Expression Of Tim-3 And PD-1 On NK Cells Of Patients With NPC And Its Clinical Significance

Objective:This study tested the expression of Tim-3 and PD-1 on NK cells of patients with NPC,and analyzes the relationship with tumor clinical stages,and discusses the effects in the occurrence and development of NPC To provide experimental basis for immunotherapy of NPC.Tim-3 and PD-1 will provide new targets for improving NPC.Methods:1?From August 2019 to December 2019,we have collected30 patients with newly diagnosed NPC in the Affiliated Hospital of Guilin Medical College.The control group consisted of 22 normal physical examiners with no significant difference in age,sex and case group.2?Using flow cytometry,we determined the percentage of NK cells and the expression of Tim-3 and PD-1 on NK cells in peripheral blood of patients with NPC.And using tissue immunofluorescence double staining,we determined the expression of Tim-3 and CD56 in NPC tumor tissue.Finally,we measured the concentrations of cytokines IFN-? and TNF-? in peripheral blood by enzyme-linked immunosorbent assay(ELISA).3?The clinical data of patients in the experimental group and normal persons in the control group were collected,including sex,age,history of smoking and drinking,clinical stage and lymph node metastasis.The clinical staging was based on the(AJCC)eighth edition staging criteria of the American Joint Commission on Cancer.4?The experimental data were processed by SPSS23.0 software and Graphpad prism 5.0statistical software.The percentage of NK cells and the expression of Tim-3 and PD-1 on NK cells in patients with NPC and normal controls were expressed by mean ± standard deviation(x ± s).T-test was used to compare the difference between groups,and non-normal distribution data were expressed by nonparametric test.Fisher's exact probability method was used to analyze the expression level of CD56 and Tim-3 of NK cells between NPC and NPE groups.The correlation between the two factors was analyzed by Spearman rank correlation analysis.Results:1?Percentage of NK cells in Peripheral Blood of patients with NPC and the relations with tumor stage.The difference between NPC patients and healthy subjects on the percentage of NK cells in peripheral blood was not statistically significant(P = 0.899).The difference between NPC patients and healthy subjects on the percentage of NK cells in peripheral blood of NPC patientsin stage ?-? and stage ? A-? B was not statistically significant(P = 0.937).2?Expression of Tim-3 in peripheral blood NK cells of patients with NPC and the relations with tumor stage: The difference between NPC patients and healthy subjects on the expression of Tim-3 on NK cells was not statistically significant(P = 0.0307);The difference between NPC patients and healthy subjects on the percentage of Tim-3+NK cells in peripheral blood of NPC patients in stage ?-? and stage ? A-? B was not statistically significant(P = 0.109).3?Expression and significance of Tim-3 on NK cells of patients with NPC tumor tissue: CD56+NK cells increase infiltration in tumor tissues of NPC(P= 0.045),and Tim-3+CD56+NK cells increase in tumor tissues of NPC(P =0.025).4?Concentration of cytokines IFN-? and TNF-? of NK cells in patients with NPC.There was significant difference in IFN-? concentration between NPC patients and healthy subjects(P < 0.0001).There was no significant difference in the concentration of TNF-? between NPC patients and healthy subjects(P = 0.4462).5 ? Concentration of cytokines IFN-? and TNF-? in NK cells after PHA stimulation in peripheral blood of patients with NPC.After PHA stimulation culture,there was significant difference in IFN-?concentration between NPC patients and healthy subjects(P = 0.0009).There was no significant difference in the concentration of TNF-?(P = 0.0525).Conclusion(s):1?The expression of Tim-3 is up-regulated on NK cells in peripheral blood and tumor tissues of patients with NPC,and the function of anti-tumor cytokines secreted by NK cells is decreased.We speculate that Tim-3 may negatively regulate the function of NK cells and participate in the progression of NPC.Therefore,targeted expression of Tim-3 on NK cells may provide a new way and target for immunotherapy of NPC.2?PD-1 is highly expressed in peripheral blood NK cells of patients with NPC,indicating that it participates in the occurrence and development of NPC and is expected to become a new target for immunotherapy of nasopharyngeal carcinoma.3?Our research was not deep enough and there were some limitations.The effects of Tim-3 and PD-1 expression on the function or NK cells needs to be further studied and determined,such as carrying out Tim-3 and PD-1 blocking experiments to effectively reverse the immunomodulatory function of NK cells in patients with NPC and provide further basis for new approaches and targets of immunotherapy for patients with NPC.