Applications and efficiency of flow cytometry for leukemia diagnostics

Principe, Maria Ilaria Del; Bellis, Eleonora De; Gurnari, Carmelo; Buzzati, Elisa; Savi, Arianna; Consalvo, María Antonietta Irno; Venditti, Adriano

doi:10.1080/14737159.2019.1691918

Cited by 16 publications

(20 citation statements)

References 81 publications

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“…Surface immunoglobulins (Ig) kappa and lambda light chain expression was evaluated on CD22-positive B cells. In agreement with the recommendations for the analysis of rare events, a cluster of 10 events was considered to define a positive result and identify a leukocyte subpopulation ( 10 ). Disease infiltration of the CSF was excluded, being negative for the lymphoma-associated phenotype identified by histopathology.…”

Section: Methodsmentioning

confidence: 99%

“…In recent years, several studies have demonstrated that CSF flow cytometry is superior to conventional cytology for detection of CNS involvement in non-Hodgkin lymphomas, acute leukemia and multiple myeloma (3)(4)(5)(6)(7)(8)(9)(10)(11). Thereafter, flow cytometry is recognized among the basic elements for the diagnosis of leptomeningeal metastasis in hematologic cancers (12,13), although the low cell count of CSF samples, combined with the rapidly declining leukocyte viability, makes CSF flow cytometry challenging (14).…”

Section: Introductionmentioning

confidence: 99%

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Major Differences in Lymphocyte Subpopulations Between Cerebrospinal Fluid and Peripheral Blood in Non-Hodgkin Lymphoma Without Leptomeningeal Involvement: Flow Cytometry Evidence of a Cerebral Lymphatic System

et al. 2021

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Cerebrospinal fluid (CSF) flow cytometry has a crucial role in the diagnosis of leptomeningeal disease in onco-hematology. This report describes the flow cytometry characterization of 138 CSF samples from patients affected by non-Hodgkin lymphoma, negative for disease infiltration. The aim was to focus on the CSF non-neoplastic population, to compare the cellular composition of the CSF with paired peripheral blood samples and to document the feasibility of flow cytometry in hypocellular samples. Despite the extremely low cell count (1 cell/µl, range 1.0–35) the study was successfully conducted in 95% of the samples. T lymphocytes were the most abundant subset in CSF (77%; range 20–100%) with a predominance of CD4-positive over CD8-positive T cells (CD4/CD8 ratio = 2) together with a minority of monocytes (15%; range 0–70%). No B cells were identified in 90% of samples. Of relevance, a normal, non-clonal B-cell population was documented in 5/7 (71%) patients with primary central nervous system lymphoma at diagnosis (p<0.0001), suggesting a possible involvement of blood-brain barrier cell permeability in the pathogenesis of cerebral B-cell lymphomas. The highly significant differences between CSF and paired peripheral blood lymphoid phenotype (p<0.0001) confirms the existence of an active mechanism of lymphoid migration through the meninges.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Major Differences in Lymphocyte Subpopulations Between Cerebrospinal Fluid and Peripheral Blood in Non-Hodgkin Lymphoma Without Leptomeningeal Involvement: Flow Cytometry Evidence of a Cerebral Lymphatic System

et al. 2021

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“…Methods to assess these parameters must be highly standardized, robust, and transferable. Multiparametric immunophenotyping by flow cytometry allows the fast, comprehensive and mostly antibodybased routine analysis of progenitor and immune cells in the peripheral blood, bone marrow or cerebrospinal fluid in healthy people and in patients with leukemia and lymphoma (13)(14)(15). However, flow cytometry is prone to variability due to individual operator handling, reagent/antibody issues and data analysis (16).…”

Section: Introductionmentioning

confidence: 99%

Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications

et al. 2021

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Adoptive immunotherapy using chimeric antigen receptor (CAR)-T cells has achieved successful remissions in refractory B-cell leukemia and B-cell lymphomas. In order to estimate both success and severe side effects of CAR-T cell therapies, longitudinal monitoring of the patient’s immune system including CAR-T cells is desirable to accompany clinical staging. To conduct research on the fate and immunological impact of infused CAR-T cells, we established standardized 13-colour/15-parameter flow cytometry assays that are suitable to characterize immune cell subpopulations in the peripheral blood during CAR-T cell treatment. The respective staining technology is based on pre-formulated dry antibody panels in a uniform format. Additionally, further antibodies of choice can be added to address specific clinical or research questions. We designed panels for the anti-CD19 CAR-T therapy and, as a proof of concept, we assessed a healthy individual and three B-cell lymphoma patients treated with anti-CD19 CAR-T cells. We analyzed the presence of anti-CD19 CAR-T cells as well as residual CD19+ B cells, the activation status of the T-cell compartment, the expression of co-stimulatory signaling molecules and cytotoxic agents such as perforin and granzyme B. In summary, this work introduces standardized and modular flow cytometry assays for CAR-T cell clinical research, which could also be adapted in the future as quality controls during the CAR-T cell manufacturing process.

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“…At present, the diagnosis of CNSL is mostly based on FCM, including FCM combined with lactate dehydrogenase, β-2 microglobulin, cystatin C and other laboratory indicators to improve the diagnosis rate [8,[20][21][22]. However, there are still some disadvantages such as false negative and inability to locate the focus.…”

Section: Discussionmentioning

confidence: 99%

The diagnostic and prognostic value of MRI in central nervous system involvement of acute myeloid leukemia: a retrospective cohort of 84 patients

et al. 2020

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Objective: To assess the diagnostic and prognostic value of magnetic resonance imaging (MRI) in Acute Myeloid Leukemia (AML) complicated with central nervous system leukemia (CNSL). Methods: A total of 84 patients with AML and confirmed of CNSL from January 2010 to September 2019 were selected and underwent MRI scan. We retrospectively analyzed their MRI findings, summarized the imaging features of AML central infiltration, and assessed the guiding significance of MRI on diagnosis and prognosis of this disease. Results: A total of 52 patients (61.90%, 52/84) had abnormal MRI findings, of which 31 cases clearly indicated intracranial infiltration of leukemia. Among the 31 patients, the most common site of infiltration is parenchyma (19/31). Most MRI of these patients showed multiple lesions with low T1 signal and high T2 signal, which were more obvious on enhanced scan. Sensitivity of MRI in diagnosing AML central infiltration was 36.90%. Despite of its low sensitivity, it still had superior diagnostic value on some patients with falsenegative CSF. The median disease-free survival (DFS) and overall survival (OS) time of patients with MRI clearly indicated central invasion were 4 and 9 months, respectively. But there was no significant difference in survival analysis compared with MRI negative patients (including abnormal but non-invasive). Conclusion: MRI manifestation of central infiltration in AML patients has certain characteristic findings, which is helpful to improve the diagnostic efficiency. Prognosis of MRI positive patients is relatively worse than that of MRI negative patients however there is no siginificant difference.

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Applications and efficiency of flow cytometry for leukemia diagnostics

Cited by 16 publications

References 81 publications

Major Differences in Lymphocyte Subpopulations Between Cerebrospinal Fluid and Peripheral Blood in Non-Hodgkin Lymphoma Without Leptomeningeal Involvement: Flow Cytometry Evidence of a Cerebral Lymphatic System

Major Differences in Lymphocyte Subpopulations Between Cerebrospinal Fluid and Peripheral Blood in Non-Hodgkin Lymphoma Without Leptomeningeal Involvement: Flow Cytometry Evidence of a Cerebral Lymphatic System

Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications

The diagnostic and prognostic value of MRI in central nervous system involvement of acute myeloid leukemia: a retrospective cohort of 84 patients

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