Comparative Study On The Immunophenotype And Proteomics Between Acute Leukemia Subtypes

Acute leukemia(AL) is hematopoietic malignant tumor with high heterogeneous whicl" caused by the both effection of hereditary and environmental factors.There is much difference at the molecular level in the process of leukemogenesis which might determine the different prognosis and response to treatment.However,the current classification system cannot reflect the molecular characteristics,and cannot give instructions to make specific treatment according to different types of AL.The current basis method for the classification of AL is French-American-British(FAB) classification system which based on cytomorphology and cytochemistry.It has low correct rate and cannot reflect the essential features of AL.Although flow cytometric immunophenotyping has remained an indispensable tool for the diagnosis,classification,staging,and monitoring of hematologic neoplasms in the last 10 years,the phenotypic aberrancies still have not been recognized completely for the availability of limited range of antibodies and fluorochromes.It is crucial to establish a new classification system that can reflect the leukemogenesis and guide treatment and prognosis.This study aimed to investigate the molecular characteristics of different types of AL from three aspects such as the protein expressed on cell membrane, cell plasma and cytokines in serum by using flow cytometry(FCM) and proteomic techniques.There-color multiparametric flow cytometry with CD45/SSC gating and a set of monoclone-antibodies were used to analyze the immunophenotypic characteristics of bone marrow cells from acute leukemia patients with different subtypes including acute myeloid leukemia(AML) subtypes M1,M2,M3,M4,M5 and acute lymphoid leukemia(ALL) classified by FAB.The results showed that flow cytometry is a reliable technique in the diagnosis,differential diagnosis and classification of acute lymphocytic leukemia,but still lacked effective marker on the classification of AML.To study the expression of leukemic stem cell(LSC) associated membrane antigens on AL cells(M1,M2,M3,M4 and ALL),antigens CD96,CD90,CD123 and CD71were measured by Flow cytometry.The results showed for the first time that the positive rate of CD96 expression in AML was significantly higher than in ALL,and M3 had the highest positive rate among all AML subtypes.So,CD96 may become a marker for AML and its high expression might contribute to the overgrowth of AML cells.The up-regulation of CD123 has been identified in all AL and has no difference between the subtypes.CD71 also may become a marker to tell AML from ALL for its higher expression in AML.After the 2-DE patterns with high resolution were obtained,the Peptide Mass Fingerprints corresponding to 38 selected proteins with high resolution and up-regulated expression levels in the groups were obtained by MALDI-TOF-TOF mass spectrometry analysis.The results showed that PFN1 had lower expression but MPO and PRDX3 were higher in granulocytic lineage of AL compared with monocytic lineage of AL and ALL. These proteins might be markers to distinguish granulocytic lineage of AL.HNRNP A1,40S ribosomal protein S12 and ARHGDIB were highly expressed in AML and ALL respectively, and they might contribute to the classification of AML and ALL.The level ofcytokines IFN,IL-6,and IL-10 in serum of different subtypes of AL was exactly evaluated by Cytometric Bead Array(CBA) method.The results showed that the levels of lFN,IL-6,and IL-10 in serum of AL were significantly higher than those in controls.Compared with AML,the levels of IFN and IL-10 in acute lymphoid leukemia (ALL) group were significantly higher(P＜0.05).The IL-6 level in M1 was the highest one among the AML subtypes.This study indicated that IFN,IL-6,and IL-10 might play important role in AL mechenism and the IL-6 level might be related with the AML subtypes.