van de Loosdrecht, A. A., Kern, W., Porwit, A., Valent, P., Kordasti, S., Cremers, E., Alhan, C., Duetz, C., Dunlop, A., Hobo, W., Preijers, F., Wagner-Ballon, O., Chapuis, N., Fontenay, M., Bettelheim, P., Eidenschink-Brodersen, L., Font, P., Johansson, U., Loken, M. R., … Ireland, R. (2023). Clinical application of flow cytometry in patients with unexplained cytopenia and suspected myelodysplastic syndrome: A report of the European LeukemiaNet International MDS-Flow Cytometry Working Group. Cytometry Part B: Clinical Cytometry, 104(1), 77–86. https://doi.org/10.1002/cyto.b.22044
In the originally published article, the Austrian Science Fund, Grant/Award Number: F4704 was incorrectly listed as a funder. This has been corrected in the online version of the article. We apologize for this error.
van de Loosdrecht, A. A., Kern, W., Porwit, A., Valent, P., Kordasti, S., Cremers, E., Alhan, C., Duetz, C., Dunlop, A., Hobo, W., Preijers, F..、Wagner-Ballon, O., Chapuis, N., Fontenay, M., Bettelheim, P., Eidenschink-Brodersen, L., Font, P., Johansson, U., Loken, M. R., ... Ireland, R. (2023).流式细胞仪在不明原因细胞减少症和疑似骨髓增生异常综合征患者中的临床应用：欧洲白血病网络国际骨髓增生异常综合征流式细胞术工作组报告》。Cytometry Part B: Clinical Cytometry, 104(1), 77-86. https://doi.org/10.1002/cyto.b.22044In 在最初发表的文章中，奥地利科学基金的资助/奖励编号为 F4704：F4704 被错误地列为资助方。在线版文章已对此进行了更正。我们对此错误深表歉意。
{"title":"Correction to “Clinical application of flow cytometry in patients with unexplained cytopenia and suspected myelodysplastic syndrome: A report of the European LeukemiaNet International MDS-Flow Cytometry Working Group”","authors":"","doi":"10.1002/cyto.b.22141","DOIUrl":"10.1002/cyto.b.22141","url":null,"abstract":"<p>van de Loosdrecht, A. A., Kern, W., Porwit, A., Valent, P., Kordasti, S., Cremers, E., Alhan, C., Duetz, C., Dunlop, A., Hobo, W., Preijers, F., Wagner-Ballon, O., Chapuis, N., Fontenay, M., Bettelheim, P., Eidenschink-Brodersen, L., Font, P., Johansson, U., Loken, M. R., … Ireland, R. (2023). Clinical application of flow cytometry in patients with unexplained cytopenia and suspected myelodysplastic syndrome: A report of the European LeukemiaNet International MDS-Flow Cytometry Working Group. <i>Cytometry Part B: Clinical Cytometry</i>, <i>104</i>(1), 77–86. https://doi.org/10.1002/cyto.b.22044</p><p>In the originally published article, the Austrian Science Fund, Grant/Award Number: F4704 was incorrectly listed as a funder. This has been corrected in the online version of the article. We apologize for this error.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"106 3","pages":"216"},"PeriodicalIF":3.4,"publicationDate":"2023-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22141","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10137882","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}
Sa A. Wang, Jeffrey L. Jorgensen, Shimin Hu, Fuli Jia, Shaoying Li, Sanam Loghavi, Chi Young Ok, Beenu Thakral, Jie Xu, L. Jeffrey Medeiros, Wei Wang
Background
Acute myeloid leukemia (AML) minimal/measurable residual disease (MRD) by multicolor flow cytometry is a complex laboratory developed test (LDT), challenging for implementation. We share our experience in the validation of a 12-color AML MRD flow cytometry assay to meet stringent regulatory requirements.
