and/or a killed canine

pneumovirus, and/or a killed feline pneumovirus. In addition, bacterins (or

subfractions of the bacterins, e.g., the pilus subfraction) of Chlamydophila felis,

and/or Bordetella bronchiseptica and/or Bartonella spp. (e.g., B. henselae) can also

be included in such multivalent vaccines.

It also is to be understood that this invention is not limited to the particular

configurations, process steps, and materials disclosed herein as such

configurations, process steps, and materials may vary somewhat. It further is to be

understood that the terminology employed herein is used for the purpose of

describing particular embodiments only and is not intended to be limiting, since the

scope of the present invention will be limited only by the appended claims and

equivalents thereof.

SEQUENCE SEQUENCE TABLE TABLE

SEQ ID NO: Description Type Description 1 Feline Calicivirus (VS-FCV) nucleic acid

DNA 2 Feline Calicivirus (VS-FCV) amino acid

3 Feline Calicivirus (F9-like) nucleic acid

DNA 4 Feline Calicivirus (F9-like) amino acid

5 FeLV viral glycoprotein (gp85) nucleic acid

DNA 6 FeLV viral glycoprotein (gp85) amino acid

7 FeLV viral glycoprotein (gp70) nucleic acid

DNA 8 FeLV viral glycoprotein (gp70) amino acid

9 Rabies virus Glycoprotein nucleic acid

DNA 10 Rabies virus Glycoprotein amino acid

11 nucleic acid GGCGCGCCGCACC 12 Feline Calicivirus (VS-FCV) nucleic acid

RNA 13 13 Feline Calicivirus (F9-like) nucleic acid

RNA 14 FeLV viral glycoprotein (gp85) nucleic acid

RNA 15 FeLV viral glycoprotein (gp70) nucleic acid

RNA 16 16 Rabies virus Glycoprotein nucleic acid

RNA nucleic acid TTAATTAA

SEQUENCES Feline Calicivirus capsid (VS-FCV) SEQ ID NO: 1 atggctgacgacggatctgtgaccaccccagaacaaggaacaatggtcggaggagtgatt atggctgacgacggatctgtgaccaccccagaacaaggaacaatggtcggaggagtgatt gccgaacccagcgctcagatgtcaactgcggcggacatggcctccggaaagtcggtggac tccgagtgggaagccttcttctcgttccacacgtccgtgaactggagcacctccgaaaco ccgagtgggaagccttcttctcgttccacacgtccgtgaactggagcacctccgaaaco caaggaaagatcctcttcaagcagtccctgggtcccctgctgaacccgtacctggagcac caaggaaagatcctcttcaagcagtccctgggtcccctgctgaacccgtacctggagcac atcagcaagctgtacgtcgcttggagcgggtcgatcgaagtgcgattttccatctcggga atcagcaagctgtacgtcgcttggagcgggtcgatcgaagtgcgattttccatctoggga agcggcgtgttcggtggtaaactggccgccatcgtcgtgccgcctggtgtcgaccctgtc agcggcgtgttcggtggtaaactggccgccatcgtcgtgccgcctggtgtcgaccctgtc cagtcaacctccatgctgcagtacccgcacgtcctgttcgacgcaagacaagtggagcc. cagtcaacctccatgctgcagtacccgcacgtcctgttcgacgcaagacaagtggagcca gtgatcttctccatcccggacctccgcaacagcctgtatcacttgatgtccgataccgat gtgatcttctccatcccggacctccgcaacagcctgtatcacttgatgtccgataccgat accacttccctcgtgatcatggtgtacaacgatctgatcaacccgtacgccaatgactce accacttccctcgtgatcatggtgtacaacgatctgatcaacccgtacgccaatgactc aacagctcgggttgcatcgtgaccgtcgaaacgaagcctggcatcgatttcaagtttcat aacagctcgggttgcatcgtgaccgtcgaaacgaagcctggcatcgatttcaagtttcat ctgctgaaaccgcccggatccatgcttactcacgggtccatcccttccgatctgatccco aagagctcctccctgtggattgggaaccgccactggaccgatattaccgatttcgtgatt aagagctcctccctgtggattgggaaccgccactggaccgatattaccgatttcgtgatt

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aggcctttcgtgttccaagccaaccggcacttcgacttcaaccaggagactgccggctgg tcaactccacggttccgcccattggccgtgactgtgtcgcagtcaaagggagccaagct ggaacggcatcgccaccgactacattgtgcctggaatccccgacggatggcctgatact accatccccaccaagctgacccctaccggagattacgccatcacctcctccgacggcaat $ gatattgaaaccaagctggaatacgagaacgcggacgtgattaagaacaacaccaactt cgctccatgtatatctgcggaagcctccagagggcttggggcgacaagaagatcagcaad ccgggttcatcactaccggagtgatttctgacaactccatcagcccttcgaaca gaccagtccaagatcgtggtgtaccaggacaaccatgtcaattcggaggtccagactag gacatcactcttgccatcctgggctacaccggaattggagaagaggccataggcgccaac OI agggactccgtcgtgagaatttccgtgcttccggaaactggagcaaggggcggaaatc accatcttctacaaaaattccatgaagctgggctacgtgatctcctccattgacgtgtt aactcccaaatcctccacacctcgcgccagctgtcactgaacaactacttgttgccccct gactccttcgcggtgtaccggattattgacagcaacggatcatggttcgacattgggatt gacagcgatgggttttcattcgtgggcgtgtcgtcatttccaaagctggagtttccgctc tccgcctcatacatgggcatccagctcgcaaagatccggctggcgtccaacatccggtca tccatgactaagctgtga

Feline Calicivirus capsid (VS-FCV) SEQ ID NO: 2 MADDGSVTTPEQGTMVGGVIAEPSAQMSTAADMASGKSVDSEWEAFFSFHTSVNWSTSET

2GKILFKQSLGPLLNPYLEHISKLYVAWSGSIEVRFSISGSGVFGGKLAAIVVPPGV QSTSMLQYPHVLFDARQVEPVIFSIPDLRNSLYHLMSDTDTTSLVMVYNDLINPYAND SSGCIVTVETKPGIDFKFHLLKPPGSMLTHGSIPSDLIPKSSSLWIGNRHWTDITDF) RPFVFQANRHFDFNQETAGWSTPRFRPLAVTVSQSKGAKLGNGIATDyIVPGIPDGWj PIPTKLTPTGDYAITSSDGNDIETKLEYENADVIKNNTNFRSMYICGSLORAWGDKKISN

GFITTGVISDNSISPSNTIDQSKIVVYQDNHVNSEVQTSDITLAILGYTGIGEEAIGAl RDSVVRISVLPETGARGGNHPIFYKNSMKLGYVISSIDVFNSQILHTSRQLSLNNYLLP: DSFAVYRIIDSNGSWFDIGIDSDGFSFVGVSSFPKLEFPLSASYMGIQLAKIRLASNIRS SMTKL

Feline Calicivirus (VS-FCV) capsid (SEQ ID NO: 12) suggcugacgacggaucugugaccaccccagaacaaggaacaauggucggaggagugar auggcugacgacggaucugugaccaccccagaacaaggaacaauggucggaggagugauu gccgaacccagcgcucagaugucaacugcggcggacauggccuccggaaagucggugga uccgagugggaagccuucuucucguuccacacguccgugaacuggagcaccuccgaal caaggaaagauccucuucaagcagucccuggguccccugcugaacccguaccuggagca

aucagcaagcuguacgucgcuuggagcgggucgaucgaagugcgauuuuccaucucggga e666ononeoonnnne656n6ee65ne6on66606e66nn363n6oen6no6ee3beane agcggcguguucggugguaaacuggccgccaucgucgugccgccuggugucgacccugu cagucaaccuccaugcugcaguacccgcacguccuguucgacgcaagacaaguggagco cagucaaccuccaugcugcaguacccgcacguccuguucgacgcaagacaaguggagcca gugaucuucuccaucccggaccuccgcaacagccuguaucacuugauguccgauaccgau nefooene6oon6ne6nnoeonenbnoobeoeeo6oonooebbooonpoononnonebnb accacuucccucgugaucaugguguacaacgaucugaucaacccguacgccaaugacue accacuucccucgugaucaugguguacaacgaucugaucaacccguacgccaaugacucc

lacagcucggguugcaucgugaccgucgaaacgaagccuggcaucgauuucaaguuud neonnn6eeonnne6oneo66no36ee6oeee6on6ooebn6oneobnn666ono6eoee cugcugaaaccgcccggauccaugcuuacucacggguccaucccuuccgaucugaucc agagcuccucccuguggauugggaaccgccacuggaccgauauuaccgauuucgugal cggccuuucguguuccaagccaaccggcacuucgacuucaaccaggagacugccggcugg 66no66oo6noe6e66eopeeonnoe6onnoeo66ooeeoofeeoonn6n6onnno0660 acaacuccacgguuccgcccauuggccgugacugugucgcagucaaagggagccaagcu gggaacggcaucgccaccgacuacauugugccuggaauccccgacggauggccugaua accauccccaccaagcugaccccuaccggagauuacgccaucaccuccuccgacggca accauccccaccaagcugaccccuaccggagauuacgccaucaccuccuccgacggcaau gauauugaaaccaagcuggaauacgagaacgcggacgugauuaagaacaacaccaacuu cgcuccauguauaucugcggaagccuccagagggcuuggggcgacaagaagaucage accggguucaucacuaccggagugauuucugacaacuccaucagcccuucgaacacaar accggguucaucacuaccggagugauuucugacaacuccaucagcccuucgaacacaauu 09 gaccaguccaagaucgugguguaccaggacaaccaugucaauucggagguccagacuago gacaucacucuugccauccugggcuacaccggaauuggagaagaggccauaggcgccaad cgggacuccgucgugagaauuuccgugcuuccggaaacuggagcaaggggcggaaaud 0e0neee6656666ee36e66n3eee66oonno6n63onnnee6e6n6on600n0e6663 cccaucuucuacaaaaauuccaugaagcugggcuacgugaucuccuccauugacguguuc onn6n6oe6nneoonoonone6nfoenof66no6eebneoonnpppeeopnonnoneoo0 aacucccaaauccuccacaccucgcgccagcugucacugaacaacuacuuguugcccc acuccuucgcgguguaccggauuauugacagcaacggaucaugguucgacauugggar nne666nneoe6onn66neone66oeeobeoebnnenneb6ooenbnf60bonnoonoe6 gacagcgauggguuuucauucgugggcgugucgucauuuccaaagcuggaguuuccgcu 6no6oonnn6e66no6eeeoonnneon6on6n6o666n6onneonnnn666ne636e3e6 accgccucauacaugggcauccagcucgcaaagauccggcuggcguccaacauccgguca uccaugacuaagcuguga

