Human purified CD8+ T cells:: Ex vivo expansion model to generate a maximum yield of functional cytotoxic cells

CD8(+) T cells are a critical component of the cellular immune response. They play an important role in the control of viral infection and eliminating cells with malignant potential. However, attempts to generate and expand human CD8(+) T cells in vitro for an adoptive immunotherapy have been conducted with limitation of the very low frequency of CD8(+) T cells in blood. Therefore, several expansion protocols have been developed to obtain large and efficient numbers of human CD8(+) T cells for use in adoptive immunotherapies. In this study various common culture conditions using different cytokines IL-2, IL-4, IL-7, IL-10, IL-12 and IL-15 and autologous feeders and sera were investigated to expand human purified CD8(+) T cells. The importance and the influence of these factors on the growth and phenotype of CD8(+) T cell were assessed by serially sampling cultures using flow cytometry. We demonstrated that combination of IL-2 (50U/ml) and autologous feeders induced maximal CD8(+) T cell proliferation (40-50 folds) compared to other cytokines. Immunophenotypic analysis of cultured cells showed that expanded CD8(+) T cells were activated and differentiated. Furthermore our expansion model also demonstrated that expanded CD8(+) T cells are functionally cytotoxic active by killing Allogeneic LCLs cells. In conclusion, we have developed a reliable, simple method that uses minimal cell numbers to generate a high yield of functional cytotoxic CD8(+) T cells, which can be used for the development of cellular immunotherapies.