Methods
We worked under the guidelines of the CLSI HL62 publication, illustrated the details of the validation process that was tailored to uniqueness of AML MRD, and tested its clinical validity in 61 patients. The “trueness” was determined by correlating with concurrent molecular genetic testing and follow-up bone marrow examinations.
Results
Under assay specificity, we shared the details of panel design, analysis, and criteria for interpretation and reporting. The assay accuracy was assessed by testing known positive and negative samples and correlating with molecular genetic testing and follow-up bone marrow examination. The limit of detection (LOD) and limit of quantification (LOQ) were validated to a level between 0.01% and 0.1%, varied from the leukemia-associated immunophenotypes (LAIP) and the numbers of events obtained for analysis. Assay linearity, precision and carry over studies all met acceptable criteria. In the clinical validity test, the concordance was 93%, specificity 98% and sensitivity 83%. The most challenging aspects of the assay were the discrimination of pre-leukemic cells (persistent clonal hematopoiesis) or underlying myelodysplastic clones from AML MRD with immunophenotypic switch or subclone selection.
Conclusion
The validation met all criteria and obtained FDA IDE (investigational device exemption) approval. This study provides ample technical and professional details in setting up the AML MRD flow cytometry assay and illustrates through the example of the “fit for purpose” validation process. We also highlight the need for further characterization of abnormal blasts bearing the potential for AML relapse.
{"title":"Validation of a 12-color flow cytometry assay for acute myeloid leukemia minimal/measurable residual disease detection","authors":"Sa A. Wang, Jeffrey L. Jorgensen, Shimin Hu, Fuli Jia, Shaoying Li, Sanam Loghavi, Chi Young Ok, Beenu Thakral, Jie Xu, L. Jeffrey Medeiros, Wei Wang","doi":"10.1002/cyto.b.22140","DOIUrl":"10.1002/cyto.b.22140","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Acute myeloid leukemia (AML) minimal/measurable residual disease (MRD) by multicolor flow cytometry is a complex laboratory developed test (LDT), challenging for implementation. We share our experience in the validation of a 12-color AML MRD flow cytometry assay to meet stringent regulatory requirements.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We worked under the guidelines of the CLSI HL62 publication, illustrated the details of the validation process that was tailored to uniqueness of AML MRD, and tested its clinical validity in 61 patients. The “trueness” was determined by correlating with concurrent molecular genetic testing and follow-up bone marrow examinations.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Under assay specificity, we shared the details of panel design, analysis, and criteria for interpretation and reporting. The assay accuracy was assessed by testing known positive and negative samples and correlating with molecular genetic testing and follow-up bone marrow examination. The limit of detection (LOD) and limit of quantification (LOQ) were validated to a level between 0.01% and 0.1%, varied from the leukemia-associated immunophenotypes (LAIP) and the numbers of events obtained for analysis. Assay linearity, precision and carry over studies all met acceptable criteria. In the clinical validity test, the concordance was 93%, specificity 98% and sensitivity 83%. The most challenging aspects of the assay were the discrimination of pre-leukemic cells (persistent clonal hematopoiesis) or underlying myelodysplastic clones from AML MRD with immunophenotypic switch or subclone selection.