Study On The Expression Of Peripheral Blood Lymphocyte Immune Checkpoints In Patients With Malignant Solid Tumors And The Effect Of Allogeneic Blood Transfusion On Their Levels

Objective1.To understand the proportion of peripheral blood lymphocyte subsets and the expression of immune checkpoint molecules PD-1,LAG-3,and TIM-3 on CD4+T,CD8+T,NK cells in patients with malignant solid tumors;2.To understand the expression of PD-1,LAG-3 and CD39 molecules on peripheral blood CD4+CD25+Foxp3+T(Treg)cells in patients with malignant solid tumors;3.To explore the correlation between different phenotypes of lymphocyte subsets and checkpoint molecules and clinicopathological parameters in patients with malignant solid tumors;4.To understand the effect of allogeneic blood transfusion on peripheral blood lymphocyte subsets and expression levels of PD-1,LAG-3,and TIM-3 molecules on CD4+T,CD8+T,NK cells in patients with malignant solid tumors;5.To understand the effect of allogeneic blood transfusion on the expression of CD39,PD-1 and LAG-3 molecules on peripheral blood CD4+CD25+Foxp3+T cells in patients with malignant solid tumors;6.To explore the correlation between the changes of different phenotypes of immune cell subsets in peripheral blood and clinical pathological parameters of patients with malignant solid tumors before and after blood transfusion;Method1.Flow cytometry(FCM)detection of peripheral blood lymphocyte subsets and immune checkpoints PD-1,LAG-3,TIM-3 expression in tumor patients and healthy controls;2.Flow cytometry to detect the expression of peripheral blood Treg cells and immune checkpoints PD-1,LAG-3,CD39 in tumor patients and healthy controls;3.Collect clinical data of patients and analyze the correlation between different phenotypes of cell subsets in peripheral blood of tumor patients and the molecular and clinical pathological parameters of immune checkpoints;4.Patient blood type identification,cross-matching,clinical transfusion;5.Flow cytometry(FCM)detection of peripheral blood lymphocyte subsets and their immune checkpoint molecules PD-1,LAG-3,TIM-3 before and after blood transfusion in tumor patients;6.Flow cytometry detection of peripheral blood Treg cells and their immune checkpoints PD-1,LAG-3,CD39 before and after blood transfusion in tumor patients;7.Collect clinical data of patients and analyze the correlation between different phenotype cell subpopulations and clinical pathological parameters in peripheral blood before and after blood transfusion in tumor patients.Result1.Patients with malignant solid tumors have an increased proportion of peripheral blood CD3-T,a decreased proportion of CD3+T cells,and a reduced proportion of NK cells,with little difference in CD4+T,CD8+T,and B cells;2.In peripheral blood of patients with malignant tumors,PD-1,LAG-3,TIM-3 negative checkpoint molecules are overexpressed on CD4+T,CD8+T,NK cells,and the proportion of "exhausted" CD4+T with double positive checkpoints is increased;3.The proportion of peripheral blood CD4+CD25+Foxp3+T cells increased in patients with malignant tumors,and the immune checkpoint molecules CD39,PD-1,LAG-3 on Treg cells were highly expressed;4.The high expression of PD-1 in tumor patients is negatively correlated with the proportion of CD4+T and NK cells,and positively correlated with the proportion of Treg cells.There was a positive correlation between the expression rates of PD-1,LAG-3,TIM-3,and CD39 molecules;5.The proportion of T lymphocyte subsets and B cells in peripheral blood of tumor patients after allogeneic blood transfusion did not change significantly,the proportion of NK cells decreased significantly,and the proportion of LAG-3+CD8+T cells and PD-1+CD8+T cells increased.High,double positive "exhausted" CD4+T cell subsets decreased;6.Allogeneic blood transfusion resulted in a significant increase in the proportion of peripheral blood Treg cells,PD-1+Treg cells,LAG-3+Treg cells,PD-1+LAG-3+Treg cells in tumor patients;7.Changes in peripheral blood lymphocyte subsets and their immune checkpoint molecules PD-1,LAG-3,TIM-3,and CD39 after blood transfusion are related to patient age,blood type,blood transfusion type,WBC change,and HB change.Conclusion1.The proportion of peripheral blood lymphocytes in patients with malignant solid tumors is unbalanced,and the degree of suppression of innate immune cells is higher than that of adaptive immune cells.PD-1,LAG-3,TIM-3 negative checkpoint molecules are highly expressed and the immunosuppression is severe.Peripheral blood lymphocytes of tumor patients are affected by multiple inhibitory pathways.For those who do not respond to single-agent immune checkpoint inhibitors,multi-target combined therapy may achieve better results.2.Peripheral blood Treg cells in tumor patients have enhanced immunosuppressive functions.Immunotherapy programs that target Treg cells to reduce their number and limit their functions will have an important role in the future.3.Highly expressed immune checkpoint molecules in tumor patients can not only regulate the proportion of lymphocyte subpopulations,but also have complex inter-regulatory relationships between the molecules.4.The proportion of peripheral blood lymphocytes in tumor patients is further imbalanced after blood transfusion.Blood transfusion treatment has a greater impact on the innate immune system of tumor patients.PD-1,LAG-3,TIM-3,and CD39 negative checkpoint molecules are highly expressed,which exacerbates immunosuppression.May cause a poor prognosis.5.Strengthen blood management of cancer patients,strictly grasp the indications for blood transfusion,and perform precise and differentiated blood transfusion.The transfusion of tumor patients needs to change from the previous "free experience blood transfusion concept" to "restrictive blood transfusion concept".