| Objective:The purpose of this study was to evaluate the changes of monocyte immunophenotype in patients with myelodysplastic syndrome by flow cytometry,and to determine whether the expression of monocyte surface antigen has clinical significance in differential diagnosis of low-risk MDS and other bone marrow failure diseases,to further explore the correlation between the changes of monocyte immunophenotype and IPSS-R and prognosis in MDS patients,to explore whether the detection of monocyte immunophenotype can provide a reference for the differential diagnosis and prognosis evaluation of MDS.Methods:This study included 98 newly diagnosed MDS patients from November 2017 to November 2019 in the Department of Hematology,Tianjin Medical University General Hospital,including 45 patients with lower-risk MDS(LR MDS)and 53 patients with higher-risk MDS(HR MDS),according to WHO(2016)revised classification of MDS: 9 cases of MDS with single lineage dysplasia(MDS-SLD),9cases of MDS with multilineage dysplasia(MDS-MLD),30 cases of MDS with ring sideroblasts(MDS-RS),3 cases of MDS with isolated del(5q),47 cases of MDS with excess blasts type(MDS-EB),including 21 cases of MDS-EB-1 and 26 cases of MDS-EB-2.There were 21 CCUS patients,24 AA patients,and 38 normal controls.FCM was used to detect the expression of CD33,CD10,CD7,CD34,CD64,CD14,CD300 e,CD13 and CD11 b on the surface of bone marrow monocytes of patients with different bone marrow failure diseases and normal controls.According to the normal bone marrow monocytes antigen differentiation rules,CD64-CD14-and CD64-CD300e-,CD64+CD14-and CD64+CD300e-,CD64+CD14+ and CD64+CD300e+ antigen combinations were selected,and the expression of antigen combinations on the surface of bone marrow monocytes in each group was analyzed.According to the IPSS-R,patients with MDS were classified into different risk grades,and FCM was used to detect the expression of CD33,CD10,CD7,CD34,CD64,CD14,CD300 e,CD13 and CD11 b.the expression of antigen combinations on the surface of bone marrow monocytes in different risk groups of MDS and normal controls were analyzed,to explore the correlation between the changes of monocyte immunophenotype and IPSS-R and prognosis in MDS patients.Results:1.(1)Expression of antigens on the surface of bone marrow monocytes in bone marrow failure diseases: The expression of CD34 on the surface of monocytes in lower-risk MDS was significantly higher than that in AA and normal controls(p=0.022;p=0.025).The expression of CD300 e on the surface of monocytes in lower-risk MDS was significantly lower than that in AA and normal controls(p=0.003;p=0.001).The expression of CD300 e on the surface of monocytes in CCUS was significantly lower than that in AA and normal controls(p=0.049;p=0.036).(2)Expression of antigen combinations on the surface of monocytes in bone marrow failure diseases: The proportion of CD64+CD300e-cells(48.08%±17.89%)in monocytes in lower-risk MDS was significantly higher than that in AA and normal controls(p=0.022;p=0.007).The proportion of CD64+CD300ecells(48.94%±16.64%)in monocytes in CCUS was significantly higher than that in AA and normal controls(p=0.047;p=0.027).The proportion of CD64+CD300e+cells(34.11%±20.45%)in monocytes in lower-risk MDS was significantly lower than that in AA and normal controls(p=0.004;p=0.001).The proportion of CD64+CD300e+ cells(35.50%±15.10%)in monocytes in CCUS was significantly lower than that in AA and normal controls(p=0.046;p=0.033).There was no significant difference in the expression of surface antigens of monocytes in the other groups.2.The correlation between monocytes immunophenotype and clinical parameters of MDS patients:(1)the expression of CD14 and CD300 e on the surface of monocytes in MDS with primary hemocytopenia was significantly higher than that in MDS with multiple hemocytopenia(p=0.020,p=0.000).The expression of CD34 on the surface of monocytes in MDS with primary hemocytopenia was significantly lower than that in MDS with multiple hemocytopenia(p=0.044).(2)The proportion of CD64+CD14-(16.69%±13.47%),CD64+CD300e-(43.11%±14.59%)cells inmonocytes in MDS with primary hemocytopenia was significantly lower than that in MDS with multiple hemocytopenia(p=0.009;p=0.000).The proportion of CD64+CD14+(64.69% ± 20.36%),CD64+CD300e+(38.68% ± 19.72%)cells in monocytes in MDS with primary hemocytopenia was significantly higher than that in MDS with multiple hemocytopenia(p=0.027;p=0.000).