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The validation met all criteria and obtained FDA IDE (investigational device exemption) approval. This study provides ample technical and professional details in setting up the AML MRD flow cytometry assay and illustrates through the example of the “fit for purpose” validation process. We also highlight the need for further characterization of abnormal blasts bearing the potential for AML relapse.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"104 5","pages":"356-366"},"PeriodicalIF":3.4,"publicationDate":"2023-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10040504","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}
<p>Diagnostic techniques within hematology are becoming more sensitive and measurable residual disease (MRD) is gaining importance as a result—for both diagnosticians and patients. MRD measurement is increasingly used as a study endpoint in clinical trials and also in routine diagnostics—and as a strong predictor of treatment outcome. Currently, more sensitive and specific methods of measurement are in development. Besides molecular assays, flow cytometry is one of the most frequently used methods for MRD assessment due to its shorter turnaround time, its cost-effectiveness, and broader applicability.</p><p>In this issue, several researchers have delved into various aspects of flow cytometry techniques being used for MRD detection within hematological malignancies. Gao et al. introduced a single tube flow cytometry assay with high sensitivity for monitoring MRD in B-lymphoblastic leukemia/lymphoma (B-ALL), independent of specific surface antigen expression like CD19 and CD22. As the authors point out, the development of such assays has become relevant due to the emergence of targeted anti-CD19 and anti-CD22 therapies (Gao, Chen, et al., <span>2023</span>; Gao, Liu, et al., <span>2023</span>). Targeted immunotherapy demonstrated encouraging results in recent years but induces significant changes in the phenotype of leukemic blasts concurrently. Therefore, alternative gating strategies have gained importance and potential CD19 substitutes have been proposed (Chen et al., <span>2023</span>; Mikhailova et al., <span>2022</span>).</p><p>The development of alternative gating strategies is also relevant independently of targeted therapies, as shown by the authors of the next article. In a rare case study, Ramalingam et al. reported a patient with CD19-negative diffuse large B-cell lymphoma (DLBCL). Since CD19 is currently the primary gating marker for B cell neoplasms, its absence may lead to erroneous results and potentially affect therapeutic strategies, so the authors (Ramalingam et al., <span>2022</span>). Challenges for flow cytometry approaches have been studied before (Gao, Chen, et al., <span>2023</span>; Gao, Liu, et al., <span>2023</span>; Huang et al., <span>2023</span>; Martig & Fromm, <span>2022</span>).</p><p>In MRD detection, the avoidance of false positives is crucial as shown by Zhou et al. The authors discussed the pitfalls in MRD detection in B-ALL following targeted immunotherapy, describing the presence of two CD22-positive non-neoplastic cell populations. One progenitor population of uncertain lineage and one mature B-cell population were both immunophenotypic mimics of B-ALL. Zhou et al. concluded that an understanding of these normal cell populations is essential to avoid misinterpretation in MRD assessments and CD19-independent gating strategies, including CD22 and CD24, are key (Zhou et al., <span>2022</span>). Optimizing MRD measurement is at the forefront of numerous studies and alternative antigens (besides CD22 and CD24