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Feline Calicivirus (F9-like) capsid (SEQ ID NO: 3) atgactgcccoggaacaaggaacgatggtcggaggagtgattgcagaacogtcagcacag tgactgccccggaacaaggaacgatggtcggaggagtgattgcagaaccgtcagcacag atgtccaccgctgccgacatggccactggaaagagcgtggactccgaatgggaagccttc atgtccaccgctgccgacatggccactggaaagagcgtggactccgaatqggaagccttc ttctccttccacacttcggtcaactggtcgactagcgaaacccaggggaagattttgtt stctccttccacacttcggtcaactggtcgactagcgaaacccaggggaagattttgtto aagcaatccctcggccctctgctgaacccctacctggagcatctggccaagctgtacgt aagcaatccctcggccctctgctgaacccctacctggagcatctggccaagctgtacgtg jcatggtcgggcagcatcgaagtgcgctttagcatttccggctccggagtgttcggggg gcatggtcgggcagcatcgaagtgcgctttagcatttccggctccggagtgttcggggga aagcttgctgccattgtcgtgccgccaggagtggacccggtgcagtccacttctatgct aagcttgctgccattgtcgtgccgccaggagtggacccggtgcagtccacttctatgcto caatacccgcatgtcctgttcgacgccagacaggtggagcctgtgatcttttgcctgccg caatacccgcatgtcctgttcgacgccagacaggtggagcctgtgatcttttgcctgccg gatctcaggtccaccctgtatcacctcatgtccgacaccgacaccacctcgctcgtga gatctcaggtccaccctgtatcacctcatgtccgacaccgacaccacctogctcgtgato atggtgtacaacgacctgatcaacccctacgctaacgacgccaacagctcaggttgcatt atggtgtacaacgacctgatcaacccctacgctaacgacgccaacagctcaggttgcatt gtgactgtcgaaaccaagccaggccctgacttcaagtttcatttgctgaagccgcccggt ccatgctgacccacggctcgatcccatccgacctgatccccaagacgagctccctgtg tccatgctgacccacggctcgatcccatccgacctgatccccaagacgagctccctgtgg atcggaaaccgctactggtccgatattaccgacttcgtgatcagaccattcgtgttccaa atcggaaaccgctactggtccgatattaccgacttcgtgatcagaccattcgtgttcca gccaaccgccatttcgacttcaaccaggaaaccgcaggatggtcgacccctcgattccgc gccaaccgccatttcgacttcaaccaggaaaccgcaggatggtcgacccctcgattccg cgatttcagtgaccatcaccgaacagaacggcgcgaagctgggaattggcgtggcgad ccgatttcagtgaccatcaccgaacagaacggcgcgaagctgggaattggcgtggcgaco gactacatcgtgccgggaatcccggatggatggcctgatacgaccattcccggggagc gactacatcgtgccgggaatcccggatggatggcctgatacgaccattcccggggagctg atccctgccggggactacgccatcaccaacggtactggaaacgacatcaccactgccaco atccctgccggggactacgccatcaccaacggtactggaaacgacatcaccactgccace gttacgacaccgccgacatcataaagaacaacaccaacttcagaggaatgtacattte ggttacgacaccgccgacatcataaagaacaacaccaacttcagaggaatgtacatttgo ggctccctgcaacgcgcttggggtgacaaaaagatctcgaacactgccttcatcacaad ggctccctgcaacgcgcttggggtgacaaaaagatctcgaacactgccttcatcacaaca gcgactctggacggcgataacaacaacaagatcaatccttgtaataccatcgaccagtc gcgactctggacggcgataacaacaacaagatcaatccttgtaataccatcgaccagtcc aaaatcgtggtgttccaggataaccacgtgggaaagaaggcgcagacctccgacgacact aaaatcgtggtgttccaggataaccacgtgggaaagaaggcgcagacctccgacgacact :tggcgctgcttggctacaccgggatcggcgagcaggccattggaagcgatcgggatcg ctggcgctgcttggctacaccgggatcggcgagcaggccattggaagcgatcgggatcgg gtcgtgcggatctccaccctccccgagactggagcaaggggaggcaaccaccccatctt gtcgtgcggatctccaccctccccgagactggagcaaggggaggcaaccaccccatcttf tacaaaaacagcattaagctcggatacgtcatccgctccatcgatgtgttcaactctca tacaaaaacagcattaagctcggatacgtcatccgctccatcgatgtgttcaactctcaa

tcctgcacacttcgcggcagctgtccctgaaccactacctcttgccgcccgactcc atcctgcacacttcgcggcagctgtccctgaaccactacctcttgccgcccgactccttc gccgtctaccggatcattgattcgaacgggagctggttcgacatcggcattgatagcgat ggcttctcgtttgtgggcgtgtcgggcttcgggaagctggagttcccactgagcgcctca acatgggtatccagctggccaagatcaggctggcctccaacatccgctcacctatgact tacatgggtatccagctggccaagatcaggctggcctccaacatccgctcacctatgact aagctgtga aagctgtga

Feline Calicivirus (F9-like) capsid (SEQ ID NO: 4) MTAPEQGTMVGGVIAEPSAQMSTAADMATGKSVDSEWEAFFSFHTSVNWSTSETQGKILJ MTAPEQGTMVGGVIAEPSAQMSTAADMATGKSVDSEWEAFFSFHTSVNWSTSETQGKILE KQSLGPLLNPYLEHLAKLYVAWSGSIEVRFSISGSGVFGGKLAAIVVPPGVDPVQSTS KQSLGPLINPYLEHLAKLYVAWSGSIEVRFSISGSGVFGGKLAAIVVPPGVDPVQSTSML )YPHVLFDARQVEPVIFCLPDLRSTLYHLMSDTDTTSLVIMVYNDLINPYANDANSSG QYPHVLFDARQVEPVIFCLPDLRSTLYHLMSDTDTTSLVIMVYNDLINPYANDANSSGCI VTVETKPGPDFKFHLLKPPGSMLTHGSIPSDLIPKTSSLWIGNRYWSDITDFVIRPFVF VTVETKPGPDFKFHLLKPPGSMLTHGSIPSDLIPKTSSLWIGNRYWSDITDFVIRPFVFC ANRHFDFNQETAGWSTPRFRPISVTITEQNGAKLGIGVATDYIVPGIPDGWPDTTIPGEL ANRHFDFNQETAGWSTPRFRPISVTITEQNGAKLGIGVATDYIVPGIPDGWPDTTIPGEL IPAGDYAITNGTGNDITTATGYDTADIIKNNTNFRGMYICGSLQRAWGDKKISNTAFI ATLDGDNNNKINPCNTIDQSKIVVFQDNHVGKKAQTSDDTLALLGYTGIGEQAIGSDRI ATLDGDNNNKINPCNTIDQSKIVVFQDNHVGKKAQTSDDTLALLGYTGIGEQAIGSDRDF VVRISTLPETGARGGNHPIFYKNSIKLGYVIRSIDVFNSQILHTSRQLSLNHYLLPPDSE VVRISTLPETGARGGNHPIFYKNSIKLGYVIRSIDVFNSQILHTSRQLSLNHYLLPPDSF AVYRIIDSNGSWFDIGIDSDGFSFVGVSGFGKLEFPLSASYMGIQLAKIRLASNIRSPMT AVYRIIDSNGSWFDIGIDSDGFSFVGVSGFGKLEFPLSASYMGIQLAKIRLASNIRSPMT KL

Feline Calicivirus (F9-like) capsid (SEQ ID NO: 13) augacugccccggaacaaggaacgauggucggaggagugauugcagaaccgucagcaca augacugccccggaacaaggaacgauggucggaggagugauugcagaaccgucagcacag suguccaccgcugccgacauggccacuggaaagagcguggacuccgaaugggaagccut auguccaccgcugccgacauggccacuggaaagagcguggacuccgaaugggaagccuuc lucuccuuccacacuucggucaacuggucgacuagcgaaacccaggggaagauuuuguuc ucuccuuccacacuucggucaacuggucgacuagcgaaacccaggggaagauuuuguue aagcaaucccucggcccucugcugaaccccuaccuggagcaucuggccaagcuguaco aagcaaucccucggcccucugcugaaccccuaccuggagcaucuggccaagcuguacguc gcauggucgggcagcaucgaagugcgcuuuagcauuuccggcuccggaguguucggggga gcauggucgggcagcaucgaagugcgcuuuagcauuuccggcuccggaguguucggggga aagcuugcugccauugucgugccgccaggaguggacccggugcaguccacuucuaugcu aagcuugcugccauugucgugccgccaggaguggacccggugcaguccacuucuaugcu caauacccgcauguccuguucgacgccagacagguggagccugugaucuuuugccugco caauacccgcauguccuguucgacgccagacagguggagccugugaucuuuugccugccg jfaucucagguccacccuguaucaccucauguccgacaccgacaccaccucgcucgugat gaucucagguccacccuguaucaccucauguccgacaccgacaccaccucgcucgugauc augguguacaacgaccugaucaaccccuacgcuaacgacgccaacagcucagguugcauu augguguacaacgaccugaucaaccccuacgcuaacgacgccaacagcucagguugcauu gugacugucgaaaccaagccaggcccugacuucaaguuucauuugcugaagccgcccgo gugacugucgaaaccaagccaggcccugacuucaaguuucauuugcugaagccgcccggu uccaugcugacccacggcucgaucccauccgaccugauccccaagacgagcucccugugg uccaugcugacccacggcucgaucccauccgaccugauccccaagacgagcucccugug aucggaaaccgcuacugguccgauauuaccgacuucgugaucagaccauucguguuccaa aucggaaaccgcuacugguccgauauuaccgacuucgugaucagaccauucguguuccaa gccaaccgccauuucgacuucaaccaggaaaccgcaggauggucgaccccucgauuccg gccaaccgccauuucgacuucaaccaggaaaccgcaggauggucgaccccucgauuccgo ccgauuucagugaccaucaccgaacagaacggcgcgaagcugggaauuggcguggcgacc ccgauuucagugaccaucaccgaacagaacggcgcgaagcugggaauuggcguggcgad gacuacaucgugccgggaaucccggauggauggccugauacgaccauucccggggagcu gacuacaucgugccgggaaucccggauggauggccugauacgaccauucccggggagcug aucccugccggggacuacgccaucaccaacgguacuggaaacgacaucaccacugccacc ucccugccggggacuacgccaucaccaacgguacuggaaacgacaucaccacugccaca

