A process for producing dendritic cells that are highly efficient in stimulating T cells to mezothelin protein.

Abstract

OCR 14 / 07 / 2566 A process for producing highly efficient denitic cells capable of stimulating T cells to mezothelin protein according to... This invention involves the design and preparation of a recombinant plasmid. A lentiviral vector containing a set of granulocyte-macrophage cytokine genes. Colony-stimulating factor (granulocyte-macrophage colony-stimulating factor; GM-CSF), Interleukin-4 (L-4) and mezothelin (MSLN) were used to isolate monocytes from blood samples. And modifying gene sets, presenting antigens, and stimulating T cells with high-efficiency dendritic cells. The mezothelin protein produced by dendritic cells is highly effective, and the mezothelin protein produced can be used in... The mesothelin antigen is introduced, stimulating T lymphocytes to effectively recognize the mesothelin antigen. Mesothelin can specifically eliminate breast cancer cells that express high levels of mesothelin protein. This invention will help reduce the steps involved in preparing and transferring antigen segments, as well as reduce variability. The quality of dendritic cells in presenting antigen in each production batch.

Claims

OCR 09WP 02 / 06 / 2569 1. A process for producing dendritic cells with high efficiency in stimulating T cells to mezothelin protein. It consists of the following steps: A. Design and preparation of recombinant plasmids (Lenti type). A lenticular vector is used to create modified monocytes. Dendritic cells expressing mesothelin (MSLN) protein. B. Isolation of monocytes from whole blood samples and gene modification by: The time required to transform dendritic cells into high-efficiency dendritic cells. Stimulating T cells to react to mesothelin protein (SmartDCs-MSLN) takes a total of 7 days. C. Antigen presentation and T-cell activation using modified monocytes at a specific cell ratio. The dendritic to T-lymphocyte ratio was 1 to 10 for 3 days. Where stimulated T cells are isolated and proliferated in a cell culture medium containing a specific type of cytokine. Interleukin-2 (IL-2), Interleukin-7 (IL-7), and Interleukin-15 (IL-15), and serum from normal humans. Blood type AB, 5% (5% Human AB serum) The resulting dendritic cells expressed mezothelin antigen (MSLN), which is specific to... Triple negative breast cancer (TNC) (TNBC) 2. A process for producing highly efficient dendritic cells capable of stimulating T cells to mezothelin protein, as per the following points: Holding the first right, where the design is a gene set design consisting of... A. The promoter portion determines the level of gene expression. Human elongation factor-1 alpha (EF-1 alpha) B. Gene set for determining cytokine expression: granulocyte-macrophage colony stimulation. Granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon. Interleukin-4 (IL-4) and mezothelin antigen (MSLN). C. A set of genes that determine the expression of the connector (P2A) and (F2A) segments.

3. A process for producing highly efficient dendritic cells capable of stimulating T cells to mezothelin protein, as per the following points: He holds the second right, which includes the gene set for the proteins GM-CSF, interleukin-4 (IL-4), and... The mezothelin antigen (MSLN) is a continuous, single-piece tri-systronic CD-DNA sequence. (tri-cistronic cDNA sequence)

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