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US 11,649,437 B2Granted
Generating HPV antigen-specific cells from a naive T cell population
AssigneeChildren's National Medical Center
InventorsCatherine M. Bollard, Conrad Russell Cruz, Patrick J. Hanley, Michael Keller
Life of this patent
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Abstract
Safe, rapid and efficient methods for producing antigen-specific T cells recognizing human papilloma virus or HPV antigens.
Claims26
A method for treating or preventing an infection by human papillomavirus (HPV) in a subject, comprising:
- administering to the subject an HPV antigen-specific T cell population, wherein the HPV antigen-specific T cell population is prepared by a method comprising:
- (a) dividing mononuclear cells from a donor source containing immune cells naïve to an HPV antigen into a CDI4-positive cell population and a CD14-negative cell population;
- (b) culturing a first portion of the CD14-positive cell population in a medium comprising granulocyte/macrophage-colony stimulating factor (GM-CSF) and IL-4 to produce a first population of dendritic cells;
- (c) culturing the first population of dendritic cells produced in step (b) in a medium comprising peptides derived from two or more HPV antigens;
- (d) culturing the first population of dendritic cells from step (c) in a medium comprising IL-6, IL-1β, TNFα, prostaglandin E2, GM-CSF, IL-4, and LPS;
- (e) culturing the CD14-negative cell population with the first population of dendritic cells from step (d) in a medium comprising IL-6, IL-7, IL-12, and IL-15 to produce an HPV-antigen specific T-cell population;
- (f) culturing the HPV-antigen specific T-cell population from step (e) with a second population of dendritic cells in a medium comprising IL-7 and IL-15, wherein the second population of dendritic cells are generated from a second portion of the CD-14 positive cell population as provided in steps (b)-(d);
- (g) culturing the HPV-antigen specific T-cell population from step (f) with a third population of dendritic cells in a medium comprising IL-2 and IL-15, wherein the third population of dendritic cells are generated from a third portion of the CD-14 positive cell population as provided in steps (b)-(d);
- and,
- (h) recovering the HPV-antigen specific T cell population from step (g);
- wherein the recovered HPV-antigen specific T-cell population comprises a mixed phenotype of CD4+ and CD8+ T-cells, and,
- wherein the recovered HPV-antigen specific T-cell population comprises CD27+ and CD28+ T-cells.
9 more elements
A composition comprising an HPV antigen-specific T cell population and a medium that maintains the viability of the T cell population, wherein the HPV antigen-specific T cell population is prepared by a method comprising:
- (a) dividing mononuclear cells from a donor source containing immune cells naïve to an HPV antigen into a CD14-positive cell population and a CD14-negative cell population;
- (b) culturing a first portion of the CD14-positive cell population in a medium comprising granulocyte/macrophage-colony stimulating factor (GM-CSF) and IL-4 to produce a first population of dendritic cells;
- (c) culturing the first population of dendritic cells produced in step (b) in a medium comprising peptides derived from two or more HPV antigens;
- (d) culturing the first population of dendritic cells from step (c) in a medium comprising IL-6, IL-1β, TNFα, prostaglandin E2, GM-CSF, IL-4, and LPS;
- (e) culturing the CD14-negative cell population with the first population of dendritic cells from step (d) in a medium comprising IL-6, IL-7, IL-12, and IL-15 to produce an HPV-antigen specific T-cell population;
- (f) culturing the HPV-antigen specific T-cell population from step (e) with a second population of dendritic cells in a medium comprising IL-7 and IL-15, wherein the second population of dendritic cells are generated from a second portion of the CD-14 positive cell population as provided in steps (b)-(d);
- (g) culturing the HPV-antigen specific T-cell population from step (f) with a third population of dendritic cells in a medium comprising IL-2 and IL-15, wherein the third population of dendritic cells are generated from a third portion of the CD-14 positive cell population as provided in steps (b)-(d);
- and,
- (h) recovering the HPV-antigen specific T cell population from step (g);
- wherein the recovered HPV-antigen specific T-cell population comprises a mixed phenotype of CD4+ and CD8+ T-cells, and,
- wherein the recovered HPV-antigen specific T-cell population comprises CD27+ and CD28+ T-cells.
