US 12,291,722 B2
Methods for manufacturing adoptive cell therapies
Richard Morgan, Center Harbor, NH (US); Kevin Friedman, Medford, MA (US); and Dawn Maier, North Reading, MA (US)
Assigned to 2seventy bio, Inc., Cambridge, MA (US)
Appl. No. 15/306,729
Filed by 2seventy bio, Inc., Cambridge, MA (US)
PCT Filed Apr. 24, 2015, PCT No. PCT/US2015/027518
§ 371(c)(1), (2) Date Oct. 25, 2016,
PCT Pub. No. WO2015/164745, PCT Pub. Date Oct. 29, 2015.
Claims priority of provisional application 61/984,558, filed on Apr. 25, 2014.
Prior Publication US 2017/0051252 A1, Feb. 23, 2017
Int. Cl. C12N 5/0783 (2010.01); A61K 35/12 (2015.01); A61K 38/17 (2006.01); A61K 39/00 (2006.01); C12N 7/00 (2006.01); C12N 15/86 (2006.01)
CPC C12N 5/0636 (2013.01) [A61K 38/177 (2013.01); A61K 39/4611 (2023.05); A61K 39/4631 (2023.05); A61K 39/464412 (2023.05); A61K 39/464417 (2023.05); C12N 7/00 (2013.01); C12N 15/86 (2013.01); A61K 2035/124 (2013.01); C07K 2319/00 (2013.01); C12N 2501/2301 (2013.01); C12N 2501/998 (2013.01); C12N 2740/10043 (2013.01); C12N 2740/15043 (2013.01); C12N 2799/021 (2013.01); C12N 2799/027 (2013.01); C12N 2810/855 (2013.01)] 35 Claims
 
1. A method for manufacturing a human T cell therapeutic for a subject comprising:
a) obtaining a population of human peripheral blood mononuclear cells (PBMCs) from the subject, wherein the PBMCs comprise CD4+ T cells, CD8+ T cells, and antigen presenting cells (APCs);
b) culturing the population of PBMCs of step a) for about 20 hours to about 24 hours prior to transduction in a cell culture medium comprising i) one or more cytokines, ii) a soluble anti-CD3 antibody or CD3-binding fragment thereof, and iii) a soluble anti-CD28 antibody or a CD28-binding fragment thereof, thereby producing activated T cells;
c) transducing the population of PBMCs of step b) comprising activated T cells with a lentiviral vector comprising a polynucleotide encoding an anti-B cell maturation antigen (BCMA) chimeric antigen receptor (CAR), thereby producing transduced T cells; and
d) culturing the population of PBMCs of step c) in a cell growth medium to expand the number of the transduced T cells,
thereby manufacturing the human T cell therapeutic for the subject.