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Objective To explore the influence of AKT inhibitors on tumor infiltrating T lymphocytes (TIL)in patients with liver metastasis of colorectal cancer.Methods The tumor tissues from the patients with liv-er metastasis of colorectal cancer in Department of General Surgery,The Affiliated Cancer Hospital of Zhengzhou University from January 2016 to December 2016 were collected.TIL and tumor cells were isolated by percoll densi-ty gradient centrifugation. The profiling of AKT inhibitors on TIL were analyzed by flow cytometry. Results AKT inhibition enhances the expansion of TIL with memory cell without affects its proliferation,also the cells obtained under AKT inhibitor with IL-2 showed higher frequency of IFN-γproducing cells than IL-2. Conclusion Add AKT inhibitors in TIL cultivation system can strengthen the proliferation of central memory T cells,and does not affect the number of CD8+T cells.This might be developed for cell-based immunotherapy of cancer.
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Objective To study the anti-tumor effects of mixed cultured B16 melanoma cells supernatant.Methods The supernatant from purely cultured B16 melanoma cells or mixedly cultured B16 melanoma cells with lymphocytes and macrophages at indicated time points were collected,respectively.The chemotaxis of the two different cell supernatants was determined by Boyden room method.The activation effects towards lymphocytes of the two different supernatants were determined by CCK-8 method.Results When the cells were mixed cultured for 0.5,1,2,4,8 and 12 h,the numbers of lymphocytes to travel from the upper well to the bottom well were (1.00±0.82) × 104,(7.00±1.63) × 104,(9.50±0.58) × 104,(11.25±2.36) ×104,(17.25±1.71) × 104 and (21.50±1.29) × 104,respectively.When the cells were purely cultured for 0.5,1,2,4,8 and 12 h,the numbers of lymphocytes to travel from the upper well to the bottom well were (0.00±0.00) ×104,(0.25±0.50) × 104,(1.75±0.96) × 104,(5.25±0.96) × 104,(5.75±1.26) × 104 and (10.75±3.20) × 104,respectively.The mixed cultured group showed higher chemotaxis effects towards lymphocytes in comparison with the purely cuhured one at the same points except for 0.5 h (P ＜ 0.05).The mixed cultured group showed higher activation effects towards lymphocytes in comparison with the purely cultured at the same points except for 0.5 and 1 h (P ＜ 0.05).Each group showed higher chemotaxis and activation effects towards lymphocytes when they were cultured for 12 h than the other time points (P ＜0.05).Conclusion The supernatant from mixed cultured cells shows much higher chemotaxis and activation effects towards lymphocytes to kill tumor cells.
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[Objective]To explore the roles of TIDC(tumor infiltrating dendritic cell)and TIL-T(tumor infiltrating T lymphocyte)in tumor immunity of EBV-associated gastric carcinoma(EBVaGC)and their clinical significance.[Methods]TIDC and TIL-T in EBVaGC(n=49)and EBVnGC(EBV-negative gastric carcinoma)(n=20)were detected by immunohistochemistry method and then their subtypes and their relationships with clinical and pathological factors were analyzed.[Results]Mild infiltration was detected in 29 of 49(59.2%)EBVaGCs and 18 of 20(90%)EBVnGCs,while marked infiltration of TIDC was detected in 20 of 49(40.8%)EBVaGCs and 2 of 20(10%)EBVnGCs.And the difference between the two groups was significant (P=0.013).The number of TIL-T in EBVaGC was hisher than that in EBVnGC(P=0.017),and most of them were CTLs.The number of TIL-T was positively correlated with that of TIDC(r=0.386,n=49,P=0.006).The degree of TIL-T infiltration in EBVaGC had negative relationship with the lymph node metastasis(P＜0.001),while TIDC had no relationship with the clinical features.[Conclusion]TIDC and TIL-T may play important roles in the tumor immunity of EBVaGC;TIDC may contribute to the TIL-T recruitment in EBVaGC.