血液学的诊断技术正变得越来越敏感，可测量的残余疾病(MRD)对诊断医生和患者都越来越重要。MRD测量越来越多地被用作临床试验和常规诊断的研究终点，并作为治疗结果的有力预测指标。目前，更敏感和具体的测量方法正在开发中。除了分子分析，流式细胞术是最常用的MRD评估方法之一，因为它的周转时间短，成本效益高，适用性广。在这一期中，几位研究人员深入研究了流式细胞术技术在血液恶性肿瘤中用于MRD检测的各个方面。Gao等人介绍了一种高灵敏度的单管流式细胞术检测方法，用于监测b淋巴细胞白血病/淋巴瘤(B-ALL)的MRD，不依赖于CD19和CD22等特异性表面抗原的表达。正如作者所指出的，由于靶向抗cd19和抗cd22疗法的出现，这种检测方法的发展已经变得相关(Gao, Chen等，2023;高，刘，等，2023)。靶向免疫治疗近年来显示出令人鼓舞的结果，但同时诱导白血病母细胞表型的显着变化。因此，替代门控策略变得越来越重要，并提出了潜在的CD19替代品(Chen et al.， 2023;Mikhailova et al.， 2022)。正如下一篇文章的作者所示，替代门控策略的发展也与靶向治疗无关。在一个罕见的病例研究中，Ramalingam等人报道了一例cd19阴性弥漫性大b细胞淋巴瘤(DLBCL)患者。由于CD19目前是B细胞肿瘤的主要门控标志物，缺乏它可能导致错误的结果，并可能影响治疗策略，因此作者(Ramalingam et al.， 2022)。之前已经研究过流式细胞术方法面临的挑战(Gao, Chen等，2023;高，刘等，2023;黄等人，2023;Martig,弗洛姆,2022)。在MRD检测中，如Zhou等人所示，避免假阳性是至关重要的。作者讨论了靶向免疫治疗后B-ALL MRD检测的缺陷，描述了两种cd22阳性非肿瘤细胞群的存在。一个谱系不确定的祖细胞群体和一个成熟的b细胞群体都是B-ALL的免疫表型模拟者。Zhou等人得出结论，了解这些正常细胞群对于避免MRD评估中的误解至关重要，而CD22和CD24等cd19独立门控策略是关键(Zhou等人，2022)。优化MRD测量是众多研究的前沿，已经提出了用于预测ZNF384重排的替代抗原(除了CD22和CD24之外，还有iCD79a)或雷达图和流式细胞术评分系统(Mikhailova等人，2022;Shopsowitz et al.， 2022;Wang et al.， 2022)。正如下一篇文章所描述的，MRD测量及其优化不仅在淋巴肿瘤中而且在髓系肿瘤中起着重要作用。由于MRD分析尚未包括在英国的常规诊断中，McMillan等人采用并验证了多发性骨髓瘤(MM) MRD的多色流式细胞术(MCF)检测方法，允许在较小的实验室中广泛使用该检测方法进行现实实践(McMillan等人，2022)。在MM中实现MRD阴性对于改善患者预后至关重要。在新诊断和复发的MM中，发现bcma阳性的异常浆细胞比例很高，支持其作为CAR-T细胞和单克隆抗体治疗靶点的潜力(Sriram et al.， 2022)。另一项研究发现多克隆浆细胞和肿瘤浆细胞治疗后的免疫调节异常，强调了在正常和反应性条件下验证MRD分析的重要性(Das等，2022)。已经提出了一个共识协议，以减少MM MRD报告的实验室间差异，从而提高一致性。这种统一的方法使MM MRD成为临床试验中评估无进展和总生存期的潜在替代临床终点(Soh等人，2022)。Hsu等人研究了唐氏综合征特异性非恶性造血再生对唐氏综合征相关髓性白血病(ML-DS) MRD检测的影响。骨髓中ds特异性髓系祖细胞的存在会使ML-DS患者的MRD解释复杂化。因此，作者假设，其意识对MRD检测至关重要(Hsu et al.， 2023)。在这期的最后一篇文章中，Jurado等人着重于优化单核细胞门控策略诊断慢性髓单细胞白血病(CMML)。 他们提出了一种10色管和计算分析来提高单核细胞亚群定量的可重复性(Jurado et al.， 2022)。进一步的研究强调了“单核细胞试验”诊断CMML的稳健性，区分CMML与母浆细胞样树突状细胞肿瘤(BPDCN)的复杂性，以及免疫表型分析在区分CMML与反应性单核细胞增多症方面的潜力(Espasa等，2021;Feng et al.， 2018;Wagner-Ballon et al.， 2023)。总之，本期的重点文章为开发MRD检测的敏感诊断工具、解决潜在的诊断挑战和实施策略以提高各种血液恶性肿瘤MRD评估的准确性提供了有价值的见解。这些进步对改善血液学领域的患者预后和指导治疗决策具有很大的希望。
{"title":"Issue highlights—July 2023","authors":"Wolfgang Kern","doi":"10.1002/cyto.b.22138","DOIUrl":"10.1002/cyto.b.22138","url":null,"abstract":"<p>Diagnostic techniques within hematology are becoming more sensitive and measurable residual disease (MRD) is gaining importance as a result—for both diagnosticians and patients. MRD measurement is increasingly used as a study endpoint in clinical trials and also in routine diagnostics—and as a strong predictor of treatment outcome. Currently, more sensitive and specific methods of measurement are in development. Besides molecular assays, flow cytometry is one of the most frequently used methods for MRD assessment due to its shorter turnaround time, its cost-effectiveness, and broader applicability.</p><p>In this issue, several researchers have delved into various aspects of flow cytometry techniques being used for MRD detection within hematological malignancies. Gao et al. introduced a single tube flow cytometry assay with high sensitivity for monitoring MRD in B-lymphoblastic leukemia/lymphoma (B-ALL), independent of specific surface antigen expression like CD19 and CD22. As the authors point out, the development of such assays has become relevant due to the emergence of targeted anti-CD19 and anti-CD22 therapies (Gao, Chen, et al., <span>2023</span>; Gao, Liu, et al., <span>2023</span>). Targeted immunotherapy demonstrated encouraging results in recent years but induces significant changes in the phenotype of leukemic blasts concurrently. Therefore, alternative gating strategies have gained importance and potential CD19 substitutes have been proposed (Chen et al., <span>2023</span>; Mikhailova et al., <span>2022</span>).