Studying Immunophenotypic Modulation at Different Time Points of Chemotherapy : A Walk through the Disease Course in 121 Pediatric B Lymphoblastic Leukemia Cases; Providing Implications for MRD Detection

• Published in: Blood 2020-11, Vol.136 (Supplement 1), p.15-16
• Main Authors: Javed, Omer, Mansoor, Neelum, Khan, Hamza, Jabbar, Naeem, Israr, Talha, Jamal, Saba, Meraj, Fatima
• Format: Article
• Language: English
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2020-140213

Introduction Disease monitoring in management of acute leukemia is essentially important in terms of risk stratification and assessing response to chemotherapy throughout the disease course for which minimal residual disease (MRD) testing is the most reliable tool. Measurement of MRD by flow cytometry is based on the principle of studying difference in antigenic expression of the leukemic cells, hematogones and residual normal B cells. Modulation of antigen expression is an established fact that occurs on leukemic blasts during treatment under the influence of chemotherapeutic drugs, which may cause difficulty in analysis and detection of MRD [H.Burnusuzov et al. Folia Medica 2016;58(1):28-35]. Literature has reported antigen expression modulation, particularly during the glucocorticoid phase of induction therapy [Dworzak et al. Cytometry Part B 2010; 78B: 147-153]. For MRD assessment, such phenotypic shifts pose a particular challenge since they can cause misinterpretations and false results. However to our knowledge there hasn't been a large cohort studied to map antigenic modulation and analyze similar antigenic trends in comparison between leukemic blasts and non-leukemic B cells which may coexist during different phases of chemotherapy. Key words: B-lymphoblastic leukemia, Flowcytometry, Antigen modulation, Minimal residual disease, Immunophenotype Methods: The study is conducted at the Hematology department of The Indus Hospital from April 2018 to March 2020. Among 445 pediatric B-lymphoblastic leukemia patients (1 month-18 years); we studied 121 patients who were MRD positive in bone marrow throughout the evaluation period. All patients received Berlin-Frankfurt-Munster (BFM) chemotherapy protocol as per National Cancer Institute (NCI) risk stratification criteria. To observe immunophenotypic modulation (IM) in antigen expression of Tdt, CD34, CD10, CD20, CD45, CD13, CD33 and CD66, compared from the time of diagnosis (day 0), post induction chemotherapy (day 35), post consolidation chemotherapy (day 52) and during maintenance chemotherapy (day78). Using eight color Flowcytometry (BD FACS CANTO-II; DIVA software version 8.0.2), the IM was assessed by comparative analyses of the changes in the mean flourescence intensity (MFI) of leukemic and non-leukemic B cells. The Wilcoxon signed rank test is used to compare median of MFI values at diagnosis and subsequent time points of MRD. Independent sample T-test/Mann-Whitney U test was applied as appropriate