COMPOSITIONS AND METHODS FOR IMMUNOTHERAPY
The present disclosure provides compositions and methods for engineered cellular compositions and methods of immunotherapy utilizing the same. Compositions of the present disclosure for immune cell regulation comprise a chimeric antigen receptor polypeptide, a T cell receptor polypeptide, and combinations thereof.
1 .- 22 . (canceled)
23 . A method of producing a population of engineered immune cells expressing a chimeric antigen receptor (CAR) and/or an engineered T cell receptor (TCR), comprising:
(a) contacting a population of immune cells obtained from a subject with an activation moiety (“population of contacted immune cells”);
(b) introducing a polynucleotide encoding the CAR or TCR in the population of contacted immune cells in (a), thereby producing a population of engineered immune cells expressing the CAR or TCR, and
(c) harvesting the population of engineered immune cells expressing the CAR or TCR for infusion to the subject;
wherein the total duration of both (a) and (b) is no more than 36 hours and wherein the method does not comprise an ex vivo expansion except (a) to (c).
24 . The method of claim 23 , wherein the harvesting comprises cryopreserving the population of engineered immune cells.
25 . The method of claim 23 , wherein the activation moiety is removed after step (b).
26 . The method of claim 23 , wherein the introducing in (b) is performed on the same day as the contacting in (a).
27 . The method of claim 26 , wherein the introducing in (b) is performed about 4 hours or less after the contacting in (a).
28 . The method of claim 23 , wherein the activation moiety and the polynucleotide together are in contact with the population of immune cells in (b) for no more than 24 hours.
29 . The method of claim 23 , wherein the population of immune cells are activated after (a).
30 . The method of claim 28 , wherein the activation moiety comprises anti-CD3 antibody and/or anti-CD28 antibody.
31 . The method of claim 23 , wherein the activation moiety is associated with the population of immune cells during the introducing in (b).
32 . The method of claim 31 , wherein the activation moiety is conjugated to a bead, a plate, and/or a matrix.
33 . The method of claim 23 , wherein the introducing in (b) comprises transducing the population of immune cells with a viral vector and/or a transposon vector.
34 . The method of claim 33 , wherein the viral vector is a retroviral vector, a lentiviral vector and/or an adeno-associated viral vector.
35 . The method of claim 23 , wherein the population of immune cells is seeded at a density from about 10 4 cells/mL to about 10 8 cells/mL.
36 . The method of claim 33 , wherein the viral vector is plated at a mean of infectivity (MOI) from about 0.1 to about 20.
37 . The method of claim 23 , further comprising contacting a cytokine to the population of immune cells.
38 . The method of claim 37 , wherein the cytokine comprises IL2, IL7, IL15 and/or IL21.
39 . The method of claim 23 , wherein the population of engineered immune cells are not cultured in the presence of human serum.
40 . The method of claim 23 , wherein the population of engineered immune cells is characterized in that cell memory T cells (TCM) in the population are more abundant than effector memory T cells (TEM).
41 . A population of engineered immune cells produced by the method of claim 23 .
42 . A point-of-care facility comprising a cell infusion equipment configured to: infuse the population of engineered immune cells of claim 41 .
43 . A method of treating a cancer in a subject in need thereof, comprising infusing the population of engineered immune cells of claim 41 in the subject.