Cytometry Part B: Clinical Cytometry最新文献

{"title":"Use of a hybrid intelligence decision tree to identify mature B-cell neoplasms.","authors":"Inès Vergnolle,&nbsp;Theo Ceccomarini,&nbsp;Alban Canali,&nbsp;Jean-Baptiste Rieu,&nbsp;François Vergez","doi":"10.1002/cyto.b.22136","DOIUrl":"10.1002/cyto.b.22136","url":null,"abstract":"<p><strong>Background: </strong>Mature B-cell neoplasms are challenging to diagnose due to their heterogeneity and overlapping clinical and biological features. In this study, we present a new workflow strategy that leverages a large amount of flow cytometry data and an artificial intelligence approach to classify these neoplasms.</p><p><strong>Methods: </strong>By combining mathematical tools, such as classification algorithms and regression tree (CART) models, with biological expertise, we have developed a decision tree that accurately identifies mature B-cell neoplasms. This includes chronic lymphocytic leukemia (CLL), for which cytometry has been extensively used, as well as other non-CLL subtypes.</p><p><strong>Results: </strong>The decision tree is easy to use and proposes a diagnosis and classification of mature B-cell neoplasms to the users. It can identify the majority of CLL cases using just three markers: CD5, CD43, and CD200.</p><p><strong>Conclusion: </strong>This approach has the potential to improve the accuracy and efficiency of mature B-cell neoplasm diagnosis.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9935370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CD19-negative B-cell precursors in bone marrow: A potential mimicker for CD19-negative B-lymphoblastic leukemia by flow cytometry in patients with anti-CD19 treatment","authors":"Weijie Li,&nbsp;Ruth Morgan,&nbsp;Roxanne Nieder,&nbsp;Sahibu Sultan M. Habeebu,&nbsp;G. Doug Myers","doi":"10.1002/cyto.b.22135","DOIUrl":"10.1002/cyto.b.22135","url":null,"abstract":"Novel therapies such as monoclonal antibody or chimeric antigen receptor (CAR) T cell therapy (CAR T) have shown promising results in the management of relapsed/refractory B-lymphoblastic leukemia (B-ALL). CD19, a transmembrane glycoprotein almost always expressed in B-ALL, is a commonly used target by these therapies. Blinatumomab, a bispecific monoclonal antibody T cell engager (BiTE), induces antibody-dependent cellular cytotoxicity against the leukemic cells by binding to CD19 on leukemic cells and CD3 on T cells simultaneously. Tisagenlecleucel (Kymriah), a CAR T targeting CD19, uses patients' own modified T cells to attack CD19+ leukemic cells. The superior efficacy of blinatumomab and tisagenlecleucel has been well demonstrated in relapsed/refractory B-ALL compared with high-dose chemotherapy. Multiparametric flow cytometry (FCM) is commonly used to monitor the therapeutic responses to these treatments. The key strategy for FCM to detect residual or relapsed B-ALL is to find an immature B-cell population phenotypically different from normal B-cell precursors (BCPs, also known as hematogones) in the bone marrow (BM). To investigate if these two anti-CD19 therapies could alter the phenotype of normal BCPs, we retrospectively studied the BCPs in the BM specimens from patients treated with blinatumomab and tisagenlecleucel. We also studied the BCPs from the patients after conventional chemotherapy and hematopoietic stem cell transplantation (HSCT) as a comparison.","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9879768","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acute leukemia of ambiguous lineage, not otherwise specified with FLT3-ITD mutation and a possible origin in the common lymphoid progenitor","authors":"Fernando Martin-Moro,&nbsp;Jose A. Garcia-Vela","doi":"10.1002/cyto.b.22134","DOIUrl":"10.1002/cyto.b.22134","url":null,"abstract":"The current major classification of acute leukemias is based on the assignment of cellular lineage to the blast population, for which an immunophenotypic analysis generally performed by multiparametric flow cytometry (FCM) is mandatory (Porwit & Béné, 2019). Furthermore, the phenotypic characterization of acute leukemias provides information about the pathogenesis of the disease, although it is sometimes difficult to correlate the blast population compartment with the normal stages of hematopoiesis. Acute leukemias of ambiguous lineage (ALAL) — category included in both the 5th edition of the World Health Organization Classification of Haematolymphoid Tumours (Khoury et al., 2022) and the International Consensus Classification of Myeloid Neoplasms and Acute Leukemias (Arber et al., 2022) — comprise a heterogeneous group of rare and aggressive diseases that includes acute undifferentiated leukemias (AUL) and mixedphenotype