(3)The expression of CD14 and CD300e on the surface of bone marrow monocytes in patients with MDS was negatively correlated with the percentage of primitive immature cells(r=-0.204,p=0.043;r=-0.230,p=0.035).(4)The proportion of CD64+CD14-,CD64+CD300ecells in monocytes in MDS was positively correlated with the percentage of primary immature cells(r=0.269,p=0.007;r=0.215,p=0.049),and the proportion of CD64+CD300e+ cells in monocytes in MDS was negatively correlated with the percentage of primary immature cells(r=-0.243,p=0.026).3.(1)Expression of antigens on the surface of monocytes in lower-risk and higher-risk MDS: The expression of CD14 on the surface of monocytes in higher-risk MDS was significantly lower than in lower-risk MDS and normal controls(p<0.05).The expression of CD300 e on the surface of monocytes in lower-risk and higher-risk MDS was significantly lower than that in normal controls(p=0.001;p=0.000),and the expression of CD300 e on the surface of monocytes in lower-risk MDS was significantly higher than that in higher-risk MDS(p=0.001).The expression of CD34 on the surface of monocytes in higher-risk MDS was significantly higher than in lower-risk MDS and normal controls(p=0.002;p=0.000).(2)Expression of antigen combinations on the surface of monocytes in lower-risk and higher-risk MDS: the proportion of CD64+CD14-cells(26.93%±18.44%)in monocytes in higher-risk MDS was significantly higher than that in lower-risk MDS and normal controls(p=0.012;p=0.038),the proportion of CD64+CD300e-cells(60.26%±19.33%)in monocytes in higher-risk MDS was significantly higher than that in lower-risk MDS and normal controls(p=0.003;p=0.000),the proportion of CD64+CD300e-cells(48.08%±17.89%)in monocytes in lower-risk MDS was significantly higher than that in normal controls(p=0.013).the proportion of CD64+CD14+ cells(53.08%±24.76%)in monocytes in higher-risk MDS was significantly lower than that in lower-risk MDS and normalcontrols(p=0.033;p=0.008),the proportion of CD64+CD300e+cells(21.17%±16.22%)in monocytes in higher-risk MDS was significantly lower than that in lower-risk MDS and normal controls(p=0.001;p=0.000),the proportion of CD64+CD300e+cells(34.11%±20.45%)in monocytes in lower-risk MDS was significantly lower than that in normal controls(p=0.001).(3)As the IPSS-R scores go from low to high,the expression of CD34 on the surface of monocytes in MDS was positively correlated with the IPSS-R scores(r=0.272,p=0.007),and the expression of CD14 and CD300 e on the surface of monocytes in MDS was negatively correlated with the IPSS-R scores(r=-0.258,p=0.010;r=-0.366,p<0.001).(4)As the IPSS-R scores go from low to high,the proportion of CD64+CD14-,CD64+CD300e-cells in monocytes in MDS was positively correlated with IPSS-R scores(r=0.292,p=0.004;r=0.356,p<0.001),and the proportion of CD64+CD14+,CD64+CD300e+ cells in monocytes in MDS was negatively correlated with IPSS-R scores(r=-0.235,p=0.020;r=-0.379,p<0.001).(5)The number of abnormal expression of antigen and antigen combination on the surface of monocytes in LR MDS was significantly lower than that in HR MDS(p=0.010).As the IPSS-R scores go from low to high,the number of abnormal expression of surface antigen and antigen combination on monocytes in MDS patients was positively correlated with the IPSS-R scores(r=0.305,p=0.002).4.Kaplan Meier survival analysis showed that MDS patients with low CD300 e expression on the surface of bone marrow monocytes had low progression free survival and AML free survival(p=0.004;p=0.049);The higher-risk MDS patients with low CD300 e expression on the surface of bone marrow monocytes had low progression free survival(p=0.024).Conclusion:1.The expression of antigen on the surface of monocytes in low-risk MDS monocytes was significantly different from that in AA and normal control groups,suggesting that the monocytes differentiation and maturation of MDS patients was impaired.There was no significant difference in the expression of antigen on the surface of monocytes in low-risk MDS and CCUS,suggesting that CCUS patients already had abnormal blood cell development.2.There is abnormal accumulation of differentiation in the progression of MDSfrom low risk to high risk.3.It is suggested that CD300 e on the surface of monocytes is a good prognostic marker in MDS patients. |
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