Quality assays for antigen presenting cells
AssigneeNorthwest Biotherapeutics, Inc.
InventorsGopi Shankar, Patricia Anne Lodge, Alan Norman Brunelle
Life of this patent
Estimating the term…
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 APCs are provided.
Claims39
A method for determining the quality of the antigen-independent, co-stimulatory activity of a population of antigen presenting cells (APCs), comprising:
- providing allogeneic 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, wherein the antigen-mimetic consists of a CD3 binding agent that is an antibody, a plant lectin, or a mitogen;
- 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 quality of the co-stimulatory activity of the population of APCs.
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A method for determining the quality of the 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, wherein the antigen-mimetic consists of a CD3 binding agent that is an anti-CD3 antibody, a plant lectin, or a mitogen, 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 quality of the co-stimulatory activity of the dendritic cell preparation.
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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 allogeneic 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, wherein the antigen-mimetic is a CD3 binding agent consisting of an anti-CD3 antibody, or an antigen binding fragment thereof, a plant lectin, or a mitogen, and with a first sample of the dendritic cell preparation;
- (c) determining the activation of the contacted allogeneic 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.
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Dependencies are inferred from the claim text.