EP4735032A2 - Compositions for prevention of cardiomyopathy syndrome - Google Patents

Compositions for prevention of cardiomyopathy syndrome

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Publication number
EP4735032A2
EP4735032A2 EP24832852.8A EP24832852A EP4735032A2 EP 4735032 A2 EP4735032 A2 EP 4735032A2 EP 24832852 A EP24832852 A EP 24832852A EP 4735032 A2 EP4735032 A2 EP 4735032A2
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Prior art keywords
seq
clause
polyprotein
combination
pmcv
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EP24832852.8A
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German (de)
French (fr)
Inventor
Alicia MACDONALD
Jonathan Snyder
Øystein EVENSEN
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Intervet International BV
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Intervet International BV
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Publication of EP4735032A2 publication Critical patent/EP4735032A2/en
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Abstract

The present disclosure provides exemplary sequences and compositions that can be used to active immunization of animals to aid in the prevention of cardiomyopathy syndrome (CMS) caused by Piscine Myocarditis Virus (PMCV). Vaccines and kits comprising the sequences and compositions are also provided, as well as methods of administering the vaccine to non-human animals.

Description

COMPOSITIONS FOR PREVENTION OF CARDIOMYOPATHY SYNDROME
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63/523,471, filed on June 27, 2023, the entire disclosure of which is incorporated herein by reference.
TECHNICAL FIELD
The disclosure relates to constructs and compositions directed to immunization of animals to aid in the prevention of cardiomyopathy syndrome (CMS) caused by Piscine Myocarditis Virus (PMCV). In one aspect, the present disclosure provides compositions and methods directed to protection of salmon.
BACKGROUND AND SUMMARY OF THE INVENTION
Cardiomyopathy syndrome (CMS) is an emerging disease of economic importance in salmon. In particular, the Piscine Myocarditis Virus (PMCV) has been identified as a causative link for CMS. PMCV is of large economic concern in the salmonid industry because it causes morbidity and mortality in adult fish that are nearly full grown and almost ready for harvest.
PMCV encodes at least three major proteins: a capsid (ORF1), a RNA- dependent RNA polymerase (ORF2), and a structural protein of unknown function (ORF3) that may be involved in virus assembly or egress. However, attempts to propagate PMCV using in vitro cell culture systems have failed. As a result, traditional live, attenuated, and inactivated vaccine approaches have not yet materialized. Thus, there exists a need for a PMCV vaccine that utilizes DNA, RNA, or recombinant approaches for design.
Accordingly, the present disclosure provides constructs and compositions capable of immunizing animals against PMCV. The exemplary embodiments of the present disclosure include two different approaches of DNA-based vaccination. Using nanoplasmid technology, a series of plasmids were designed as DNA vaccine vectors. The series includes exemplary plasmids such as a plasmid including ORF1, a plasmid including ORF3, and a dual expression plasmid including both ORF1 and ORF3. The constructs and compositions of the present disclosure can provide beneficial effects against CMS for salmon. BRIEF DESCRIPTIONS OF THE DRAWINGS
The detailed description particularly refers to the accompanying figures in which:
FIGURE 1 shows atrium scores (0-4) for the different groups at 12 weeks postchallenge.
FIGURE 2 shows ventricle scores (0-4) for the different groups at 12 weeks post-challenge.
FIGURE 3 shows the mean ±95% CI for the sum of scores (severity) for atrium + ventricle is shown for all groups at 12 weeks post-challenge. A distinct separation was observed between groups at 12 weeks post-challenge.
FIGURE 4 shows Ct values for heart in different groups at 12 weeks postchallenge. Results are shown as mean and 95% CI (spikes).
DETAILED DESCRIPTION
Various embodiments are described herein as follows. In an illustrative aspect, an expression vector is provided. The expression vector encodes i) a salmonid alphavirus (SAV) polyprotein and ii) a piscine myocarditis virus (PMCV) polyprotein. Salmonid alphavirus (SAV) is known in the art as an enveloped, single-stranded, positive-sense RNA virus with a genome of about 12 kb. For instance, Clynav™ is a DNA vaccine that can be used to protect Atlantic salmon against pancreatic disease caused by SAV. Certain aspects regarding Clynav™ are found, for example, in U.S. Patent No. 9,303,071, which is herein incorporated by reference in its entirety.
In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein. In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3. As described herein, SEQ ID NO: 3 corresponds to the ORF_3 (wild type) sequence. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. As described herein, SEQ ID NO: 2 corresponds to the ORF_3 (mutant) sequence. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein encoded by the expression vector is fused with the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector. Methods of fusing are generally known in the art to a skilled artisan.
In an embodiment, the SAV polyprotein comprises one or more regions selected from the group consisting of El, E2, E3, 6K, and any combination thereof. In an embodiment, the PMCV polyprotein encoded by the expression vector is fused to a N-terminus of the E2 region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector. In an embodiment, the PMCV polyprotein encoded by the expression vector is fused to a C-terminus of the E2 region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector. In an embodiment, the PMCV polyprotein encoded by the expression vector is fused to a N- terminus of the 6K region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector. In an embodiment, the PMCV polyprotein encoded by the expression vector is fused to a C-terminus of the 6K region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector.
In an illustrative aspect, a composition is provided. The composition comprises i) a first expression vector encoding a salmonid alphavirus (SAV) polyprotein and ii) a second expression vector encoding a piscine myocarditis virus (PMCV) polyprotein.
In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein. In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein encoded by the first expression vector is fused with the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors. In an embodiment, the SAV polyprotein comprises one or more regions selected from the group consisting of El, E2, E3, 6K, and any combination thereof. In an embodiment, the PMCV polyprotein encoded by the first expression vector is fused to a N-terminus of the E2 region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors. In an embodiment, the PMCV polyprotein encoded by the first expression vector is fused to a C- terminus of the E2 region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors. In an embodiment, the PMCV polyprotein encoded by the first expression vector is fused to a N-terminus of the 6K region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors. In an embodiment, the PMCV polyprotein encoded by the first expression vector is fused to a C-terminus of the 6K region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors.
In an illustrative aspect, a polypeptide is provided. The polypeptide comprises i) a salmonid alphavirus polyprotein and ii) a PMCV polyprotein.
In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein. In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein. In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is fused with the SAV polyprotein, and wherein the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector.
In an embodiment, the PMCV polyprotein is fused to a N-terminus of the E2 region of the SAV polyprotein, and In an embodiment, the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector. In an embodiment, the PMCV polyprotein is fused to a C-terminus of the E2 region of the SAV polyprotein, and In an embodiment, the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector. In an embodiment, the PMCV polyprotein is fused to a N- terminus of the 6K region of the SAV polyprotein, and In an embodiment, the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector. In an embodiment, the PMCV polyprotein is fused to a C-terminus of the 6K region of the SAV polyprotein, and In an embodiment, the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector.
In an illustrative aspect, an immunogenic composition is provided. The immunogenic composition comprises the expression vector or the expression vectors of any embodiment described herein.
In an embodiment, the immunogenic composition further comprises an adjuvant. In an embodiment, the immunogenic composition further comprises salmonid interferon type A. As described herein, SEQ ID NO: 4 corresponds to the Salmonid Interferon A sequence. In an embodiment, the salmonid interferon type A is encoded in a DNA vector.
In an embodiment, the immunogenic composition further comprises salmonid interferon type C. As described herein, SEQ ID NO: 5 corresponds to the Salmonid Interferon C sequence. In an embodiment, the salmonid interferon type C is encoded in a DNA vector.
In an embodiment, the immunogenic composition further comprises salmonid interferon type A and salmonid interferon type C. In an embodiment, the salmonid interferon type A is encoded in a DNA vector. In an embodiment, the salmonid interferon type C is encoded in a DNA vector.
