Formulation and method for preparing specific T cell, and method for preparing the formulation
The present invention provides a formulation and method for preparing specific T cells, and a method for fabricating the formulation is also disclosed. The formulation can induce specific T cell responses, and comprises at least a cell population of dendritic killer cells presenting specific antigens. In addition, the method mentioned above for preparing specific T cells comprises following steps. A T cell population is provided, and then a formulation of preparing specific T cells is mixing with the T cell population. After cultivating, the specific T cells are harvested. Furthermore, the method for fabricating the above formulation comprises the following steps. First, a cell population of dendritic killer cells is provided. A target sample is then provided, and a step of making the cell population of the dendritic killer cells is co-cultivated with the target sample. After co-cultivating, a cell population of dendritic killer cells presenting specific antigens is harvested.
1. A method for preparing antigen specific T cells comprising the following steps:
(a) obtaining human peripheral blood mononuclear cells;
(b) culturing the cells with IL-15 and 0.5-20 ng/mL IL-12;
(c) sorting dendritic killer cells, wherein the dendritic killer cells are cells with the cell surface marker HLA-G − CD14 − CD19 − CD3 − CD56 + HLA-DR + ;
(d) co-culturing the dendritic killer cells with a plurality of target cells, wherein the target cells express a specific antigen;
(e) harvesting the dendritic killer cells presenting the specific antigen;
(f) mixing the dendritic killer cells presenting the specific antigen with a cell population of T cells and co-culturing for a period; and
(g) harvesting the antigen specific T cells.
2. The method according to claim 1 , wherein in the step (d) and (e), the dendritic killer cells kill the target cells and present the specific antigen of the target cells on their surface to form the dendritic killer cells presenting the specific antigen.
3. The method according to claim 1 , wherein in the step (d), the target cells are cancer cells.
4. The method according to claim 3 , wherein in the step (d), the specific antigen is a cancer-specific antigen.
5. The method according to claim 4 , wherein the specific T cells are cancer-specific T cells.
6. The method according to claim 4 , wherein the target cells in the step (d) are cancer cells, the specific antigen in the step (d) is a cancer antigen, the dendritic killer cells presenting the specific antigen in the step (e) and (f) are the dendritic killer cells presenting the cancer antigen, the cell population of T cells in the step (f) are a cell population of CD8 + T cells, and the specific T cells in the step (g) are cancer antigen-specific CD8 + T cells.
7. The method according to claim 6 , wherein the cancer cells, the dendritic killer cells, and the cell population of CD8 + T cells are obtained from a cancer patient.
8. The method according to claim 3 , wherein in the step (d), the target cells and the dendritic killer cells are both obtained from a cancer patient.
9. The method according to claim 1 , wherein in the step (b), the concentration of IL-15 is 10 ng/mL, or the concentration of IL-12 is 2 ng/mL.
10. The method according to claim 1 , wherein in the step (e), the dendritic killer cells presenting the specific antigen are capable of activating specific CD8 + T cells.
11. The method according to claim 1 , wherein the T cells in the step (f) are isolated CD8 + T cells, and the specific T cells in the step (g) are specific CD8 + T cells.
12. The method according to claim 1 , wherein in the step (f) and (g), the cell population of T cells is activated by the dendritic killer cells presenting the specific antigen to form the specific T cells.
13. The method according to claim 1 , wherein the target cells, the dendritic killer cells, and the cell population of T cells are obtained from a cancer patient.
14. A cancer treatment method for a human subject in need thereof, the method comprising the following steps:
(A) preparing a plurality of cancer antigen-specific CD8 + T cells, comprising the following steps:
(a″) obtaining human peripheral blood mononuclear cells from the human subject;
(b″) culturing the cells with IL-15 and 0.5-20 ng/mL IL-12;
(c″) sorting dendritic killer cells, wherein the dendritic killer cells are cells with the cell surface marker HLA-G − CD14 − CD19 − CD3 − CD56 + HLA-D12 + ;
(d″) co-culturing the dendritic killer cells with a plurality of cancer cells obtained from the human subject, wherein the cancer cells express a cancer antigen;
(e″) harvesting the dendritic killer cells presenting the cancer antigen;
(f″) mixing the dendritic killer cells presenting the cancer antigen with a cell population of CD8 + T cells obtained from the human subject and co-culturing for a period; and
(g″) harvesting a plurality of cancer antigen-specific CD8 + T cells; and
(B) delivering the cancer antigen-specific CD8 + T cells to the human subject.