</p><p>The development of alternative gating strategies is also relevant independently of targeted therapies, as shown by the authors of the next article. In a rare case study, Ramalingam et al. reported a patient with CD19-negative diffuse large B-cell lymphoma (DLBCL). Since CD19 is currently the primary gating marker for B cell neoplasms, its absence may lead to erroneous results and potentially affect therapeutic strategies, so the authors (Ramalingam et al., <span>2022</span>). Challenges for flow cytometry approaches have been studied before (Gao, Chen, et al., <span>2023</span>; Gao, Liu, et al., <span>2023</span>; Huang et al., <span>2023</span>; Martig & Fromm, <span>2022</span>).</p><p>In MRD detection, the avoidance of false positives is crucial as shown by Zhou et al. The authors discussed the pitfalls in MRD detection in B-ALL following targeted immunotherapy, describing the presence of two CD22-positive non-neoplastic cell populations. One progenitor population of uncertain lineage and one mature B-cell population were both immunophenotypic mimics of B-ALL. Zhou et al. concluded that an understanding of these normal cell populations is essential to avoid misinterpretation in MRD assessments and CD19-independent gating strategies, including CD22 and CD24, are key (Zhou et al., <span>2022</span>). Optimizing MRD measurement is at the forefront of numerous studies and alternative antigens (besides CD22 and CD24","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"104 4","pages":"277-278"},"PeriodicalIF":3.4,"publicationDate":"2023-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22138","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10028473","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}
Arun Kumar Arunachalam, Sushil Selvarajan, Thenmozhi Mani, Nancy Beryl Janet, Madhavi Maddali, Sharon Anbumalar Lionel, Uday Kulkarni, Anu Korula, Fouzia N. Aboobacker, Aby Abraham, Biju George, Poonkuzhali Balasubramanian, Vikram Mathews
The assessment of measurable residual disease (MRD) has emerged as a powerful prognostic tool for both pediatric and adult acute lymphoblastic leukemia (ALL). This retrospective study aimed to evaluate the prognostic relevance of the end of induction MRD in B-cell acute lymphoblastic leukemia (B ALL) patients. The study included 481 patients who underwent treatment for B ALL between August 2012 and March 2019 and had their MRD at the end of induction assessed by flow cytometry. Baseline demographic characteristics were collected from the patient's clinical records. Event free survival (EFS) and relapse free survival (RFS) were calculated using Kaplan–Meier analysis and survival estimates were compared using the log-rank test. End of induction MRD and baseline karyotype were the strongest predictors of EFS and RFS on multivariate analysis. The EFS was inversely related to the MRD value and the outcomes were similar in patients without morphological remission at the end of induction and patients in remission with MRD ≥1.0%. Even within the subgroups of ALL based on age, karyotype, BCR::ABL1 translocation and the treatment protocol, end of induction MRD positive patients had poor outcomes compared to patients who were MRD negative. The study outcome would help draft end of induction MRD-based treatment guidelines for the management of B ALL patients.
