# US 5,989,805 A · Immortal avian cell line to grow avian and animal viruses to produce vaccines > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,989,805 A - Document ID: US5989805A - Status: Granted - Kind: A - Application number: 08/967,716 - Filed: 1997-11-10 - Granted: 1999-11-23 - Earliest claimed: 1995-10-27 - Assignees: Board of Trustees operating Michigan State University - Inventors: John David Reilly; Daniel C. Taylor; Roger Maes; Paul M. Coussens - Web page: https://patentgrep.com/patent/US5989805A ## CPC - G01N 33/56983: {Viruses} - A61K 39/12: Viral antigens - A61K 39/245: Herpetoviridae, e.g. herpes simplex virus - A61K 39/255: Marek's disease virus - A61P 31/12: Antivirals - C07K 14/005: from viruses - C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)} - A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) - A61K 2039/5252: {inactivated (killed)} - A61K 2039/5254: {avirulent or attenuated} - A61K 2039/552: {Veterinary vaccine} - C12N 2510/02: Cells for production - C12N 2510/04: Immortalised cells - C12N 2710/16322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2710/16334: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2710/22034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2710/24034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2720/10034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2720/12234: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2760/18134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - 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 - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Method of propagating viruses that replicate in embryonated eggs or in primary cultures of chicken embryo cells on an immortal, virus-free, contact-inhibited, and non-oncogenic chicken embryo cell line. The method supports replication of avian viruses of the Birnaviridae, Coronaviridae, Herpesviridae, Orthomyxoviridae, Paramyxoviridae, Picornaviridae, Polyomaviridae, Poxviridae, and Reoviridae families. The immortal cell line of the present invention supports replication of swine influenza virus, for instance. The cell line is useful for virus isolation diagnostic assays and for propagating virus suitable for live or killed vaccines. ## Claims ### Claim 1 An avian vaccine, comprising: an immortal chicken embryo cell line infected with a vaccine strain of Marek's disease virus, wherein the vaccine induces a protective immune response when administered to an avian. ### Claim 2 (depends on 1) The avian vaccine according to claim 1, wherein the cell line is derived from chicken embryo cells that had been treated with a chemical mutagen to render the immortal cell line. ### Claim 3 (depends on 2) The avian vaccine according to claim 2, wherein the cell line comprises CHCC-OU2 cells deposited as ATCC CRL 12302. ### Claim 4 (depends on 2) The avian vaccine of claim 2, wherein the chemical mutagen is MNNG. ### Claim 5 A method for propagating an avian virus, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line said cell line being virus free; (b) infecting said immortal cell line with an avian virus selected from the group of virus families consisting of Adenoviradae, Birnaviradae, Coronaviradae, Herpesviradae Orthomyxoviradae, Picornaviradae, Poxviradae, Reoviradae, and Polyomaviradae; (c) cultivating the infected immortal cell line to produce the immortal cell line infected with the avian virus; and (d) removing the virus from the immortal cell line. ### Claim 6 (depends on 5) The method according to claim 5, wherein the chemical mutagen is MNNG. ### Claim 7 (depends on 5) The method according to claim 5, wherein the cell line comprises CHCC-OU2 cells deposited as ATCC CRL 12302. ### Claim 8 (depends on 5) The method according to claim 5, wherein the avian virus is selected from the group consisting of avian influenza virus, avian reovirus, fowl pox virus, pigeon pox virus, canary pox virus, psittacine herpesvirus, pigeon herpesvirus, falcon herpesvirus, infectious bursal disease virus, infectious bronchitis virus, avian encephalomyelitis virus, chicken anemia virus, avian adenovirus, and avian polyomavirus. ### Claim 9 A method for providing an avian vaccine, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line; (b) infecting said cell line with a virus; (c) cultivating said infected cell line to produce the virus; (d) harvesting the virus from the cultivated cell line; and (e) subjecting the harvested virus to at least one of the following treatments: (i) inactivating the virus, (ii) adding a pharmaceutically acceptable carrier or diluent, (iii) adding an adjuvant, (iv) adding a stabilizer, (v) lyophilizing, wherein the vaccine induces protective immunity in avians. ### Claim 10 (depends on 9) The method according to claim 9, wherein the chemical mutagen is MNNG. ### Claim 11 (depends on 9) The method according to claim 9, wherein the cell line comprises CHCC-OU2 deposited as ATCC CRL 12302. ### Claim 12 A method for propagating swine influenza virus, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line said cell line being virus free; (b) infecting said cell line with swine influenza