acute leukemias (MPAL), thus, ALAL are those that either fail to show evidence of either myeloid, B, or T-lymphoid lineages, or show evidence of commitment to more than one lineage, respectively. The so-called subgroup ALAL, not otherwise specified (NOS) is a catch-all category of diseases that express a combination of markers that do not allow their classification as either other ALAL, and in which a definitive classification along a single lineage is difficult. All those infrequent entities are considered as high-risk diseases and show a poor prognosis when treated with either conventional chemotherapy used for acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). More aggressive approaches such as the FLAG-IDA regimen (fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin) have been explored in patients with ALAL, and","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9546342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Monocyte HLA-DR expression as an enrollment biomarker in sepsis clinical trials: Evaluation of two sampling tubes and definition of respective clinical thresholds","authors":"Muzhda Haem Rahimi,&nbsp;Filippo Conti,&nbsp;Jean-Francois Llitjos,&nbsp;Aurore Fleurie,&nbsp;Valérie Cerro,&nbsp;Fabienne Venet,&nbsp;Anne-Claire Lukaszewicz,&nbsp;Guillaume Monneret","doi":"10.1002/cyto.b.22133","DOIUrl":"10.1002/cyto.b.22133","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9522535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Resolving 31 colors on a standard 3-laser full spectrum flow cytometer for immune monitoring of human blood samples","authors":"Linda Hammerich,&nbsp;Yaroslava Shevchenko,&nbsp;Jana Knorr,&nbsp;Wiebke Werner,&nbsp;Alix Bruneau,&nbsp;Frank Tacke","doi":"10.1002/cyto.b.22126","DOIUrl":"10.1002/cyto.b.22126","url":null,"abstract":"<p>Immune monitoring of patients on a single-cell level is becoming increasingly important in various diseases. Due to the often very limited availability of human specimens and our increased understanding of the immune systems there is an increasing demand to analyze as many markers as possible simultaneously in one panel. Full spectrum flow cytometry is emerging as a powerful tool for immune monitoring since 5-laser instruments enable characterization of 40 parameters or more in a single sample. Nevertheless, even if only machines with fewer lasers are available, development of novel fluorophore families enables increasing panel sizes. Here, we demonstrate that careful panel design enables the use of 31-color panels on a 3-laser Cytek® Aurora cytometer for analyzing human peripheral blood leukocytes, without the need for custom configuration and using only commercially available fluorochromes. The panel presented here should serve as an example of a 31-fluorochrome combination that can be resolved on a 3-laser full spectrum cytometer and that can be adapted to comprise other (and possibly more) markers of interest depending on the research focus.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22126","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9489135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Automation in flow cytometry: Guidelines and review of systems","authors":"Ahmad Al-Attar,&nbsp;Kirthi R. Kumar,&nbsp;Dorian Untersee,&nbsp;Marci O'Driscoll,&nbsp;Miguel Francoise S. Ventura,&nbsp;Leo Lin","doi":"10.1002/cyto.b.22125","DOIUrl":"10.1002/cyto.b.22125","url":null,"abstract":"<p>Automation in flow cytometry has recently advanced from the partial laboratory automation and robotics islets, to more fully integrated systems. This article reviews three manufacturers' newest sample preparation systems: the Beckman CellMek, the Sysmex PS-10, and the BD FACSDuet. These three instruments are capable of performing many of the manual steps in flow cytometry sample processing (pipetting, staining, lysing, washing, fixing). General description, capabilities, advantages, and disadvantages of each system are compared. Overall, these systems have the potential to become mainstay items in today's busy clinical flow cytometry laboratories, and save a significant amount of hands-on time for laboratory staff.