Nk cell compositions and preparations for immunotherapy and methods for their production
AssigneeDeverra Therpaeutics Inc.
InventorsColleen Delaney, Carrie Stoltzman, Elizabeth Dam, Mary Prieve
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Abstract
The present invention provides NK cell compositions and/or preparations and methods of using such NK cell compositions and/or preparations for immunotherapy. The NK cell compositions and/or preparations can be used in therapies of a broad range of viral infections, bacterial infections, cancer and leukemia malignancies, and other diseases.
Claims51
A method of preparing a natural killer (NK) cell preparation for use in immunotherapy, comprising:
- selecting a plurality of umbilical cord blood or placental blood cells without immunological matching to each other;
- preparing enriched CD34+ hematopoietic stem and progenitor cells (HSPCs) that are depleted of red blood cells and T cells;
- culturing the CD34+ enriched HSPCs in an expansion culture medium comprising interleukin-3 (IL-3), interleukin-6 (IL-6), thrombopoietin (TPO), Flt-3 ligand (Flt-3L), and stem cell factor (SCF) on a solid phase coated with a Notch ligand and recombinant human fibronectin or fragments thereof, and in the absence of exogenous feeder cells, for a sufficient time to produce expanded HSPCs, wherein the expanded HSPCs do not substantially differentiate into CD56+ cells during the expansion;
- and
- culturing the expanded HSPCs in a differentiation culture medium comprising effective amounts of IL-2 and IL-15, a non-animal sourced serum replacement, and in the absence of the exogenous feeder cells for a sufficient time to produce an NK cell composition and/or preparation comprising about 50 to about 80% CD56+ cells and about 50 to about 20% endogenous CD56− cells, or about 50 to about 85% CD56+ cells and about 50 to about 15% endogenous CD56− cells;
- wherein the CD56+ cells express a high frequency of NKp30, NKp46, NKp44, NKG2A, and granzyme B, a moderate to high frequency of perforin and CD107a, a low to moderate frequency of CD16, and substantially no KIRs;
- and wherein the CD56− cells express a moderate to high frequency of granzyme B, a high frequency of CD107a, and a low frequency of perforin.
The method of any of the preceding claims, wherein the differentiation culture medium does not contain added Flt-3L, FGF-2, IL-6, IL-7, IL-12, IL-3, GM-CSF, G-CSF, LIF, MIP-1alpha, SCF, IL-21, IL-18, and 4-1BBL.
The method of any of the preceding claims, wherein IL-2 and IL-15 are the only added cytokines to the differentiation culture medium.
The method of any of the preceding claims, wherein the differentiation culture medium does not contain exogenous antigen presenting cells.
The method of any of the preceding claims, wherein the non-animal serum replacement is human AB serum, fresh frozen human plasma, or human platelet lysate.
The method of any of the preceding claims, wherein the HSPCs are not derived from somatic cells, embryonic stem cells, peripheral blood mononuclear cells or induced pluripotent stem cells.
The method of any of the preceding claims, wherein the NK cell composition and/or preparation comprises less than 2% CD3+ cells, less than 2% CD19+ cells, and/or less than 2% CD34+ cells.
The method of any of the preceding claims, wherein the CD56+ cells further express a high frequency of KIR2DL4.
The method of any of the preceding claims, wherein the Notch ligand is DXI or an antibody specific for Notch.
The method of any of the preceding claims, wherein cells of the NK cell composition and/or preparation are genetically modified.
The method of any of the preceding claims, wherein the NK cell composition and/or preparation further comprises a cryoprotective agent.
The method of any of the preceding claims, further comprising formulating the NK cell preparation to form a NK cell composition for infusion into a subject.
A natural killer (NK) cell composition for use in immunotherapy, comprising:
- about 50 to about 80% CD56+ cells and about 50 to about 20% endogenous CD56− cells, or about 50 to about 85% CD56+ cells and about 50 to about 15% endogenous CD56− cells, wherein the CD56+ cells express a high frequency of NKp30, NKp46, NKp44, NKG2A, NKG2D, and granzyme B, a moderate to high frequency of perforin and CD107a, a low to moderate frequency of CD16 and substantially no KIRs;
- and wherein the CD56− cells express a moderate to high frequency of granzyme B, a high frequency of CD107a, and a low frequency of perforin, and a pharmaceutically acceptable carrier.
Dependencies are inferred from the claim text.