In an embodiment, the salmonid interferon type A is encoded in a first DNA vector and the salmonid interferon type C is encoded in a second DNA vector. In an embodiment, the salmonid interferon type A and the salmonid interferon type C are encoded in a single DNA vector. In an illustrative aspect, a vaccine is provided. The vaccine comprises the expression vector or the expression vectors of any embodiment described herein.
In an embodiment, the vaccine further comprises an adjuvant. In an embodiment, the vaccine further comprises salmonid interferon type A. In an embodiment, the vaccine further comprises salmonid interferon type C. In an embodiment, the vaccine further comprises salmonid interferon type A and salmonid interferon type C.
In an illustrative aspect, a vaccine is provided. The vaccine comprises the immunogenic composition of any embodiment described herein.
In an embodiment, the vaccine further comprises an adjuvant. In an embodiment, the vaccine further comprises salmonid interferon type A. In an embodiment, the vaccine further comprises salmonid interferon type C. In an embodiment, the vaccine further comprises salmonid interferon type A and salmonid interferon type C.
In an illustrative aspect, a method for inducing an immune response in a host against a PMCV is provided. The method comprises the step of administering to the host the expression vector or the expression vectors of any embodiment described herein.
In an embodiment, the expression vector or the expression vectors is a plasmid which is administered by injection into muscle tissue of the host. In an embodiment, the expression vector or the expression vectors is a plasmid which is administered by injection intraperitoneally into the host. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
In an embodiment, the step of administering the immunogenic composition to the host comprises administering a single dose. In an embodiment, the method further comprises administration of a second therapeutic agent. In an embodiment, the second therapeutic agent comprises an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, a method for inducing an immune response in a host against a PMCV is provided. The method comprises the step of administering to the host the immunogenic composition of any embodiment described herein. In an embodiment, the immunogenic composition comprises a plasmid comprising the expression vector or the expression vectors of any embodiment described herein which is administered by injection into muscle tissue of the host. In an embodiment, the expression vector or the expression vectors of any embodiment described herein is a plasmid which is administered by injection intraperitoneally into the host. In an embodiment, the method is a prophylactic administration to the host.
In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
In an embodiment, the step of administering the immunogenic composition to the host comprises administering a single dose. In an embodiment, the method further comprises administration of a second therapeutic agent. In an embodiment, the second therapeutic agent comprises an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, a method for inducing an immune response in a host against a PMCV is provided. The method comprises the step of administering to the host the vaccine of any embodiment described herein.
In an embodiment, the method is a prophylactic administration to the host. In an embodiment, the method is an intramuscular administration to the host. In an embodiment, the method is an intraperitoneal administration to the host. In an embodiment, the method comprises administering a single dose.
In an embodiment, the vaccine further comprises an adjuvant. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
In an embodiment, the step of administering the immunogenic composition to the host comprises administering a single dose. In an embodiment, the method further comprises administration of a second therapeutic agent. In an embodiment, the second therapeutic agent comprises an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, an expression vector encoding a piscine myocarditis virus (PMCV) polyprotein is provided.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence selected from a group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. As described herein, SEQ ID NO: 1 corresponds to the ORF_1 (wild type) sequence. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 1. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 1.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 1 . In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 1. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 1.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2.
In an embodiment, the PMCV polyprotein comprises a first polyprotein at least 90% identical to SEQ ID NO: 1 and a second polyprotein, wherein the second polyprotein is at least 90% identical to SEQ ID NO: 2 or is at least 90% identical to SEQ ID NO: 3.
In an embodiment, the first polyprotein is at least 90% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 91% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 92% identical with SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 93% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 94% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 95% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 96% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 97% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 98% identical to SEQ ID NO: 1. In an embodiment, the first polyprotein is at least 99% identical to SEQ ID NO: 1.
In an embodiment, the second polyprotein is at least 90% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 91% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 92% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 93% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 94% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 95% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 96% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 97% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 98% identical to SEQ ID NO: 2. In an embodiment, the second polyprotein is at least 99% identical to SEQ ID NO: 2.
In an embodiment, the second polyprotein is at least 90% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 91% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 92% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 93% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 94% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 95% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 96% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 97% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 98% identical to SEQ ID NO: 3. In an embodiment, the second polyprotein is at least 99% identical to SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 1 and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 1 and SEQ ID NO: 3. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 1 and SEQ ID NO: 3.
In an embodiment, the PMCV polyprotein comprises SEQ ID NO: 1 and SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists of SEQ ID NO: 1 and SEQ ID NO: 2. In an embodiment, the PMCV polyprotein consists essentially of SEQ ID NO: 1 and SEQ ID NO: 2.
In an illustrative aspect, a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and any combination thereof.
In an embodiment, the amino acid sequence comprises SEQ ID NO: 1. In an embodiment, the amino acid sequence consists of SEQ ID NO: 1. In an embodiment, the amino acid sequence consists essentially of SEQ ID NO: 1.
In an embodiment, the amino acid sequence is at least 90% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 91% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 92% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 93% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 94% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 95% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 96% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 97% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 98% identical to SEQ ID NO: 1. In an embodiment, the amino acid sequence is at least 99% identical to SEQ ID NO: 1.
In an embodiment, the amino acid sequence comprises SEQ ID NO: 2. In an embodiment, the amino acid sequence consists of SEQ ID NO: 2. In an embodiment, the amino acid sequence consists essentially of SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 90% identical to SEQ ID
NO: 2. In an embodiment, the amino acid sequence is at least 91% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 92% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 93% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 94% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 95% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 96% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 97% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 98% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence is at least 99% identical to SEQ ID NO: 2.
In an embodiment, the amino acid sequence comprises SEQ ID NO: 3. In an embodiment, the amino acid sequence consists of SEQ ID NO: 3. In an embodiment, the amino acid sequence consists essentially of SEQ ID NO: 3.
In an embodiment, the amino acid sequence is at least 90% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 91% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 92% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 93% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 94% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 95% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 96% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 97% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 98% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence is at least 99% identical to SEQ ID NO: 3.
In an embodiment, the amino acid sequence comprises a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence consists of a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 3. In an embodiment, the amino acid sequence consists essentially of a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 3.
In an embodiment, the amino acid sequence comprises a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence consists of a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 2. In an embodiment, the amino acid sequence consists essentially of a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 2.
In an embodiment, the first sequence is at least 90% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 91% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 92% identical to SEQ ID NO: 1 . In an embodiment, the first sequence is at least 93% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 94% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 95% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 96% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 97% identical to SEQ ID NO: 1. In an embodiment, the first sequence is at least 98% identical to SEQ ID NO:
1. In an embodiment, the first sequence is at least 99% identical to SEQ ID NO: 1.
In an embodiment, the second sequence is at least 90% identical to SEQ ID NO:
2. In an embodiment, the second sequence is at least 91% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 92% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 93% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 94% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 95% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 96% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 97% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 98% identical to SEQ ID NO: 2. In an embodiment, the second sequence is at least 99% identical to SEQ ID NO: 2.
In an embodiment, the second sequence is at least 90% identical to SEQ ID NO:
3. In an embodiment, the second sequence is at least 91% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 92% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 93% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 94% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 95% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 96% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 97% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 98% identical to SEQ ID NO: 3. In an embodiment, the second sequence is at least 99% identical to SEQ ID NO: 3.
In an illustrative aspect, an immunogenic composition comprising the expression vector of any embodiment described herein is provided. In an embodiment, the immunogenic composition further comprises an adjuvant. In an embodiment, the immunogenic composition further comprises salmonid interferon type A. In an embodiment, the salmonid interferon type A is encoded in a DNA vector. In an embodiment, the immunogenic composition further comprises salmonid interferon type C. In an embodiment, the salmonid interferon type C is encoded in a DNA vector. In an embodiment, the immunogenic composition further comprises salmonid interferon type A and salmonid interferon type C. In an embodiment, the salmonid interferon type A is encoded in a DNA vector. In an embodiment, the salmonid interferon type C is encoded in a DNA vector. In an embodiment, the salmonid interferon type A is encoded in a first DNA vector and the salmonid interferon type C is encoded in a second DNA vector. In an embodiment, the salmonid interferon type A and the salmonid interferon type C are encoded in a single DNA vector.
