# US 10,619,169 B2 · EHV insertion site ORF70 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,619,169 B2 - Document ID: US10619169B2 - Status: Granted - Kind: B2 - Application number: 15/706,889 - Filed: 2017-09-18 - Published: 2018-03-22 - Granted: 2020-04-14 - Earliest claimed: 2016-09-20 - Applicants: Boehringer Ingelheim Vetmedica GmbH - Inventors: Alice Mundt; Andreas Gallei; Kristina Rehmet - Web page: https://patentgrep.com/patent/US10619169B2 ## CPC - C12N 15/86: Viral vectors - A61K 39/12: Viral antigens - A61K 39/145: Orthomyxoviridae, e.g. influenza virus - A61P 31/16: for influenza or rhinoviruses - A61K 2039/5256: {expressing foreign proteins} - A61K 2039/54: {characterised by the route of administration} - A61K 2039/544: {to the airways} - A61K 2039/545: {characterised by the dose, timing or administration schedule} - A61K 2039/552: {Veterinary vaccine} - A61K 2039/575: {humoral response} - A61K 2039/60: {characteristics by the carrier linked to the antigen} - A61K 2039/70: {Multivalent vaccine} - C12N 2710/16734: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2710/16743: viral genome or elements thereof as genetic vector - C12N 2760/12034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2760/12071: Demonstrated in vivo effect - C12N 2760/16111: Influenzavirus A, i.e. influenza A virus - C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2760/16171: Demonstrated in vivo effect - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract The present invention relates to the field of (vector) vaccines, and especially to the novel EHV insertion site ORF70. The present invention further concerns related expression cassettes and vectors, which are suitable to express genes of interest, especially antigen encoding sequences. The viral vectors of the present invention are useful for producing an immunogenic composition or vaccine. ## Claims ### Claim 1 An Equid Alphaherpesvirus 1 (EHV-1) vector, strain RacH, comprising an expression cassette comprising an exogenous nucleotide sequence of interest, wherein said nucleotide sequence of interest inserted into ORF70 in an EHV-1 vector (ORF70) and operably linked to a promoter sequence, and a left ORF70 flanking region selected from the group consisting of sequences having at least 95% homology with any of SEQ ID Nos: 13, 15, and 17, and a right ORF70 flanking region selected from the group consisting of sequences having at least 95% homology with any of SEQ ID Nos: 14, 16, and 18. ### Claim 2 (depends on 1) The EHV-1 vector of claim 1, wherein the ORF70 has a partial deletion, truncation, substitution, or modification, and wherein ORF71 remains functional. ### Claim 3 (depends on 1) The EHV-1 vector of claim 1, wherein ORF70 possesses an approximately 801 bp deletion corresponding to SEQ ID NO.: 20 or a sequence having at least 95% sequence homology with SEQ ID NO.: 20. ### Claim 4 (depends on 1) The EHV-1 vector of claim 1, wherein the EHV-1 vector comprises (i) at least one left ORF70 flanking region selected from the group consisting of: SEQ ID NO.: 13, SEQ ID NO.: 15, and SEQ ID NO.: 17, and (ii) at least one right ORF70 flanking region selected from the group consisting of: SEQ ID NO.: 14, SEQ ID NO.: 16, and SEQ ID NO.: 18. ### Claim 5 (depends on 1) The EHV-1 vector of claim 1, wherein said nucleotide sequence of interest is recombinant. ### Claim 6 (depends on 1) The EHV-1 vector of claim 1, wherein said nucleotide sequence of interest is recombinant and/or heterologous. ### Claim 7 (depends on 1) The EHV-1 vector of claim 1, wherein said nucleotide sequence of interest is an antigen encoding sequence related to a pathogen infecting a food producing animal such as swine or cattle. ### Claim 8 (depends on 1) The EHV-1 vector of claim 1, further comprising additional regulatory sequences such as a termination signal or polyadenylation sequence. ### Claim 9 (depends on 1) The EHV-1 vector of claim 1, wherein the promoter sequence operably linked to the nucleotide sequence of interest is selected from the group consisting of: SV40 large T, HCMV and MCMV immediate early gene 1, human elongation factor alpha promoter, baculovirus polyhedrin promoter, a sequence having at least 95% sequence identity with any one of SEQ ID Nos.: 1, 2, 3, and 4 or the complementary sequences thereof. ### Claim 10 (depends on 1) A mammalian host cell characterized in that it comprises a vector according to claim 1. ### Claim 11 (depends on 1) A method comprising manufacturing an immunogenic composition or vaccine using the vector according to claim 1. ### Claim 12 (depends on 1) A method