Expression Levels And Significance Of LAG-3,PD-1 And Related Cytokines In Peripheral Blood Of Patients With Myelodysplastic Syndrome

Research background:Myelodysplastic syndrome(MDS)is a hematological malignancy that is highly heterogeneous and at high risk of transformation to acute myeloid leukemia(AML).Disturbances in the immune microenvironment play a crucial role in the development of MDS.Immunotherapy has revolutionised the treatment of malignant tumours.Lymphocyte activation gene-3(LAG-3),a novel negatively regulated immune checkpoint,has been initially reported to be expressed at altered levels in MDS patients.However,it has been studied limitedly in the risk stratification of MDS patients,and its synergistic mechanism of action with programmed cell death protein-1(PD-1)has been rarely reported.This study is an attempt to build on the existing research to explore the role of LAG-3 in the development and transformation of MDS and other immune abnormalities,and to provide a new way of targeting immunotherapy for MDS.Objective:By detecting the expression of LAG-3 and PD-1 on CD4~+T cells,CD8~+T cells and regulatory T cell(Treg)in the peripheral blood of patients with primary MDS,and detecting the changes in the levels of interleukin(IL)-6,IL-8,IL-10,TNF-αand IFN-γ,we investigated the changes in the expression of LAG-3 and PD-1 in the peripheral blood of patients with primary MDS,the correlation between them and the relationship between them and cytokines.Methods:A total of 21 patients with primary diagnosis of MDS were collected from December 2021 to January 2023 at the Second Hospital of Lanzhou University,including 14 patients in the Higher Risk(HR)group,7 patients in the Lower Risk(LR)group,and 11 patients in the healthy control group.The expression levels of LAG-3and PD-1 on CD4~+T cells,CD8~+T cells and Tregs in the peripheral blood were detected by flow cytometry,and the expression levels of IL-6,IL-8,IL-10,TNF-αand IFN-γwere detected by cytometric bead array.Results:1.Compared with healthy controls,the expression of LAG-3 on Treg was elevated in the LR-MDS group(P<0.05);the expressions of PD-1 on CD4~+T cells,CD8~+T cells,and Treg were elevated(P<0.001).And the expressions of LAG-3on CD8~+T cells,and Treg were elevated in the HR-MDS group(P<0.05,P<0.001);the expressions of PD-1 on CD4~+T cells,CD8~+T cells,and Treg were elevated(P<0.001);the expression levels of LAG-3 and PD-1 on Treg were significantly higher in the HR-MDS group than in the LR-MDS group(P<0.05).2.Compared with healthy controls,the levels of IL-6,IL-8,IL-10 and TNF-αwere increased in the LR-MDS group(P<0.05);the levels of IL-10,TNF-αand IFN-γwere increased in the HR-MDS group(P<0.05),and the levels of IL-6 and IL-8 were significantly higher in the HR-MDS group(P<0.001);the levels of IL-6,IL-8,IL-10,TNF-α,and IFN-γlevels were significantly higher in the HR-MDS group than in the LR-MDS group.3.Correlation analysis showed that IL-6 was positively correlated with LAG-3expression on CD4~+T cells,TNF-αwas negatively correlated with LAG-3 expression on CD4~+T cells,IFN-γwas positively correlated with PD-1 expression on Treg;LAG-3~+CD8~+T was positively correlated with PD-1~+CD4~+T,PD-1~+CD8~+T and PD-1~+Treg expression,and PD-1~+CD4~+T was positively correlated with PD-1~+CD8~+T expression.Conclusion:1.LAG-3 and PD-1 expressions were increased and correlated with MDS stratification,and IL-6,IL-8,IL-10,TNF-α,and IFN-γlevels were significantly increased.It is suggested that LAG-3,PD-1,and related cytokines may be involved in disease development,progression and disturbance of the immune microenvironment in MDS patients.2.Targeting related molecules such as LAG-3、PD-1 may be a new way for the treatment of MDS.