In an illustrative aspect, a vaccine comprising the expression vector of any embodiment described herein is provided.
In an embodiment, the vaccine further comprises an adjuvant. In an embodiment, the vaccine further comprises salmonid interferon type A. In an embodiment, the vaccine further comprises salmonid interferon type C. In an embodiment, the vaccine further comprises salmonid interferon type A and salmonid interferon type C.
In an illustrative aspect, a vaccine comprising the immunogenic composition of any embodiment described herein is provided.
In an embodiment, the vaccine further comprises an adjuvant. In an embodiment, the vaccine further comprises salmonid interferon type A. In an embodiment, the vaccine further comprises salmonid interferon type C. In an embodiment, the vaccine further comprises salmonid interferon type A and salmonid interferon type C.
In an illustrative aspect, a kit comprising a container comprising the vaccine of any embodiment described herein is provided.
In an embodiment, the vaccine is a parenteral formulation for administration to a non-human animal via a route of administration selected from the group consisting of intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intracerebroventricular, intraurethral, intrastemal, intracranial, intratumoral, intramuscular, and subcutaneous. In an embodiment, the container comprises an effective amount of the vaccine.
In an embodiment, the vaccine in the container is a ready-to-use liquid composition. In an embodiment, the vaccine in in the container is a lyophilized composition. In an embodiment, the vaccine in the container is a single unit dose. In an embodiment, the vaccine in the container is a multi-unit dose. In an embodiment, the kit further comprises comprising a diluent.
In an embodiment, the kit further comprises a second container comprising a salmonid alphavirus polyprotein (SAV) vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
In an embodiment, the kit further comprises a second container comprising an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, a method for inducing an immune response in a host against a PMCV is provided. The method comprises the step of administering to the host the expression vector of any embodiment described herein.
In an embodiment, the expression vector is a plasmid which is administered by injection into muscle tissue of the host. In an embodiment, the expression vector is a plasmid which is administered by injection intraperitoneally into the host. In an embodiment, the method is a prophylactic administration to the host.
In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
In an embodiment, the step of administering the immunogenic composition to the host comprises administering a single dose. In an embodiment, the method further comprises administration of a second therapeutic agent. In an embodiment, the second therapeutic agent is a salmonid alphavirus polyprotein (SAV) vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
In an embodiment, the second therapeutic agent comprises an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, a method for inducing an immune response in a host against a PMCV is provided. The method comprises the step of administering to the host the immunogenic composition of any embodiment described herein.
In an embodiment, the immunogenic composition comprises a plasmid comprising the expression vector of any embodiment described herein which is administered by injection into muscle tissue of the host. In an embodiment, the expression vector of any embodiment described herein is a plasmid which is administered by injection intraperitoneally into the host. In an embodiment, the method is a prophylactic administration to the host.
In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C. In an embodiment, the step of administering the immunogenic composition to the host comprises administering a single dose.
In an embodiment, the method further comprises administration of a second therapeutic agent. In an embodiment, the second therapeutic agent is a SAV polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
In an embodiment, the second therapeutic agent comprises an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, a method for inducing an immune response in a host against a PMCV is provided. The method comprises the step of administering to the host the vaccine of any embodiment described herein.
In an embodiment, the method is a prophylactic administration to the host. In an embodiment, the method is an intramuscular administration to the host. In an embodiment, the method is an intraperitoneal administration to the host. In an embodiment, the method comprises administering a single dose. In an embodiment, the vaccine further comprises an adjuvant.
In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. In an embodiment, the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
In an embodiment, the method further comprises administration of a second therapeutic agent. In an embodiment, the second therapeutic agent is a SAV polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alpha virus subtype 3 (SAV3) polyprotein vaccine.
In an embodiment, the second therapeutic agent comprises an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
In an illustrative aspect, a kit comprising a container comprising the vaccine of any embodiment described herein is provided.
In an embodiment, the vaccine is a parenteral formulation for administration to a non-human animal via a route of administration selected from the group consisting of intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intracerebroventricular, intraurethral, intrastemal, intracranial, intratumoral, intramuscular, and subcutaneous. In an embodiment, the container comprises an effective amount of the vaccine.
In an embodiment, the vaccine in the container is a ready-to-use liquid composition. In an embodiment, the vaccine in in the container is a lyophilized composition.
In an embodiment, the vaccine in the container is a single unit dose. In an embodiment, the vaccine in the container is a multi-unit dose. In an embodiment, the kit further comprises a diluent.
In an embodiment, the kit further comprises a second container comprising a SAV polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine. In an embodiment, the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine. In an embodiment, the kit further comprises a second container comprising an immunogenic material. In an embodiment, the immunogenic material comprises a material of viral origin or a material of bacterial origin. In an embodiment, the immunogenic material comprises a material of viral origin. In an embodiment, the immunogenic material comprises a material of bacterial origin.
The following numbered embodiments are contemplated and are non-limiting. In any of the following embodiments, SEQ ID NO: 2 can be used instead of SEQ ID NO: 3.
1. An expression vector encoding i) a salmonid alphavirus (SAV) polyprotein and ii) a piscine myocarditis virus (PMCV) polyprotein.
2. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein.
3. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein.
4. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3.
5. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3.
6. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3.
7. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3.
8. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3.
9. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. 10. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3.
11. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3.
12. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3.
13. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
14. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 3.
15. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 3.
16. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 3.
17. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2.
18. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2.
19. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. 0. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. 1. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. 22. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2.
23. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2.
24. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2.
25. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2.
26. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2.
27. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 2.
28. The expression vector of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 2.
29. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 2.
30. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the expression vector is fused with the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector.
31. The expression vector of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein comprises one or more regions selected from the group consisting of El, E2, E3, 6K, and any combination thereof.
32. The expression vector of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the expression vector is fused to a N-terminus of the E2 region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector.
33. The expression vector of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the expression vector is fused to a C-terminus of the E2 region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector.
34. The expression vector of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the expression vector is fused to a N-terminus of the 6K region of the SAV polyprotein encoded hy the expression vector when a polyprotein is formed by expression of the expression vector.
35. The expression vector of clause 31, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the expression vector is fused to a C-terminus of the 6K region of the SAV polyprotein encoded by the expression vector when a polyprotein is formed by expression of the expression vector.
36. A composition comprising i) a first expression vector encoding a salmonid alphavirus (SAV) polyprotein and ii) a second expression vector encoding a piscine myocarditis virus (PMCV) polyprotein.
37. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein.
38. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein.
39. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3.
40. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3.
41. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3.
42. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3.
43. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. 44. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3.
45. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3.
46. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3.
47. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3.
48. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
49. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 3.
50. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 3.
51. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 3.
52. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2.
53. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2.
54. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2.
55. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 2. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the first expression vector is fused with the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors. The composition of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein comprises one or more regions selected from the group consisting of El, E2, E3, 6K, and any combination thereof. The composition of clause 66, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the first expression vector is fused to a N-terminus of the E2 region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors.
68. The composition of clause 66, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the first expression vector is fused to a C-terminus of the E2 region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors.
69. The composition of clause 66, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the first expression vector is fused to a N-terminus of the 6K region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors.
70. The composition of clause 66, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein encoded by the first expression vector is fused to a C-terminus of the 6K region of the SAV polyprotein encoded by the second expression vector when a polyprotein is formed by expression of the expression vectors.
71. A polypeptide comprising i) a salmonid alphavirus polyprotein and ii) a PMCV polyprotein.
72. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein.
73. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein.
74. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3.
75. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3.
76. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3.
77. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 3. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 2. The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 2. The polypeptide of clause 71 , any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 2. . The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is fused with the SAV polyprotein, and wherein the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector. . The polypeptide of clause 71, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein comprises one or more regions selected from the group consisting of El, E2, E3, 6K, and any combination thereof. 102. The polypeptide of clause 101, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is fused to a N-terminus of the E2 region of the SAV polyprotein, and wherein the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector.
103. The polypeptide of clause 101, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is fused to a C-terminus of the E2 region of the SAV polyprotein, and wherein the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector.
104. The polypeptide of clause 101, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is fused to a N-terminus of the 6K region of the SAV polyprotein, and wherein the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector.
105. The polypeptide of clause 101, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is fused to a C-terminus of the 6K region of the SAV polyprotein, and wherein the PMCV polyprotein and the SAV polyprotein are encoded by an expression vector, when a polyprotein is formed by expression of the expression vector.
106. An immunogenic composition comprising the expression vector or the expression vectors of any one of clauses 1 to 70.
107. The immunogenic composition of clause 106, any other suitable clause, or any combination of suitable clauses, further comprising an adjuvant.
108. The immunogenic composition of clause 106, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A.
109. The immunogenic composition of clause 108, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A is encoded in a DNA vector.
110. The immunogenic composition of clause 106, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type C.
111. The immunogenic composition of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type C is encoded in a DNA vector. 112. The immunogenic composition of clause 106, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A and salmonid interferon type C.
113. The immunogenic composition of clause 112, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A is encoded in a DNA vector.
114. The immunogenic composition of clause 112, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type C is encoded in a DNA vector.
115. The immunogenic composition of clause 112, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A is encoded in a first DNA vector and the salmonid interferon type C is encoded in a second DNA vector.
116. The immunogenic composition of clause 112, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A and the salmonid interferon type C are encoded in a single DNA vector.
117. A vaccine comprising the expression vector or the expression vectors of any one of clauses 1 to 70.
118. The vaccine of clause 117, any other suitable clause, or any combination of suitable clauses, further comprising an adjuvant.
119. The vaccine of clause 117, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A.
120. The vaccine of clause 117, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type C.
121. The vaccine of clause 117, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A and salmonid interferon type C.
122. A vaccine comprising the immunogenic composition of any one of clauses 106 to 116.
123. The vaccine of clause 122, any other suitable clause, or any combination of suitable clauses, further comprising an adjuvant.
124. The vaccine of clause 122, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A. 125. The vaccine of clause 122, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type C.
126. The vaccine of clause 122, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A and salmonid interferon type C.
127. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the expression vector or the expression vectors of any one of clauses 1 to 70.
128. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the expression vector or the expression vectors is a plasmid which is administered by injection into muscle tissue of the host.
129. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the expression vector or the expression vectors is a plasmid which is administered by injection intraperitoneally into the host.
130. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
131. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
132. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
133. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the step of administering the immunogenic composition to the host comprises administering a single dose.
134. The method of clause 127, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administration of a second therapeutic agent.
135. The method of clause 134, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent comprises an immunogenic material. 136. The method of clause 135, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
137. The method of clause 135, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.
138. The method of clause 135, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
139. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the immunogenic composition of any one of clauses 106 to 116.
140. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic composition comprises a plasmid comprising the expression vector or the expression vectors of any one of clauses 1 to 70 which is administered by injection into muscle tissue of the host.
141. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the expression vector or the expression vectors of any one of clauses 1 to 70 is a plasmid which is administered by injection intraperitoneally into the host.
142. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the method is a prophylactic administration to the host.
143. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
144. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
145. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
146. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the step of administering the immunogenic composition to the host comprises administering a single dose. 147. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administration of a second therapeutic agent.
148. The method of clause 147, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent comprises an immunogenic material.
149. The method of clause 148, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
150. The method of clause 148, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.
151. The method of clause 148, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
152. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the vaccine of any one of clauses 117 to 126.
153. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method is a prophylactic administration to the host.
154. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method is an intramuscular administration to the host.
155. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method is an intraperitoneal administration to the host.
156. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a single dose.
157. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the vaccine further comprises an adjuvant.
158. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
159. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C. 160. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
161. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the step of administering the immunogenic composition to the host comprises administering a single dose.
162. The method of clause 152, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administration of a second therapeutic agent.
163. The method of clause 162, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent comprises an immunogenic material.
164. The method of clause 163, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
165. The method of clause 163, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.
166. The method of clause 163, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
167. An expression vector encoding a piscine myocarditis virus (PMCV) polyprotein.
168. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence selected from a group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
169. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
170. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. 171. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
172. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
173. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
174. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
175. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
176. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
177. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
178. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 1.
179. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 1 . . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. 213. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3.
214. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3.
215. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3.
216. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3.
217. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises a first polyprotein at least 90% identical to SEQ ID NO: 1 and a second polyprotein, wherein the second polyprotein is at least 90% identical to SEQ ID NO: 2 or is at least 90% identical to SEQ ID NO: 3.
218. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 90% identical to SEQ ID NO: 1.
219. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 91% identical to SEQ ID NO: 1.
220. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 92% identical with SEQ ID NO: 1.
221. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 93% identical to SEQ ID NO: 1.
222. The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 94% identical to SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 95% identical to SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 96% identical to SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 97% identical to SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 98% identical to SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the first polyprotein is at least 99% identical to SEQ ID NO: 1. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 90% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 91% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 92% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 93% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 94% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 95% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 96% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 97% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 98% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 99% identical to SEQ ID NO: 2. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 90% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 91% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 92% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 93% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 94% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 95% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 96% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 97% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 98% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the second polyprotein is at least 99% identical to SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein comprises i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists of i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3. . The expression vector of clause 167, any other suitable clause, or any combination of suitable clauses, wherein the PMCV polyprotein consists essentially of i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3. . A polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and any combination thereof. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists of SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists essentially of SEQ ID NO: 1.. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 91% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 92% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 93% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 94% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 96% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 97% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 98% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 99% identical to SEQ ID NO: 1. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists of SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists essentially of SEQ ID NO: 2.. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 2. 269. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 91% identical to SEQ ID NO: 2.
270. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 92% identical to SEQ ID NO: 2.
271. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 93% identical to SEQ ID NO: 2.
272. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 94% identical to SEQ ID NO: 2.
273. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 2.
274. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 96% identical to SEQ ID NO: 2.
275. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 97% identical to SEQ ID NO: 2.
276. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 98% identical to SEQ ID NO: 2.
277. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 99% identical to SEQ ID NO: 2.
278. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises SEQ ID NO: 3.
279. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists of SEQ ID NO: 3.
280. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists essentially of SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 91 % identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 92% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 93% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 94% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 96% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 97% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 98% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence is at least 99% identical to SEQ ID NO: 3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence comprises a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence, wherein the second sequence is at least 90% identical to SEQ ID NO: 2 or is at least 90% identical to SEQ ID NO: 3. 292. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists of a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 3.
293. The polypeptide of clause 251 , any other suitable clause, or any combination of suitable clauses, wherein the amino acid sequence consists essentially of a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence at least 90% identical to SEQ ID NO: 3.
294. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 90% identical to SEQ ID NO: 1.
295. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 91% identical to SEQ ID NO: 1.
296. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 92% identical to SEQ ID NO: 1.
297. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 93% identical to SEQ ID NO: 1.
298. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 94% identical to SEQ ID NO: 1 .
299. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 95% identical to SEQ ID NO: 1.
300. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 96% identical to SEQ ID NO: 1.
301. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 97% identical to SEQ ID NO: 1.
302. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 98% identical to SEQ ID NO: 1.
303. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the first sequence is at least 99% identical to SEQ ID NO: 1
304. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 90% identical to SEQ ID NO: 2
305. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 91% identical to SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 92% identical to SEQ ID NO:
2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 93% identical to SEQ ID NO:
2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 94% identical to SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 95% identical to SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 96% identical to SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 97% identical to SEQ ID NO: 2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 98% identical to SEQ ID NO:
2. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 99% identical to SEQ ID NO: 2. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 90% identical to SEQ ID NO: 3. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 91% identical to SEQ ID NO:
3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 92% identical to SEQ ID NO:
3. . The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 93% identical to SEQ ID NO:
3. 318. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 94% identical to SEQ ID NO: 3.
319. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 95% identical to SEQ ID NO: 3.
320. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 96% identical to SEQ ID NO: 3.
321. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 97% identical to SEQ ID NO: 3.
322. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 98% identical to SEQ ID NO: 3.
323. The polypeptide of clause 251, any other suitable clause, or any combination of suitable clauses, wherein the second sequence is at least 99% identical to SEQ ID NO: 3.
324. An immunogenic composition comprising the expression vector of any one of clauses 167 to 250.
325. The immunogenic composition of clause 324, any other suitable clause, or any combination of suitable clauses, further comprising an adjuvant.
326. The immunogenic composition of clause 324, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A.
327. The immunogenic composition of clause 326, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A is encoded in a DNA vector.
328. The immunogenic composition of clause 324, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type C.
329. The immunogenic composition of clause 328, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type C is encoded in a DNA vector. 330. The immunogenic composition of clause 324, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A and salmonid interferon type C.
331. The immunogenic composition of clause 330, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A is encoded in a DNA vector.
332. The immunogenic composition of clause 330, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type C is encoded in a DNA vector.
333. The immunogenic composition of clause 330, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A is encoded in a first DNA vector and the salmonid interferon type C is encoded in a second DNA vector.
334. The immunogenic composition of clause 324, any other suitable clause, or any combination of suitable clauses, wherein the salmonid interferon type A and the salmonid interferon type C are encoded in a single DNA vector.
335. A vaccine comprising the expression vector of any one of clauses 167 to 250.
336. The vaccine of clause 335, any other suitable clause, or any combination of suitable clauses, further comprising an adjuvant.
337. The vaccine of clause 335, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A.
338. The vaccine of clause 335, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type C.
339. The vaccine of clause 335, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A and salmonid interferon type C.
340. A vaccine comprising the immunogenic composition of any one of clauses 324 to 334.
341. The vaccine of clause 340, any other suitable clause, or any combination of suitable clauses, further comprising an adjuvant.
342. The vaccine of clause 340, any other suitable clause, or any combination of suitable clauses,
343. The vaccine of clause 340, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type C. 344. The vaccine of clause 340, any other suitable clause, or any combination of suitable clauses, further comprising salmonid interferon type A and salmonid interferon type C.
345. A kit comprising a container comprising the vaccine of any one of clauses 335 to 344.
346. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is a parenteral formulation for administration to a nonhuman animal via a route of administration selected from the group consisting of intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intracerebroventricular, intraurethral, intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.
347. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the container comprises an effective amount of the vaccine.
348. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in the container is a ready-to-use liquid composition.
349. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in in the container is a lyophilized composition.
350. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in the container is a single unit dose.
351. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in the container is a multi-unit dose.
352. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, further comprising a diluent.
353. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the kit further comprises a second container comprising a salmonid alphavirus polyprotein (SAV) vaccine.
354. The kit of clause 353, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine.
355. The kit of clause 353, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine. 356. The kit of clause 345, any other suitable clause, or any combination of suitable clauses, wherein the kit further comprises a second container comprising an immunogenic material.
357. The kit of clause 356, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
358. The kit of clause 356, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.
359. The kit of clause 356, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
360. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the expression vector of any one of clauses 167 to 250.
361. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the expression vector is a plasmid which is administered by injection into muscle tissue of the host.
362. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the expression vector is a plasmid which is administered by injection intraperitoneally into the host.
363. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the method is a prophylactic administration to the host.
364. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
365. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
366. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
367. The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the step of administering the immunogenic composition to the host comprises administering a single dose. . The method of clause 360, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administration of a second therapeutic agent. . The method of clause 368, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent is a salmonid alphavirus polyprotein (SAV) vaccine. . The method of clause 369, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype
2 (SAV2) polyprotein vaccine. . The method of clause 369, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype
3 (SAV3) polyprotein vaccine. . The method of clause 368, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent comprises an immunogenic material. . The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin. . The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.. The method of clause 372, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin. . A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the immunogenic composition of any one of clauses 324 to 334. . The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic composition comprises a plasmid comprising the expression vector of any one of clauses 167 to 250 which is administered by injection into muscle tissue of the host. . The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the expression vector of any one of clauses 167 to 250 is a plasmid which is administered by injection intraperitoneally into the host. 379. The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the method is a prophylactic administration to the host.
380. The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
381. The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
382. The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
383. The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the step of administering the immunogenic composition to the host comprises administering a single dose.
384. The method of clause 376, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administration of a second therapeutic agent.
385. The method of clause 384, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent is a SAV polyprotein vaccine.
386. The method of clause 385, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype
2 (SAV2) polyprotein vaccine.
387. The method of clause 385, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype
3 (SAV3) polyprotein vaccine.
388. The method of clause 384, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent comprises an immunogenic material.
389. The method of clause 388, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
390. The method of clause 388, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin. 391. The method of clause 388, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
392. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the vaccine of any one of clauses 335 to 344.
393. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method is a prophylactic administration to the host.
394. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method is an intramuscular administration to the host.
395. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method is an intraperitoneal administration to the host.
396. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a single dose.
397. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the vaccine further comprises an adjuvant.
398. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
399. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
400. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
401. The method of clause 392, any other suitable clause, or any combination of suitable clauses, wherein the method further comprises administration of a second therapeutic agent.
402. The method of clause 401, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent is a SAV polyprotein vaccine.
403. The method of clause 402, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine. 404. The method of clause 402, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
405. The method of clause 401, any other suitable clause, or any combination of suitable clauses, wherein the second therapeutic agent comprises an immunogenic material.
406. The method of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
407. The method of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.
408. The method of clause 405, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
409. A kit comprising a container comprising the vaccine of any one of clauses 335 to 344.
410. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is a parenteral formulation for administration to a nonhuman animal via a route of administration selected from the group consisting of intravenous, intraarterial, intraperitoneal, intrathecal, intradermal, epidural, intracerebroventricular, intraurethral, intrasternal, intracranial, intratumoral, intramuscular, and subcutaneous.
411. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the container comprises an effective amount of the vaccine.
412. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in the container is a ready-to-use liquid composition.
413. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in in the container is a lyophilized composition.
414. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in the container is a single unit dose.
415. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the vaccine in the container is a multi-unit dose.
416. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, further comprising a diluent. 417. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the kit further comprises a second container comprising a SAV polyprotein vaccine.
418. The kit of clause 417, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine.
419. The kit of clause 417, any other suitable clause, or any combination of suitable clauses, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
420. The kit of clause 409, any other suitable clause, or any combination of suitable clauses, wherein the kit further comprises a second container comprising an immunogenic material.
421. The kit of clause 420, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
422. The kit of clause 420, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of viral origin.
423. The kit of clause 420, any other suitable clause, or any combination of suitable clauses, wherein the immunogenic material comprises a material of bacterial origin.
EXAMPLE 1
Exemplary DNA Vaccine Vectors
Exemplary DNA vaccine vectors can be constructed using nanoplasmid technology to create various plasmids. Various polypeptide sequences as shown in Table 1 can be utilized for the plasmids.
Table 1.
