| ObjectiveThe expression of immunosuppressation-related molecules on peripheral blood T cells of colorectal cancer patients with different stages was detected,the correlation between each indicator and clinical information was analyzed,and the immunosuppressive status of patients was evaluated and analyzed,which provided a theoretical basis for the application of immunotargeted drugs for colorectal cancer.MethodsIn the first part of this study,20 patients with stage I+II,III+IV colorectal cancer were collected from the second Department of general surgery,Yunnan first people’s Hospital,Affiliated Hospital of Kunming University of science and technology from November 2018 to February 2019,and 18 cases in the control group.The peripheral venous blood of patients before operation and the control group was drawn and anticoagulated,after adding antibody markers,the expression levels of PD-1,2B4,Tim-3,LAG-3,KLRG1,BTLA,CD160,CTLA-4 on peripheral blood CD8~+T cells were detected by flow cytometry.The second part continued to collect a total of 17 patients with stageⅢ+Ⅳcolorectal cancer who were hospitalized from March 2019 to December 2019,The peripheral venous blood was drawn and anticoagulated before operation,The expression levels of CD38,HLA-DR,CD69,CD27 and CD28 on CD8~+T cells expressing PD-1 and2B4 in peripheral blood were detected by flow cytometry.Results1.There was no significant difference in the expression of Tim-3,LAG-3,KLRG1,BTLA and CD160 on CD8~+T cells in peripheral blood between the colorectal cancer group and the control group,nor was there any significant difference in the expression of CTLA-4 on CD4~+T cells(P>0.05).The expression of PD-1 and 2B4 on CD8~+T cells in peripheral blood of stageⅢ+Ⅳcolorectal cancer group was significantly higher than that in the control group:the expression rate of PD-1 on peripheral blood CD8~+T cells in the control group was 34.95±2.479%,The expression rate of CD8~+T cells in the peripheral blood of the stageⅢ+Ⅳcolorectal cancer group was47.47±3.954;the expression rate of CD8~+T cells in the peripheral blood of the control group was 48.68±4.479%.The expression rate of blood CD8~+T cells was66.32±5.984%,and the difference was statistically significant(P<0.05)2.The expression rates of CD38~+HLA-DR~+double positive in CD8~+T cells with high-expression of PD-1 and low-expression of PD-1 were 5.80%and 2.65%,respectively,and the expression rates of CD38~+HLA-DR~+double positive in CD8~+T cells with high-expression of 2B4 and low-expression of 2B4 were 5.23%and 0.93%,respectively,with statistically significant differences(P<0.001).3.MCF of CD69 in CD8~+T cells with high-expression of PD-1 and low-expression of pd-1 were 154 and 115,and MCF of CD69 in CD8~+T cells with high-expression of2B4 and low-expression of 2B4 were 158 and 88,respectively,in the peripheral blood of the group with advanced colorectal cancer(P<0.001).4.MCF of CD27 in CD8~+T cells with high-expression of pd-1 and low-expression of pd-1 were 405 and 331,and MCF of CD27 in CD8~+T cells with high-expression of2B4 and low-expression of 2B4 were 188 and 853,respectively,in the peripheral blood of the group with advanced colorectal cancer(P<0.001).5.MCF of CD28 in CD8~+T cells with high-expression of PD-1 and low-expression of PD-1 were 162 and 140,and MCF of CD28 in CD8~+T cells with high-expression of2B4 and low-expression of 2B4 were 106 and 240,respectively,in the peripheral blood of the group with advanced colorectal cancer(P<0.001).Conclusion1.PD-1,2B4 is highly expressed on CD8~+T cells in peripheral blood of patients with advanced colorectal cancer,suggesting that peripheral CD8~+T cells in patients with advanced colorectal cancer are immunosuppressed,and there may be synergistic effects of PD-1 and 2B4.2.By analyzing the expression levels of CD27 and CD28 in CD8~+T cells expressing2B4 and PD-1 in peripheral blood of the advanced colorectal cancer group,it is suggested that CD8~+T cells with high expression of 2B4 May be the real depleted cells。3.PD-1/PD-L1 antagonists have achieved considerable efficacy in the diagnosis and treatment of different solid tumors,such as malignant melanoma,non-small cell lung cancer,renal cell carcinoa etc.The results of this study suggest that PD-1 and 2B4have certain predictive effects on the application of immunosuppressive agents in colorectal cancer,especially 2B4 may be a biomarker for colorectal cancer and a new target for immunotherapy. |
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