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US 2006/0234309 A1Published application
Quality assays for antigen presenting cells
AssigneeNorthwest Biotherapeutics, Inc.
InventorsGopi Shankar, Patricia Lodge, Alan Brunelle
Life of this patent
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Abstract
The present invention provides methods for evaluating the quality of a preparation of antigen presenting cells, such as dendritic cells. Assays for antigen-independent co-stimulation of T cells and for presentation of predetermined antigen by APC's are provided.
Claims47
A method for determining antigen-independent, co-stimulatory activity of antigen presenting cells (APCs), comprising:
- providing T cells having a known functional activity and being substantially free of co-stimulatory activity;
- providing a sample of APCs of unknown co-stimulatory activity;
- contacting the T cells with a sub-optimal concentration of an antigen-mimetic agent;
- contacting the T cells with the sample of APCs of unknown co-stimulatory activity;
- determining the activation of the T cells contacted with the antigen-mimetic agent and the sample of APCs;
- and
- comparing the determined activation of the T cells with a standard activation index for the T cells to determine the co-stimulatory activity of the APCs.
4 more elements
A method for determining antigen-independent co-stimulatory activity of a preparation of dendritic cells, comprising:
- contacting a first quantity of T cells, which are substantially free of co-stimulatory activity and have a known functional activity, with a suboptimal quantity of an antigen-mimetic agent and with a first sample of a dendritic cell preparation of unknown co-stimulatory activity;
- determining a first activation value for the first quantity of T cells;
- contacting a second quantity of T cells with a second sample of the dendritic cell preparation or the suboptimal quantity of the antigen-mimetic agent;
- determining a second activation value for the second quantity of T cells;
- and
- comparing the first and second activation values to determine the co-stimulatory activity of the dendritic cell preparation.
3 more elements
A method for determining the quality of a preparation of dendritic cells, comprising:
- (1) providing a dendritic cell preparation of unknown co-stimulatory activity and unknown antigen presenting ability for a predetermined antigen;
- (2) determining the co-stimulatory activity of the dendritic cell preparation, said determination of co-stimulatory activity comprising
- (a) providing T cells of known functional activity and substantially free of co-stimulatory activity;
- (b) contacting the T cells with a suboptimal quantity of an antigen-mimetic agent and with a first sample of the dendritic cell preparation;
- (c) determining the activation of the contacted T cells;
- and
- (d) comparing the determined activity of the contacted T cells with the standard activation index for the T cells to the determined co-stimulatory activity of the dendritic cell preparation;
- (3) determining presentation of the predetermined antigen by the preparation of dendritic cells, said determination of presentation comprising:
- (a) contacting a second sample of the dendritic cell preparation with the predetermined antigen;
- and
- (b) determining the amount of predetermined antigen presented by the dendritic cells;
- and
- (4) determining the quality of the dendritic cell preparation based on the determined co-stimulatory activity and determined antigen-specific presentation of the predetermined antigen.
10 more elements
Dependencies are inferred from the claim text.