Expression And Clinical Significance Of TIM3,PD-1 And B7-H3 In Malignant Hematological Diseases

Background:Malignant hematologic diseases are a group of malignant clonal diseases originating from the hematopoietic system,mainly including leukemia,myelodysplastic syndrome,lymphoma,multiple myeloma,etc.Their etiology and pathogenesis are complex,with high malignancy,difficult treatment,short patient survival,poor prognosis,easy relapse and drug resistance formation being the challenges faced today,and negative costimulatory molecules such as TIM3,PD-1,B7-H3,etc.are Negative costimulatory molecules such as TIM3,PD-1,and B7-H3 are important in the development of disease,and they restrict,terminate,or attenuate T cell responses through negative regulatory signals,altering the immune surveillance status of tumor cells and promoting tumor development.This study focused on the expression of T lymphocyte subsets and the surface expression of TIM3,PD-1 and B7-H3 in primary cells from patients with haematological malignancies using flow cytometry to analyze its expression in relation to the pathogenesis,clinical manifestations and prognosis,to further explore their role in disease pathogenesis and prognosis,as well as to provide theoretical basis for targeted immunotherapy of hematologic malignancies.The purpose of this study is to provide a theoretical basis for targeted immunotherapy of hematologic malignancies.Methods:(1)Collection:a total of 88 patients diagnosed with hematologic malignancies from June 2022 to January 2023 attending the Department of Hematology,Affiliated Hospital of Southwest Medical University,including 54 primary patients,13 relapsed refractory patients,and 21 patients in complete remission,and anticoagulated bone marrow,peripheral blood,medical history data and clinical outcomes of patients were collected from the study population,and 22patients with iron deficiency anemia were also collected Peripheral blood was collected as control.(2)To measure the expression of the T-cell subsets and PD-1,TIM3 on the surface as well as of B7-H3 in primary cells of patients with haematological malignancies by Flow cytometry.(3)To detect cytokine levels before and after treatment and analysis of changes in the immune status of patients.(4)Analyze the expression levels of T cell subsets and their surface TIM3,PD-1 and B7-H3 of primitive cells,and analyze the relationship between their expression and clinical features,disease status and prognosis.(5)Data were collected in Excel 2007 and analysed with SPSS 22.0.Data were matched to a normal distribution by the t-test,otherwise the Wilcoxon rank sum test was used;the X~2test was used to compare data between the two groups,with P<0.05 being considered a statistically significant difference.Result:(1)The total number of patients with hematologic malignancies was 88,50(57%)males and 38(43%)females,with a median age of 56(14-92)years,There were 53 patients(60%)with myeloid tumours,including 42 cases of acute myeloid leukaemia(AML),9 cases of myelodysplastic syndrome(MDS),and 2 cases of primary myelofibrosis(PMF);35 patients(40%)with lymphocytic tumors,including 18 cases of lymphoma,13 cases of acute lymphoblastic leukaemia(ALL),3 cases of multiple myeloma(MM),and 1 case of chronic lymphocytic leukaemia(CLL).(2)The expression of PD-1 and TIM3 in T cells and B7H3 in primary cells of patients was measured by flow cytometry.The results showed that the expression levels of CD4+PD1+T,CD4+TIM3+T,CD8+TIM3+T and CD8+PD1+T was higher in the newly diagnosed,complete remission and non-remission groups than in healthy controls,TIM3 and PD1 expression levels were higher in non-remission and newly diagnosed group than in complete remission(P<0.05);PD-1 and TIM3 expression levels were higher in the T-cell subgroups of the myeloid and lymphoid tumour groups than in the healthy controls.The median B7H3 expression in primary cells was 5.22%,with a cut-off value of 5.22%,higher than this being the high-expression group and the rest being the low-expression group.There were 8 cases(80%)with high and 2 cases(20%)with low expression in the primary group,3 cases(21%)with high and 11 cases(79%)with low expression in the complete remission group and 2 cases(67%)with high and 1 case(33%)with low expression in the non-remission group.The frequency of high B7H3 expression in the newly diagnosed group and non-remission group was higher than in the complete remission group,and the difference was statistically significant(P<0.05).(3)Flow cytometry was used to detect T cell subsets and their ratios in the bone marrow of patients with hematological malignancies.The CD4+T lymphocyte levels in the initial diagnosis group,complete remission group,and non remission group were lower than those in the control group(P<0.05);The levels of CD8+T lymphocytes in the initial diagnosis group(P<0.05),non remission group(P<0.05),and remission group(P>0.05)were all higher than those in the control group.The CD4+T/CD8+T ratios in the initial diagnosis group,non remission group,and complete remission group were all lower than those in the control group,and the differences were statistically significant(P<0.05)(4)Analysis of changes in patients’immune status by measuring cytokines in their peripheral blood before and after treatment,the levels ofγ-Interferon and IL-2 increased,and the levels of IL-6,IL-10,IL-4andα-tumor necrosis factor were significantly lower(P<0.05),suggesting that the th1/th2 subpopulation was dominated by th1 response,and the immune function was partially restored and anti-tumor effect was enhanced.Conclusion:(1)PD-1,TIM3 and B7H3 are co-expressed in hematologic malignancies,there is no significant difference between diseases,the expression levels correlate with disease status and prognosis,there is positive correlation between PD-1and TIM3 expression levels and poor prognosis,there is synergistic expression between various immunosuppressive molecules,high expression represents immunosuppressive status,suggesting involvement in the development and recurrence of malignant hematologic diseases.Involved in immune escape of tumor cells and promote the development of disease.(2)An increase in the CD4+T/CD8+T ratio of T lymphocyte subsets in patients with haematological malignancies indicates restored immune function and enhanced anti-tumour activity,while a decrease in this ratio indicates reduced immune function.In treated patients,the th1/th2 subpopulation is dominated by the th1 response,suggesting partial recovery of immune function,T lymphocyte and cytokine levels is key to maintaining a balanced cellular immune response and effective anti-tumour activity.