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gguuacgacaccgccgacaucauaaagaacaacaccaacuucagaggaauguacauu guuacgacaccgccgacaucauaaagaacaacaccaacuucagaggaauguacauuugo ggcucccugcaacgcgcuuggggugacaaaaagaucucgaacacugccuucaucacaad ggcucccugcaacgcgcuuggggugacaaaaagaucucgaacacugccuucaucacaaca gcgacucuggacggcgauaacaacaacaagaucaauccuuguaauaccaucgaccaguca gcgacucuggacggcgauaacaacaacaagaucaauccuuguaauaccaucgaccagucc aaaaucgugguguuccaggauaaccacgugggaaagaaggcgcagaccuccgacgacad aaaaucgugguguuccaggauaaccacgugggaaagaaggcgcagaccuccgacgacact cuggcgcugcuuggcuacaccgggaucggcgagcaggccauuggaagcgaucgggauc cuggcgcugcuuggcuacaccgggaucggcgagcaggccauuggaagcgaucgggaucgg jucgugcggaucuccacccuccccgagacuggagcaaggggaggcaaccaccccaucuu gucgugcggaucuccacccuccccgagacuggagcaaggggaggcaaccaccccaucuuu acaaaaacagcauuaagcucggauacgucauccgcuccaucgauguguucaacucud lacaaaaacagcauuaagcucggauacgucauccgcuccaucgauguguucaacucucaa auccugcacacuucgcggcagcugucccugaaccacuaccucuugccgcccgacuccuu uccugcacacuucgcggcagcugucccugaaccacuaccucuugccgcccgacuccuuc gcgucuaccggaucauugauucgaacgggagcugguucgacaucggcauugauagga gccgucuaccggaucauugauucgaacgggagcugguucgacaucggcauugauagcgau ggcuucucguuugugggcgugucgggcuucgggaagcuggaguucccacugagcgccuca uacauggguauccagcuggccaagaucaggcuggccuccaacauccgcucaccuaugacu uacauggguauccagcuggccaagaucaggcuggccuccaacauccgcucaccuaugacu aagcuguga

Feline Leukemia Virus envelope glycoprotein (gp85) SEQ ID NO: 5 tggagtcaccaacacaccctaaaccttctaaagacaaaaccctctcgtggaato atggagtcaccaacacaccctaaaccttctaaagacaaaaccctctcgtggaatctcgccttccttgt gggcatcctgttcacaatcgacatcggatggccaacccttcgccgcatcagatctacaatgtgacat gggcatcctgttcacaatcgacatcggcatggccaacccttcgccgcatcagatctacaatgtgacat gggtcattactaatgtgcagacaaacacccaggcaaatgctacttctatgcttggtactctgactgat gggtcattactaatgtgcagacaaacacccaggcaaatgctacttctatgcttggtactctgactgat= gcttatccaaccctgcacgtcgacctttgcgatctcgtcggtgacacatgggagcccatcgtgctgaa gcttatccaaccctgcacgtcgacctttgcgatctcgtcggtgacacatgggagcccatogtgctgaa tccaactaatgtcaaacatggtgccaggtattcttctagcaaatacgggtgtaagaccactgatcgga tccaactaatgtcaaacatggtgccaggtattcttctagcaaatacgggtgtaagaccactgatcgga agaaacagcaacaaacctacccattctacgtgtgcccgggtcacgcaccgtccctgggtccgaaggga agaaacagcaacaaacctacccattctacgtgtgcccgggtcacgcaccgtccctgggtccgaaggga cacattgtgggggagcccaagacggtttttgcgctgcttggggttgtgaaacaaccggagaagccto acacattgtgggggagcccaagacggtttttgcgctgcttggggttgtgaaacaaccggagaagcctq gtggaagcctacctcatcttgggactacattactgtgaaaagaggctctagccaggataacagctgcg gtggaagcctacctcatcttgggactacattactgtgaaaagaggctctagccaggataacagctgco aaggaaagtgtaatcccctggtgcttcaattcacccagaaaggccggcaggcatcatgggatggaccg aaggaaagtgtaatcccctggtgcttcaattcacccagaaaggccggcaggcatcatgggatggacco laaatgtggggacttagactctatcgcaccggatacgaccccatcgctctgtttactgtgtcacgcca aaaatgtggggacttagactctatcgcaccggatacgaccccatcgctctgtttactgtgtcacgcca agtctccaccattactccgccacaggccatggggccgaatctggtcctccccgatcagaagccaccct agtctccaccattactccgccacaggccatggggccgaatctggtcctccccgatcagaagccaccct cacggcaaagtcaaaccggctcaaaagtggccacccaacggccccagacaaatgagtccgcacctagg cacggcaaagtcaaaccggctcaaaagtggccacccaacggccccagacaaatgagtccgcacctagg tcagtggcacctacaacaatgggtccaaagcggatcggaaccggagacaggctcattaacctcgtgca tcagtggcacotacaacaatgggtccaaagcggatcggaaccggagacaggctcattaacctcgtgca gggacttatctggcccttaacgctactgaccccaacaagaccaaggattgctggctctgccttgtg agggacttatotggccottaacgctactgaccccaacaagaccaaggattgctggctctgccttgtga gcagacctccttactatgaggggatcgccattctcggaaactactcaaatcagaccaacccccctccg. gcagacctccttactatgaggggatcgccattctcggaaactactcaaatcagaccaacccccctccg

agtgtctgagcaccccccagcacaagcttactatttcagaagtcagtggacagggaatgtgcatcg tcgtgtctgagcaccccccagcacaagcttactatttcagaagtcagtggacagggaatgtgcatc99 aaccgtgccaaagactcatcaagccctttgcaacaaaactcaacaagggcacactggagctcattato aaccgtgccaaagactcatcaagccctttgcaacaaaactcaacaagggcacactggagctcattatc tcgccgcacctaacgggacctactgggcttgcaatactggattgaccccgtgtatctctatggccgtg. tcgccgcacctaacgggacctactgggcttgcaatactggattgaccccgtgtatctctatggccgtg ctgaattggacttccgacttctgcgtgcttattgagctttggcctagagtgacataccatcagcctgal ctgaattggacttccgacttctgcgtgcttattgagctttggcctagagtgacataccatcagcctga gtacgtctatacccatttcgccaaggcagtcagattccggcgggagcctatctccctgactgtggcc gtacgtctatacccatttcgccaaggcagtcagattccggcgggagcctatctccctgactgtggcct tgatgctcggtggactgacagtgggaggaattgcagctggagtcggaactggaaccaaggccctgo tgatgctcggtggactgacagtgggaggaattgcagctggagtoggaactggaaccaaggccctgctc gaaactgctcagttccggcagctgcagatggccatgcacactgacatccaggctctggaggaatcaat gaaactgctcagttccggcagctgcagatggccatgcacactgacatccaggctctggaggaatcaat ttcagcccttgagaaaagcttgacctcgctgtctgaagtggtcctccaaaacaggcgcggtttggaca ttcagcccttgagaaaagcttgacctcgctgtctgaagtggtcctccaaaacaggcgcggtttggaca tcctgttccttcaagagggtggtctgtgcgccgctctcaaggaggaatgctgtttctacgctgaccat tcctgttccttcaagagggtggtctgtgcgccgctctcaaggaggaatgctgtttctacgctgaccat accgggctggtgcgcgataacatggcaaagctgcgggaacgcttgaaacagaggcagcaactgttcga accgggctggtgcgcgataacatggcaaagctgcgggaacgcttgaaacagaggcagcaactgttcga

tctcagcagggatggttcgagggctggtttaacaagagcccatggtttaccactctgatctcttc ctctcagcagggatggttcgagggctggtttaacaagagcccatggtttaccactctgatctcttcaa tcatgggtccactgctcatcctgcttctgattcttctcttcggaccgtgtattctcaacaggctggtg tcatgggtccactgctcatcctgcttctgattcttctcttcggaccgtgtattctcaacaggctggt0 cagtttgtcaaggacagaatctcggtggtccaggccctgattcttactcagcagtatcagcagattaa cagtttgtcaaggacagaatctoggtggtccaggccctgattcttactcagcagtatcagcagattaa

