Methods for transduction and cell processing

Abstract

Provided are methods, systems, and kits for cell processing, e.g., for therapeutic use, such as for adoptive cell therapy. The provided methods include transduction methods, in which cells and virus are incubated under conditions that result in transduction of the cells with a viral vector. The incubation in some embodiments is carried out in an internal cavity of a generally rigid centrifugal chamber, such as a cylindrical chamber made of hard plastic, the cavity of which may have a variable volume. The methods include other processing steps, including those carried out in such a chamber, including washing, selection, isolation, culture, and formulation. In particular, the disclosure relates to method providing advantages over available processing methods, such as available methods for large-scale processing. Such advantages include, for example, reduced cost, streamlining, increased efficacy, increased safety, and increased reproducibility among different subjects and conditions.

1 . A composition comprising at least 1×10 7  T cells, wherein at least 10% of the T cells in the composition are transduced with a recombinant viral vector and express a chimeric antigen receptor (CAR) encoded by the recombinant viral vector, and among the T cells in the composition that are transduced with the recombinant viral vector and express the CAR, the average copy number of the recombinant viral vector is no more than 10. 
     
     
         2 . The composition of  claim 1 , wherein the composition comprises at least 5×10 7  T cells. 
     
     
         3 . The composition of  claim 1 , wherein at least 30% of the T cells in the composition express CD69 or TGF-beta-II. 
     
     
         4 . The composition of  claim 1 , wherein at least 30%, of the T cells in the composition do not express CD62L. 
     
     
         5 . The composition of  claim 1 , wherein at least 30% of the T cells in the composition express CD25, ICAM, GM-CSF, IL-8, and/or IL-2. 
     
     
         6 . The composition of  claim 1 , wherein the composition comprises at least 1×10 8  cells. 
     
     
         7 . The composition of  claim 1 , wherein the T cells are unfractionated T cells, isolated CD8 +  T cells, or isolated CD4 +  T cells. 
     
     
         8 . The composition of  claim 1 , wherein the cells in the composition are primary cells. 
     
     
         9 . The composition of  claim 1 , wherein at least 20% of the T cells in the composition are transduced with the recombinant viral vector and express the CAR. 
     
     
         10 . The composition of  claim 1 , wherein at least 40% of the T cells in the composition are transduced with the recombinant viral vector and express the CAR. 
     
     
         11 . The composition of  claim 1 , wherein at least 50% of the T cells in the composition are transduced with the recombinant viral vector and express the CAR. 
     
     
         12 . The composition of  claim 1 , wherein the recombinant viral vector is a lentiviral vector. 
     
     
         13 . The composition of  claim 1 , wherein the CAR comprises an extracellular portion that specifically binds to a ligand and an intracellular signaling portion comprising an activating domain and a costimulatory domain. 
     
     
         14 . The composition of  claim 1 , wherein among the T cells in the composition that are transduced with the recombinant viral vector and express the CAR, the average copy number of the recombinant viral vector is no more than 6. 
     
     
         15 . A composition comprising at least 1×10 7  T cells, wherein at least 20% of the T cells in the composition are transduced with a recombinant lentiviral vector and express a chimeric antigen receptor (CAR) encoded by the recombinant lentiviral vector, and among the T cells in the composition that are transduced with the recombinant lentiviral vector and express the CAR, the average copy number of the recombinant lentiviral vector is no more than 6. 
     
     
         16 . The composition of  claim 15 , wherein the CAR comprises an extracellular portion that specifically binds to a ligand and an intracellular signaling portion comprising an activating domain and a costimulatory domain. 
     
     
         17 . The composition of  claim 1 , comprising a pharmaceutically acceptable excipient. 
     
     
         18 . The composition of  claim 1 , wherein the composition is cryopreserved. 
     
     
         19 . The composition of  claim 15 , comprising a pharmaceutically acceptable excipient. 
     
     
         20 . The composition of  claim 15 , wherein the composition is cryopreserved.