Objectives Assessment for minimal/measurable residual disease (MRD) is a powerful prognostic factor in B lymphoblastic leukemia/lymphoma (B-LL/L) that is quickly becoming standard of care in assessing patients with B-LL/L posttherapy. MRD can be assessed using methodologies including flow cytometry and molecular genetics, with the former being rapid, relatively inexpensive, and widely applicable in many hematopathology/flow cytometry laboratories. Methods This article presents an approach to MRD detection in B-LL/L by flow cytometry through case presentations with illustration of several potential pitfalls. We review normal maturation patterns, antigens used for assessment, flow panels that can be utilized, considerations to be made during therapy, and clinical impact. The benefits and drawbacks when using the "different from normal" and "leukemia associated phenotype" approaches are considered. Results Evaluation for MRD in B-LL/L by flow cytometry relies on a knowledge of normal immunophenotypic patterns associated with B-cell maturation in states of rest and marrow regeneration so that one can identify patterns of antigen expression that differentiate abnormal, leukemic populations from regenerating hematogones or B-cell precursors. The nature of therapy can affect normal patterns, a phenomenon especially important to take into consideration given the increased use of targeted therapies in the treatment of B-LL/L. Conclusions Flow cytometry is widely available in many laboratories and is a cost-effective way to evaluate for B-LL/L MRD. However, panel validation and interpreter education are crucial for accurate assessment.

How I Diagnose Minimal/Measurable Residual Disease in B Lymphoblastic Leukemia/Lymphoma by Flow Cytometry.

Clinical features and outcome of pediatric acute lymphoblastic leukemia with low peripheral blood blast cell count at diagnosis.

Platelet-Acute Leukemia Interactions.

Combined use of peripheral blood blast count and platelet count during and after induction therapy to predict prognosis in children with acute lymphoblastic leukemia.

ackground: Minimal residual disease (MRD) detection has been shown to be the best prognostic factor in B-acute lymphoblastic leukaemia (B-ALL). Multicolour flow cytometry (FCM) and specific molecular aberrations (MOL) are the classic techniques used to assess MRD. The former is faster and less costly. Aim: This study compares morphology and FCM to MOL in detecting MRD. Setting: The study was conducted at Inkosi Albert Luthuli Central Hospital (IALCH). Methods: A retrospective review of children with B-ALL managed at IALCH from January 2013 to January 2018 was conducted. Multicolour flow cytometry was performed using Euroflow® panels. Molecular aberrations looked at common cytogenetic markers. Presentation and post-induction morphology (May–Grunwald Giemsa stain), FCM and MOL data for MRD were analysed. Results: Eleven patients were excluded (6-demised, 5-incomplete records), leaving 64 to be analysed (54% female, median age 5 years). Five post-induction aspirates were unsuitable but the rest (92%) were in morphological remission. At diagnosis and post-induction, 62 (95%) and 61 (94%) children, respectively, had FCM performed. A positive MOL result was found in 39 (60%) patients. MOL turn-around times (TATs) averaged 14 days compared with those of FCM’s average of 3 days. MRD was found in 9 patients (FCM) and 7 patients (MOL). Of these patients, 4 had a good correlation between the two and 2 patients with negative FCM had positive MOL MRD post-induction. Conclusion: Morphology is insensitive in MRD assessment. FCM correlated well with molecular MRD and has the shortest turn-around time. FCM has major benefit in the 40% of patients with negative MOL. It can also be safely used to guide treatment escalation in those patients awaiting molecular results.

/pdf/comparing-morphology-flow-cytometry-and-molecular-genetics-17ickmy4ie.pdf

Comparing morphology, flow cytometry and molecular genetics in the assessment of minimal residual disease in children with B-acute lymphoblastic leukaemia (B-ALL)

No prognostic significance of immunophenotypic changes at the end of remission induction therapy in children with B-lineage acute lymphoblastic leukemia.

OBJECTIVE
To evaluate the prognosis value of average daily platelet amount increase in children with B-cell acute lymphoblastic leukemia(B-ALL) treated by CCCG-ALL-2015 regimen.


METHODS
106 children with primary B-ALL were retrospective analyzed, standardized MRD test protocol was used to detect the MRD level (19 d and 46 d) after chemotherapy. The platelet count was measured by Sysmex XE-2100. Kaplan-Meier survival curve statistics was used to analyze the event free survival(EFS) rate of the children.


RESULTS
The trend of negative correlation existed between PPC and TPR (rs=-0.519, P=0.021). The 3-year EFS rate of the patients in Ap>5.4×109/L group was 95.7%, which was significantly higher than those in Ap≤5.4×109/L group(79.5%) (χ2=5.236, P=0.035); multivariate analysis showed that Ap≤5.4×109/L was the independent prognostic factor affecting survival of the patients (RR=3.978; 95%CI: 1.336-11.523, P=0.041). With both MRD and Ap≤5.4×109/L as candidate variables, Ap≤5.4×109/L lost its independent prognostic value (RR=1.225; 95%CI: 0.892-13.696, P=0.089), the correlation between d 19/d 46 MRD levels and Ap>5.4×109/L (χ2=4.318, P=0.038) could explain the phenomenon.


CONCLUSION
Ap can reflect the effect of B-ALL chemotherapy and can be used to monitor the curative effect and prognosis of B-ALL children.

[Prognostic Value of Average Daily Platelet Increase in Childhood B-cell Acute Lymphoblastic Leukemia Patients].

OBJECTIVE
We aimed to establish appropriate review criteria for blood cell analysis in a specialized women's and children's hospital. Also, the CellaVision DI-60, was developed as one of the automated digital cell morphology analyzer, we evaluated if it was shown to be most effective under the certain review criteria.


METHODS
A total of 2890 blood samples were detected to optimize the previously established review criteria for women and children with the Sysmex XE-2100. A total of 623 samples were used to validate the criteria.


RESULTS
The microscopic-review rate based on the initial review criteria was 51.0%. After optimization, it was reduced to 17.3% and the false-negative rate was 3.85%. There was > 80% consistency between manual review results and CellaVision DI-60 preclassification when samples triggered the platelet- or red cell-related rules. The sensitivity for abnormalities (immature granulocytes, nucleated red blood cells) of reclassification was 90% to 100% and the false-negative rate was < 5%. However, direct microscopic review was required when the "Blasts/AbnLympho?" and "Atypical Lympho?" flags were triggered.


CONCLUSION
Specialized review criteria are needed for women and children. An automated morphology identification system might help to improve the review criteria.

Establishment of Review Criteria Coordinating With the Automated Digital Cell Morphology Identification System in a Specialized Women's and Children's Hospital.

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Papers

No prognostic significance of immunophenotypic changes at the end of remission induction therapy in children with B-lineage acute lymphoblastic leukemia.

[Prognostic Value of Average Daily Platelet Increase in Childhood B-cell Acute Lymphoblastic Leukemia Patients].

Establishment of Review Criteria Coordinating With the Automated Digital Cell Morphology Identification System in a Specialized Women's and Children's Hospital.

[Prognostic Value of Average Daily Platelet Increase in Childhood B-cell Acute Lymphoblastic Leukemia Patients].