Compositions and Methods for Inducing the Activation of Immature Monocytic Dendritic Cells
The present invention provides methods for inducing the maturation of immature dendritic cells (DC) and for activating those cells without the use of a dendritic cell maturation agent. The activated DC can be used for inducing an antigen specific T cell response. Methods of the invention can also comprise the addition of a directional maturation agent, such as interferon gamma, to induce a Th-I and/or Th-2 bias in the response obtained. The present invention also provides dendritic cell populations useful for activating and for inducing antigen specific T cells. Similarly, activated antigen specific T cell populations, and methods of making the same are provided.
1 . A method for producing a cell population comprising an enriched population of activated dendritic cells comprising: contacting a cell population comprising an enriched population of immature dendritic cells with a tissue culture substrate and a culture medium having a dendritic cell differentiation agent under conditions suitable for adhesion of the immature dendritic cells with the tissue culture substrate and for in vitro culture of the dendritic cells for a time period sufficient for maturation of the activated dendritic cells.
2 . The method according to claim 1 , wherein a predetermined antigen is contacted with the cell population prior to, simultaneous with, or subsequent to contact with the tissue culture substrate.
3 . The method according to claim 2 , wherein the predetermined antigen is a tumor specific antigen, a tumor associated antigen, a viral antigen, a bacterial antigen, tumor cells, bacterial cells, recombinant cells expressing an antigen, a cell lysate, a membrane preparation, a recombinantly produced antigen, a peptide, or an isolated antigen.
4 . The method according to claim 3 , wherein the cell lysate or membrane preparation is obtained from a brain tumor, a prostate tumor, prostate tissue, an ovarian tumor, a breast tumor, breast tissue, a leukemic cell population, a lung tumor, a melanoma, a bladder tumor, or a tumor cell line.
5 . The method according to claim 4 , wherein the brain tumor is glioblastoma multiforme or an oligoastrocytoma.
6 . The method according to any of claims 1 through 5 , wherein the dendritic cell differentiation agent is GM-CSF, IL-4, IL-13, IL-15 or combinations thereof.
7 . The method according to any of claims 1 through 6 , wherein the immature dendritic cells are derived from a cell population enriched for monocytic dendritic cell precursors.
8 . The method according to claim 7 , wherein the cell population enriched for monocytic dendritic cell precursors is contacted with a dendritic cell differentiation inducing agent.
9 . The method according to claim 8 , wherein the dendritic cell differentiation inducing agent is GM-CSF, IL-4, IL-13, or IL-15, and combinations thereof.
10 . The method according to claims 7 through 9 , wherein the monocytic dendritic cell precursors are from a patient or from an HLA-matched individual.
11 . The method according to any of claims 3 through 9 , wherein the tumor cells and the dendritic cells are from a patient.
12 . The method according to any of claims 1 through 11 , wherein the immature dendritic cells are further contacted with a directional maturation agent.
13 . The method according to claim 12 , wherein the directional maturation agent is interferon gamma.
14 . A method for activating immature dendritic cells, comprising contacting the immature dendritic cells with a tissue culture substrate and a dendritic cell differentiation inducing agent under conditions suitable for in vitro culture of immature dendritic cells and for adhesion of the immature dendritic cells to the tissue culture substrate.
15 . The method according to claim 14 , further comprising contacting the immature dendritic cells with a predetermined antigen prior to, simultaneous with, or subsequent to contacting the immature dendritic cells with the tissue culture substrate.
16 . The method according to claim 15 , wherein the predetermined antigen is a tumor specific antigen, a tumor associated antigen, a viral antigen, a bacterial antigen, tumor cells, bacterial cells, recombinant cells expressing an antigen, a cell lysate, a membrane preparation, a recombinantly produced antigen, a peptide, or an isolated antigen.
17 . The method according to claim 16 , wherein the cell lysate or membrane preparation is derived from tumor tissue or a tumor cell line.
18 . The method according to claim 17 , wherein the tumor tissue or tumor cell line is obtained from a brain tumor, a prostate tumor, an ovarian tumor, a breast tumor, a lung tumor, a melanoma, a bladder tumor, or a leukemic cell population.
19 . The method according to claim 18 , wherein the brain tumor is glioblastoma multiforme or an oligoastrocytoma.
20 . The method according to any of claims 14 through 19 , wherein the dendritic cell differentiation inducing agent is GM-CSF, IL-4, IL-13, IL-15, or combinations thereof.
21 . The method according to any of claims 14 through 20 , wherein the immature dendritic cells are further contacted with a directional maturation agent.
22 . The method according to claim 21 , wherein the directional maturation agent is interferon gamma.
23 . A method for activating T cells, comprising:
contacting T cells with a dendritic cell population matured by contact with a tissue culture substrate and a dendritic cell differentiation inducing agent under conditions conducive for adhesion of immature dendritic cells to the tissue culture substrate; and
a predetermined antigen.
24 . A method for producing activated antigen specific T cells, comprising:
contacting isolated immature dendritic cells with a predetermined antigen;
contacting the isolated immature dendritic cells with a tissue culture substrate and a dendritic cell differentiation inducing agent under conditions conducive for adhesion of the immature dendritic cells to the tissue culture substrate to activate tie immature dendritic cells; and
contacting the activated dendritic cells with naïve T cells to form activated antigen specific T cells.
25 . The method according to claim 24 , wherein the predetermined antigen is a tumor specific antigen, a tumor associated antigen, a viral antigen, a bacterial antigen, tumor cells, bacterial cells, recombinant cells expressing an antigen, a cell lysate, a membrane preparation, a recombinantly produced antigen, a peptide antigen, or an isolated antigen.
26 . The method according to claim 25 , wherein the cell lysate or membrane preparation is obtained from a brain tumor, a prostate tumor, prostate tissue, an ovarian tumor, a leukemic cell population, a lung tumor, a breast tumor, or a bladder tumor.
27 . The method according to claim 26 , wherein the brain tumor is glioblastoma multiforme or an oligoastrocytoma.
28 . The method according to any of claims 24 through 27 , further comprising as a first step:
culturing monocytic dendritic cell precursors in the presence of a differentiation inducing agent to form the immature dendritic cells.
29 . The method according to claim 28 , wherein the differentiation inducing agent is GM-CSF, Interleukin 4, a combination of GM-CSF and Interleukin 4, or Interleukin 13.
30 . The method according to claim 28 and 29 , wherein the monocytic dendritic cell precursors are isolated from a human subject.
31 . The method according to claim 30 , wherein the immature dendritic cells and T cells are autologous to each other.