Generation of CTL lines with specificity against multiple tumor antigens or multiple viruses
AssigneesBaylor College of Medicine, Wilson Wolf Manufacturing
InventorsAnn M. Leen, Ulrike Gerdemann, Cliona M. Rooney, Juan Vera, John R. Wilson
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Abstract
The present invention encompasses methods and compositions for the generation and use of cytotoxic T lymphocytes that target multiple viruses or that are specific for multiple tumor antigens. In specific embodiments, the generation methods employ use of certain cytokines to promote proliferation and reduce cell death in an activated T cell population and/or that employ a particular bioreactor having a gas permeable membrane.
Claims28
A method of generating cytotoxic T-lymphocytes (CTLs) that target at least one antigen from two or more viruses, comprising the steps of:
- contacting a population of dendritic cells (DCs) from an individual with more than one library of peptides, which combine to represent at least two different antigens and at least two different viruses, wherein the peptides in each library overlap in sequence to span at least a part of a viral antigen, thereby producing a population of DCs that present at least one epitope from at least two different viruses, wherein the produced DCs present at least one epitope through major histocompatibility complex (MHC) class I and present at least one epitope through MHC class II;
- contacting peripheral blood mononuclear cells (PBMCs) from the individual with the produced DCs, wherein the PBMCs comprise antigen-specific T lymphocytes that recognize at least one said epitope from at least two different viruses;
- and
- culturing the antigen-specific T lymphocytes in medium in a vessel, wherein the medium comprises one or more cytokines, to produce cytotoxic T-lymphocytes (CTLs) that target at least one antigen from two or more viruses.
A method of expanding the quantity of antigen specific T-lymphocytes that target at least one antigen from two or more viruses, comprising the steps of:
- contacting a population of DCs from an individual with more than one library of peptides, which combine to represent at least two different antigens and at least two different viruses, wherein the peptides in each library overlap in sequence to span at least a part of a viral antigen, thereby producing a population of DCs that present at least one epitope from at least two different viruses;
- and
- contacting PBMCs from the individual with the produced DCs, wherein the PBMCs comprise antigen-specific T lymphocytes that recognize said at least one epitope from at least two different viruses;
- and
- culturing the antigen-specific T lymphocytes in medium in a vessel, wherein the medium comprises one or more cytokines, to increase the quantity of T-lymphocytes that target at least one antigen from two or more viruses, wherein the T lymphocytes comprise CD4 + T-lymphocytes and CD8+ T-lymphocytes.
A method of generating cytotoxic T-lymphocytes that target at least one antigen from two or more viruses, comprising the steps of:
- nucleofecting DCs from an individual with plasmids that encode at least one epitope from at least two antigens and at least two viruses, thereby producing a population of DCs that present at least one epitope from at least two different antigens and at least two different viruses;
- wherein the nucleofected DCs present at least one epitope through major histocompatibility complex (MHC) class I and present at least one epitope through MHC class II;
- and
- contacting PBMCs from the individual with the produced population of DCs, wherein the PBMCs comprise antigen-specific T lymphocytes that recognize said at least one epitope from at least two different viruses;
- and
- culturing the antigen-specific T lymphocytes in medium in a vessel, wherein the medium comprises one or more cytokines, to produce cytotoxic T-lymphocytes that target at least one antigen from two or more viruses.
A method of expanding the quantity of antigen specific T-lymphocytes that target at least one antigen from two or more viruses, comprising the steps of:
- nucleofecting DCs from an individual with plasmids that encode at least one epitope from at least two antigens and at least two viruses, thereby producing a population of DCs that present at least one epitope from at least two different antigens and at least two different viruses;
- and
- contacting PBMCs from the individual with the produced population of DCs, wherein the PBMCs comprise antigen-specific T lymphocytes that recognize said at least one epitope from at least two different viruses;
- and
- culturing the antigen-specific T lymphocytes in medium in a vessel, wherein the medium comprises one or more cytokines, to increase the quantity of T-lymphocytes that target at least one antigen from two or more viruses, wherein the T lymphocytes comprise CD4+ T-lymphocytes and CD8+ T-lymphocytes.
Dependencies are inferred from the claim text.