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US 2006/0140922 A1Published application
Universal GM-CSF expressing bystander human K562 cell line
AssigneeThe Johns Hopkins University School of Medicine
InventorsHyam Levitsky, Ivan Borrello
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Abstract
The present invention provides a universal immunomodulatory cytokine-expressing bystander cell line, a composition comprising such a cell line and a cancer antigen, a method of making such a cell line, and a method of using such a composition.
Claims21
A universal bystander cell line, which:
- (i) is the human cell line K562 or a subclone thereof, which is modified to lack major histocompatibility class I (MHC-I) antigens and major histocompatibility class II (MHC-II) antigens, and
- (ii) is modified by introduction of a nucleic acid molecule comprising a nucleic acid sequence encoding granulocyte macrophage-colony stimulating factor (GM-CSF) operably linked to a promoter, wherein the universal bystander cell line expresses about 500 ng or greater GM-CSF/106 cells/24 hours.
A method of making a universal GM-CSF-expressing bystander cell line, which method comprises:
- (i) obtaining the human cell line K562 or a subclone thereof, which is modified to lack MHC-I antigens and MHC-II antigens;
- (ii) modifying the human cell line by introducing into the human cell line a nucleic acid molecule comprising a nucleic acid sequence encoding GM-CSF operably linked to a promoter and a nucleic acid sequence encoding a selectable marker operably linked to a promoter;
- and
- (iii) using the selectable marker to isolate cells that produce about 500 ng or greater of said GM-CSF/106 cells/24 hours.
1 more element
Dependencies are inferred from the claim text.