A novel flow cytometric assay for detection of residual disease in patients with B‐lymphoblastic leukemia/lymphoma post anti‐CD19 therapy

Cherian, Sindhu; Miller, Valerie; McCullouch, Vivian; Dougherty, Katy J.; Fromm, Jonathan R.; Wood, Brent L.

doi:10.1002/cyto.b.21482

Cited by 89 publications

(119 citation statements)

References 23 publications

(35 reference statements)

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“…Although alternative gating strategies can be applied (eg, based on CD10, CD34, and/or CD45), one could also decide to add CD24 and/or CD22 to the current tubes (transforming it into a 10-color tube) in MRD-based trials involving Blinatumomab. 43 Addition of CD24 and CD22 will also have the advantage that the earliest BCP cells, expressing CD24 and/or CD22 but not yet CD19, 44 can be identified; this may be of relevance for the identification of all BCP cells in regenerating BM samples.…”

Section: Discussionmentioning

confidence: 99%

Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia

Theunissen

Mejstříková

Sędek

et al. 2017

Blood

338

296

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Key Points• Standardized flow cytometry allows highly sensitive MRD measurements in virtually all BCP-ALL patients.• If sufficient cells are measured (.4 million), flow cytometric MRD analysis is at least as sensitive as current PCR-based MRD methods.A fully-standardized EuroFlow 8-color antibody panel and laboratory procedure was stepwise designed to measure minimal residual disease (MRD) in B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) patients with a sensitivity of £10 25, comparable to real-time quantitative polymerase chain reaction (RQ-PCR)-based MRD detection via antigen-receptor rearrangements. Leukocyte markers and the corresponding antibodies and fluorochromes were selected based on their contribution in separating BCP-ALL cells from normal/regenerating BCP cells in multidimensional principal component analyses. After 5 multicenter design-test-evaluate-redesign phases with a total of 319 BCP-ALL patients at diagnosis, two 8-color antibody tubes were selected, which allowed separation between normal and malignant BCP cells in 99% of studied patients. These 2 tubes were tested with a new erythrocyte bulk-lysis protocol allowing acquisition of high cell numbers in 377 bone marrow follow-up samples of 178 BCP-ALL patients.Comparison with RQ-PCR-based MRD data showed a clear positive relation between the percentage concordant cases and the number of cells acquired. For those samples with >4 million cells acquired, concordant results were obtained in 93% of samples. Most discordances were clarified upon high-throughput sequencing of antigen-receptor rearrangements and blind multicenter reanalysis of flow cytometric data, resulting in an unprecedented concordance of 98% (97% for samples with MRD < 0.01%). In conclusion, the fully standardized EuroFlow BCP-ALL MRD strategy is applicable in >98% of patients with sensitivities at least similar to RQ-PCR (£10 25 ), if sufficient cells (>4 3 10 6 , preferably more) are evaluated. (Blood. 2017;129(3):347-357)

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Section: Discussionmentioning

confidence: 99%

Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia

Theunissen

Mejstříková

Sędek

et al. 2017

Blood

338

296

View full text Add to dashboard Cite

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“…A comparison of B-ALL diagnostic IP vs. relapse IP showed that CD19 is most stable, while CD34 and CD20 differ significantly in nearly half of the cases (31). Utilization of CD22 and CD24, according the method of Cherian et al is a useful approach for MRD detection of CD19 negative/silent disease (15). In our study, we observed a slight down-regulation of CD19 and CD20 at relapse after the second CAR-T cell infusion in Patient 5.…”

Section: Discussionmentioning

confidence: 99%

“…PB and BM were collected pre-and post-CAR-T, and processed for 8-color FCM within 24 h of collection with cocktailed antibodies (see Supporting information), based on Children's Oncology Group guidelines (14), and modification of the Cherian et al approach (15). FCM procedure (16) included NH 4 Cl whole blood lysis, phosphate-buffered saline wash and antibody incubation.…”

Section: Flow Cytometrymentioning

confidence: 99%

Early recovery of circulating immature B cells in B‐lymphoblastic leukemia patients after CD19 targeted CAR T cell therapy: A pitfall for minimal residual disease detection

Xiao

Salem

McCoy

et al. 2017

Cytometry Part B Clinical

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These circulating CD10 + B-cells are compatible with immature B-cells, and are a reflection of B-cell recovery within the marrow. They are immunophenotypically distinguishable from residual B-ALL. Expression of light chain sIg and key surface antigens characterizing regenerating B-cell precursors can distinguish immature B-cells from B-ALL MRD and prevent misdiagnosis. © 2017 International Clinical Cytometry Society.

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“…Immunotherapy has also been associated with decreased expression of non-targeted antigens, such as decreased CD19 following anti-CD20 monoclonal antibody therapy, decreased CD19, CD21, and CD79a by trogocytosis following anti-CD22 therapy (68) and decreased CD20 during therapy with the B-cell receptor inhibitor ibrutinib (69). It is also essential to consider post-treatment changes during panel design for MRD detection (63), and demonstrate that the selected antibody combination meets desired assay performance characteristics through appropriate validation studies (70,71). It is recommended that panels include antibodies to several lineage-associated antigens and a gating strategy is adopted that accounts for all events and highlights antigen negative cells (70) (Fig.…”

Section: Role Of Flow Cytometry In Therapeutic Decisionmentioning

confidence: 99%

The current role of clinical flow cytometry in the evaluation of mature B‐cell neoplasms

Seegmiller

Hsi

Craig

2018

Cytometry Part B Clinical

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Flow cytometry (FC) has a well-established role in the diagnostic evaluation of mature B-cell neoplasms. Effective assessment for lineage associated antigens, aberrant antigen expression, and immunoglobulin light chain restriction requires a welldesigned, optimized, and controlled FC assay. However, it is important for hematopathologists to know when flow cytometry has a more limited role, and other modalities, such as immunohistochemistry, cytogenetic and molecular testing, are more important. This review will discuss the features of an optimal FC assay for the evaluation of mature B-cell neoplasms, and the current role of FC in the diagnosis and sub-classification, prognostic assessment, identification of therapeutic targets, and assessment for disease response to therapy.

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A novel flow cytometric assay for detection of residual disease in patients with B‐lymphoblastic leukemia/lymphoma post anti‐CD19 therapy

Cited by 89 publications

References 23 publications

Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia

Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia

Early recovery of circulating immature B cells in B‐lymphoblastic leukemia patients after CD19 targeted CAR T cell therapy: A pitfall for minimal residual disease detection

The current role of clinical flow cytometry in the evaluation of mature B‐cell neoplasms

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