The invention discloses a preparation method of a high-efficiency killer
cell preparation adopting an immunodetection point dual-block CTL (cytotoxic
lymphocyte). The preparation method comprises steps as follows: step 1), collecting mononuclear cells in
peripheral blood; step 2), performing DC (
dendritic cell) separation and
T cell separation on the separated mononuclear cells with an adherence method; step 3), adding adhering DCs to a serum-free culture medium containing GM-CSF (
granulocyte-macrophage colony-stimulating factor) and IL-4 (
interleukin-4) for preparation of a
DC vaccine; step 4), adding suspending T cells to CD3 and IL-2 for
T cell culture and amplification; step 5), adding
antigen-supported DCs to the amplified T cells for preparation of the CTL, and adding PD-1 (
Programmed death 1) and CTLA-4 (cytotoxic
t lymphocyte associated
antigen-4) antibodies for preparation of the
cell preparation with the dual-block CTL effect. According to the novel technology and the novel method provided by the invention, an immune
brake effect is effectively regulated and
T cell depletion is retarded by blocking
inhibitory molecules on the
tumor specific CTL surfaces, so that the
tumor cytotoxicity of CTL
effector cells is substantially improved.