Methods for testing an immune response using cultures of T cells, B cells, dendritic cells and follicular dendritic cells
Inventors
Warren, William L. • Drake, III, Donald • Moser, Janice • Singh, Inderpal • Song, Haifeng • Mishkin, Eric • Tew, John G.
Assignees
Virginia Commonwealth University • Sanofi Pasteur VaxDesign Corp
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Publication Number
US-8962319-B2
Publication Date
2015-02-24
Expiration Date
Abstract
The present invention relates to methods for preparing an artificial immune system. The artificial immune system comprises a cell culture comprising T cells, B cells and antigen-primed dendritic cells. The artificial immune system of the present invention can be used for in vitro testing of vaccines, adjuvants, immunotherapy candidates, cosmetics, drugs, biologics and other chemicals.
Core Innovation
The invention provides an artificial immune system for testing an immune response to an antigen by using antigen-primed dendritic cells and defined enriched populations of blood-derived T cells and blood-derived B cells, optionally with follicular dendritic cells. A cell culture is prepared as a lymphoid tissue equivalent model, and dendritic cells are primed with an antigen prior to being introduced to the culture. The system is designed to assess T cell and/or B cell activity in a controlled in vitro co-culture environment.
The method focuses on using enriched blood-derived T cells together with enriched blood-derived B cells in a more physiological T:B composition, rather than relying on PBMCs. The claimed approach includes a preferred approximately 1:1 composition, with a variable range described in the disclosed rationale. This defined T:B configuration is used alongside antigen-primed dendritic cells to generate antigen-specific immune responses in the model.
The documented test framework assaying for immune response includes measuring T cell and/or B cell activity, and the disclosure reports stronger antigen-specific T/B activation, proliferation, and antibody secretion when dendritic cells are pulsed with antigen. Example antigens referenced include tetanus toxoid, Candida albicans, and influenza, including H1N1, in a 2D lymphoid tissue equivalent model. Readouts referenced in the disclosure include ELISPOT and flow cytometry, including CFSE loss, and characterization of antigen-presenting and cell activation markers such as HLA-DRhigh and CD86high, with additional memory phenotype and related immune markers referenced in the document context.
The invention is stated to be applicable to testing vaccines, adjuvants, immunotherapy candidates, cosmetics, drugs, biologics, and other chemicals by assaying antigen-specific T cell and/or B cell activity in the described immune system format. By providing an artificial immune system that supports antigen-primed dendritic cell co-culture with enriched T and B cell populations, the approach targets evaluation of antigen-specific immune response strength and character in a lymphoid tissue equivalent model.
Claims Coverage
The provided independent claims cover two method formulations for testing an immune response to an antigen: (i) a method where antigen-primed dendritic cells are added to a prepared culture, and (ii) a method where dendritic cells primed with a selected antigen are included in the prepared culture prior to assaying. Across these independent claims, the inventive features center on enriched blood-derived T and B cell populations, the presence of follicular dendritic cells, antigen priming of dendritic cells, and assaying T cell and/or B cell activity, with dependent refinements adding specific measurement/readout and antigen-type selection constraints.
Enriched T, B, and follicular dendritic cell culture for immune testing
Preparing a cell culture consisting of an enriched population of blood-derived T cells, an enriched population of blood-derived B cells, and follicular dendritic cells.
Antigen-primed dendritic cells co-culture
Priming a population of dendritic cells with an antigen; and adding to the cell culture the population of antigen-primed dendritic cells.
Assaying T cell and/or B cell activity to test immune response
Assaying for T cell and/or B cell activity in the cell culture of the adding step, thereby testing an immune response to an antigen.
Culture including dendritic cells primed with a selected antigen
Preparing a cell culture consisting of an enriched population of blood-derived T cells, an enriched population of blood-derived B cells, follicular dendritic cells, and dendritic cells primed with a selected antigen.
Assaying in a culture containing primed dendritic cells
Assaying for T cell and/or B cell activity in the cell culture of preparing step, thereby testing an immune response to an antigen.
The independent claims broadly cover antigen-primed dendritic cell co-culture with enriched blood-derived T cells and enriched blood-derived B cells, optionally including follicular dendritic cells, followed by assaying T cell and/or B cell activity as a measure of antigen-specific immune response. Dependent claim refinements described in the provided claim set further specify assay readouts (activation, proliferation, and antibody production), constrain the T:B ratio to approximately 1:1, and define the antigen as one selected from specified categories such as vaccines, adjuvants, immunotherapy candidates, cosmetics, drugs, biologics, or chemical compounds.
Stated Advantages
Stronger antigen-specific T/B activation, proliferation, and antibody secretion in the described model when dendritic cells are pulsed with antigen (as reported in the disclosure context).
Documented Applications
Testing vaccines (as an antigen category in the disclosed applicability).
Testing adjuvants (as an antigen category in the disclosed applicability).
Testing immunotherapy candidates (as an antigen category in the disclosed applicability).
Testing cosmetics (as an antigen category in the disclosed applicability).
Testing drugs (as an antigen category in the disclosed applicability).
Testing biologics (as an antigen category in the disclosed applicability).
Testing other chemicals (as an antigen category in the disclosed applicability).
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