{"title":"Clinical significance of end of induction measurable residual disease monitoring in B-cell acute lymphoblastic leukemia: A single center experience","authors":"Arun Kumar Arunachalam, Sushil Selvarajan, Thenmozhi Mani, Nancy Beryl Janet, Madhavi Maddali, Sharon Anbumalar Lionel, Uday Kulkarni, Anu Korula, Fouzia N. Aboobacker, Aby Abraham, Biju George, Poonkuzhali Balasubramanian, Vikram Mathews","doi":"10.1002/cyto.b.22139","DOIUrl":"10.1002/cyto.b.22139","url":null,"abstract":"<p>The assessment of measurable residual disease (MRD) has emerged as a powerful prognostic tool for both pediatric and adult acute lymphoblastic leukemia (ALL). This retrospective study aimed to evaluate the prognostic relevance of the end of induction MRD in B-cell acute lymphoblastic leukemia (B ALL) patients. The study included 481 patients who underwent treatment for B ALL between August 2012 and March 2019 and had their MRD at the end of induction assessed by flow cytometry. Baseline demographic characteristics were collected from the patient's clinical records. Event free survival (EFS) and relapse free survival (RFS) were calculated using Kaplan–Meier analysis and survival estimates were compared using the log-rank test. End of induction MRD and baseline karyotype were the strongest predictors of EFS and RFS on multivariate analysis. The EFS was inversely related to the MRD value and the outcomes were similar in patients without morphological remission at the end of induction and patients in remission with MRD ≥1.0%. Even within the subgroups of ALL based on age, karyotype, <i>BCR::ABL1</i> translocation and the treatment protocol, end of induction MRD positive patients had poor outcomes compared to patients who were MRD negative. The study outcome would help draft end of induction MRD-based treatment guidelines for the management of B ALL patients.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"104 6","pages":"440-452"},"PeriodicalIF":3.4,"publicationDate":"2023-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10014115","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":"An ultra-rapid screening method for acute leukemias","authors":"Olof Axler, Filippa Bild, Åsa C. M. Johansson","doi":"10.1002/cyto.b.22137","DOIUrl":"10.1002/cyto.b.22137","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"104 6","pages":"474-477"},"PeriodicalIF":3.4,"publicationDate":"2023-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9957921","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}
Inès Vergnolle, Theo Ceccomarini, Alban Canali, Jean-Baptiste Rieu, François Vergez
Background: 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.
Methods: 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.
Results: 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.
Conclusion: This approach has the potential to improve the accuracy and efficiency of mature B-cell neoplasm diagnosis.
{"title":"Use of a hybrid intelligence decision tree to identify mature B-cell neoplasms.","authors":"Inès Vergnolle, Theo Ceccomarini, Alban Canali, Jean-Baptiste Rieu, 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":" ","pages":""},"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}
Weijie Li, Ruth Morgan, Roxanne Nieder, Sahibu Sultan M. Habeebu, G. Doug Myers
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.
{"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, Ruth Morgan, Roxanne Nieder, Sahibu Sultan M. Habeebu, 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":"104 5","pages":"392-396"},"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}
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
{"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, 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":"104 5","pages":"400-403"},"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, Filippo Conti, Jean-Francois Llitjos, Aurore Fleurie, Valérie Cerro, Fabienne Venet, Anne-Claire Lukaszewicz, 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":"104 6","pages":"468-470"},"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}
Linda Hammerich, Yaroslava Shevchenko, Jana Knorr, Wiebke Werner, Alix Bruneau, Frank Tacke
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.
{"title":"Resolving 31 colors on a standard 3-laser full spectrum flow cytometer for immune monitoring of human blood samples","authors":"Linda Hammerich, Yaroslava Shevchenko, Jana Knorr, Wiebke Werner, Alix Bruneau, 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":"104 5","pages":"367-373"},"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}
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