COMPOSITIONS AND METHODS FOR PRIMING MONOCYTIC DENDRITIC CELLS AND T CELLS FOR TH-1 RESPONSE
The present invention provides compositions and methods for inducing maturation of immature dendritic cells (DC) and for priming those cells for inducing a type 1 immune response. The present invention also provides dendritic cell populations useful for activating and for preparing T cells polarized towards production of type 1 cytokines and/or a type 1 response. Similarly, activated, polarized T cell populations, and methods of making the same are provided.
1 . A method for producing a mature dendritic cell population, comprising:
providing immature dendritic cells; and
contacting the immature dendritic cells with an effective amount of BCG and Interferon gamma (IFNγ) under culture conditions suitable for maturation of the immature dendritic cells to form a mature dendritic cell population;
wherein the mature dendritic cell population produces an increased ratio of Interleukin 12 to Interleukin 10 than an immature dendritic cell population not contacted with BCG and IFNγ during maturation.
2 . The method of claim 1 , further comprising contacting the immature dendritic cells with a predetermined antigen prior to contacting with BCG and IFNγ.
3 . The method of claim 1 , further comprising simultaneously contacting the immature dendritic cells with a predetermined antigen, BCG and IFNγ.
4 . The method of claim 2 or 3 , wherein the predetermined antigen is a tumor specific antigen, a tumor associated antigen, a viral antigen, a bacterial antigen, tumor cells, bacterial cells, a cell lysate, a membrane preparation, a recombinantly produced antigen, a peptide antigen, or an isolated antigen.
5 . The method of claim 1 , further comprising:
isolating monocytic dendritic cell precursors; and
culturing the precursors in the presence of a differentiating agent to form the immature dendritic cells.
6 . The method of claim 5 , wherein the differentiating agent is GM-CSF, Interleukin 4, a combination of GM-CSF and Interleukin 4, or Interleukin 13.
7 . The method of claim 5 , wherein the monocytic dendritic cell precursors are isolated from a human subject.
8 . The method of claim 1 , wherein the mature dendritic cells produce a ratio of IL-12 to IL-10 of at least about 1:1, at least about 10:1, at least about 100:1.
9 . A method for producing a mature dendritic cell population, comprising:
providing immature dendritic cells; and
contacting the immature dendritic cells with an effective amount of BCG and Interferon gamma (IFNγ) under culture conditions suitable for maturation of the immature dendritic cells to form a mature dendritic cell population;
wherein the mature dendritic cell population produces a type 1 immune response.
10 . The method of claim 9 , further comprising contacting the immature dendritic cells with a predetermined antigen prior to contacting with BCG and IFNγ.
11 . The method of claim 9 , further comprising simultaneously contacting the immature dendritic cells with a predetermined antigen, BCG and IFNγ.
12 . The method of claim 10 or 11 , 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.
13 . The method of claim 9 , further comprising:
isolating monocytic dendritic cell precursors; and
culturing the precursors in the presence of a differentiating agent to form the immature dendritic cells.
14 . The method of claim 13 , wherein the differentiating agent is GM-CSF, Interleukin 4, a combination of GM-CSF and Interleukin 4, or Interleukin 13.
15 . The method of claim 9 , wherein the monocytic dendritic cell precursors are isolated from a human subject.
16 . The method of claim 9 , wherein the mature dendritic cells produce a ratio of IL-12 to IL-10 of at least about 1:1, at least about 10:1, or at least about 100:1.
17 . A composition for activating T cells, comprising:
a dendritic cell populations matured with an effective concentration of BCG and IFNγ under suitable conditions for maturation; and
a predetermined antigen;
wherein the dendritic cell population produces an increased ratio of Interleukin 12 (IL-12) to Interleukin 10 (IL-10) than a mature dendritic cell population contacted with BCG without IFNγ during maturation.
18 . The composition of claim 17 , wherein the dendritic cell population produces IL-12 to IL-10 in a ratio of at least about 10:1.