[cagtacgaccccgatcggccttga gcagtacgaccccgatcggccttga

Feline Leukemia Virus envelope glycoprotein (gp85) SEQ ID NO: 6 MESPTHPKPSKDKTLSWNLAFLVGILFTIDIGMANPSPHQIYNVTWVITNVQTNTQAN MESPTHPKPSKDKTLSWNLAFLVGILFTIDIGMANPSPHQIYNVTWVITNVOTNTQANAT- SMLGTLTDAYPTLHVDLCDLVGDTWEPIVLNPTNVKHGARYSSSKYGCKTTDRKKQQQ7 SMLGTLTDAYPTLHVDLCDLVGDTWEPIVLNPTNVKHGARYSSSKYGCKTTDRKKQQQT PFYVCPGHAPSLGPKGTHCGGAQDGFCAAWGCETTGEAWWKPTSSWDYITVKRGSSC PFYVCPGHAPSLGPKGTHCGGAQDGFCAAWGCETTGEAWWKPTSSWDYITVKRGSSODNS CEGKCNPLVLQFTQKGRQASWDGPKMWGLRLYRTGYDPIALFTVSRQVSTITPPQAMGPN. CEGKCNPLVLQFTQKGRQASWDGPKMWGLRLYRTGYDPIALFTVSRQVSTITPPOAMGPF LVLPDQKPPSRQSQTGSKVATQRPQTNESAPRSVAPTTMGPKRIGTGDRLINLVQGTYLA LVLPDQKPPSRQSQTGSKVATORPQTNESAPRSVAPTTMEPKRIGTGDRLINLVOGTYLA LNATDPNKTKDCWLCLVSRPPYYEGIAILGNYSNQTNPPPSCLSTPQHKLTISEVSGQC LNATDPNKTKDCWLCLVSRPPYYEGIAILGNYSNQTNPPPSCLSTPOHKLTISEVSGQGM CIGTVPKTHQALCNKTQQGHTGAHYLAAPNGTYWACNTGLTPCISMAVLNWTSDFCVL CIGTVPKTHQALCNKTQQGHTGAHYLAAPNGTYWACNTGLTPCISMAVLNWTSDFCVLIE LWPRVTYHQPEYVYTHFAKAVRFRREPISLTVALMLGGLTVGGIAAGVGTGTKALLETAQ LWPRVTYHQPEYVYTHFAKAVRFRREPISLTVALMLGGLTVGGIAAGVGTGTKALLETAG FRQLQMAMHTDIQALEESISALEKSLTSLSEVVLQNRRGLDILFLQEGGLCAALKEECO RQLQMAMHTDIQALEESISALEKSLTSLSEVVLONRRGLDILFLQEGGLCAALKEECCF YADHTGLVRDNMAKLRERLKQRQQLFDSQQGWFEGWFNKSPWFTTLISSIMGPLLILLLI YADHTGLVRDNMAKLRERLKQRQQLFDSQQGWFEGWFNKSPWFTTLISSIMGPLLILLL LLFGPCILNRLVQFVKDRISVVQALILTQQYQQIKQYDPDRP* LLFGPCILNRLVQFVKDRISVVQALILTQQYQQIKQYDPDRP*

Feline Leukemia Virus envelope glycoprotein (gp85) SEQ ID NO: 14

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auggagucaccaacacacccuaaaccuucuaaagacaaaacccucucguggaaucucgccuuccuugus auggagucaccaacacacccuaaaccuucuaaagacaaaacccucucguggaaucucgccuuccuugu gggcauccuguucacaaucgacaucggcauggccaacccuucgccgcaucagaucuacaaugugaca gggcauccuguucacaaucgacaucggcauggccaacccuucgccgcaucagaucuacaaugugacau gggucauuacuaaugugcagacaaacacccaggcaaaugcuacuucuaugcuugguacucugacugau gggucauuacuaaugugcagacaaacacccaggcaaaugcuacuucuaugcuugguacucugacugau gcuuauccaacccugcacgucgaccuuugcgaucucgucggugacacaugggagcccaucgugcugaa gcuuauccaacccugcacgucgaccuuugcgaucucgucggugacacaugggagcccaucgugcugaa succaacuaaugucaaacauggugccagguauucuucuagcaaauacggguguaagaccacugaucgga uccaacuaaugucaaacauggugccagguauucuucuagcaaauacggguguaagaccacugaucgga agaaacagcaacaaaccuacccauucuacgugugcccgggucacgcaccgucccuggguccgaaggg agaaacagcaacaaaccuacccauucuacgugugcccgggucacgcaccgucccuggguccgaaggga acacauugugggggagcccaagacgguuuuugcgcugcuugggguugugaaacaaccggagaag. acacauugugggggagcccaagacgguuuuugcgcugcuugggguugugaaacaaccggagaagccug guggaagccuaccucaucuugggacuacauuacugugaaaagaggcucuagccaggauaacagcugcg aaggaaaguguaauccccuggugcuucaauucacccagaaaggccggcaggcaucaugggauggaccg. aaggaaaguguaauccccuggugcuucaauucacccagaaaggccggcaggcaucaugggauggaccg aaaauguggggacuuagacucuaucgcaccggauacgaccccaucgcucuguuuacugugucacgcca aaaauguggggacuuagacucuaucgcaccggauacgaccccaucgcucuguuuacugugucacgcca agucuccaccauuacuccgccacaggccauggggccgaaucugguccuccccgaucagaagccacccu cacggcaaagucaaaccggcucaaaaguggccacccaacggccccagacaaaugaguccgcaccuago cacggcaaagucaaaccggcucaaaaguggccacccaacggccccagacaaaugaguccgcaccuagg ucaguggcaccuacaacaauggguccaaagcggaucggaaccggagacaggcucauuaaccucgug ucaguggcaccuacaacaauggguccaaagcggaucggaaccggagacaggcucauuaaccucgugca agggacuuaucuggcccuuaacgcuacugaccccaacaagaccaaggauugcuggcucugccuugug agggacuuaucuggcccuuaacgcuacugaccccaacaagaccaaggauugcuggcucugccuuguga gcagaccuccuuacuaugaggggaucgccauucucggaaacuacucaaaucagaccaaccccccuco gcagaccuccuuacuaugaggggaucgccauucucggaaacuacucaaaucagaccaaccccccuccg agugucugagcaccccccagcacaagcuuacuauuucagaagucaguggacagggaaugugcaud ucgugucugagcaccccccagcacaagcuuacuauuucagaagucaguggacagggaaugugcaucgg aaccgugccaaagacucaucaagcccuuugcaacaaaacucaacaagggcacacuggagcucauua aaccgugccaaagacucaucaagcccuuugcaacaaaacucaacaagggcacacuggagcucauuauc ucgccgcaccuaacgggaccuacugggcuugcaauacuggauugaccccguguaucucuauggccgug ecugaauuggacuuccgacuucugcgugcuuauugagcuuuggccuagagugacauaccaucagccuga cugaauuggacuuccgacuucugcgugcuuauugagcuuuggccuagagugacauaccaucagccuga juacgucuauacccauuucgccaaggcagucagauuccggcgggagccuaucucccugacuguggo guacgucuauacccauuucgccaaggcagucagauuccggcgggagccuaucucccugacuquggccu ugaugcucgguggacugacagugggaggaauugcagcuggagucggaacuggaaccaaggcccugcuo ugaugcucgguggacugacagugggaggaauugcagcuggagucggaacuggaaccaaggcccugcuc gaaacugcucaguuccggcagcugcagauggccaugcacacugacauccaggcucuggaggaauca gaaacugcucaguuccggcagcugcagauggccaugcacacugacauccaggcucuggaggaaucaal uucagcccuugagaaaagcuugaccucgcugucugaagugguccuccaaaacaggcgcgguuuggad uucagcccuugagaaaagcuugaccucgcugucugaagugguccuccaaaacaggcgcgguuuggaca accuguuccuucaagaggguggucugugcgccgcucucaaggaggaaugcuguuucuacgcugacca: uccuguuccuucaagaggguggucugugcgccgcucucaaggaggaaugcuguuucuacgcugaccalu accgggcuggugcgcgauaacauggcaaagcugcgggaacgcuugaaacagaggcagcaacuguucg accgggcuggugcgcgauaacauggcaaagcugcgggaacgcuugaaacagaggcagcaacuguucga ucucagcagggaugguucgagggcugguuuaacaagagcccaugguuuaccacucugaucucuucaa cucucagcagggaugguucgagggcugguuuaacaagagcccaugguuuaccacucugaucucuucaa auggguccacugcucauccugcuucugauucuucucuucggaccguguauucucaacaggcuggu ucauggguccacugcucauccugcuucugauucuucucuucggaccguguauucucaacaggcuggug aguuugucaaggacagaaucucggugguccaggcccugauucuuacucagcaguaucagcagauual caguuugucaaggacagaaucucggugguccaggcccugauucuuacucagcaguaucagcagauuaa gcaguacgaccccgaucggccuuga gcaguacgaccccgaucggccuuga

