Method for preparing antigen-specific cytotoxic t-cells by using activated b-cells and use thereof

A method of producing antigen-specific cytotoxic T cells, comprising the steps of:

• providing human peripheral blood mononuclear cells;
• separating the peripheral blood mononuclear cells into B cells and other mononuclear cells;
• treating the B cells with any one of biological response modifiers selected from the group consisting of alpha-galactosylceramide, alpha-glucuronosylceramide, phosphatidylinositol tetramannoside, isoglobotrihexylceramide, ganglioside GD3, phosphatidylcholine, beta-galactosylceramide, lipophosphoglycan, glycoinositol phospholipid, alpha-galactosylceramide analogs, including beta-anomer galactosylceramide and alpha-anomer galactosylceramide to produce an activated B cell;
• introducing into the activated B cell an antigen polypeptide or a nucleic acid encoding an antigen to produce a activated B cells loaded with an antigen;
• mixing and a first batch of the antigen-loaded activated B cell and the other mononuclear cells separated from the peripheral blood mononuclear cells to produce a first mixture of the cells;
• subjecting the first mixture of the cells to a first incubation;
• adding a second batch of the antigen-loaded activated B cells to the first mixture of the cells after the first incubation to produce a second mixture;
• and
• subjecting the second mixture to a second incubation, wherein the antigen-specific cytotoxic T cell is generated during the first and the second incubation steps.