Vector production
An enveloped viral particle producer or packaging cell, wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell.
1. An enveloped viral particle producer or packaging cell, wherein:
the producer or packaging cell is genetically engineered to decrease expression of MHC-I on the surface of the producer or packaging cell such that the number of surface-exposed MHC-I molecules is less than 50% of the number of surface-exposed MHC-I molecules that are displayed in the absence of the genetic engineering, and
wherein the producer or packaging cell comprises one or more nucleic acid sequences encoding one or more viral particle structural proteins; and wherein:
(a) the packaging cell lacks a vector encoding a viral genome; whereas
(b) the producer cell further comprises a vector encoding a viral genome comprising a therapeutic nucleotide of interest.
2. The enveloped viral particle producer or packaging cell of claim 1 wherein the cell comprises a genetically engineered disruption of a gene encoding β2-microglobulin.
3. The enveloped viral particle producer or packaging cell of claim 1 wherein the cell comprises a genetically engineered disruption of one or more genes encoding an MHC-I α chain.
4. The enveloped viral particle producer or packaging cell of claim 1 , wherein the cell is a HEK-293 cell or a derivative thereof.
5. The enveloped viral particle producer or packaging cell of claim 4 , wherein the cell is a HEK-293T or a HEK-293 T-REx cell.
6. The enveloped viral particle producer or packaging cell of claim 1 , wherein the enveloped viral particle is a retrovirus, herpes simplex virus or vaccinia virus, hepadnavirus, togavirus, flavivirus, arenavirus, coronavirus, orthomyxovirus, paramyxovirus, bunyavirus, bornavirus, rhabdovirus, filovirus or derived therefrom.
7. The enveloped viral particle producer or packaging cell of claim 1 , wherein the enveloped viral particle is derived from a retrovirus or herpes simplex virus or vaccinia virus.
8. The enveloped viral particle producer or packaging cell of claim 1 , wherein the enveloped viral vector is derived from a lentivirus.
9. The enveloped viral particle producer or packaging cell of claim 1 , wherein the cell comprises one or more nucleic acid sequences encoding a viral capsid protein and/or a viral envelope protein.
10. The enveloped viral particle producer or packaging cell of claim 1 , wherein the viral particle is a retroviral particle, and wherein the cell comprises one or more nucleic acid sequences encoding one or more proteins selected from the group consisting of Gag, Pol and Env.
11. The enveloped viral particle producer or packaging cell of claim 10 , wherein the viral particle is a lentiviral particle.
12. The enveloped viral particle producer or packaging cell of claim 11 , wherein the cell further comprises a nucleic acid sequence encoding Rev.
13. The enveloped viral particle producer or packaging cell of claim 1 , wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell such that the number of surface-exposed MHC-I molecules is less than 40% of the number of surface-exposed MHC-1 molecules that are displayed in the absence of the genetic engineering.
14. The enveloped viral particle producer or packaging cell of claim 1 , wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell such that the number of surface-exposed MHC-I molecules is 0% of the number of surface-exposed MHC-I molecules that are displayed in the absence of the genetic engineering.
15. The enveloped viral particle producer or packaging cell of claim 1 , wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell such that the cell is substantially devoid of surface-exposed MHC-I molecules.
16. A method of producing enveloped viral particles comprising the steps of:
a) providing an enveloped viral particle producer cell according to claim 1 ; and
b) culturing the cell under conditions suitable for the production of the enveloped viral particles.
17. The method of claim 12 , wherein the enveloped viral particles are substantially devoid of surface-exposed MHC-1 molecules.
18. An enveloped viral particle derived from a retrovirus, herpes simplex virus, vaccinia virus, hepadnavirus, togavirus, flavivirus, arenavirus, coronavirus, orthomyxovirus, paramyxovirus, bunyavirus, bornavirus, rhabdovirus or filovirus, wherein the particle is produced according to the method of claim 12 and is substantially devoid of surface-exposed MHC-I molecules.
19. A cell transduced by the enveloped viral particle of claim 18 .
20. A pharmaceutical composition comprising the enveloped viral particle of claim 18 , or comprising a cell transduced by said enveloped viral particle, and a pharmaceutically acceptable carrier, diluent or excipient.
21. A method of gene therapy comprising transducing a cell with the enveloped viral particle of claim 18 .
22. The method of claim 21 wherein the transduction is carried out ex vivo.
23. A method of gene therapy comprising administering the enveloped viral particle of claim 18 , or a cell transduced by said enveloped viral particle, to a subject in need thereof.
24. A method of vaccination comprising administering the enveloped viral particle of claim 18 to a subject in need thereof.