Correlation Of TIM-3,TIGIT And 2B4 Expression In T Cells Of Locally Advanced Nasopharyngeal Carcinoma With Clinical Prognosis

Objective: To investigate the correlation between TIM-3,TIGIT and 2B4 expression and their co-expression in peripheral blood lymphocytes of locally advanced nasopharyngeal carcinoma and their clinical features and prognosis,and to provide a basis for immunotherapeutic combination strategies for locally advanced nasopharyngeal carcinoma.Methods: Peripheral Blood Mononuclear Cells(PBMCS)were collected from83 patients with primary nasopharyngeal carcinoma in the Cancer Hospital of Xinjiang Medical University,and PBMCS from 44 healthy individuals were collected as controls.Flow cytometry was applied to detect the expression and co-expression of TIM-3,TIGIT and 2B4 in different cell subpopulations of peripheral blood;the differences in the expression and co-expression of the three inhibitory receptors in different cell subpopulations between patients with locally advanced nasopharyngeal carcinoma and healthy controls were analyzed;the changes in the expression and co-expression of the three inhibitory receptors in different cell subpopulations before and during treatment were analyzed,and the diagnostic value of the three Finally,the correlation between the expression and co-expression of the three inhibitory receptors in different cell subpopulations in peripheral blood and their clinical characteristics and prognosis was analyzed.Results:(1)Compared with healthy controls,the expression levels of TIM-3,TIGIT,TIM-3+TIGIT,TIM-3+2B4 were higher in CD4+ T cells of patients with locally advanced nasopharyngeal carcinoma(all P<0.05);among CD8+ T cells,TIM-3,TIM-3+TIGIT,TIM-3+2B4 were higher(all P<0.05).2)Clinical characteristics analysis showed that among CD4+ T cells,TIM-3+TIGIT was associated with age and TIGIT+2B4 was associated with pathological staging(all P<0.05);among CD8+ T cells,TIM-3 and TIGIT were associated with age,TIM-3+TIGIT,2B4 was associated with pathological staging,TIM-3+TIGIT,TIM-3+2B4 were related to gender,and TIGIT,TIGIT+2B4 were related to T-stage(all P<0.05).3)The area under the ROC curve(AUC)for TIGIT+2B4 was 0.723 and for TIM-3+TIGIT was 0.738(all P<0.05).4)Patients with locally advanced nasopharyngeal carcinoma had pre-treatment and 3 inhibitory receptor expression as well as co-expression after treatment showed that in CD4+T,the expression level of TIM-3 was higher than that before treatment at March and June after treatment;the expression level of TIGIT was higher than that before treatment at March after treatment;the expression level of 2B4 was higher than that before treatment at January,March,June and December after radiotherapy;the expression level of TIM-3+TIGIT was higher than that before treatment at March and June after radiotherapy were higher than before treatment;the expression levels of TIM-3+2B4 were lower than before treatment at December after radiotherapy(all P<0.05).In CD8+T,the expression levels of TIGIT were higher at 3 months after treatment than before treatment;the expression levels of 2B4 were higher at 1 month,3 months,6 months,and 12 months after radiotherapy than before treatment(all P<0.05).5)Univariate analysis of clinical characteristics showed that age,sex,and smoking history were risk factors for OS;age and smoking history were risk factors for PFS.TIM-3,TIGIT,2B4 expression and their co-expression in the univariate COX proportional model and K-M analysis showed that TIM-3+CD8+T,TIGIT+CD8+T,TIM-3+TIGIT+CD4+T,TIM-3+2B4+CD4+T,TIM-3+TIGIT+CD8+T,TIM-3+2B4+CD8+T,TIGIT+2B4+CD8+ T were risk factors for OS.2B4+CD4+T,TIGIT+CD8+T,TIM-3+TIGIT+CD4+T,TIM-3+2B4+CD4+T,and TIM-3+TIGIT+CD8+T were risk factors for PFS(all P<0.05).Multifactorial analysis showed that TIM-3+2B4+CD4+T was an independent prognostic factor for OS;smoking was an independent prognostic factor for PFS(all P<0.05).Conclusions: 1)The expression level of TIM-3 in peripheral blood CD4/CD8 T cells is upregulated in patients with nasopharyngeal carcinoma when expressed alone and in combination with co-expression;2)TIM-3+TIGIT and TIGIT+2B4 are more diagnostic in CD8+T;(3)Immunoblocking therapy alters the immunosuppressive environment of T cells and then enhances the release of immunogenic tumor antigens by radiotherapy,which may be a more effective mode of combination therapy;4)TIM-3 mono-and co-expression is an important prognostic factor in locally advanced nasopharyngeal carcinoma.