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Cytometry Part B: Clinical Cytometry最新文献

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Automation in flow cytometry: Guidelines and review of systems 流式细胞仪自动化:系统指南与回顾。
IF 2.3 3区 医学 Q3 MEDICAL LABORATORY TECHNOLOGY Pub Date : 2023-05-13 DOI: 10.1002/cyto.b.22125
Ahmad Al-Attar, Kirthi R. Kumar, Dorian Untersee, Marci O'Driscoll, Miguel Francoise S. Ventura, Leo Lin

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.

最近,流式细胞仪的自动化已从部分实验室自动化和机器人系统发展到更全面的集成系统。本文评述了三家制造商的最新样品制备系统:Beckman CellMek、Sysmex PS-10 和 BD FACSDuet。这三种仪器能够执行流式细胞仪样本处理过程中的许多手动步骤(移液、染色、裂解、洗涤、固定)。对每种系统的一般描述、功能、优缺点进行了比较。总之,这些系统有可能成为当今繁忙的临床流式细胞仪实验室的主要设备,为实验室工作人员节省大量的动手时间。
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引用次数: 0
Issue highlights—May 2023 问题亮点- 2023年5月
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-05-08 DOI: 10.1002/cyto.b.22122
Sa Wang MD

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. (2023) provided an overview of MRD detection in B-lymphoblastic leukemia/lymphoma in the era of immunotherapy, and Gao et al. (2023) 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., 2022) were studied as well as VS38, CD229, and CD319 (Pojero et al., 2016; Soh et al., 2021) 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.

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., 2002). 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. (2023) 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

在个体化医疗时代,靶向治疗药物的清单得到了极大的扩展,其中许多药物被用于治疗血淋巴肿瘤。这些药物包括专门针对免疫检查点信号表位的药物,如程序性细胞死亡1 (PD1)及其配体PDL1,以抑制t细胞活化。或者,这些药物可能靶向肿瘤细胞表达的表面抗原,如b细胞肿瘤中的CD19和CD22。在后者中,靶向免疫治疗包括单克隆抗体(含或不含药物偶联物)、双特异性t细胞接合物或嵌合抗原受体(CAR) t细胞治疗。这些治疗性抗体的结合位点通常与诊断性抗体的结合位点相同或接近。因此,使用这些疗法对临床细胞实验室在评估治疗后样本时提出了巨大的挑战,特别是在检测可测量/微小残留疾病(MRD)时。在这一期中,Chen、Gao等人(2023)概述了免疫治疗时代B淋巴母细胞白血病/淋巴瘤的MRD检测,Gao等人(2023)利用流式细胞术评估了靶向治疗对成熟B细胞和浆细胞肿瘤的影响。在这两篇综述中,作者都阐述了挑战,确定了问题,并提供了可用选项和解决方案的列表。对于上述每种疾病类别,通过文献综述和作者的经验和见解,说明了最佳的门控和分析策略。研究了除CD19和CD20以外的其他b细胞标志物(如CD22、CD24和cCD79a)在成熟或未成熟b细胞肿瘤中的应用和解释(Mikhailova等人,2022),以及VS38、CD229和CD319 (Pojero等人,2016;Soh et al., 2021)对浆细胞肿瘤的影响。作者建议,在靶向治疗时代,流式细胞术检测必须在抗体组中包含重要的冗余,以便检测感兴趣的细胞,此外还应准备好利用几种门控策略来准确和一致地进行群体识别。肿瘤成熟b细胞常表现出kappa或lambda轻链表达受限;然而,在大约5%-10%的成熟b细胞淋巴瘤中可能没有轻链表达(Li et al., 2002)。提示缺乏表面轻链表达的成熟b细胞可作为诊断成熟b细胞淋巴瘤的替代标志物。Huang等(2023)报道了89例表面轻链阴性b细胞淋巴瘤,主要包括慢性淋巴细胞白血病/小淋巴细胞淋巴瘤(CLL/SLL)。有趣的是,作者还报告了从14例无临床和病理证据的淋巴瘤/白血病患者的体液和囊性液中收集的正常/反应性成熟B细胞中未检测到轻链表达。体液通常是蛋白质;因此,通常需要在kappa/lambda染色之前进行额外的洗涤,以获得足够的轻链染色,并且在解释从体液中收集的b细胞中的轻链表达时需要谨慎。众所周知,除慢性髓性白血病(CML)外(Soma et al., 2016),其他慢性髓性肿瘤,如骨髓增生异常综合征(MDS) (Xie et al., 2019)和费城阴性骨髓增生性肿瘤(MPN),淋巴母细胞危象极为罕见。Chan等人(2023)报道,在1262例MDS或ph阴性MPN中,有9例检测到低水平的异常b淋巴细胞,范围为0.012%至3.6%。细胞分选结合下一代测序表明,髓系肿瘤中异常的B淋巴母细胞通常(但并非总是)与髓系室克隆相关,分子遗传学结果表明,具有不同谱系输出的突变多能祖细胞可能由特定突变和突变发生的特定细胞阶段决定。重要的是,与CML不同,MDS和ph阴性mpn中存在异常b淋巴细胞并不一定表明母细胞危象,密切监测潜在髓系肿瘤的治疗被认为是一种合理的方法。这期的最后一篇文章(Chen, Zhang, et al., 2023)研究了再生障碍性贫血(AA)中的粘膜相关不变T细胞(MAIT) (Godfrey et al., 2019),再生障碍性贫血是一种与异常免疫反应直接相关的骨髓衰竭。MAIT细胞表现出效应记忆表型,表达多种NK受体,如NKG2D和CD161,并在自身免疫性疾病的发展中发挥重要作用。作者发现再生障碍性贫血患者中具有激活和效应功能相关的细胞表面标记谱的MAIT细胞的频率增加。 NKG2D在MAIT细胞上表达的特异性提示这些细胞可能在AA免疫发病机制中具有独特的参与机制。
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引用次数: 0
Extranodal NK/T-cell lymphoma with isolated central nervous system relapse: A defiant disease and the role of flow cytometry in monitoring 孤立性中枢神经系统复发的结外NK/T细胞淋巴瘤:一种挑战性疾病和流式细胞术在监测中的作用。
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-05-04 DOI: 10.1002/cyto.b.22124
Devasis Panda, Amardeep Pathak, Narender Tejwani, Anurag Mehta
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
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引用次数: 1
Summary of validation considerations with real-life examples using both qualitative and semiquantitative flow cytometry assays 使用定性和半定量流式细胞术分析的真实实例的验证注意事项摘要。
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-05-03 DOI: 10.1002/cyto.b.22123
Katherine A. Devitt, Teri Oldaker, Kalpesh Shah, Andrea Illingworth

