Dendritic cell assay for innate immunogenicity to gene therapy agents
AssigneeGenzyme Corporation
InventorsSourav Roy CHOUDHURY, Mona MOTWANI
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Abstract
Provided herein are methods for determining the innate immunogenicity of a gene therapy agent in an individual. In particular, the methods involve the use of isolated dendritic cells to detect innate immunogenicity to a gene therapy agent. Exemplary gene therapy agents include adeno-associated virus (AAV) vectors, adenovirus vectors, lentivirus vectors, Herpes simplex virus (HSV) vectors or a lipid nanoparticles.
Claims88
A method of determining the innate immunogenicity to a gene therapy agent in an individual, the method comprising
- a) incubating an innate immune cell from the individual with the gene therapy agent,
- b) analyzing the innate immune cell for altered expression of one or more cytokines compared to a suitable control, whereby altered expression of the one or more cytokines produces a cytokine signature,
- wherein expression of a cytokine signature following incubation with the gene therapy agent indicates innate immunogenicity to the gene therapy agent in the individual.
A method of determining the innate immunogenicity to a viral gene therapy agent in an individual, the method comprising
- a) incubating monocytes from the individual in dendritic cell culture media under conditions in which the monocytes differentiate into dendritic cells,
- b) incubating the dendritic cells with the viral gene therapy agent at an MOI of about 1×103 to about 1×105 for about 12 to about 36 hours,
- c) analyzing the dendritic cells for altered expression of one or more cytokines compared to a suitable control, whereby altered expression of the one or more cytokines produces a cytokine signature,
- wherein expression of the cytokine signature following incubation with the viral gene therapy agent indicates innate immunogenicity to the viral gene therapy agent in the individual, wherein the cytokine signature comprises increased expression of IL6, TNFα, and IL-1β.
A method of determining the innate immunogenicity to a non-viral gene therapy agent in an individual, the method comprising
- a) incubating monocytes from the individual in dendritic cell culture media under conditions in which the monocytes differentiate into dendritic cells,
- b) incubating the dendritic cells with the non-viral vector at a concentration of about 1 ng/mL to about 1 mg/mL,
- c) analyzing the dendritic cells for altered expression of one or more cytokines compared to a suitable control, whereby altered expression of the one or more cytokines produces a cytokine signature,
- wherein expression of the cytokine signature following incubation with the non-viral gene therapy agent indicates innate immunogenicity to the non-viral gene therapy agent in the individual, wherein the cytokine signature comprises increased expression of IL6, TNFα, and IL-1β.
A method of determining the innate immunogenicity to a lipid nanoparticle (LNP) in an individual, the method comprising
- a) incubating monocytes from the individual in dendritic cell culture media under conditions in which the monocytes differentiate into dendritic cells,
- b) incubating the dendritic cells with the LNP,
- c) analyzing the dendritic cells for altered expression of one or more cytokines compared to a suitable control, whereby altered expression of the one or more cytokines produces a cytokine signature,
- wherein expression of the cytokine signature following incubation with the LNP indicates innate immunogenicity to the non-viral gene therapy agent in the individual.
A method of determining a cytokine signature of a gene therapy agent comprising
- a) incubating one or more innate immune cell from one or more individuals with the gene therapy agent,
- b) analyzing the one or more innate immune cells for altered expression of one or more cytokines compared to a suitable control,
- wherein the altered expression of one or more cytokines in step b) indicates the cytokine signature of the gene therapy agent.
Dependencies are inferred from the claim text.