Correlation Between Regulatory T Cell,Cytokine And CTLA-4 Levels And Efficacy In Diffuse Large B-cell Lymphoma

OBjective(s):Diffuse large B-cell lymphoma(DLBCL)is the most common aggressive lymphoma,accounting for 30-40%of non-Hodgkin lymphoma in adults.At present,the response rate of CD20 mab combined with chemotherapy is greatly increased,but some people still have relapse progression.To further improve the efficacy,new prognostic indicators need to be added.In recent years,the role of tumor microenvironment in the development and progression of diffuse large B-cell lymphoma has been further studied,but the mechanism of regulatory T cells and CTLA-4 on CTL surface still needs to be further clarified.The purpose of this study was to analyze the relationship between the expression of CTLA4,Tregs,lymphocyte subsets and cytokines on the surface of CTL in newly diagnosed diffuse large B-cell lymphoma and after 4 cycles of chemotherapy and the efficacy,and to find new indicators related to the efficacy evaluation to better guide the choice of treatment plan for patients,so that DLBCL patients can get better survival benefits.Methods:According to the inclusion and exclusion criteria,clinical data of 76 patients with initial diffuse large B-cell lymphoma admitted to the First People’s Hospital of Yunnan province from January 2021 to January 2023 were collected.According to IPI score,the patients were divided into low-risk group,low-medium-risk group,medium-high risk group and high-risk group.Peripheral blood of DLBCL patients was collected before chemotherapy and after 4 cycles of chemotherapy,and CTLA-4 level on peripheral blood CTL surface was detected by flow cytometry.SPSS 25.0 software was used for statistical analysis.Count data is represented by[n(%)].Shapiro-Wilk test was used to test the normality of measurement data,and[M(P25,P75)]was used to represent non-normality data.Mann-Whitney U test or Kruskal-Wails H test were used to compare and analyze the differences of each index between two or three groups.Spearman correlation analysis was used to analyze the correlation among indicators.There are differences in said(P<0.05).Regulatory T cells,lymphocyte subsets,cytokines and CTLA-4 levels of different risk stratified groups were analyzed.The difference of each index after 4 cycles of regular treatment;After four cycles of treatment,the difference of each index between different therapeutic effect groups;The difference of indexes before and after treatment among different therapeutic effect groups;The correlation between prognostic indicators and clinical features was analyzed.Results:1.The number of regulatory T cells in low-risk group was significantly higher than that in high-risk group(P<0.05).The number of CD4~+T cells in high-risk group was significantly lower than that in low-risk group(P<0.05)and low-medium-risk group(P<0.05).The number of CD19~+cells in low-risk group was significantly higher than that in high-risk group(P<0.05).IL-6 in high-risk group was significantly higher than that in low-risk group(P<0.05)and low-medium-risk group(P<0.05).IL-8 in low-risk group was significantly lower than that in high-risk group(P<0.05).There was no difference in other indexes among groups(P>0.05).2.After 4 cycles of regular chemotherapy,the number of CD4~+T cells,B lymphocytes,NK cells,IL-6,IL-8,IL-10,IL-17F and IL-22 were all decreased compared with the initial diagnosis(P<0.05).Regulatory T cells,CD8~+T cells,IL-4,IL-5,IL-12P70,IL-1β,IL-2,IFN-γ,TNF-α,TNF-β,IL-17A and CTLA-4 were not significantly different before and after treatment(P>0.05).3.In different therapeutic groups after 4 cycles of treatment,CD8~+T cells in CR group were significantly higher than those in PR group(P<0.05)and poor therapeutic effect group(P<0.05),and there were no significant differences in other indexes among the three groups (P>0.05).In the complete response group,CD4~+T cells,B lym Phocytes,IL-6,and IL-8were decreased after treatment compared with those at initial diagnosis(P<0.05).Regulatory T cells,CD8~+T cells,NK cells,IL-4,IL-5,IL-10,IL-12P70,IL-1β,IL-2,IFN-γ,TNF-α,TNF-β,IL-17A,IL-17F,IL-22 and CTLA-4 were not significantly different before and after treatment(P>0.05).In the partial remission group,NK cells,B lymphocytes,IL-6,IL-8,IL-10,and IL-22 were decreased after treatment compared with those at initial diagnosis(P<0.05).Regulatory T cells,CD8~+T cells,CD4~+T cells,IL-4,IL-5,IL-12P70,IL-1β,IL-2,IFN-γ,TNF-α,TNF-β,IL-17A,IL-17F and CTLA-4 were not significantly different before and after treatment(P>0.05).In the poor treatment group,CD4~+T cells,B lymphocytes,NK cells,IFN-γ,IL-17A decreased after treatment compared with the initial diagnosis(P<0.05).Regulatory T cells,CD8~+T cells,NK cells,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12P70,IL-1β,IL-2,TNF-α,TNF-β,IL-17F,IL-22 and CTLA-4 were not significantly different before and after treatment(P>0.05).4.Regulatory T cells were positively correlated with CD4~+T cells,CD8~+T cells,B lymphocytes and NK cells in newly diagnosed diffuse large B-cell lymphoma patients.Regulatory T cells were negatively correlated with IL-6,IL-8 and IL-10.5.LDH and ferritin were positively correlated with IL-6,IL-8 and IL-10 at initial diagnosis.Conclusion(s):1.High levels of IL-6,IL-8,and IL-10 in newly diagnosed DLBCL patients suggest high tumor load.2.Persistently high levels of IL-6,IL-8,and IL-10 may be predictors of poor outcomes in DLBCL patients.3.The low level of peripheral blood regulatory T cells in newly diagnosed DLBCL patients may be a risk factor for poor prognosis of DLBCL patients.4.CTLA-4 on CTL surface in peripheral blood of newly diagnosed DLBCL patients showed no significant difference before and after treatment,so it could not be used as a factor in the evaluation of efficacy.