Investigation Into The Determination Of Surface Antigens In Multiple Myeloma Cells With Flow Cytometry

BackgroundMultiple myeloma(MM)is a kind of tumor characteristics itself with plasma cells dyscrasia in bone marrow secreting monoclonal immunoglobulin or light chain,as well as normal immunoglobulin decrease and widespread osteolysis lesion and(or)bone rarefaction.It is familiar in middle-aged and aged people, along with the age growth,its disease incidence also goes up obviously.MM once be treated as a kind of seldom disease.In recent years,along with the aging of population and people known more about this disease and the diagnosis level improves,the MM incidence shows observably increment.There is no MM epidemiology data in the our country currently,but along with the development of aging in our country,the MM incidence tend to increase high trend year by year,it will be certainly threat the health diathesis of our country's population. Bone marrow cellular morphology detection is indeed one of the main final diagnosis evidences to MM,but myeloma cells usually have focal hyperplasia,a part of borderline cases hard to achieve the proportion of diagnostic criteria,and some myeloma cells' morphous are quite similar with the normal plasma cells', it is difficult to discriminate them.In recent years along with the utendus of monoclonal antibody(McAb)and the fast development of flow cytometry (FCM),FCM-Imm method has already become the significant evidence to diagnosis and Immunophenotype leukemias,it can reflect the biology characteristic of leukemia cells in a quick,accurate and objective way,guide the therapy and hint the prognosis.Immunophenotyping of MM with FCM yet is still place in a research and investigates stage.There are less domestic reports about the researches in Immunophenotyping of MM,most of these researches used CD45/SSC-the classics gating method of Immunophenotyping leukemias, this gating method is liable to be influenced by the variability of CD45 on plasma cells and nucleated red blood cells.Therefore,it is necessary to find out a more reliable gating way in FCM analysis.ObjectiveThe objective of this test is to determining surface antigen of multiple myeloma cells with CD38/SSC gating method,explore a better method to determine MM,provide the clinic a correct evidence that can help earlier diagnosis,guide therapy and judge prognostic.MethodsResearch 37 cases of MM which have been final diagnosed by bone marrow cellular morphology detection,clinical situation,imageology and serology standards,with 20 reactive plasmacytosis(RP)patients to be control group. Extract bone marrow of patients,determine surface antigens of multiple myeloma cells with FCM,main McAbs:CD19,CD56,CD13,CD138,HLA-DR, CD33,CD64,CD34,CD 8,CD3,CD5,CD10,CD1a,CD14,CD40,CD4,CD25,CD20, CD7,CD45and CD49e.All of the numerical data is handled with SPSS13.0 statistics software.Results1.With CD38/SSC gating method,all the samples can be seen a cluster of CD38 positive cells in the scatterplot,the proportion range from 0.5%to 80%.2.The most phenotype of MM is CD38~+CD19~-CD56~+(89.2%),the expression level of CD56 in initial treatment and non-remission group is obviously higher than that in remission group,while obviously descend in extramedullary encroachment group.3.The expression of CD40 and CD138 in myeloma cells is obviously higher than that in normal plasma cells of control group.4.The ratio of immature plasma cell/myeloma cells and the CD45~+CD49e~+ cells proportion present a negative relation,we can judge approximately the cell differentiation inside marrow according to the comparison of the CD45~+CD49e~+cells.Conclusions1,Use FCM to immunophenotype MM with CD38/SSC gating method,base on the heterogeneity of cell surface antigens,we can discriminate myeloma cells from plasma cells,provide evidence for targeted therapy and judge prognostic.2,With the phenotype of CD38~+CD19~-CD56~+ to be the positive detection of MM with FCM method,have a high sensitivity,specificity,positive predictive and negative predictive,it needs a short examine time,and have a strong clinical practicability,is hoped to be a new MM diagnose test and early screening test,be generally used in big hospital which has a flow cytometry.