METHODS OF USING IL-21 FOR ADOPTIVE IMMUNOTHERAPY AND IDENTIFICATION OF TUMOR ANTIGENS
Methods for preparing ex vivo T cell cultures using IL-21 compositions for use in adoptive immunotherapy are described. Addition of IL-21 to cultures of non-terminally differentiated T cells population, either isolated or present in peripheral blood mononuclear cells are exposed to one or more tumor antigens, and in the presence of IL-21 compositions and antigen presenting cells (APCs), the resulting T cell population has an enhanced antigen-specificity; and can be reintroduced into the patient. Methods are also disclosed for identifying tumor antigens by culturing T cell populations exposed to IL-21 compositions and APCs in the presence of tumor material.
1 . A method of preparing a T cell population comprising:
obtaining the T cell population from PBMCs,
activating the obtained T cell population,
transducing the activated T cell population with a viral vector comprising a nucleic acid encoding a protein,
expanding the transduced T cell population, and
wherein the transducing is performed in the presence of IL-21.
2 . The method of claim 1 , wherein the PBMCs are obtained from a patient or a healthy donor.
3 . The method of claim 1 , wherein the transducing is further performed in the presence of at least one cytokine selected from IL-2, IL-12, and IL-15.
4 . The method of claim 1 , wherein the protein is a T cell receptor.
5 . The method of claim 1 , further comprising restimulating the expanded transduced T cell population in the presence of a feeder cell and an anti-CD3 antibody.
6 . The method of claim 5 , wherein the feeder cell is an irradiated allogenic feeder cell.
7 . The method of claim 6 , wherein the feeder cell comprises a PBL and an LCL.
8 . The method of claim 7 , further comprising culturing the restimulated cells in the presence of IL-2.
9 . A method of preparing a T cell population comprising:
obtaining the T cell population from PBMCs,
activating the obtained T cell population,
transducing the activated T cell population with a viral vector comprising a nucleic acid encoding a protein,
expanding the transduced T cell population, and
wherein the activating and the transducing are performed in the presence of IL-21.
10 . The method of claim 9 , wherein the PBMCs are obtained from a patient or a healthy donor.
11 . The method of claim 9 , wherein the activating and the transducing are further performed in the presence of at least one cytokine selected from IL-2, IL-12, and IL-15.
12 . The method of claim 9 , wherein the protein is a T cell receptor.
13 . The method of claim 9 , further comprising restimulating the expanded transduced T cell population in the presence of a feeder cell and an anti-CD3 antibody.
14 . The method of claim 13 , wherein the feeder cell is an irradiated allogenic feeder cell.
15 . A method of preparing a T cell population comprising:
obtaining the T cell population from PBMCs,
activating the obtained T cell population,
transducing the activated T cell population with a viral vector comprising a nucleic acid encoding a protein,
expanding the transduced T cell population, and
wherein the transducing and the expanding are performed in the presence of IL-21.
16 . The method of claim 15 , wherein the PBMCs are obtained from a patient or a healthy donor.
17 . The method of claim 15 , wherein the transducing and the expanding are further performed in the presence of at least one cytokine selected from IL-2, IL-12, and IL-15.
18 . The method of claim 15 , wherein the protein is a T cell receptor.
19 . The method of claim 15 , further comprising restimulating the expanded transduced T cell population in the presence of a feeder cell and an anti-CD3 antibody.
20 . The method of claim 19 , wherein the feeder cell is an irradiated allogenic feeder cell.