Replication-defective arenavirus vectors
The invention relates to an infectious arenavirus particle that is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells. One or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells, and foreign genes coding for an antigen or other protein of interest or nucleic acids modulating host gene expression are expressed under control of the arenavirus promoters, internal ribosome entry sites or under control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III. The modified arenaviruses are useful as vaccines and therapeutic agents for a variety of diseases.
1. An infectious arenavirus particle comprising a genome wherein one or more of the three arenavirus open reading frames coding for the glycoprotein (GP), matrix protein Z and RNA-dependent RNA polymerase L are removed or functionally inactivated, wherein the arenavirus particle is infectious, but unable to produce further infectious progeny particles in non-complementing cells, wherein the arenavirus is Pichinde virus or Junin virus, and wherein the genome is engineered to have the ability to amplify and express its genetic information in arenavirus infected cells and comprises a foreign ribonucleic acid sequence coding for:
(a) a viral antigen selected from a hepatitis B antigen, a respiratory syncytial virus antigen, a human immunodeficiency virus antigen, an influenza antigen, a hepatitis C virus antigen, a varicella zoster virus antigen, a herpes simplex virus antigen, or a cytomegalovirus antigen;
(b) a bacterial antigen selected from a Haemophilus spp. antigen, a Pneumoccoccus spp. antigen, or a Mycobacterium tuberculosis antigen;
(c) a parasite antigen selected from the group consisting of a plasmodia antigen, an amebia antigen, and a philaria antigen; or
(d) a tumor antigen.
2. The arenavirus particle according to claim 1 , wherein the foreign ribonucleic acid sequence codes for a hepatitis B antigen, a respiratory syncytial virus antigen, a human immunodeficiency virus antigen, an influenza antigen, a hepatitis C virus antigen, a varicella zoster virus antigen, a herpes simplex virus antigen, or a cytomegalovirus antigen.
3. The arenavirus particle according to claim 1 , wherein the foreign ribonucleic acid sequence codes for a human immunodeficiency virus antigen.
4. The arenavirus particle according to claim 1 , wherein the foreign ribonucleic acid sequence is under the control of:
i) one or more of the arenavirus promoters within a 5′ UTR or a 3′ UTR of an arenavirus S segment or L segment, and/or;
ii) a regulatory element that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III, and
wherein the foreign ribonucleic acid sequence is expressed either by itself or as read-through by fusion to an arenavirus protein open reading frame.
5. The arenavirus particle according to claim 1 wherein the arenavirus open reading frame coding for the glycoprotein (GP) is removed or functionally inactivated.
6. The arenavirus particle according to claim 1 wherein the arenavirus open reading frame glycoprotein (GP) of the genome is removed and replaced by a foreign ribonucleic acid sequence.
7. The arenavirus particle according to claim 1 wherein the foreign ribonucleic acid sequence codes for a Haemophilus spp. antigen, a Pneumococcus spp. antigen, or a Mycobacterium tuberculosis antigen.
8. The arenavirus particle according to claim 1 wherein the foreign ribonucleic acid sequence codes for a plasmodia antigen, an amebia antigen, or a philaria antigen.
9. A vaccine or pharmaceutical preparation comprising the arenavirus particle according to claim 1 .
10. A method of vaccination against an infection caused by a virus in a patient comprising administering a therapeutically effective amount of a vaccine comprising the arenavirus particle according to claim 2 to a patient in need thereof, wherein the virus is hepatitis B virus, respiratory syncytial virus, human immunodeficiency virus, influenza virus, hepatitis C virus, varicella zoster virus, herpes simplex virus, or a cytomegalovirus.
11. A method of vaccination against an infection caused by a bacteria in a patient comprising administering a therapeutically effective amount of a vaccine comprising the arenavirus particle according to claim 7 to a patient in need thereof, wherein the bacteria is a Haemophilus spp., a Pneumoccoccus spp., or Mycobacterium tuberculosis.
12. A method of expressing a protein or peptide of interest comprising infecting cells in a cell culture with the arenavirus particle according to claim 1 .
13. The arenavirus particle according to claim 1 , wherein the foreign ribonucleic acid sequence codes for a tumor antigen.
14. The arenavirus particle according to claim 1 , wherein an internal ribosome entry site is introduced in the genome.
15. The arenavirus particle according to claim 5 , wherein the open reading frames coding for the nucleoprotein (NP) and the RNA-dependent RNA polymerase L of the arenavirus are intact.
16. The arenavirus particle according to claim 8 , wherein the plasmodia antigen is a malarial circumsporozoite protein.
17. The arenavirus particle according to claim 1 , wherein the arenavirus is Pichinde virus.
18. The arenavirus particle according to claim 1 , wherein the arenavirus is Junin virus.
19. A method of vaccination against an infection caused by a parasite in a patient comprising administering a therapeutically effective amount of a vaccine comprising the arenavirus particle according to claim 8 to a patient in need thereof, wherein the parasite is plasmodia, amebia, or philaria.
20. A method of treating a neoplastic disease in a patient comprising administering a therapeutically effective amount of a pharmaceutical composition comprising the arenavirus particle according to claim 13 to a patient in need thereof.