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2023-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9453519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Issue highlights—May 2023","authors":"Sa Wang MD","doi":"10.1002/cyto.b.22122","DOIUrl":"10.1002/cyto.b.22122","url":null,"abstract":"<p>In the era of personalized medicine, the list of targeted therapeutic drugs has been greatly expanded, many of which are employed to treat hematolymphoid neoplasms. These include drugs specifically targeting immune checkpoint signaling epitopes such as programmed cell death 1 (PD1) and its ligand PDL1 to inhibit T-cell activation. Alternatively, these drugs might target the surface antigens expressed by tumor cells such as CD19 and CD22 in B-cell neoplasms. Of the latter, targeted immunotherapies include monoclonal antibodies with or without drug conjugates, bispecific T-cell engagers, or chimeric antigen receptor (CAR) T-cell therapy. The binding sites of these therapeutic antibodies are often identical or in proximity with the binding sites of the diagnostic antibodies. Therefore, use of these therapies pose great challenges for clinical cytometry labs in the assessment of post-treatment samples, especially in the detection of measurable/minimal residual disease (MRD). In this issue, Chen, Gao, et al. (<span>2023</span>) provided an overview of MRD detection in B-lymphoblastic leukemia/lymphoma in the era of immunotherapy, and Gao et al. (<span>2023</span>) focused their review on the impact of targeted therapy on mature B- and plasma cell neoplasms utilizing flow cytometry assessments. In both reviews, the authors illustrated the challenges, identified the problems, and provided a list of available options and solutions. For each of the above-mentioned disease categories, optimal gating and analysis strategies were illustrated with literature review and inputs from the authors' experience and insights. The utility and interpretation of additional B-cell markers other than CD19 and CD20 for mature or immature B-cell neoplasms such as CD22, CD24, and cCD79a (Mikhailova et al., <span>2022</span>) were studied as well as VS38, CD229, and CD319 (Pojero et al., <span>2016</span>; Soh et al., <span>2021</span>) for plasma cell neoplasms. The authors recommended that the flow cytometry assays in the era of targeted therapies must contain significant redundancy in the antibody panels allowing the detection of the cells of interest and additionally should be ready to utilize several gating strategies for accurate and consistent population identification.</p><p>Neoplastic mature B-cells often show restricted kappa or lambda light chain expression; however, light chain expression may be absent in around 5%–10% of mature B-cell lymphoma (Li et al., <span>2002</span>). It is suggested that mature B-cells lacking surface light chain expression can be used as a surrogate marker to diagnose mature B-cell lymphomas. Huang et al. (<span>2023</span>) reported a series of 89 cases of surface light chain negative B-cell lymphoma which consisted primarily of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). Interestingly, the authors also reported no detectable light chain expression in normal/reactive mature B cells collected from body fluids an","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22122","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9431290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extranodal NK/T-cell lymphoma with isolated central nervous system relapse: A defiant disease and the role of flow cytometry in monitoring","authors":"Devasis Panda,&nbsp;Amardeep Pathak,&nbsp;Narender Tejwani,&nbsp;Anurag Mehta","doi":"10.1002/cyto.b.22124","DOIUrl":"10.1002/cyto.b.22124","url":null,"abstract":"Central nervous system (CNS) involvement in extranodal NK/T cell lymphoma (ENKTCL) is rare and confers a dismal prognosis. Though newer treatment modalities like L-asparaginase based chemotherapy, immunotherapy, and hematopoietic stem cell transplant (HSCT) have yielded encouraging results, treatment failure with relapse of primary disease continues to be a major a concern (Yamaguchi et al., 2018). Herein, we report a rare phenomenon of isolated CNS relapse in a patient with ENKTCL within 3 months of allogenic HSCT subsequent to SMILE (dexamethasone, methotrexate, ifosfamide, L-asparaginase, and etoposide) and nivolumab induced disease remission. Our patient, a 51-year-old male, was a known case of ENKTCL with simultaneous involvement of both nasal and right testes 1 year back. Initial diagnosis was made on histopathology and EBER in situ hybridization was positive on tissue sections. No signs or symptoms of CNS involvement was present at initial diagnosis and CSF was clear of any disease. He had undergone right sided orchiectomy and received six cycles of CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) with gemcitabine and prophylactic intrathecal methotrexate. He experienced a relapse 6 months into therapy with a left testicular hypoechoic mass lesion and required left sided orchiectomy. Subsequently, he was treated with low dose nivolumab and SMILE regimen for six cycles and achieved remission. This was followed by allo-HSCT after a 10/10 HLA match with his blood brother. The