19 . The composition of claim 17 , wherein the dendritic cell population produces IL-12 to IL-10 in a ratio of at least about 100:1.
20 . An isolated, immature dendritic cell population, comprising:
isolated immature monocytic dendritic cells, and an effective concentration of BCG and IFNγ to induce maturation of the immature dendritic cells;
wherein the resulting mature dendritic cells produce more Interleukin 12(IL-12) to Interleukin 10 (IL-10).
21 . The method of claim 20 , further comprising a predetermined antigen.
22 . The cell population of claim 20 , further comprising isolated T cells.
23 . The cell population of claim 22 , wherein the T cells are naïve T cells.
24 . The composition of claim 20 , further comprising isolated lymphocytes.
25 . A method for producing T cells, comprising:
providing immature dendritic cells;
contacting the immature dendritic cells with a predetermined antigen;
contacting the immature dendritic cells with an effective concentration of BCG and IFNγ under culture conditions suitable for maturation of the immature dendritic cells to form mature dendritic cells; and
contacting the mature dendritic cells with naïve T cells to form an activated T cells producing IFNγ.
26 . The method of claim 25 , 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.
27 . The method of claim 25 , wherein the immature dendritic cells are contacted simultaneously with the predetermined antigen, BCG and IFNγ.
28 . The method of claim 25 , further comprising:
isolating monocytic dendritic cell precursors; and
culturing the precursors in the presence of a differentiating agent to form the immature dendritic cells.
29 . The method of claim 28 , wherein the differentiating agent is GM-CSF, Interleukin 4, a combination of GM-CSF and Interleukin 4, or Interleukin 13.
30 . The method of claim 28 , wherein the monocytic dendritic cell precursors are isolated from a human subject.
31 . The method of claim 25 , wherein the immature dendritic cells and T cells are autologous to each other.
32 . Isolated mature dendritic cells producing more Interleukin 12 (IL-12) to Interleukin 10 (IL-10) prepared by maturation of immature dendritic cells with a composition comprising effective concentrations of BCG and IFNγ under conditions suitable for the maturation of the dendritic cells.
33 . The isolated, mature dendritic cells of claim 32 , further comprising a predetermined antigen.
34 . Isolated mature dendritic cells loaded with a predetermined antigen, the dendritic cells producing more Interleukin 12 (IL-12) than Interleukin 10 (IL-10).
35 . The isolated mature dendritic cells of claim 34 , wherein the cells produce at least 10-fold more IL-12 than IL-10.
36 . A method for producing a type 1 immune response in an animal, comprising:
providing immature dendritic cells;
contacting the immature dendritic cells with effective amounts of BCG and Interferon gamma (IFNγ) and a predetermined antigen under culture conditions suitable for maturation of the immature dendritic cells to form mature dendritic cells;
contacting the mature dendritic cells with naïve T cells to form activated T cells producing more Interleukin 12 (IL-12) to Interleukin 10 (IL-10); and
administering the activated T cells to the animal.
37 . The method of claim 36 , 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.
38 . The method of claim 36 , wherein the immature dendritic cells are simultaneously contacted with the predetermined antigen, BCG and IFNγ.
39 . The method of claim 36 , wherein the immature dendritic cells are contacted with the predetermined antigen prior to contacting with BCG and IFNγ.
40 . The method of claim 36 , further comprising:
isolating monocytic dendritic cell precursors from the animal; and
culturing the precursors in the presence of a differentiating agent to form the immature dendritic cells.
41 . The method of claim 40 , wherein the differentiating agent is GM-CSF, Interleukin 4, a combination of GM-CSF and Interleukin 4, or Interleukin 13.
42 . The method of claim 36 , wherein the immature dendritic cells and T cells are autologous to the animal.
43 . The method of claim 36 , wherein the immature dendritic cells and T cells are allogenic to the animal.
44 . The method of claim 36 , wherein the immature dendritic cells and T cells have the same MHC haplotype as the animal.
45 . The method of claim 36 , wherein the animal is human.