US 12,448,604 B2
Methods of producing modified natural killer cells and methods of use
Richard W. Childs, Rockville, MD (US); and David S. J. Allan, Rockville, MD (US)
Assigned to The United States of America, as represented by the Secretary, Department of Health and Human Services, Bethesda, MD (US)
Filed by The United States of America, as represented by the Secretary, Dept. of Health and Human Services, Bethesda, MD (US)
Filed on Mar. 7, 2022, as Appl. No. 17/688,435.
Application 17/688,435 is a division of application No. 15/801,085, filed on Nov. 1, 2017, granted, now 11,293,010.
Application 15/801,085 is a continuation in part of application No. PCT/US2017/043774, filed on Jul. 25, 2017.
Claims priority of provisional application 62/366,493, filed on Jul. 25, 2016.
Prior Publication US 2022/0228117 A1, Jul. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0783 (2010.01); A61K 40/15 (2025.01); A61K 40/42 (2025.01); C12N 5/00 (2006.01); C12N 15/867 (2006.01)
CPC C12N 5/0646 (2013.01) [A61K 40/15 (2025.01); A61K 40/42 (2025.01); C12N 5/0068 (2013.01); C12N 15/867 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); C12N 2501/2302 (2013.01); C12N 2501/505 (2013.01); C12N 2501/515 (2013.01); C12N 2501/599 (2013.01); C12N 2501/65 (2013.01); C12N 2501/999 (2013.01); C12N 2502/1135 (2013.01); C12N 2502/30 (2013.01); C12N 2502/99 (2013.01); C12N 2510/00 (2013.01); C12N 2510/04 (2013.01); C12N 2740/15041 (2013.01); C12N 2740/15043 (2013.01); C12N 2799/027 (2013.01)] 14 Claims
 
1. An expanded population of transduced natural killer (NK) cells comprising one or more heterologous nucleic acids produced by a method comprising:
culturing a population of isolated NK cells in the presence of 500 IU/ml interleukin-2 (IL-2) and in the absence of irradiated feeder cells for 2-3 days to produce a population of activated NK cells;
transducing the population of activated NK cells with a viral vector comprising one or more heterologous nucleic acids to produce a population of transduced NK cells, wherein the heterologous nucleic acid encodes CXCR4, CXCR3, CD34, double negative TGFβ type II receptor, VLA-4 or LFA-1 or encodes a chimeric antigen receptor that specifically binds to CD19, CD20, CD33, CD138, CS1, GD2, HER2, erbB2, CEA, EpCAM, NKG2D-L, or TRAIL-R1; and
culturing the population of transduced NK cells in the presence of 500 IU/ml IL-2 and irradiated feeder cells to produce the expanded population of transduced NK cells.