In recent years, CAR-T therapy has emerged as one of the most promising approaches for treating cancer. Despite all its positive attributes, the therapy also has some drawbacks, such as cytokine release syndrome and neurotoxicity syndrome. However, the most significant limitation is the limited efficacy in treating solid tumors. NK cells have shown potential as candidates for CAR therapy because they cause fewer side effects. In this master's thesis, we aimed to further enhance their activity in destroying tumor cells. Using molecular cloning methods, we prepared various genetic constructs to transduce the cells. The resulting NK-92 cells expressed a chimeric antigen receptor along with either Neo-2/15, which was added to the CAR architecture, or membrane-bound IL-2 fused with the IL-2 receptor α subunit. We demonstrated that the growth of NK-92 cells is dependent on exogenously added IL-2. However, we demonstrated that growth and proliferation in cells expressing Neo-2/15 or membrane-bound IL-2 was independent of added IL-2. We also showed that NK-92 cells expressing CAR and Neo-2/15 or mbIL-2/IL2Rα are effective in killing cancer cells in an IL-2-free environment. They produce several key molecules such as granzyme B and IFN-γ that play an important role in exerting cytotoxicity. The results indicate that CAR-NK-92 cells with the addition of Neo-2/15 or IL-2 are not only capable of surviving in an environment devoid of IL-2, typical of the tumor microenvironment, but also effectively act against cancer cells.
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