8 more elements
A method for treating or preventing an infection by human papillomavirus (HPV) in a subject, comprising:
- administering to the subject an HPV antigen-specific T cell population, wherein the HPV antigen-specific T cell population is prepared by a method comprising:
- (a) dividing mononuclear cells from a donor source containing immune cells naïve to an HPV antigen into a CD14-positive cell population and a CD14-negative cell population;
- (b) culturing a first portion of the CD14-positive cell population in a medium comprising granulocyte/macrophage-colony stimulating factor (GM-CSF) and IL-4 to produce a first population of dendritic cells;
- (c) culturing the first population of dendritic cells produced in step (b) in a medium comprising the peptides derived from two or more HPV antigens;
- (d) culturing the first population of dendritic cells from step (c) in a medium comprising IL-6, IL-1β, TNFα, prostaglandin E2, GM-CSF, IL-4, and LPS;
- (e) culturing a first portion of the CD14-negative cell population with the first population of dendritic cells from step (d) in a medium comprising IL-6, IL-7, IL-12, and IL-15 to produce an HPV-antigen specific T-cell population;
- (f) culturing a second portion of the CD14-negative cell population in a medium with phytohemagglutinin and IL-2 to produce a first population of activated T-cells (ATCs);
- (g) culturing the HPV-antigen specific T-cell population from step (e) with the first population of ATCs from step (f) in a medium comprising the two or more HPV antigens, IL-2 or IL-15, or IL-2 and IL-15;
- (h) culturing the HPV-antigen specific T-cell population from step (g) with a second population of ATCs in a medium comprising the two or more HPV antigens, IL-2 or IL-15, or IL-2 and IL-15;
- wherein the second population of ATCs is generated from a third portion of the CD-14 negative cell population as provided in step (f);
- and,
- (i) recovering the HPV-antigen specific T cell population from step (h);
- wherein the recovered HPV-antigen specific T-cell population comprises a mixed phenotype of CD4+ and CD8+ T-cells, and,
- wherein the recovered HPV-antigen specific T-cell population comprises CD27+ and CD28+ T-cells.
11 more elements
A composition comprising an HPV antigen-specific T cell population and a medium that maintains the viability of the T cell population, wherein the HPV antigen-specific T cell population is prepared by a method comprising:
- (a) dividing mononuclear cells from a donor source containing immune cells naïve to an HPV antigen into a CD14-positive cell population and a CD14-negative cell population;
- (b) culturing a first portion of the CD14-positive cell population in a medium comprising granulocyte/macrophage-colony stimulating factor (GM-CSF) and IL-4 to produce a first population of dendritic cells;
- (c) culturing the first population of dendritic cells produced in step (b) in a medium comprising the peptides derived from two or more HPV antigens;
- (d) culturing the first population of dendritic cells from step (c) in a medium comprising IL-6, IL-1β, TNFα, prostaglandin E2, GM-CSF, IL-4, and LPS;
- (e) culturing a first portion of the CD14-negative cell population with the first population of dendritic cells from step (d) in a medium comprising IL-6, IL-7, IL-12, and IL-15 to produce an HPV-antigen specific T-cell population;
- (f) culturing a second portion of the CD14-negative cell population in a medium with phytohemagglutinin and IL-2 to produce a first population of activated T-cells (ATCs);
- (g) culturing the HPV-antigen specific T-cell population from step (e) with the first population of ATCs from step (f) in a medium comprising the two or more HPV antigens, IL-2 or IL-15, or IL-2 and IL-15;
- (h) culturing the HPV-antigen specific T-cell population from step (g) with a second population of ATCs in a medium comprising the two or more HPV antigens, IL-2 or IL-15, or IL-2 and IL-15;
- wherein the second population of ATCs is generated from a third portion of the CD-14 negative cell population as provided in step (f);
- and,
- (i) recovering the HPV-antigen specific T cell population from step (h);
- wherein the recovered HPV-antigen specific T-cell population comprises a mixed phenotype of CD4+ and CD8+ T-cells, and, wherein the recovered HPV-antigen specific T-cell population comprises CD27+ and CD28+ T-cells.