The sequences include the following:
MEPNTSVIATEQQQAAMREVEAEAAARDEVVEKIAFAEGAMMVQTRRLPSGKSSVGG FLGELAQNIRAMNRSLHTDTNMLTEGAMVDRARAKVHKIIREGNLDSRVFSNTGSNT MLSLWVPAVPGPPAVPEHWDVAPSWFVCRPGKKGGIKITQSASMAALNPLFRGADVG PIGTAVRADVNAFSMNAVLGALRAGGFNTEHSLVSFVEPLIRILLMGVQTQDRGTSPW DWVGGMSSRIVNPLVFTTSGNFFPGGPNLRVWGANDTVARIVNVEDYMREAAGEGRF
DAGWGPEFWGGTGDDAVAVVPIRAVEAGLGEVNAGWTLAHMEYPVKVRLLDVDDR TIGPGGSLPLNANREYTAAGATHVPGPYARVLYVVVDQNADRCVGVRVQGQGAVIDV DPALNYVIGGADLGMLPLIQWSVGLGAEDMAQGSIAQTQRWVRMYGNEDDWESAW HLVSSAYTVYSPAFRRSGVAVEGGFWAQPAAGAAPFPLGGLAGWVRYDNQARAAQV ALCRERADMAECPWGGYRERGVRPGSVANWQYVRFDPTVAVGVAAHFWSVVKVM VAPVPDRAAALADMAWGKGKVQAMGEDVINGQMGQPESMMRGVALNENQGLAAA
TVRRVVGLENESMQTTHWSTTEVAMNGYYGRAGATAHHAAFPLSEGGTMRKRIPAIE MRENGVEGDLMNDDLYSIGTA AGYLA VEGMAG AQGGIWDVVQYQLPGPDDEARGV MNTVGAMGGWTRAVTPVDNVATMRDNGVEGEPCGIVMSLPTSGTAVVDRLANFGLP
PARAELREVPFGGYQRSVTNTNHRVKVSVSGGRAVVQKGNKAEMNPVFVNRTPGQTT LGQPTTDTTGMTTADFLDI (SEQ ID NO: 1) MSNKMKSFLLVLLCLCVGEGIVPMFRREWCLCTAGNARVPLVGEGRAEKIELFNQSAT CRKKELIITWKGKRWCYDIGSKRGKVLVQTLSGGGHLEQEGKGYKLVRNGFHLASFG GKKEEIQDSSHIEKVNGKDAIVKKGQHVEHLPGGNDLIVTEGGNLCGNVGFFDNTQCT YNSVINTGGGSVDNSQEAKNDKSNTIDNLIDMLPLVVGIAGGCLIVIVILYLTIKYCKCK
KKRTNPEPVEPEEHEMRDLRRRLEPRPPYQRQMGVEFEINEALEFMGVEGSESPDSGCQ SDEEGFRVGV (SEQ ID NO: 2)
MSNKMKSFLLVLLCLCVGEGIVPMFRREWCLCTAGNARVPLVGEGRAEKIELFNQSAT CGKKELIITWKGKRWCYDIESKRGKILVKTLSGGGHLEREGKGYKLVRNGFHLASFGG
KKEEIQDSSHIEKVNGKDAIVKKGQHVEHLPGGNDLIVTEGGNLCGNVGFFDNTQCTY NSVINIGGGSVDNSQEAKNDKSNTIDNLIDMLPLVVGIAGGCLIVIVVLYLTIKYCKCKK KRTNPEPAEPEEHEMRDLRRRLEPRPPYQRQMGVEFEINEALEFMGVEGSESPDSGCQS DEEGFRVGV (SEQ ID NO: 3)
MYTVQSWTCICLIICSMQSVCHCCDWIRHHYGHLSSEYLSLLDQMGGDITKQDAPVFF PTSLYRHIDDAEVEDQVRFLKETIYQITKLFDGNMKSVTWDKKKLDDFLNILERQLENL KSCVSPAMKPEKRLKRYFKKLNKNVLRKMNYSAQAWELIRKETKRHLQRLDILAAQM Y (SEQ ID NO: 4)
MALQTITWMSAFLCVAHVCSMPMPCQLQGQLVRITHNLLRDMGGNFPLECLQENVFV AFPATAFASSGAPQLGSSGAKAIYETLKNIDILFEADDLPTQWDQQKLKNFQNIVYRQIE ESKCMMGSVDTSDYLIRTEGLNTYFGNIAAVLKEKNFSYCAWEVVRKELLYTLQFILE HNSDSLLWANRT (SEQ ID NO: 5)
Plasmid Construct #1 is a traditional nanoplasmid including Interferon A (SEQ ID NO: 4) and Interferon C (SEQ ID NO: 5). The final formulation can be sterile filtered to 0.3mg/mL in PBS with an endotoxin level of < 15 EU/mg.
Plasmid Construct #2 is a traditional nanoplasmid including ORF_1 (wild type) (SEQ ID NO: 1) and ORF_3 (wild type) (SEQ ID NO: 3). The final formulation can be sterile filtered to 0.3mg/mL in PBS with an endotoxin level of < 15 EU/mg.
Plasmid Construct #3 is a traditional nanoplasmid including ORF_1 (wild type) (SEQ ID NO: 1) and 0RF_3 (mutant) (SEQ ID NO: 2). The final formulation can be sterile filtered to 0.3mg/mL in PBS with an endotoxin level of < 15 EU/mg. Plasmid Construct #4 is a traditional nanoplasmid including ORF_1 (wild type) (SEQ ID NO: 1). The final formulation can be sterile filtered to 0.3mg/mL in PBS with an endotoxin level of < 15 EU/mg.
Plasmid Construct #5 is a traditional nanoplasmid including 0RF_3 (wild type) (SEQ ID NO: 3). The final formulation can be sterile filtered to 0.3mg/mL in PBS with an endotoxin level of < 15 EU/mg.
Plasmid Construct #6 is a traditional nanoplasmid including 0RF_3 (mutant) (SEQ ID NO: 2). The final formulation can be sterile filtered to 0.3mg/mL in PBS with an endotoxin level of < 15 EU/mg.
The various plasmids can be utilized according to the description and methods of the present disclosure.
EXAMPLE 2
Exemplary DNA Vaccine Vectors
Exemplary DNA vaccine vectors can be constructed using fusion technology to create various plasmids. For instance, starting with an existing plasmid, a variant plasmid can be designed to include a PMCV polyprotein of the present disclosure. The various plasmids can be utilized according to the description and methods of the present disclosure.
EXAMPLE 3
Efficacy Evaluation
Plasmids as described herein can be formulated as immunogenic compositions and/or vaccine compositions and then evaluated for their efficacy in preventing PMCV in salmon such as Atlantic salmon. The compositions can induce neutralizing antibody and cellular immune responses which reduce clinical signs and mortality in the fish.
The immunogenic compositions and/or vaccine compositions can be administered to a salmon and then the fish can be subsequently challenged with a PMCV challenge strain, for instance a PMCV challenge strain that is genetically distant to the sequence of the plamid in the composition.
Efficacy can be measured by collection, identification to group and enumeration of mortalities post-challenge with one or more gross pathological lesions and/or confirmed via being RT-PCR positive for PMCV. For instance, a reduced occurance of pathological changes in the heart can be indicative of efficacy, optionally combined with identifcaiton of reduced PMCV viral load in the heart.
EXAMPLE 4
Methods for Atlantic Salmon Efficacy Evaluation
For the instant example, Atlantic salmon (AquaGen strain) from Fossing Storsmolt, Valle, Norway were evaluated using the immunogenic compositions of the present disclosure. Fish were vaccinated before study initiation against bacterial diseases with AlphaJect Micro 6 (furunculosis, vibriosis, cold-water vibriosis, winter ulcer) and infectious pancreatic necrosis (IPN).
The fish were kept in the laboratory freshwater stage at approximately 13°C. Fish were maintained in 12 tanks of 90-270 L with a stocking density of approximately 15 kg/m3 (freshwater period). The fish were transferred from the laboratory to the seawater facility during week 41 in oxygenated plastic bags by car.