Feline Leukemia Virus envelope glycoprotein (gp70) SEQ ID NO: 7 hatcctagtccacaccaaatatataatgtaacttgggtaataaccaatgtacaaactaacacc aatcctagtccacaccaaatatataatgtaacttgggtaataaccaatgtacaaactaacacc Baagctaacgccacctctatgttaggaaccttaaccgatgcctaccctaccctacatgttga caagctaacgccacctctatgttaggaaccttaaccgatgcctaccctaccctacatgttgac :tatgtgacctagtgggagacacctgggaacctatagtcctaaacccaaccaatgtaaaacal ttatgtgacctagtgggagacacctgggaacctatagtcctaaacccaaccaatgtaaaacac ggggcacgttactcctcctcaaaatatggatgtaaaactacagatagaaaaaaacagcaacag acataccccttttacgtctgccccggacatgccccctcgttggggccaaagggaacacattgt acataccccttttacgtctgccccggacatgccccctcgttggggccaaagggaacacattgt ggaggggcacaagatgggttttgtgccgcatggggatgtgagaccaccggagaagcttggt ggaggggcacaagatgggttttgtgccgcatggggatgtgagaccaccggagaagcttggtgg aagcccacctcctcatgggactatatcacagtaaaaagagggagtagtcaggacaatagc aagcccacctcctcatgggactatatcacagtaaaaagagggagtagtcaggacaatagctgt jagggaaaatgcaaccccctggttttgcagttcacccagaagggaagacaagcctcttggo gagggaaaatgcaaccccctggttttgcagttcacccagaagggaagacaagcctcttgggac gacctaagatgtggggattgcgactataccgtacaggatatgaccctatcgctttattca ggacctaagatgtggggattgcgactataccgtacaggatatgaccctatcgctttattcacg tgtcccggcaggtatcaaccattacgccgcctcaggcaatgggaccaaacctagtcttad gtgtcccggcaggtatcaaccattacgccgcctcaggcaatgggaccaaacctagtcttacct gatcaaaaacccccatcccgacaatctcaaacagggtccaaagtggcgacccagaggccccaa gatcaaaaacccccatcccgacaatctcaaacagggtccaaagtggcgacccagaggccccaa cgaatgaaagcgccccaaggtctgttgcccccaccaccatgggtcccaaacggattggga acgaatgaaagcgccccaaggtctgttgcccccaccaccatgggtcccaaacggattgggacc ggagataggttaataaatttagtacaagggacatacctagccttaaatgccaccgaccccaad ggagataggttaataaatttagtacaagggacatacctagccttaaatgccaccgaccccaac laaactaaagactgttggctctgcctggtttctcgaccaccctattacgaagggattgcaatc aaaactaaagactgttggctctgcctggtttctcgaccaccctattacgaagggattgcaatc. taggtaactacagcaaccaaacaaacccccccccatcctgcctatctactccgcaacaca. ttaggtaactacagcaaccaaacaaacccccccccatcctgcctatctactccgcaacacaaa ctaactatatctgaagtatcagggcaaggaatgtgcatagggactgttcctaaaacccad ctaactatatctgaagtatcagggcaaggaatgtgcatagggactgttcctaaaacccaccag gctttgtgcaataagacacaacagggacatacaggggcgcactatctagccgcccccaacggo gctttgtgcaataagacacaacagggacatacaggggcgcactatctagccgcccccaacggc ctattgggcctgtaacactggactcaccccatgcatttccatggcggtgctcaattgga acctattgggcctgtaacactggactcaccccatgcatttccatggcggtgctcaattggacc tctgatttttgtgtcttaatcgaattatggcccagagtgacttaccatcaacccgaatatgtg tctgatttttgtgtcttaatcgaattatggcccagagtgacttaccatcaacocgaatatgtg tacacacattttgccaaagctgtcaggttccgaaga Feline Leukemia Virus envelope glycoprotein (gp70) SEQ ID NO: 8 8 JPSPHQIYNVTWVITNVQTNTQANATSMLGTLTDAyPTLHVDLCDLVGDTWEPIVLNPTNVKHGARYSSS NPSPHQIYNVTWVITNVQTNTQANATSMLGTLTDAYPTLHVDLCDLVGDTWEPIVLNPTNVKHGARYSSS 7GCKTTDRKKQQQTYPFYVCPGHAPSLGPKGTHCGGAQDGFCAAWGCETTGEAWWKPTSSWDYItVK KYGCKTTDRKKQQQTYPFYVCPGHAPSLGPKGTHCGGAQDGFCAAWGCETTGEAWWKPTSSWDYITVKR1 SQDNSCEGKCNPLVLQFTQKGRQASWDGPKMWGLRLYRTGYDPIALFTVSRQVSTITPPQAMGPNLVLP SSQDNSCEGKCNPLVLQFTQKGRQASWDGPKMWGLRLYRTGYDPIALFTVSRQVSTITPPQAMGPNLVLE DQKPPSRQSQTGSKVATQRPQTNESAPRSVAPTTMGPKRIGTGDRLINLVQGTYLaLNatDPNKTKDCWL DQKPPSRQSQTGSKVATQRPQTNESAPRSVAPTTMGPKRIGTGDRLINLVQGTYLALNATDPNKTKDCWIL CLVSRPPYYEGIAILGNYSNQTNPPPSCLSTPQHKLTISEVSGQGMCIGTVPKTHQALCNKTQQGHTGAH CLVSRPPYYEGIAILGNYSNQTNPPPSCLSTPQHKLTISEVSGQGMCIGTVPKTHQALCNKTQQGHTGAL YLAAPNGTYWACNTGLTPCISMAVLNWTSDFCVLIELWPRVTYHQPEYVYTHFAKAVRFRR YLAAPNGTYWACNTGLTPCISMAVLNWTSDFCVLIELWPRVTYHOPEYVYTHFAKAVRFRR

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29

Feline Leukemia Virus envelope glycoprotein (gp70) SEQ ID NO: 15 aauccuaguccacaccaaauauauaauguaacuuggguaauaaccaauguacaaacuaacaco auccuaguccacaccaaauauauaauguaacuuggguaauaaccaauguacaaacuaacac caagcuaacgccaccucuauguuaggaaccuuaaccgaugccuacccuacccuacauguugac caagcuaacgccaccucuauguuaggaaccuuaaccgaugccuacccuacccuacauguugac uaugugaccuagugggagacaccugggaaccuauaguccuaaacccaaccaauguaaaaca uuaugugaccuagugggagacaccugggaaccuauaguccuaaacccaaccaauguaaaacac ggggcacguuacuccuccucaaaauauggauguaaaacuacagauagaaaaaaacagcaacag ggggcacguuacuccuccucaaaauauggauguaaaacuacagauagaaaaaaacagcaacaq cauaccccuuuuacgucugccccggacaugcccccucguuggggccaaagggaacacauug acauaccccuuuuacgucugccccggacaugcccccucguuggggccaaagggaacacauugu ggaggggcacaagauggguuuugugccgcauggggaugugagaccaccggagaagcuuggu ggaggggcacaagauggguuuugugccgcauggggaugugagaccaccggagaagcuuggug aagcccaccuccucaugggacuauaucacaguaaaaagagggaguagucaggacaauagcuqu aagcccaccuccucaugggacuauaucacaguaaaaagagggaguagucaggacaauagcugu gagggaaaaugcaacccccugguuuugcaguucacccagaagggaagacaagccucuuggga gagggaaaaugcaacccccugguuuugcaguucacccagaagggaagacaagccucuugggac ggaccuaagauguggggauugcgacuauaccguacaggauaugacccuaucgcuuuauuca ggaccuaagauguggggauugcgacuauaccguacaggauaugacccuaucgcuuuauucacg gugucccggcagguaucaaccauuacgccgccucaggcaaugggaccaaaccuagucuuaccu gugucccggcagguaucaaccauuacgccgccucaggcaaugggaccaaaccuagucuuaccu lucaaaaacccccaucccgacaaucucaaacaggguccaaaguggcgacccagaggccco gaucaaaaacccccaucccgacaaucucaaacaggguccaaaguggcgacccagaggccccaa acgaaugaaagcgccccaaggucuguugcccccaccaccaugggucccaaacggauugggad acgaaugaaagcgccccaaggucuguugcccccaccaccaugggucccaaacggauugggacc ggagauagguuaauaaauuuaguacaagggacauaccuagccuuaaaugccaccgaccccaac ggagauagguuaauaaauuuaguacaagggacauaccuagccuuaaaugccaccgaccccaac laacuaaagacuguuggcucugccugguuucucgaccacccuauuacgaagggauugcaa aaaacuaaagacuguuggcucugccugguuucucgaccacccuauuacgaagggauugcaauc uagguaacuacagcaaccaaacaaacccccccccauccugccuaucuacuccgcaacac iuagguaacuacagcaaccaaacaaacccccccccauccugccuaucuacuccgcaacacaaã cuaacuauaucugaaguaucagggcaaggaaugugcauagggacuguuccuaaaacccaccag cuaacuauaucugaaguaucagggcaaggaaugugcauagggacuguuccuaaaacccaccag cuuugugcaauaagacacaacagggacauacaggggcgcacuaucuagccgcccccaacg cuuugugcaauaagacacaacagggacauacaggggcgcacuaucuagccgcccccaacggc accuauugggccuguaacacuggacucaccccaugcauuuccauggaggugcucaauuggaco accuauugggccuguaacacuggacucaccccaugcauuuccauggcggugcucaauuggacc cugauuuuugugucuuaaucgaauuauggcccagagugacuuaccaucaacccgaauaugug icugauuuuugugucuuaaucgaauuauggcccagagugacuuaccaucaacccgaauauqug uacacacauuuugccaaagcugucagguuccgaaga e6ee6oonn66eon6no6eeeoo6nnnneopopoen