9 more elements
A method for treating or preventing an infection by human papillomavirus (HPV) in a subject, comprising:
- administering to the subject an HPV antigen-specific T cell population, wherein the HPV antigen-specific T cell population is prepared by a method comprising:
- (a) dividing mononuclear cells from a naïve donor source containing immune cells naïve to an HPV antigen into a CD14-positive cell population and a CD14-negative cell population;
- (b) culturing a first portion of the CD14-positive cell population in a medium comprising granulocyte/macrophage-colony stimulating factor (GM-CSF) and IL-4 to produce a first population of dendritic cells;
- (c) culturing the first population of dendritic cells produced in step (b) in a medium comprising peptides derived from two or more HPV antigens;
- (d) culturing the first population of dendritic cells from step (c) in a medium comprising IL-6, IL-1β, TNFα, prostaglandin E2, GM-CSF, IL-4, and LPS;
- (e) culturing the CD I4-negative cell population with the first population of dendritic cells from step (d) in a medium comprising IL-6, IL-7, IL-12, and IL-15 to produce an HPV-antigen specific T-cell population;
- (f) culturing the HPV-antigen specific T-cell population from step (e) with a second population of dendritic cells in a medium comprising IL-7 and IL-15, wherein the second population of dendritic cells are generated from a second portion of the CD-14 positive cell population as provided in steps (b)-(d);
- (g) culturing the HPV-antigen specific T-cell population from step (I) with a third population of dendritic cells in a medium comprising IL-2 and IL-15, wherein the third population of dendritic cells are generated from a third portion of the CD-14 positive cell population as provided in steps (b)-(d);
- and,
- (h) recovering the HPV-antigen specific T cell population from step (g);
- wherein the recovered HPV-antigen specific T-cell population comprises a mixed phenotype of CD4+ and CD8+ T-cells, and,
- wherein the recovered HPV-antigen specific T-cell population comprises CD27+ and CD28+ T-cells.
9 more elements
A composition comprising an HPV antigen-specific T cell population, wherein the HPV antigen-specific T cell population is prepared by a method comprising:
- (a) dividing mononuclear cells from a naïve donor source containing immune cells naïve to an HPV antigen into a CD14-positive cell population and a CD14-negative cell population;
- (b) culturing a first portion of the CD14-positive cell population in a medium comprising granulocyte/macrophage-colony stimulating factor (GM-CSF) and IL-4 to produce a first population of dendritic cells;
- (c) culturing the first population of dendritic cells produced in step (b) in a medium comprising peptides derived from two or more HPV antigens;
- (d) culturing the first population of dendritic cells from step (c) in a medium comprising IL-6, IL-1β, TNFα, prostaglandin E2, GM-CSF, IL-4, and LPS;
- (e) culturing the CD14-negative cell population with the first population of dendritic cells from step (d) in a medium comprising IL-6, IL-7, IL-12, and IL-15 to produce an HPV-antigen specific T-cell population;
- (f) culturing the HPV-antigen specific T-cell population from step (e) with a second population of dendritic cells in a medium comprising IL-7 and IL-15, wherein the second population of dendritic cells are generated from a second portion of the CD-14 positive cell population as provided in steps (b)-(d);
- (g) culturing the HPV-antigen specific T-cell population from step (f) with a third population of dendritic cells in a medium comprising IL-2 and IL-15, wherein the third population of dendritic cells are generated from a third portion of the CD-14 positive cell population as provided in steps (b)-(d);
- and,
- (h) recovering the HPV-antigen specific T cell population from step (g);
- wherein the recovered HPV-antigen specific T-cell population comprises a mixed phenotype of CD4+ and CD8+ T-cells, and,
- wherein the recovered HPV-antigen specific T-cell population comprises CD27+ and CD28+ T-cells.
8 more elements
Dependencies are inferred from the claim text.