initial 2 months post-transplant time period was uneventful and chimerism was 100%. On day 62 of post allo-HSCT, he presented to our outpatient department with complaints of headache since last 4 days, blurring of vision in the right eye and difficulty in chewing solid food. Physical examination revealed diminished distant vision in the right eye, weakness in the left side of face, altered taste sensation and dysarthria in speech. MRI of brain and orbits showed borderline enhancement of bilateral optic nerves at optic disc level along with right facial and trigeminal nerve enhancement. Laboratory investigation showed normal complete blood counts with no atypical cells on peripheral smear. CSF cell count on was 15 cells/μL and biochemistry showed high protein (138 mg/dL) and low sugar (26 mg/dL) levels. Meningitis panel through multiplex PCR for viral, bacterial, and parasitic infection was negative; however, the panel did not include EBV target. Though CSF cytology was suspicious, a definite opinion could not be furnished due to scant number of cells and in the absence of obvious atypia. But, flow cytometry immunophenotyping (FCMI) through stain-no lyse-no wash technique on CSF revealed an abnormal NK cell population with CD45 bright, surfaceCD3 , CD7 , CD56 bright+, CD16 , CD2dim+, CD4 , CD8 , CD5 (Figure 1a–e) confirming CNS relapse of the primary disease. PET scan did not show any other lesion elsewhere in the body and bone marrow examination was unre","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9405267","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Summary of validation considerations with real-life examples using both qualitative and semiquantitative flow cytometry assays","authors":"Katherine A. Devitt,&nbsp;Teri Oldaker,&nbsp;Kalpesh Shah,&nbsp;Andrea Illingworth","doi":"10.1002/cyto.b.22123","DOIUrl":"10.1002/cyto.b.22123","url":null,"abstract":"<p>In the clinical laboratory, flow cytometry assays are critical to providing diagnostic and prognostic information to the treating clinicians. A validation or verification provides confidence that the assay will yield reliable results that can be trusted to make critical medical decisions. The following performance specifications should be included in a validation for laboratory developed tests as needed: accuracy (or trueness), precision (reproducibility and repeatability), detection capability, selectivity, reference range, and sample and reagent stability. We define these terms and present our approach to validation of several common flow cytometry assays, including examples of a leukemia/lymphoma assay and a paroxysmal nocturnal hemoglobinuria (PNH) assay.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9832512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acquired RUNX1::CBFA2T2 fusion at extramedullary relapse in a patient of PDGFRA rearranged acute myeloid leukemia post allogenic HSCT","authors":"Devasis Panda,&nbsp;Amardeep Pathak,&nbsp;Narender Tejwani,&nbsp;Pooja Pandey,&nbsp;Anurag Mehta","doi":"10.1002/cyto.b.22121","DOIUrl":"10.1002/cyto.b.22121","url":null,"abstract":"FIP1L1::PDGFRA rearranged myeloid neoplasms encompass a broad range of malignancies including chronic eosinophilic leukemia, MDS, MPN, MDS/MPN, AML and myeloid sarcoma. A little data are available pertaining to their antigen expression pattern till now due to low disease incidence. We hereby, describe a post-transplant case of FIP1L1::PDGFRA rearranged AML that had a typical extramedullary relapse with an additional RUNX1::CBFA2T2 fusion within the first year of post-transplant period and presented with a peculiar CD45 negative immunophenotype. A 25-year-old male patient of FIP1L1::PDGFRA rearranged AML, on follow-up day 280 of post allogenic hematopoietic stem cell transplant (allo-HSCT) presented with palpable left cervical lymphadenopathy and multiple subcutaneous nodules over chest, abdomen and bilateral lower limbs averaging from 0.5 to 1 cm in maximum dimen-sion. High-resolution computed tomography of thorax revealed bilateral pleural effusion and a solitary right lung upper lobe nodule measuring 1.0 (cid:1) 0.9 cm and multiple discrete mediastinal lymph nodes. Complete blood counts showed hemoglobin 11.5 g/dL, total leukocyte count 5.61 (cid:1) 10 3 / μ L, platelet count 110 (cid:1) 10 3 / μ L and no blast on peripheral blood (PB) differential leukocyte count. Bone marrow (BM) morphology was unremarkable and multicolor flow cytometry (MFC) measurable residual disease was negative. Neither PB nor BM showed any increase in eosinophil count. Liver function test showed","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9343339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}