After transfer to the seawater facility, the vaccine groups were split into 8 tanks (approximately WOOL final volume), with 108 fish in each tank (i.e., 9 groups with 12 fish per group). The temperature was ambient during the challenge (approximately 11±1 °C) and stocking density ranged from 12-35 kg/m3.
The fish and tanks were tended and monitored daily. The fish were fed according to standard routines using automatic feeders using a daily feeding ratio of 1% of live weight. The fish was taken off feed for 24 hours before challenge or sampling. The fish weighted approximately 80 g at challenge (week 43).
Composition preparation and storage
DNA constructs and molecular adjuvants were prepared before administration by diluting the appropriate quantity in sterile saline to formulate concentrations as follows:
• Group 2: 15 pg DNA plasmid (antigen/fish) in 0.05 mL saline injection volume
• Groups 3, 4, 5, 6: 15 ug DNA plasmid (antigen) + 15 ug Molecular adjuvant (30 pg/fish total), in 0.05 mL saline injection volume
The DNA-based compositions were stored in glass vials at 2-8°C after mixing with saline. Oil-emulsion formulations for group G7 were prepared for for i.p. administration and were stored from 2-8 °C until use.
Vaccination procedure The DNA-based compositions were stored in glass vials at 2-8°C after mixing with saline. Injection was performed as follows. Intramuscular (i.m.) was performed with the needle positioned at 60° to the right epaxial muscle, midways lateral to the anterior base of the dorsal fin, and above the mid-line. Intraperitoneal (i.p.) was performed one fin length ahead of the pelvic fins along the midline. The fish weight allowed for use of standard needle sizes: for i.m. of 0.5 x 3 mm for fish, and for oil-emulsion delivery, i.p. of 0.6 x 5 mm.
Table 2. Study groups, IVP composition, and injection locations. I.M. injection was done in the epaxial muscle below the dorsal fin.
Group Antigen / Molecular Adjuvant Dose Injection Fish no.
No. Vaccine description (Dose/fish) volume(s) location(s)*
(Dose/fish) (mb)
G1 None, None 0.05 i.m. 108
(Saline -injected)
G2 Orfl (DNA) None 0.05 i.m. 108
(15 pg)
G3 Ofrl (DNA) IFNa/IFNc (DNA) 0.05 i.m. 108
(15 pg) (15 pg)
G4 Orf3 wt (DNA) IFNa/IFNc (DNA) 0.05 i.m. 108
(15 pg) (15 pg)
G5 0rfl-0rf3 (DNA) IFNa/IFNc (DNA) 0.05 i.m. 108
( 15 pg) ( 15 pg)
G6 Orf3 mutant (DNA) IFNa/IFNc (DNA) 0.05 i.m. 108
(15 pg) (15 pg) * intramuscular (i.m.)
Challenge isolates
PMCV inoculum was used for the challenge. The inoculum originated from a clinical outbreak of CMS (in Norway), and originating fish tissues used for inoculum preparation were negative for PRV, or PRV was present at a very low level (Ct values —35 j. Preparation of inoculum
The challenge inoculums were prepared from the heart of Atlantic salmon and centrifuged at 3500 rpm following homogenization to spin down debris. The supernatant was collected and pooled from the different hearts, aliquoted to 2 ml Eppendorf tubes, and frozen at -80°C. A pre-challenge vial was thawed, fluid was aspirated and prepared for the challenge where dilutions were made in sterile PBS. The ampoule used for the pre-challenge was frozen again, then thawed, and the content was used for the main challenge study.
Challenge
The fish were not fed 24 hours before the challenge. Fish were anesthetized with Benzoak® (10 ml/lOOL of water). The challenge was performed by i.p. injection of 0.1 ml/fish with disposable 1 ml syringes at dilution of the original inoculum. Allocation of fish in tanks was done randomly.
EXAMPLE 5
Atlantic Salmon Histopathology Evaluation
For the instant example, the fish of Example 4 were evaluated using histopathology. Samples for histopathology (heart) and PCR were collected from 108 unchallenged fish on the day of the challenge (time 0). The number of fish sampled per group in each tank at each sampling post-challenge is provided in Table 2. After the fish were anesthetized and euthanized by use of an overdose of Benzoak®, blood samples in heparinized containers were collected before the abdominal cavity was opened. The head and kidney was dissected using a scalpel and forceps and transferred to RNAlater for qPCR analysis, 2x2x2 mm. The heart was then excised using a forceps clamped around the Bulbus arteriosus. The atrium was carefully released from the Sinus venosus and removed for further processing. Then, the heart was placed with the Bulbus arteriosus pointing to the left, and a scalpel was used to cut through the ventricle and the atrium simultaneously, and the right cut-side was transferred to formalin tubes. The area around the apex of the heart (left cut-side) was cut from the heart (max 2x2x2 mm) and transferred to RNAlater.
The sample tubes were labeled with study number (CMS01-22), tank no., marking (Groups 1-6), fish number (as for PCR samples), organs, and date. Pre-marked formalin-filled tubes were used.
Tissue for histopathology was submerged in 10% phosphate-buffered formalin for a minimum of 24 hours and then embedded in paraffin, sectioned at 2-3 pm, and stained with hematoxylin and eosin and then examined. Histopathology scores were recorded. First, the scores were recorded as ordinal values, from 0 to 4 for atrium (‘ao’) and ventricle (‘vo’), in each fish. Then, a sum of atrium and ventricle scores was calculated (‘ao’ + ‘vo’ = ‘sumav’). Further, since the scores in the heart are not equidistant, meaning that the “difference” between a ‘0’ and ‘ F is not similar to a ‘3’ and ‘4’, and thus violates the assumptions for the ordinal regression statistics, a binary outcome was calculated (‘bin’), where ‘bin’ is set at 0 when ‘sumav’ <3 (‘ao’ or ‘vo’ being = 1 or 2), and ‘bin’ =1 when assumptions for ‘bin’=0 not being met. A logistic regression model was used to analyze the latter conversion's outcome. In addition, epicarditis was scored separately (to distinguish from possible effects of Piscine orthoreovirus (PRV).
The atrium scores (ordinal values) are shown in Figure 1, where different score levels are colored as indicated. At 12 weeks post-challenge, heart scores were observed to increase for all groups, but Group 5 was shown to have a higher proportion of 0 and 1 scores than the other groups.
The findings for ventricle scores (Figure 2) were observed to those found in the atrium, with Group 5 having a significantly lower score than non-vaccinated controls at 12 weeks post-challenge. Group 5 was shown to have a significantly lower score than all groups apart from Groups 4 and 6.
While pathological changes in different heart compartments are of interest, from a clinical viewpoint, the sum of scores in both compartments (‘sumav’) can be more relevant for the clinical outcome of infection. The general trend from observation of field samples is that the changes start in the atrium, and the severity (degree of pathological changes) and dispersion (or spatial distribution of changes) increases over time. There is a similar development in the ventricle but delayed in time (post-challenge) relative to atrium changes.
The results for the different groups for ‘sumav’ are shown in Figure 3. At 12 weeks post-challenge, ‘sumav’ is decreased in Groups 4-6, with Group 5 having the lowest sum of scores in the atrium and ventricle. The summary of scores presented above is only based on severity (e.g., degree of change).
EXAMPLE 6
Atlantic Salmon PCR Evaluation
For the instant example, the fish of Example 4 were evaluated using PCR. Samples from 108 fish were collected on the day of the challenge (time 0). The number of fish sampled per group in each tank at each sampling post-challenge is provided in Table 2. Samples in RNA-Later were labeled with study number (CMS01-22), tank no., group no., fish no. (as for histology samples), organs and date. The organ samples from each fish were split and transferred to pre-marked tubes. The tissues collected for qPCR analysis were the heart, kidney, and spleen in RNAlater tubes. Plasma samples were frozen. PCR assays included a test for PMCV (all time points) in the heart and head kidney for each fish postchallenge.
