Cytotoxic T cells are one of the most specialized immune cells to defend the body against foreign invaders in the best scenario, but failure in the function of these cells, leading to decrease defense of the body against cancer. We aims to address the functionality of cytotoxic T lymphocytes (CTLs), Natural killer (NK) and Natural killer T-cells (NKT) cells in lung cancer patient’s pre and post-stimulation of these cells in-vitro using laboratory activation protocol. Five healthy persons as volunteers and fifteen lung cancer patients were subjected to this study. The count of CTLs, NK and NKT cells and the expression of their intracellular granzyme B (GzB) pre and post-stimulation for 72 hrs in-vitro was determined. The plasma level of cytokines and chemokines was correlated with the patient’s prognosis. Data showed that after culture, there were highly significantly increased in the fold change of the percentage of GzB expression on CD3, CD4, CD8 and NKCD8 T cells in lung cancer patients before induction of chemotherapy when compared to healthy control and other lung cancer patients. The level of Proinflammatory cytokines in patients before and during induction of chemotherapy showed IL-1 and CCL-2 in a 13000, 250 fold increase respectively while it decreases in patients after induction of chemotherapy compared with the control group. Patients before the induction of chemotherapy have the ability to improve their CTL and NK functionality under activation conditions. The use of immune activation mechanisms is needed before the induction of chemotherapy.