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.