for the preparation of an immunogenic composition or a vaccine for reducing the incidence or the severity of one or more clinical signs associated with or caused by an infection, comprising the following steps: a. Infecting a mammalian host cell with the vector according to claim 1, b. cultivating the infected cells under suitable conditions, c. collecting infected cell cultures, d. optionally purifying the collected infected cell cultures of step c) e. optionally mixing said collected infected cell culture with a pharmaceutically acceptable carrier. ### Claim 13 (depends on 1) An immunogenic composition comprising the EHV-1 RacH vector according to claim 1. ### Claim 14 (depends on 1) An immunogenic composition comprising a. the vector according to claim 1, and/or b. a polypeptide expressed by the vector according to claim 1, such as a virus, a modified live virus, a virus like particle (VLP), and c. optionally a pharmaceutical- or veterinary-acceptable carrier or excipient, wherein said carrier is suitable for oral, intradermal, intramuscular or intranasal application, and said immunogenic composition comprises a virus, such as an infectious virus. ### Claim 15 (depends on 1) A vaccine or pharmaceutical composition comprising a. the vector according to claim 1, and/or b. a polypeptide expressed by the vector according to claim 1, selected from the group consisting of a modified live virus, a virus like particle (VLP), and c. a pharmaceutical- or veterinary-acceptable carrier or excipient, wherein said carrier is suitable for oral, intradermal, intramuscular or intranasal application, and optionally said vaccine further comprises an adjuvant. ### Claim 16 An Equid Alphaherpesvirus 1 (EHV-1) vector, strain RacH, comprising an exogenous nucleotide sequence of interest, wherein said nucleotide sequence of interest is operably linked to a promoter sequence, and a left ORF70 flanking region selected from the group consisting of: SEQ ID NOs.: 13, 15, and 17, and a right ORF70 flanking region selected from the group consisting of: SEQ ID Nos.: 14, 16, and 18. ### Claim 17 An Equid Alphaherpesvirus 1 (EHV-1) vector, strain RacH, comprising a nucleotide sequence of interest inserted into ORF70. ### Claim 18 An Equid Alphaherpesvirus 1 (EHV-1) vector, strain RacH, comprising a first and a second nucleotide sequence or gene of interest inserted into ORF70. ### Claim 19 (depends on 17) The EHV-1 vector of claim 17, wherein the gene of interest is operably linked to a regulatory sequence, such as a promoter sequence. ### Claim 20 (depends on 18) The EHV-1 vector of claim 18, wherein promoter sequences are operably linked to the first and second nucleotide sequence or gene of interest, and wherein said promoter sequences are different. ### Claim 21 (depends on 18) The EHV-1 vector of the claim 18, wherein the promoter sequence operably linked to the first nucleotide sequence or gene of interest of interest is p455 (SEQ ID NO. 4) or a sequence having at least 95% sequence identity with SEQ ID Nos.: 4 or the complementary sequence thereof, and wherein the promoter sequence operably linked to the second nucleotide sequence or gene of interest of interest is p430 (SEQ ID NO. 3) or a sequence having at least 95% sequence identity with SEQ ID Nos.: 3 or the complementary sequence thereof. ### Claim 22 (depends on 14) A method of reducing or preventing the clinical signs or disease caused by an infection with a pathogen in an animal or for treating or preventing an infection with a pathogen in an animal, comprising administering to the animal the immunogenic composition according to claim 14, wherein said animal is a food producing animal such as swine. ### Claim 23 (depends on 14) A method of immunizing an animal such as a food producing animal including swine against a clinical disease caused by a pathogen in said animal, said method comprising the step of administering to the animal the immunogenic composition according to claim 14, whereby said immunogenic composition or vaccine fails to cause clinical signs of infection but is capable of inducing an immune response that immunizes the animal against pathogenic forms of said pathogen. ### Claim 24 (depends on 14) A kit for vaccinating an animal, such as swine or cattle, against a disease associated with and/or reducing the incidence or the severity of one or more clinical signs associated with or caused by a pathogen in an animal comprising: a) a dispenser capable of administering a vaccine to said animal; and b) the immunogenic composition according to claim 14, and c) optionally an instruction leaflet.