Detection of Minimal Residual Disease in Childhood B-Acute Lymphoblastic Leukemia by 4-Color Flow Cytometry

Monitoring of minimal residual disease (MRD) is currently considered the most powerful predictor of outcome in acute lymphoblastic leukemia (ALL). Achievement of a negative MRD state assessed by multicolor flowcytometry (MFC) is an important predictor of disease-free survival (DFS) and overall survival (OS) in ALL patients. We sought to determine whether panels of antibodies combination are more suitable for detection of MRD in Childhood ALL. Eighty-four (84) patients with ALL (B-lineage subtype) were enrolled in this study. Normal template for B cell precursors was established in 15 control participants using 4-four panels of monoclonal Antibodies (Mo Abs),{CD22, CD45, CD58 and CD97 in combination with CD10, CD19, CD34}. At diagnosis, CD22 exhibited the lowest incidence of expression in only 50% of all patients, while CD45, CD58, and CD97 were expressed in 80.9, 59.5 and 92.8%, respectively. Analysis of MRD was performed for each Mo Abs combination at day 0 and day 14 post-induction of chemotherapy by 4-color (FCM). The incidence of MRD was 61.9, 70.6, 60.0 and 55.1% for CD22, CD45, CD58 and CD97, respectively. In B-ALL patients, (CD10/CD19/CD34/CD45) + (CD10/CD19/CD34/CD97) represented the highest incidence of expression of leukemic cells markers with a significant correlation with blasts count, suggesting that these are more specific for MRD detection. Also FCM is relatively cost effective for detection of MRD in ALL patients and its applicability in routine leukemia lab is valuable. MRD evaluation at the end of the induction therapy (i.e. day 35 or 42 according to the different schedules) is advised. Also, Ig/T cell receptor gene rearrangements and gene fusions analyzed by polymerase chain reaction (PCR) are preferred.