Compositions and methods for NK-92 cells expressing native CD16
AssigneeImmunityBio, Inc.
InventorsBarry J. Simon, Laurent H. Boissel, Prachi Jain
Life of this patent
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Abstract
A recombinant NK-92 cell has a constitutive active promotor that effects expression of native CD16, and most preferably homogenous CD16 158V. Further contemplated recombinant NK-92 cells also include a recombinant nucleic acid that encodes an intracellularly retained interleukin (e.g., IL-2 or er-IL-2), wherein the recombinant NK-92 cell will secrete no more than 5,000 pg/mL IL-2 into a culture medium. The recombinant NK-92 cells presented herein have a significantly improved signal-to-noise ratio and exhibit reduced non-ADCC cytotoxicity.
Claims17
An isolated recombinant NK-92 cell that is natively heterozygous for CD16, comprising:
- a native CD16 gene that is natively heterozygous for CD16, wherein the native CD16 gene has a CD16 158V allele and a CD16 158F allele;
- a recombinant constitutive promotor sequence upstream of the native CD16 gene that is functionally in place of a native promotor sequence of the native CD16 gene;
- wherein the recombinant promotor sequence is operably coupled to the native CD16 gene to thereby enable expression of the native CD16;
- and
- wherein the recombinant promoter sequence leads to detectable expression of the CD16 158V allele, but not the 158F allele.
A method of making a recombinant NK-92 cell, comprising:
- providing a NK-92 cell;
- introducing to the NK-92 cell a recombinant constitutive promotor sequence upstream of a gene encoding a native CD16 allele to thereby make the recombinant NK-92 cell;
- wherein the step of introducing the recombinant promotor sequence comprises functional replacement of a native promotor sequence of the native CD16 allele by homologous recombination with the recombinant promotor sequence;
- wherein the recombinant promotor sequence is operably coupled to the native CD16 allele to thereby enable expression of the native CD16 allele;
- and
- wherein the recombinant promoter sequence leads to detectable expression of the CD16 158V allele, but not the 158F allele.
Dependencies are inferred from the claim text.