Efficient amplification method of TILs serving as sources of cancerous pleural effusion

Abstract

The invention aims to provide an efficient amplification method of TILs serving as sources of cancerous pleural effusion. The TILs prepared with the method have the advantages of being high in multiplication speed, large in number, high in cell activity and the like. The method is mainly characterized in that by utilizing a cultivation bottle coated by laminin protein and fibronectin, the TILs are cultivated. The adsorption effect of the two kinds of protein can make cells in half-adherence state, therefore, the cells are favorable for being stimulated by various factors, and the two kinds of protein has the multiplication promoting effect on the TILs; by using a resistant OX-40 monoclone antibody, co-stimulatory signals of CD3+CD8+T cell subgroups in the TILs can be stimulated, cell activation is promoted, the expression of cell perforin and granular enzyme is improved, and the killing capacity of the TILs is enhanced. So far, the study report that the laminin protein, the fibronectin and the resistant OX-40 monoclone antibody are combined and used for preparing the TILs has not been given, and the method for increasing the cell number and improving the killing activity by adding the three factors in preparation of the TILs is put forward for the first time.

Description

technical field

[0001] The invention belongs to the field of cell culture, and in particular relates to the culture of tumor infiltrating lymphocytes (Tumor Infiltrating lymphocytes, TILs), that is, the combination of various cytokines is used to stimulate lymphocytes derived from pleural and ascites, so that they can be efficiently expanded. Background technique

[0002] Tumor infiltrating lymphocytes (Tumor Infiltrating lymphocytes, TIL) refer to lymphocytes that infiltrate into the surrounding tumor tissue, and the cell phenotype is CD3 + CD8 + T and CD3 + CD4 + T cells are predominant, and there are also a small amount of NKT cells and NK cells. Since most TIL cells have been in contact with tumor cells, they can theoretically have a specific killing effect on tumor cells. However, due to the presence of strong immunosuppressive factors in the tumor microenvironment, the function of TIL cells is actually greatly inhibited and cannot effectively kill tumor cells. [...