Multiparament Flow Cytometry To Detect The Minimal Residual Disease In People With Blineage Acute Lymphoblastic Leukemia

Objective:We set up a sensitive,volant,economic method to detect the minimal residual disease in people with B-lineage acute lymphoblastic leukemia by multiparament flow cytometry with two panels of 4-color monoclonal antibodies.And we detected a few bone marrow samples.The patients who were detected should be followed up.The relationship between the prognosis and the results are analyzed.At the same time,we can discuss the value of the method which is used to detecte the minimal residual disease in people with B lineage acute lymphoblastic leukemia.Method:We detected 25bone marrow samples from 22 patients with Blineage acute lymphoblastic leukemia by multiparament flow cytometry with two panelsof 4-color monoclonal antibodies. (CD45CD20CD10CD19;CD45CD20 CD34CD19).All of the bone marrow samples were took from the patients who were in hospital or in clinic in the departments of hematology or peadiatrics of the second hospital of Hebei medical university.The patients were took a definite diagnosis by morphology an immunology,and all of them were treated by the chemicals acluding vincristine,cyclophosphamide, daunorubicin, L- asparaginase and decaesadril.Cyclophosphamide were not used to treat the children.There are 11 men and 11women were detected.They are from 2 to 60 years old,and the average is 20.The bone marrow samples were took from the patients after the patients were treated and detected by multiparament flow cytometry with the two panels of 4-color monoclonal antibodies.The 25 bone marrow samples were divided into three groups according to the results.If there are some leukemia-associated aberrant immunophenotype cell can be detected—the results of minimal residual disease are positive—we call them groupA,otherwise we call it group B.If there are some primitive lymphocyte that don't express aberrant immunophenotype we call them group C.The bone marrow samples of the group A and B were detected by morphological method.The percentage of the primitive lymphocyte be recorded.And then analyze the results of the bone marrow smear to understand if there are statistical significance between the two groups.The patients were followed up,and the dates on which the patients relapse were recorded.Results:1 the results of the detectiong:25 bone marrow samples from 22 patients were detected by multiparament flow cytometry with the two panels of 4-color monoclonal antibodies(CD45CD20CD10CD19;CD45CD20CD34CD19)we found that 12 samples can be found abnormal cells.MRD is 0.009%,0.026 %,0.030%,0.060%,0.063%,0.068%,0.080%,0.096%,0.137%,0.230%,0.230%,0.270% respectively,and 7 samples can be found primitive lymphocyte which express CD34 and other primitive cell's immunophenotypes but don't express abnormal immunophenotypes.No minimal residual disease or primitive lymphocyte was found in the other 6 samples.MRD is negative.2 6 samples express CD45dim/CD20+/CD10+/CD34+/CD19+.It is the most probable value.3 compare with the morphology:The percentage of the primitive and immature lymphocyte is 2%,3%,3%,4%,1%,3%,2%,3%,4%,3%,7%,5% respectively in the bone marrow smear which belongs to groupA, x±S=3.33%±0.449%,and the percentage is 0%,2%,1%,2%,3%,1% respectively in the bone marrow smear which belongs to group B. x±S=1.50%±0.428%. The data of the group A and B was analyzed through soft ware SPSS.The percentage of the primitive and immature lymphocyte in group A is obviously higher than the one in group B(P<0.05).4 the relationgship between MRD and the relapse:There are 2 patients relapsed in group A,no one have relapsed up to now in group B,and 2 patients relapsed in group C.5 the camparision of the MRD of the patients who have relapse and the ones who are in remission:a patient who have in remission for 4 years and 2 months relapsed in the detection. The percentage of the primitive and immature lymphocyte is 7% and the MRD is 0.23%.Conclusions:1 The method of detection of MRD in people with lineage Bacute lymphoblastic leukemia by FCM with two panels of 4-color monoclonal antibodies(CD45CD10CD20CD19;CD45CD20CD34CD19)and the method of counting the percentage of the primitive and immature lymphocyte are consistent.It can be used to detect the MRD in B-ALL.There are a lot of advantages in using FCM to detect MRD such as fastness,sensitiveness,convenience,economy and so on.It can predict the relapse before the morphologocal relapse.So the patients can be cured intensively to avoid relapse.According to the results,we can predict the prognosis exactly,while the method of the morphology cannot.2 But as to the patients who can not be detected abnormal cells or primitive cells,we should ask them to recheck after a few days ,add other antibodies or use 6 or 8-color monoclonal antibodies in order to avoide the absences of the false-negative.3 We should detected MRD statedly in order to discover the relapse in time.We can adopt a measure to prevent the relapse and extend the disease free survival.