RNA was isolated from the head/kidney and liver using an RNA-Easy kit (Qiagen), and the concentration of RNA was measured using Nanodrop (230/280 nm). Before PCR analysis, RNA concentration was normalized to 10 ng/pl, and 2 pl (20ng) was used for the PCR reaction. Primers were designed for ORF2, amplicons were detected using CYBR green, and the melting curves were examined for all fish to ensure the specificity of the PCR reaction.
Virus replication levels in the heart and head/kidney were examined in individual fish in all groups. The results are presented as mean ± 95% CI in Figure 4 for 12 weeks post-challenge. At 12 weeks post-challenge, Group 5 was shown to have lower virus levels in the heart compared to the control group and demonstrated a significantly lower replication level than all other vaccine groups (p=0.014 compared to controls).

Claims

WHAT IS CLAIMED IS:
1. An expression vector encoding a piscine myocarditis virus (PMCV) polyprotein.
2. The expression vector of claim 1, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence selected from a group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
3. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
4. The expression vector of claim 1, wherein the PMCV polyprotein comprises SEQ ID NO: 1.
5. The expression vector of claim 1, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 1.
6. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 1.
7. The expression vector of claim 1, wherein the PMCV polyprotein comprises SEQ ID NO: 2.
8. The expression vector of claim 1, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2.
9. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2.
10. The expression vector of claim 1 , wherein the PMCV polyprotein comprises SEQ ID NO: 3.
11. The expression vector of claim 1, wherein the PMCV polyprote in is at least 90% identical with an amino acid sequence of SEQ ID NO: 3.
12. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3.
13. The expression vector of claim 1, wherein the PMCV polyprotein comprises a first polyprotein at least 90% identical to SEQ ID NO: 1 and a second polyprotein, wherein the second polyprotein is at least 90% identical to SEQ ID NO: 2 or is at least 90% identical to SEQ ID NO: 3.
14. The expression vector of claim 1, wherein the PMCV polyprotein comprises i)
SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3.
15. The expression vector of claim 1, wherein the PMCV polyprotein consists of i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3.
16. The expression vector of claim 1, wherein the PMCV polyprotein consists essentially of i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3.
17. A polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and any combination thereof.
18. The polypeptide of claim 17, wherein the amino acid sequence comprises SEQ ID NO: 1.
19. The polypeptide of claim 17, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 1.
20. The polypeptide of claim 17, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 1.
21. The polypeptide of claim 17, wherein the amino acid sequence comprises SEQ ID NO: 2.
22. The polypeptide of claim 17, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 2.
23. The polypeptide of claim 17, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 2.
24. The polypeptide of claim 17, wherein the amino acid sequence comprises SEQ ID NO: 3.
25. The polypeptide of claim 17, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 3.
26. The polypeptide of claim 17, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 3.
27. The polypeptide of claim 17, wherein the amino acid sequence comprises a first sequence at least 90% identical to SEQ ID NO: 1 and a second sequence, wherein the second sequence is at least 90% identical to SEQ ID NO: 2 or is at least 90% identical to SEQ ID NO: 3.
28. An immunogenic composition comprising the expression vector of any one of claims 1 to 16.
29. The immunogenic composition of claim 28, any other suitable claim, or any combination of suitable claims, further comprising an adjuvant.
30. The immunogenic composition of claim 28, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type A.
31. The immunogenic composition of claim 30, wherein the salmonid interferon type A is encoded in a DNA vector.
32. The immunogenic composition of claim 28, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type C.
33. The immunogenic composition of claim 32, wherein the salmonid interferon type C is encoded in a DNA vector.
34. The immunogenic composition of claim 28, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type A and salmonid interferon type C.
35. The immunogenic composition of claim 34, wherein the salmonid interferon type A is encoded in a DNA vector.
36. The immunogenic composition of claim 34, wherein the salmonid interferon type C is encoded in a DNA vector.
37. The immunogenic composition of claim 34, wherein the salmonid interferon type A is encoded in a first DNA vector and the salmonid interferon type C is encoded in a second DNA vector.
38. The immunogenic composition of claim 34, wherein the salmonid interferon type A and the salmonid interferon type C are encoded in a single DNA vector.
39. A vaccine comprising the expression vector of any one of claims 1 to 16.
40. The vaccine of claim 39, any other suitable claim, or any combination of suitable claims, further comprising an adjuvant.
41. The vaccine of claim 39, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type A.
42. The vaccine of claim 39, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type C.
43. The vaccine of claim 39, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type A and salmonid interferon type C.
44. A vaccine comprising the immunogenic composition of any one of claims 28 to 38.
45. The vaccine of claim 44, any other suitable claim, or any combination of suitable claims, further comprising an adjuvant.
46. The vaccine of claim 44, any other suitable claim, or any combination of suitable claims,
H . The vaccine of claim 44, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type C.
48. The vaccine of claim 44, any other suitable claim, or any combination of suitable claims, further comprising salmonid interferon type A and salmonid interferon type C.
49. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the expression vector of any one of claims 1 to 16.
50. The method of claim 49, wherein the expression vector is a plasmid which is administered by injection into muscle tissue of the host.
51. The method of claim 49, wherein the expression vector is a plasmid which is administered by injection intraperitoneally into the host.
52. The method of claim 49, wherein the method is a prophylactic administration to the host.
53. The method of claim 49, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
54. The method of claim 49, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
55. The method of claim 49, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
56. The method of claim 49, wherein the step of administering the immunogenic composition to the host comprises administering a single dose.
57. The method of claim 49, wherein the method further comprises administration of a second therapeutic agent.
58. The method of claim 57, wherein the second therapeutic agent is a salmonid alphavirus polyprotein (SAV) vaccine.
59. The method of claim 58, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine.
60. The method of claim 58, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
61. The method of claim 57, wherein the second therapeutic agent comprises an immunogenic material.
62. The method of claim 61, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
63. The method of claim 61, wherein the immunogenic material comprises a material of viral origin.
64. The method of claim 61, wherein the immunogenic material comprises a material of bacterial origin.
65. A method for inducing an immune response in a host against a PMCV, the method comprising the step of administering to the host the vaccine of any one of claims 39 to 48.
66. The method of claim 65, wherein the method is a prophylactic administration to the host.
67. The method of claim 65, wherein the method is an intramuscular administration to the host.
68. The method of claim 65, wherein the method is an intraperitoneal administration to the host.
69. The method of claim 65, wherein the method comprises administering a single dose.
70. The method of claim 65, wherein the vaccine further comprises an adjuvant.
71. The method of claim 65, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A.
72. The method of claim 65, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type C.
73. The method of claim 65, wherein the method comprises administering a DNA vaccine, wherein the DNA vaccine encodes salmonid interferon type A and salmonid interferon type C.
74. The method of claim 65, wherein the method further comprises administration of a second therapeutic agent.
75. The method of claim 74, wherein the second therapeutic agent is a SAV polyprotein vaccine.
76. The method of claim 75, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 2 (SAV2) polyprotein vaccine.
77. The method of claim 75, wherein the SAV polyprotein vaccine is a salmonid alphavirus subtype 3 (SAV3) polyprotein vaccine.
78. The method of claim 74, wherein the second therapeutic agent comprises an immunogenic material.
79. The method of claim 78, wherein the immunogenic material comprises a material of viral origin or a material of bacterial origin.
80. The method of claim 78, wherein the immunogenic material comprises a material of viral origin.
81. The method of claim 78, wherein the immunogenic material comprises a material of bacterial origin.
EP24832852.8A 2023-06-27 2024-06-26 Compositions for prevention of cardiomyopathy syndrome Pending EP4735032A2 (en)

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PCT/US2024/035580 WO2025006577A2 (en) 2023-06-27 2024-06-26 Compositions for prevention of cardiomyopathy syndrome

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