Evaluation Of The Clinical Significance Of Immunophenotyping In Myelodysplastic Syndrome And Aplastic Anemia

OBJECTIVE To detect the immunophenotypic features of blasts in the patients with myelodysplastic syndromes(MDS)and aplastic anemia(AA),investigate the clinical usage of immunophenotyping in the pathogenesis,diagnosis,classification and differential diagnosis of MDS and AA.METHODS Take 2 ml bone marrow from 23 patients with MDS,14 patients with AA and 9 healthy controls by routine bone marrow puncture.A panel of monoclonal antibodies(including CD25/CD34/CD45,HLA-DR/CD3/CD45,CD14/CD13/CD45,CD22/CD33/CD45,CD15/CD7/CD45,CD5/CD19/CD45,CD10/CD20/CD45 and IgG1 cntrols),direct immunofluorescence and flow cytometry using CD45/SSC gating strategies were used for the immunophenotype in the bone marrow nucleated cells.Medullary system differentiation index(DI) was calculated.Correlation between CD34 and HLA-DR was analyzed.RESULTS 1.Compared to the control group,the positivity of hemopoietic stem/progenitor cell surface markers CD34,HLA-DR,early medullary system surface markers CD13,CD33,monocyte system surface marker CD14,T lymphocyte cell surface marker CD7 were significantly increased in MDS patients (P＜0.05);the positiv ity of later medullary system surface maker CD15,B lymphocyte cell surface markers CD19,CD20 were significantly decreased in MDS patients(P＜0.05).There were no significant difference(P＞0.05)in the expression of CD3,CD5,CD25,CD10,CD22.DI were significantly increased in MDS patients(P＜0.05).2.As RA progressing to RAEB,the positivity of CD34,HLA-DR,CD13,CD33,CD14,CD7 were significantly increased(P＜0.05),the positivity of CD15,CD3,CD19,CD20 were significantly decreased(P＜0.05);There were no significant difference in the expression of CD5,CD25,CD10,CD22(P＞0.05). DI were significantly increased(P＜0.05).3.The positivity of CD34,CD13,CD33,CD15 in AA patients were significantly lower than control group(P＜0.05),CD3,CD7,CD25,CD22 higher than control group(P＜0.05);There were no significant difference in the expression of HLA-DR,CD14,CD5,CD10,CD19,CD20(P＞0.05).4.Compared to AA group,the positivity of CD34,HLA-DR,CD13,CD33,CD14 were significantly increased in MDS(P＜0.05),the positivity of CD3,CD5,CD7,CD15,CD19,CD20,CD22,CD25 were significantly decreased(P＜0.05). There were no significant differencein the expression of CD10(P＞0.05).5.Correlation between CD34 and HLA-DR was analyzed.Correlation coefficients were 0.909,0.834,0.777 and 0.755 in RA,RAEB,AA and healthy control group,all P＜0.01.There was significant difference between CD34 and HLA-DR.CONCLUSIONS 1.Compared to the control group,the positivity of haemopoietic stem/progenitor cell surface markers CD34,HLA-DR,early medullary system surface markers CD13,CD33,monocyte system surface marker CD14,T lymphocyte cell surface marker CD7 were significantly increased in MDS patients.It indicates that abnomal clone of MDS derives from haemopoietic stem cells or hemopoietic progenitor cells,most belonging to medullary system.As RA progressing to RAEB,malignant clone becomes dominant and inhibits normal haematopoiesis.Or as disease progressing, differentiation ability of malignant clone hematopoietic cells becomes lower,so that haemopoietic stem/progenitor cell surface maker is higher in RAEB than in RA,and later medullary system surface maker is lower in RAEB.2.The positivity of CD34,CD13,CD33,CD15 in AA patients were significantly lower than control group,which indicates the pathogenesis of AA is the decrease of hematopoietic cells in stead of differentiatiate disturbance. However,medullary system differentiation index of MDS being significantly higher indicates differentiatiate disturbance of haemopoietic stem/progenitor cell, so MDS is malignant clonal haemopoietic stem/progenitor cell disease.3.DI was used to invest MDS and AA in this study,and was confirmed significantly higher in MDS.This result is not only helpful in reveal the pathogenesis difference,but also provides convenient,shortcut and reliable method for diagnosis and differential diagnosis.4.The positivity of monocyte system surface maker CD14 was significantly higher in MDS patients than in control group.It indicates that monocyte system is abnormal and the expression of CD14 is disorder.5.The positivity of T lymphocyte cell surface markers CD3,CD5,CD25 was significantly higher in AA patients than in control group.It indicates that there is T lymphocyte cell hyperfunction in AA.6.The positivity of B lymphocyte cell surface markers CD19,CD20 was significantly lower in MDS patients than in AA patients and control group.It indicates that later B lymphocyte cells are decreased.It may be associate with amplification of malignant clone supressing the increase of B lymphocyte cell,or the excess apoptosis of B lymphocyte cell,or abonormal haematogenesis of B lymphocyte cell itself in MDS.