RABIES VIRUS G (SEQ ID NO: 9) atggtgccgcaggctctcctgtttgtcccccttctggtctttccattgtgttttgggaaattccctatctacacaattc atggtgccgcaggctctcctgtttgtcccccttctggtctttccattgtgttttgggaaattccctatctacacaatto eggacaagttgggaccctggagcccaattgacattcatcatctcagctgcccgaacaatttggtcgte cggacaagttgggaccctggagcccaattgacattcatcatctcagctgcccgaacaatttggtogtggaggaogaagg atgcaccaacctgtcggggttctcctacatggaattgaaagtcggatacatcagtgccattaag atgcaccaacctgtcggggttctcctacatggaattgaaagtcggatacatcagtgccattaagatgaacgggttcact tgcacaggcgtcgtgactgaagctgagacatacactaacttcgtgggatatgtcactaccactttcaaaagaaagcat tgcacaggcgtcgtgactgaagctgagacatacactaacttogtgggatatgtcactaccactttcaaaagaaagcatt tccgccctactcctgatgcttgtagggccgcatacaactggaagatggccggtgaccccagatatgaggaatcacttca sccgccctactcctgatgcttgtagggccgcatacaactggaagatggcoggtgaccccagatatgaggaatcacttca caatccgtaccctgactaccactggcttcggactgtcaaaaccaccaaggagtcactcgtgatcattagto aatccgtaccctgactaccactggcttcggactgtcaaaaccaccaaggagtcactcgtgatcattagtccaagtgtg gctgatcttgacccatacgaccggtcacttcactcacgggtgttcccgggggggaattgctctggtgtcgcagtgtcg. gctgatcttgacccatacgaccggtcacttcactcacgggtgttcccgggggggaattgctctggtgtcgcagtgtcgt caacctactgctccacaaaccacgattacaccatttggatgccagaaaatcctcggcttggtatgto caacctactgctccacaaaccacgattacaccatttggatgccagaaaatcctcggcttggtatgtcatgtgacatttt caccaattctcgggggaagagggcttccaaagggtctgaaacttgcggctttgtcgatgagcggggcttgtataagtca accaattctcgggggaagagggcttccaaagggtctgaaacttgcggctttgtcgatgagcggggcttgtataagtca cttaaaggtgcttgcaaactcaagctttgtggtgtcttgggattgagattgatggatggaacttgggtcgcaatgcage cttaaaggtgcttgcaaactcaagctttgtggtgtcttgggattgagattgatggatggaacttgggtcgcaatgcaga httctaacgaaaccaaatggtgccctcccggacagcttgtgaatttgcatgactttcgctctgacgaaattg. cttctaacgaaaccaaatggtgccctcccggacagcttgtgaatttgcatgactttcgctctgacgaaattgagoatct tgtcgtcgaggagttggtcaagaagcgggaagagtgtctggatgctttggaatcaatcatgaccaccaagtcagtgtct tgtcgtcgaggagttggtcaagaagçgggaagagtgtctggatgctttggaatcaatcatgaccaccaagtcagtgtct :tcagacggctctcacatcttaggaaattggtgccaggttttggaaaagcatataccattttcaacaagacccttatgo ttcagacggctctcacatcttaggaaattggtgccaggttttggaaaagcatataccattttcaacaagacccttatgg aagccgatgctcactacaagtctgtcaggacttggaatgagatcatcccgtctaaagggtgtcttagggtcggagggag aagccgatgctcactacaagtctgtcaggacttggaatgagatcatcccgtctaaagggtgtcttagggtcggagggag atgtcatcctcatgtcaacggagtctttttcaatggtatcattcttggacctgacggaaatgtccttatccctgagat atgtcatcctcatgtcaacggagtctttttcaatggtatcattcttggacctgacggaaatgtccttatccctgagatg caatcttccctcctccagcaacacatggaacttcttgtctcatcggtcatcccccttatgcaccccctggctg aatcttccctcctccagcaacacatggaacttcttgtctcatcggtcatcccccttatgcaccccctggctgacccat caaccgtgttcaagaacggtgacgaggcagaggattttgtcgaggtccaccttcccgatgtgcatgaacggatctctgo aaccgtgttcaagaacggtgacgaggcagaggattttgtcgaggtccaccttcccgatgtgcatgaacggatctctgg gtcgaccttggactccctaactggggaaagtatgtccttctgtcggcaggagccctgactgccttgatgttgattato jtcgaccttggactccctaactggggaaagtatgtccttctgtcggcaggagccctgactgccttgatgttgattato :tcctgatgacttgttggaggagagtcaatcggtcggagccaacacaacataatctcagaggaacaggaagggaggtg. ctcctgatgacttgttggaggagagtcaatcggtcggagccaacacaacataatctcagaggaacaggaagggaggtgt agtcacaccccaaagcgggaagatcatttcgtcttgggagtcatacaagagcggaggtgaaaccggactgtga cagtcacaccccaaagcgggaagatcatttcgtcttgggagtcatacaagagcggaggtgaaaccggactgtga

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30

RABIES VIRUS G (SEQ ID NO: 10) MVPQALLFVPLLVFPLCFGKFPIYTIPDKLGPWSPIDIHHLSCPNNLVVEDEGCTNLS MVPQALLFVPLLVFPLCFGKFPIYTIPDKLGPWSPIDIHHLSCPNNLVVEDEGCTNLSGE SYMELKVGYISAIKMNGFTCTGVVTEAETYTNFVGYVTTTFKRKHFRPTPDACRAAYNWE SYMELKVGYISAIKMNGFTCTGVVTEAETYTNFVGYVTTTFKRKHFRPTPDACRAAYNWK MAGDPRYEESLHNPYPDYHWLRTVKTTKESLVIISPSVADLDPYDRSLHSRVFPGGNCS MAGDPRYEESLHNPYPDYHWLRTVKTTKESLVIISPSVADLDPYDRSLHSRVFPGGNCSG

VAVSSTYCSTNHDYTIWMPENPRLGMSCDIFTNSRGKRASKGSETCGFVDERGLYKSLKG VAVSSTYCSTNHDYTIWMPENPRLGMSCDIFTNSRGKRASKGSETCGFVDERGLYKSLKG ACKLKLCGVLGLRLMDGTWVAMQTSNETKWCPPGQLVNLHDFRSDEIEHLVVEELVKKR ACKLKLCGVLGLRLMDGTWVAMQTSNETKWCPPGQLVNLHDFRSDEIEHLVVEELVKKRE ECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKTLMEADAHYKSVRTWNEIIPS ECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKTLMEADAHYKSVRTWNEIIP3 KGCLRVGGRCHPHVNGVFFNGIILGPDGNVLIPEMOSSLLQQHMELLVSSVIPLMHPLA KGCLRVGGRCHPHVNGVFFNGIILGPDGNVLIPEMQSSLLQQHMELLVSSVIPLMHPLAD PSTVFKNGDEAEDFVEVHLPDVHERISGVDLGLPNWGKYVLLSAGALTALMLIIFLMTCW PSTVFKNGDEAEDFVEVHLPDVHERISGVDLGLPNWGKYVLLSAGALTALMLIIFLMTCM RRVNRSEPTQHNLRGTGREVSVTPQSGKIISSWESYKSGGETGL* RRVNRSEPTQHNLRGTGREVSVTPQSGKIISSWESYKSGGETGL*

RABIES VIRUS G (SEQ ID NO: 16) auggugccgcaggcucuccuguuugucccccuucuggucuuuccauuguguuuugggaaauucccuaucuacacaaur auggugccgcaggcucuccuguuugucccccuucuggucuuuccauuguguuuugggaaauucccuaucuacacaauu cggacaaguugggacccuggagcccaauugacauucaucaucucagcugcccgaacaauuuggucguggaggace cggacaaguugggacccuggagcccaauugacauucaucaucucagcugcccgaacaauuuggucguggaggacgaagg augcaccaaccugucgggguucuccuacauggaauugaaagucggauacaucagugccauuaagaugaacggguucacu augcaccaaccugucgggguucuccuacauggaauugaaagucggauacaucagugccauuaagaugaacggguucaci ugcacaggcgucgugacugaagcugagacauacacuaacuucgugggauaugucacuaccacuuucaaaagaaagcaut igcacaggcgucgugacugaagcugagacauacacuaacuucgugggauaugucacuaccacuuucaaaagaaagcauu uccgcccuacuccugaugcuuguagggccgcauacaacuggaagauggccggugaccccagauaugaggaaucacuuca uccgcccuacuccugaugcuuguagggccgcauacaacuggaagauggccggugaccccagauaugaggaaucacuuc caauccguacccugacuaccacuggcuucggacugucaaaaccaccaaggagucacucgugaucauuaguccaagug caauccguacccugacuaccacuggcuucggacugucaaaaccaccaaggagucacucgugaucauuaguccaaguqug gcugaucuugacccauacgaccggucacuucacucacggguguucccgggggggaauugcucuggugucgcagugucgu gcugaucuugacccauacgaccggucacuucacucacggguguucccgggggggaauugcucuggugucgcagugucgu caaccuacugcuccacaaaccacgauuacaccauuuggaugccagaaaauccucggcuugguaugucaugugacauuu caaccuacugcuccacaaaccacgauuacaccauuuggaugccagaaaauccucggcuugguaugucaugugacauuuu caccaauucucgggggaagagggcuuccaaagggucugaaacuugcggcuuugucgaugagcggggcuuguauaaguca caccaauucucgggggaagagggcuuccaaagggucugaaacuugcggcuuugucgaugagcggggcuuquauaaguca uuaaaggugcuugcaaacucaagcuuuguggugucuugggauugagauugauggauggaacuugggucgcaaugaga cuuaaaggugcuugcaaacucaagcuuuguggugucuugggauugagauugauggauggaacuugggucgcaaugcaga uucuaacgaaaccaaauggugcccucccggacagcuugugaauuugaugacuuucgcucugacgaaauugagcau uucuaacgaaaccaaauggugcccucccggacagcuugugaauuugcaugacuuucgcucugacgaaauugagcauct ugucgucgaggaguuggucaagaagagggaagagugucuggaugcuuuggaaucaaucaugaccaccaagucagugucu non6nfeon6ee0oeooe6neoneeonee66nnno6ne66non6n6e6ee66656ee6eeon66nnbe66e6on6on6n uucagacggcucucacaucuuaggaaauuggugccagguuuuggaaaagcauauaccauuuucaacaagacccuuaugg ucagacggcucucacaucuuaggaaauuggugccagguuuuggaaaagcauauaccauuuucaacaagacccuuaugg aagccgaugcucacuacaagucugucaggacuuggaaugagaucaucccgucuaaagggugucuuagggucggagggag aagccgaugcucacuacaagucugucaggacuuggaaugagaucaucccgucuaaagggugucuuagggucggagggag sugucauccucaugucaacggagucuuuuucaaugguaucauucuuggaccugacggaaauguccuuaucccugagaus augucauccucaugucaacggagucuuuuucaaugguaucauucuuggaccugacggaaauguccuuaucccugagau caaucuucccuccuccagcaacacauggaacuucuugucucaucggucaucccccuuaugcacccccuggcugacccar caaucuucccuccuccagcaacacauggaacuucuugucucaucggucaucccccuuaugcacccccuggcugacccat caaccguguucaagaacggugacgaggcagaggauuuugucgagguccaccuucccgaugugcaugaacggaucucugg. 66nonone66oee6neo6n6ne6ooonnooeoon66e6on6nnnne66e6eo66e6oe6n660ee6eeonnfn60oee3 gucgaccuuggacucccuaacuggggaaaguauguccuucugucggcaggagcccugacugccuugauguugauuar ugucgaccuuggacucccuaacuggggaaaguauguccuucugucggcaggagcccugacugccuugauguugauuauç uuccugaugacuuguuggaggagagucaaucggucggagccaacacaacauaaucucagaggaacaggaagggaggugu uuccugaugacuuguuggaggagagucaaucggucggagccaacacaacauaaucucagaggaacaggaagggaggugt cagucacaccccaaagcgggaagaucauuucgucuugggagucauacaagagcggaggugaaaccggacuguga cagucacaccccaaagcgggaagaucauuucgucuugggagucauacaagagcggaggugaaaccggacuquga

The following examples serve to provide further appreciation of the invention

but are not meant in any way to restrict the effective scope of the invention.

