Methods and materials for expanding antigen-specific T cells in culture
AssigneesMayo Foundation for Medical Education and Research, University of Washington
InventorsPeter A. Cohen, Sandra J. Gendler, Latha B. Pathangey, Dustin B. McCurry, Jessica E. Gorman, Mary L. Disis
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Abstract
This document provides methods and materials for expanding antigen-specific T cells (e.g., antigen-specific CD4+ T cells and/or antigen-specific CD8+ T cells) in culture. For example, methods and materials for performing a polyclonal stimulation step for a particular duration (e.g., from about 1 hour to about 48 hours) to increase the expansion of T cells having a desired antigen specificity are provided.
Claims18
A method for producing a cell population comprising T cells having specificity for an antigen of interest, wherein said method comprises:
- (a) culturing a first cell population comprising PBMCs in the presence of beads comprising anti-CD3 antibodies and anti-CD28 antibodies for 1 hour to 48 hours to form a treated cell population, wherein said first cell population comprises T cells that were exposed to GM-CSF, resiquimod, E. coli lipopolysaccharide, and said antigen of interest for a first period of time and IL-7 for a second period of time, and
- (b) culturing said treated cell population in the absence of said beads for at least 3 days to form said cell population.
A method for producing a cell population comprising T cells having specificity for an antigen of interest, wherein said method comprises:
- (a) culturing a first cell population comprising PBMCs in the presence of immobilized anti-CD3 antibodies and soluble anti-CD28 antibodies for 1 hour to 48 hours to form a treated cell population, wherein said first cell population comprises T cells that were exposed to GM-CSF, resiquimod, E. coli lipopolysaccharide, and said antigen of interest for a first period of time and IL-7 for a second period of time, and
- (b) culturing said treated cell population in the absence of said immobilized anti-CD3 antibodies and said soluble anti-CD28 antibodies for at least 3 days to form said cell population.
A method for producing a cell population comprising T cells having specificity for an antigen of interest, wherein said method comprises:
- (a) exposing a cell population comprising PBMCs to GM-CSF, resiquimod, E. coli lipopolysaccharide, and an antigen of interest for a first period of time,
- (b) exposing said cell population to IL-7 for a second period of time,
- (c) exposing said cell population to (i) immobilized anti-CD3 antibodies and soluble anti-CD28 antibodies or (ii) beads comprising anti-CD3 antibodies and anti-CD28 antibodies for a third time period to form a treated cell population, and
- (d) culturing said treated cell population in the absence of (i) said immobilized anti-CD3 antibodies and soluble anti-CD28 antibodies and (ii) said beads for a fourth period of time to produce a final cell population comprising T cells having specificity for said antigen of interest.
A method for producing a cell population comprising T cells having specificity for an antigen of interest, wherein said method comprises:
- (a) exposing a cell population comprising PBMCs to GM-CSF, an antigen of interest, resiquimod, and E. coli lipopolysaccharide for a first period of time,
- (b) exposing said cell population to IL-7 for a second period of time,
- (c) while optionally continuing IL-7 exposure, exposing said cell population to (i) immobilized anti-CD3 antibodies and soluble anti-CD28 antibodies or (ii) beads or matrix comprising anti-CD3 antibodies and anti-CD28 antibodies for a third period of time to expand antigen-specific T cells, thereby forming a treated cell population, and
- (d) culturing said treated cell population in the absence of (i) said immobilized anti-CD3 antibodies and soluble anti-CD28 antibodies and (ii) said beads for at least 5 days to produce a final cell population comprising T cells having specificity for said antigen of interest.
Dependencies are inferred from the claim text.