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US 12,453,773 B2Granted
Expansion of natural killer and chimeric antigen receptor-modified cells
AssigneesRutgers, The State University of New Jersey, Houston Methodist Hospital
InventorDongfang Liu
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Abstract
Disclosed herein are methods and compositions for generating immunotherapeutic cells (e.g., NK and T cells) with enhanced cytotoxicity and capacity for expansion thereof. The methods and compositions disclosed herein can further be used for enhanced expansion of CAR-modified NK and T cells with increased cytotoxicity.
Claims20
A modified 721.221 cell expressing membrane-bound IL-21 (mIL-21), wherein the mIL-21 comprises an amino acid sequence comprising at least 90% or at least 95% sequence identity to SEQ ID NO:
- 2, and/or is encoded by a nucleic acid comprising a nucleic acid sequence comprising at least 90% or at least 95% sequence identity to SEQ ID NO:
- 1,
- wherein the mIL-21 is present in a construct further comprising an extracellular fragment from an IgG sequence, a transmembrane fragment from a CD28 sequence, an intracellular fragment from a CD28 sequence, an intracellular fragment from a 4-1BB sequence, and an intracellular fragment from a CD3ζ sequence.
A method of producing a modified 721.221 cell, comprising:
- transducing or transfecting a population of 721.221 cells with a nucleic acid encoding mIL-21, wherein the mIL-21 is encoded by a nucleic acid sequence comprising at least 90% or at least 95% sequence identity to SEQ ID NO:
- 1, and/or the mIL-21 comprises an amino acid sequence comprising at least 90% or at least 95% sequence identity to SEQ ID NO:
- 2;
- isolating the cells that express the mIL-21;
- and
- irradiating the isolated cells,
- thereby producing the modified 721.221 cell,
- wherein the mIL-21 is present in a construct further comprising an extracellular fragment from an IgG sequence, a transmembrane fragment from a CD28 sequence, an intracellular fragment from a CD28 sequence, an intracellular fragment from a 4-1BB sequence, and an intracellular fragment from a CD32 sequence.
5 more elements
Dependencies are inferred from the claim text.