EXAMPLES

EXAMPLE 1

INCORPORATION OF THE CODING SEQUENCES FOR FCV CAPSID PROTEINS INTO THE ALPHAVIRUS RNA REPLICON PARTICLES

INTRODUCTION RNA viruses have been used as vector-vehicles for introducing vaccine

antigens, which have been genetically engineered into their genomes. However,

their use to date has been limited primarily to incorporating viral antigens into the

RNA virus and then introducing the virus into a recipient host. The result is the

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induction of protective antibodies against the incorporated viral antigens. Alphavirus

RNA replicon particles have been used to encode pathogenic antigens. Such

alphavirus replicon platforms have been developed from several different

alphaviruses, including Venezuelan equine encephalitis virus (VEE) [Pushko et al.,

Virology 239:389-401 (1997)], Sindbis (SIN) [Bredenbeek et al., Journal of Virology

67:6439-6446 (1993) the contents of which are hereby incorporated herein in their

entireties], and Semliki Forest virus (SFV) [Liljestrom and Garoff, Biotechnology

(NY) 9:1356- 1361 (1991), the contents of which are hereby incorporated herein in

their entireties]. Moreover, alphavirus RNA replicon particles are the basis for

several USDA-licensed vaccines for swine and poultry. These include: Porcine

Epidemic Diarrhea Vaccine, RNA Particle (Product Code 19U5.P1 ), Swine

Influenza Vaccine, RNA (Product Code 19A5.D0), Avian Influenza Vaccine, RNA

(Product Code 1905.D0), 19O5.D0), and Prescription Product, RNA Particle (Product Code

9PP0.00).

ALPHAVIRUS RNA REPLICON PARTICLE CONSTRUCTION Amino acid sequences for FCV capsid proteins were used to generate

codon-optimized (feline codon usage) nucleotide sequences in silico. Optimized

sequences were prepared as synthetic DNA by a commercial vendor (ATUM,

Newark, CA). Accordingly, synthetic genes were designed based on the amino acid

sequences of a VS-FCV capsid protein and an FCV F9-like capsid protein,

respectively. The constructs that encode the amino acid sequence for the VS-FCV

capsid protein [SEQ ID NO: 2], or for the FCV F9-like capsid protein [SEQ ID

NO: 4], were codon-optimized for feline, with flanking sequence appropriate for

cloning into the alphavirus replicon plasmid.

The VEE replicon vectors designed to express FCV capsid proteins were

constructed as previously described [see, U.S. 9,441,247 B2; the contents of which

are hereby incorporated herein by reference], with the following modifications. The

TC-83-derived replicon vector "pVEK" [disclosed and described in

U.S. 9,441,247 B2] was digested with restriction enzymes Ascl and Pacl. A DNA

plasmid containing the codon-optimized open reading frame nucleotide sequence of

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one of the FCV capsid protein genes (either FCV F9-Like or VS-FCV), with 5'

flanking sequence (5'-GGCGCGCCGCACC-3') [SEQ ID NO: 11] and 3' flanking

sequence (5'-TTAATTAA-3'), were similarly digested with restriction enzymes Ascl

and Pacl. The synthetic gene cassette was then ligated into the similarly digested

pVEK vector, and the resulting clones were re-named "pVHV-F9" and "pVHV-

Kalem", encoding the FCV F9-Like and the VS-FCV capsid proteins respectively.

The "pVHV" vector nomenclature was chosen to refer to pVEK-derived replicon

vectors containing transgene cassettes cloned via the Ascl and Pacl sites in the

multiple cloning site of pVEK.

To create the dual construct, the pVHV vector region encoding the VEE

subgenomic promoter and FCV Kalem (VS-FCV) capsid sequences was removed by PCR and ligated into the pVHV-F9 vector between the 3' end of the F9 FCV

capsid sequence and the VEE 3' UTR sequence. The duplication of the

subgenomic promoter sequence and confirmation of the FCV Kalem capsid

sequence were achieved by sequencing of the final vector clone, termed "pVHV-F9-

Kalem".

Production of TC-83 RNA replicon particles (RP) was conducted according to

methods previously described [U.S. 9,441,247 B2 and U.S. 8,460,913 B2; the

contents of which are hereby incorporated herein by reference in their entireties].

Briefly, pVHV replicon vector DNA and helper DNA plasmids were linearized with

Notl restriction enzyme prior to in vitro transcription using MegaScript T7 RNA

polymerase and cap analog (Promega, Madison, WI). Importantly, the helper RNAs

used in the production lack the VEE subgenomic promoter sequence, as previously

described [Kamrud described [Kamrud et et al., al., J Gen J Gen Virol. Virol. (Pt 91 (Pt 7):1723-1727 7):1723-1727 (2010)]. (2010)]. Purified Purified RNA for RNA for

the replicon and helper components were combined and mixed with a suspension of

Vero cells, Vero cells,electroporated electroporatedin 4inmm4 cuvettes, and returned mm cuvettes, to OptiPro® and returned SFM cellSFM cell to OptiPro

culture media (Thermo Fisher, Waltham MA). Following overnight incubation,

alphavirus RNA replicon particles were purified from the cells and media by passing

the suspension through a ZetaPlus BioCap depth filter (3M, Maplewood, MN),

washing with phosphate buffered saline containing 5% sucrose (w/v), and finally

eluting the retained RP with 400 mM NaCI NaCl buffer. Eluted RP were formulated to a

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final 5% sucrose (w/v), passed through a 0.22 micron membrane filter, and

dispensed into aliquots for storage. Titer of functional RP was determined by

immunofluorescence assay on infected Vero cell monolayers.

EXAMPLE 2

EVALUATION OF EFFICACY AND SAFETY OF A DUAL CONSTRUCT FCV VACCINE IN CATS

A dual-construct vaccine comprising a propagation defective RNA particle

(RP) encoding the capsid proteins from two different strains of FCV, a virulent

systemic strain (VS-FCV) and a classical vaccine strain (FCV F9-Like) along with

the capsid protein and glycoproteins of the avirulent TC-83 strain of Venezuelan

Equine Encephalitis Virus (VEE) was formulated in 5% sucrose and stored frozen.

This dual-construct vaccine was used to evaluate the effectiveness against

challenge by two FCV strains, as shown in Table 1 below. Two groups of 10 cats

each were vaccinated with the dual-construct FCV vaccine in a prime/ boost

regimen at 13-14 weeks of age and then 21 days later. Two groups of control cats

were vaccinated by the same regimen with a placebo vaccine consisting of cell

culture media (Minimal Essential Media with Earle's salts, EMEM).

TABLE 1 VACCINATION PROTOCOL Treatment No. of Test Product Vaccine Challenge Challenge Group Animals Dose Strain

Dual-construct Classical FCV (Strain 255) 1 9 RP-FCV 6.1 6.1 XX 107 10

Classical FCV (Strain 255) 2 7 Placebo NA Virulent Dual-construct Systemic FCV 3 9 RP-FCV 6.1 6.1 XX 107 10 (Kalem* strain)

Virulent

4 7 Placebo NA Systemic FCV

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(Kalem strain)

"Internal reference *Internal reference

Following the vaccinations the cats were observed for adverse reactions to

the vaccines by observing the cats for any local or systemic reactions to the

vaccines as well as clinical signs of FCV infection. No adverse reactions were

observed for any of the vaccinated cats.

Three weeks following the booster vaccination, cats in Groups 1 and 2 were

challenged intranasally with a virulent culture of FCV strain 255 (classical FCV

challenge strain). Three weeks after booster vaccination cats in Groups 3 and 4

were challenged intranasally with a virulent culture of virulent systemic FCV

challenge strain (FCV strain Kalem).

Cats were observed for clinical signs of FCV infection for 14 days following

challenge as follows: cats were observed and scored daily for clinical signs

including: death, depression/ lethargy, body temperature, nasal and oral ulcers,

nasal and ocular discharge, lameness, dehydration and sneezing. Body weight was

measured on four days spaced throughout the 14 day post-challenge period. Each

of the clinical signs observed was given a weighted numerical score based on

severity and the number of days it was observed. Each cat was then given a total,

weighted score based on the sum of the daily weighted scores. A mean and

median weighted score was then calculated for each treatment group. For the

challenge to be considered valid, 80% of the control cats must show clinical signs of

FCV infection (other than fever). The results of the challenge are summarized in

Table 2 below:

TABLE 2 CHALLENGE RESULTS Treatment Test Product Challenge Challenge Median Mean Group Strain Weighted Weighted Clinical Score Clinical Score Classical FCV 1 Dual-construct (Strain 255) RP-FCV 5 6.3

2 Classical FCV

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Placebo (Strain 255) 16 14.3

Virulent Dual-construct 10 3 Systemic FCV 8.8 8.8 RP-FCV (Kalem strain) Virulent

4 Placebo Systemic FCV 38 36.9 (Kalem strain)

The challenges for both strains were considered valid as 100% of placebo-

vaccinated control cats exhibited clinical signs of FCV infection (other than fever).

The dual-construct RP-FCV vaccine encoding a virulent systemic and classical

vaccine strain of FCV protected cats against two distinct strains of FCV: a classical

FCV strain as well as a virulent systemic FCV strain. The experimental vaccine was

found safe in cats.

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EXAMPLE 3

EVALUATION OF INTERFERENCE WITH ADMINISTRATION OF TWO DIFFERENT RNA PARTICLE VACCINES

The study was conducted to evaluate multiple aspects of the alphavirus RNA

replicon particle FCV vaccine including serological response, efficacy against

challenge, and interference. A RP-FCV construct vaccine encoding the capsid

protein of a classical FCV vaccine strain (F9) was formulated in stabilizer consisting

of gelatin, NZ-amine, and sucrose and lyophilized. Two groups of five cats were

vaccinated with the RP-FCV vaccine at 17 weeks of age. Twenty-one days later,

cats in Group 1 were administered a booster dose of the RP-FCV vaccine only, cats

in Group 2 were administered a booster dose of the RP-FCV vaccine and

administered a dose of an RP-Rabies vaccine at the same time as shown in Table 3

below. The RP-Rabies virus vaccine is a construct encoding the rabies virus

glycoprotein (G) in the same TC-83 VEE alphavirus platform.