virus; (c) cultivating the infected cell line to produce the swine influenza virus; and removing the virus from the cell line. ### Claim 13 (depends on 12) The method according to claim 12, wherein the chemical mutagen is MNNG. ### Claim 14 (depends on 12) The method according to claim 12, wherein the cell line comprises CHCC-OU2 cells deposited as ATCC CRL 12302. ### Claim 15 A method for preparing a swine influenza vaccine, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line; (b) infecting said cell line with the virus; (c) cultivating said infected cell line to produce the virus; (d) harvesting the virus from the cultivated cell line; and (e) subjecting the harvested virus to at least one of the following treatments: (i) inactivating the virus, (ii) adding a pharmaceutically acceptable carrier or diluent, (iii) adding an adjuvant, (iv) lyophilizing, wherein the vaccine induces protective immunity in swine. ### Claim 16 (depends on 15) The method according to claim 15, wherein the chemical mutagen is MNNG. ### Claim 17 (depends on 15) The method according to claim 15, wherein the cell line comprises CHCC-OU2 deposited as ATCC CRL 12302. ### Claim 18 A method for propagating a virus that replicates in embryonated eggs, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line said cell line being virus free; (b) infecting said cell line with a virus selected from the group of virus families consisting of Adenoviradae, Birnaviradae, Coronaviradae, Herpesviradae Orthomyxoviradae, Picornaviradae, Poxviradae, Reoviradae, and Polyomaviradae; (c) cultivating the infected cell line to produce an immortal cell line infected with the virus; and (d) removing the virus from the cell line. ### Claim 19 A method for preparing a vaccine from a virus that replicates in embryonated eggs, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line; (b) infecting said cell line with the virus; (c) cultivating said infected cell line to produce the virus; (d) harvesting the virus from the cultivated cell line; and (e) subjecting the harvested virus to at least one of the following treatments: (i) inactivating the virus, (ii) adding a pharmaceutically acceptable carrier or diluent, (iii) adding an adjuvant, (iv) lyophilizing, wherein the vaccine induces protective immunity in a host animal. ### Claim 20 A method for preparing a vaccine from a virus that replicates in chicken embryo fibroblasts, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line; (b) infecting said cell line with the virus; (c) cultivating said infected cell line to produce the virus; (d) harvesting the virus from the cultivated cell line; and (e) subjecting the harvested virus to at least one of the following treatments: (i) inactivating the virus, (ii) adding a pharmaceutically acceptable carrier or diluent, (iii) adding an adjuvant, (iv) lyophilizing, wherein the vaccine induces protective immunity in a host animal. ### Claim 21 A method for propagating a virus that has been modified to replicate in chicken embryo fibroblasts, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line said cell line being virus free; (b) infecting said cell line with the virus; (c) cultivating the infected cell line to produce the virus; and (d) removing the virus from the chicken embryo cell line. ### Claim 22 A method for preparing a vaccine from a virus that has been modified to replicate in chicken embryo fibroblasts, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line; (b) infecting said cell line with the virus; (c) cultivating said infected cell line to produce the virus; (d) harvesting the virus from the cultivated cell line; and (e) subjecting the harvested virus to at least one of the following treatments: (i) inactivating the virus, (ii) adding a pharmaceutically acceptable carrier or diluent, (iii) adding an adjuvant, (iv) lyophilizing, wherein the vaccine induces protective immunity in a host animal. ### Claim 23 A method for identifying a virus, comprising: (a) providing a cell line generated from chicken embryo cells that have been treated with a chemical mutagen to create an immortal cell line said cell line being virus free; (b) infecting said cell line with the virus; (c) cultivating the infected cell line to produce the virus; (d) reacting said infected cell line with an antibody specific for the virus; and (e) visualizing infected cells reacted with the antibody. ### Claim 24 (depends on 23) The method according to claim 23, wherein the virus is selected from the group consisting of avian influenza virus, avian reovirus, fowl pox virus, pigeon pox virus, canary pox virus, psittacine herpesvirus, pigeon herpesvirus, falcon herpesvirus, Newcastle disease virus, infectious bursal disease virus, infectious bronchitis virus, avian encephalomyelitis virus, chicken anemia virus, avian adenovirus, swine influenza virus, equine influenza virus, and avian polyomavirus. ### Claim 25 (depends on 23) The method according to claim 23, wherein the chemical mutagen is MNNG. ### Claim 26 (depends on 23) The method according to claim 23 wherein the cell line comprises CHCC-OU2 cells deposited as ATCC CRL 12302.