TABLE 33 TABLE VACCINATION PROTOCOL Test Product(s)-Booster Test Product -Initial Vaccination Treatment Group Vaccination (Day 0) (Day 21)

1 RP-FCV (F9) RP-FCV (F9)

RP-FCV (F9) 2 RP-FCV (F9) RP-Rabies virus

Following each vaccination the cats were observed for adverse reactions to

the vaccines by observing the cats for any local or systemic reactions to the

vaccines. No adverse reactions were observed for any of the vaccinated cats.

Cats were bled for serum collection on the day of initial vaccination (Study

Day 0), the day of booster vaccination (Study Day 21) and six weeks after the initial

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vaccination (Study Day 42). The serum was tested for antibody titer to FCV F9 by a

serum neutralization assay. Serum was also tested for antibody titer to rabies virus

by the Rapid Fluorescent Focus Inhibition Test (RFFIT). RIFFT results are reported

as International Units per milliliter (IU/ mL). Serology results are summarized in

Tables 4 and 5 below.

TABLE 4 FCV F9 SEROLOGY RESULTS Vaccination FCV (F9) Antibody Titer (Geometric Mean) Treatment Regimen Day 21 Group Day 0 Day 42 (Day 0/ Day 21) RP-FCV (F9)/ 1 RP-FCV (F9) <2 3 34

RP-FCV (F9)/ 2 RP-FCV (F9)+ <2 3 38 RP-Rabies virus

Based on comparison of FCV (F9) antibody titers (on serum samples

collected post-booster) concurrent vaccination with an RP-Rabies virus vaccine

does not interfere with the antibody response to an RP-FCV (F9) vaccine.

TABLE 55 TABLE RABIES VIRUS SEROLOGY RESULTS Vaccination Rabies Antibody Titer (Geometric Mean IU/ mL) Treatment Regimen Day 21 Group Day 0 Day 42 (Day 0/ Day 21) RP-FCV (F9)/ 1 1 RP-FCV (F9) 2 Not Tested

RP-FCV (F9)/ 1 2 RP-FCV (F9)+ <1 39 RP-Rabies virus

Although this study did not include a control group, which was vaccinated

with only with onlyananRP-Rabies virus RP-Rabies vaccine, virus for the vaccine, forpurpose of comparing the purpose the post-booster of comparing the post-booster

rabies titer of Group 2, historical data from other studies is presented below in

Table 6. Table 6.

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TABLE 66 TABLE RABIES VIRUS SEROLOGY RESULTS, MULTIPLE STUDIES Rabies Antibody Titer Vaccination (Geometric Mean IU/ mL) Treatment RP-Rabies Regimen Approximately Group Group virus Potency Pre- 1 month post- vaccination vaccination* vaccination* RP-FCV (F9)/ 1.3 xX 107 1.3 10 1 2 RP-FCV (F9)+ 39 RP-Rabies virus RP-Rabies virus Study RUS- alone 2.7 xX 107 2.7 10 <0.1 <0.1 42.7 006 Single Vaccination

RP-Rabies virus Study RUS- alone 2.6 2.6 XX 106 10 <0.1 <0.1 17.6 17.6 006 Single Vaccination

*Study RUS-006 cats were bled for serum 30 days after vaccination

Based on the potency of the RP-Rabies virus vaccines and the post-

vaccination rabies antibody titers, a prior vaccination with an RP-FCV vaccine does

not interfere with the antibody response to an RP-Rabies virus vaccine. Vector

immunity is a concern with platform-based vaccines however, the results of this

study suggest that multiple RP-based vaccines can be used in an animal without

compromising efficacy.

In order to confirm the lack of interference by concurrent vaccination with

RP-Rabies virus on RP-FCV (F9) efficacy the study was amended and continued. A

group of five age-matched cats were added to the study to serve as non-vaccinated

controls. All cats were challenged intranasally with a virulent, classical strain of FCV

(FCV 255) 79 days after the initial vaccination of Groups 1 and 2.

For 14 days following challenge cats were observed and scored daily for the

following clinical signs of FCV infection: death, depression/ lethargy, body

temperature, nasal and oral ulcers, nasal and ocular discharge, lameness,

dehydration, and sneezing. Body weight was measured on four days spaced

throughout the 14-day post-challenge period. Each of the clinical signs observed

was given a weighted numerical score based on severity and the number of days it

was observed. Each cat was then given a total, weighted score based on the sum

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of the daily weighted scores. A mean and median weighted score was then

calculated for each treatment group. For the challenge to be considered valid, 80%

of the control cats must show clinical signs of FCV infection (other than fever). The

results of the challenge are summarized in the table below:

TABLE 7 INTERFERENCE STUDY CHALLENGE RESULTS Treatment Vaccination Regimen Mean Weighted Median Group Group (Day 0/ Day 21) Clinical Score Weighted Clinical Score

RP-FCV (F9)/ 1 RP-FCV (F9) 3 4.0

RP-FCV (F9)/ RP-FCV RP-FCV (F9)/RP-FCV 2 2 4.4 (F9)+ RP-Rabies virus

3 Non-Vaccinated 20 19.6

The challenge was considered valid as 100% of non-vaccinated control cats

exhibited clinical signs of FCV infection (other than fever). Both vaccinate groups

(Groups 1 and 2) were significantly protected from virulent FCV challenge (p values

0.012 for both groups).

Based on comparison of clinical scores, concurrent vaccination with an RP-

Rabies virus vaccine does not interfere with the efficacy of an RP-FCV (F9) vaccine.

The experimental vaccine was found safe in cats.

EXAMPLE 4

EVALUATION OF VACCINE EFFICACY IN CATS OF A RP CONSTRUCT ENCODING A SINGLE FCV F9-LIKE CAPSID PROTEIN

This study was conducted to further evaluate the efficacy of a feline vaccine

comprising an alphavirus RNA replicon particle that encoded a single FCV F9-Like

capsid protein (RP-FCV F9). The vaccine was compared to a placebo control group

against a classical FCV challenge. Cats were inoculated with either this monovalent

vaccine or a placebo and subsequently challenged with a classical FCV. The

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clinical scores are based on the typical signs of FCV infection, mainly oral and

external ulcers and rhinitis, scored over a period of 14 days following the FCV

challenge. The scoring system is the same as that described in Examples 2 and 3

above. Whereas, the mean clinical score for the placebo controls was 92, the mean

clinical score for the RP-FCV F9 vaccine was only 2. Surprisingly the score for the

RP-FCV F9 vaccine also was significantly lower than that obtained with two

vaccines that individually comprised a single attenuated-live FCV F9-Like virus.

This experiment further demonstrates that an RP-FCV vaccine encoding a classical

vaccine strain of FCV protects cats against an FCV F9-Like virus.

The present invention is not to be limited in scope by the specific

embodiments embodimentsdescribed herein. described Indeed, herein. various Indeed, modifications various of the invention modifications in of the invention in

addition to those described herein will become apparent to those skilled in the art

from the foregoing description. Such modifications are intended to fall within the

scope of the appended claims.

It is further to be understood that all base sizes or amino acid sizes, and all

molecular weight or molecular mass values, given for nucleic acids or polypeptides

are approximate, and are provided for description.

Claims (15)

Claims:

1. An immunogenic composition comprising a Venezuelan Equine Encephalitis (VEE) alphavirus RNA replicon particle that encodes a feline calicivirus (FCV) antigen; wherein the FCV antigen is a capsid protein. 2018380582

2. The immunogenic composition of Claim 1, wherein the capsid protein is selected from the group consisting of an FCV F9-Like capsid protein and a virulent systemic FCV (VS-FCV) capsid protein.

3. The immunogenic composition of Claim 2, wherein the capsid protein is a VS- FCV capsid protein.

4. The immunogenic composition of Claim 2, wherein the capsid protein is a FCV F9-Like capsid protein.

5. The immunogenic composition of any one of Claims 1, 2, 3, or 4, wherein said FCV is of a first strain; and wherein the immunogenic composition further comprises an additional VEE alphavirus RNA replicon particle encoding a second FCV antigen that originates from a second strain of FCV.

6. The immunogenic composition of Claim 3, that comprises an additional VEE alphavirus RNA replicon particle that encodes an FCV F9-Like capsid protein.

7. The immunogenic composition of Claim 3, wherein the VEE alphavirus RNA replicon particle also encodes an FCV F9-Like capsid protein.

8. The immunogenic composition of any one of Claims 2, 3, 5, 6, or 7, wherein the VS-FCV capsid protein comprises an amino acid sequence comprising at least 95% identity with the amino acid sequence of SEQ ID NO: 2.

9. The immunogenic composition of any one of Claims Claim 2, 4, 5, 6, 7, or 8, wherein the FCV F9-Like capsid protein comprises an amino acid sequence comprising at least 95% identity with the amino acid sequence of SEQ ID NO: 4.

10. A vaccine comprising the immunogenic composition of any one of Claims 1, 2, 2018380582

3, 4, 5, 6, 7, 8, or 9 and a pharmaceutically acceptable carrier, wherein the vaccine elicits protective immunity to FCV infection upon administration to a feline subject.

11. The vaccine of Claim 10, that further comprises at least one non-FCV antigen for eliciting protective immunity to a non-FCV feline pathogen.

12. The vaccine of Claim 10 or 11, that further comprises a VEE alphavirus RNA replicon particle comprising a nucleotide sequence encoding at least one protein antigen that originates from a non-FCV antigen.

13. The vaccine of any one of Claims 10, 11, or 12, that is a nonadjuvanted vaccine.

14. A method of immunizing a feline against a pathogenic FCV comprising administering to the feline an immunologically effective amount of the vaccine of any one of Claims 10, 11, 12, or 13.

15. Use of the immunogenic composition of any one of Claims 1, 2, 3, 4, 5, 6, 7, 8, or 9, in the manufacture of a medicament for immunizing a feline against a pathogenic FCV.