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.

在临床实验室中,流式细胞术检测对于向治疗临床医生提供诊断和预后信息至关重要。验证或验证提供了这样的信心,即该测定将产生可靠的结果,可以信赖该结果来做出关键的医疗决策。根据需要,实验室开发测试的验证应包括以下性能规范:准确性(或真实性)、精密度(再现性和可重复性)、检测能力、选择性、参考范围以及样品和试剂的稳定性。我们定义了这些术语,并介绍了几种常见的流式细胞术检测的验证方法,包括白血病/淋巴瘤检测和阵发性夜间血红蛋白尿(PNH)检测的例子。
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引用次数: 1
Acquired RUNX1::CBFA2T2 fusion at extramedullary relapse in a patient of PDGFRA rearranged acute myeloid leukemia post allogenic HSCT 一例PDGFRA重排急性髓系白血病患者异基因造血干细胞移植后骨髓外复发时获得性RUNX1::CBFA2T2融合。
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-04-13 DOI: 10.1002/cyto.b.22121
Devasis Panda, Amardeep Pathak, Narender Tejwani, Pooja Pandey, Anurag Mehta
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
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引用次数: 1
The impact of Down syndrome-specific non-malignant hematopoietic regeneration in the bone marrow on the detection of leukemic measurable residual disease 唐氏综合征特异性骨髓非恶性造血再生对白血病可测量残留病检测的影响
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-04-04 DOI: 10.1002/cyto.b.22118
Fan-Chi Hsu, Chad Hudson, Elisabeth R. Wilson, Laura M. Pardo, Timothy P. Singleton, Dongbin Xu, Barbara K. Zehentner, Johann Hitzler, Jason Berman, Denise A. Wells, Michael R. Loken, Lisa Eidenschink Brodersen

Background

Detection of measurable residual disease detection (MRD) by flow cytometry after the first course of chemotherapy is a standard measure of early response in patients with acute myeloid leukemia (AML). Myeloid leukemia associated with Down Syndrome (ML-DS) is a distinct form of AML. Differences in steady-state and regenerating hematopoiesis between patients with or without DS are not well understood. This understanding is essential to accurately determine the presence of residual leukemia in patients with ML-DS.

Methods

A standardized antibody panel defined quantitative antigen expression in 115 follow-up bone marrow (BM) aspirates from 45 patients following chemotherapy for ML-DS or DS precursor B-cell acute lymphoblastic leukemia (B-ALL-DS) with the “difference from normal (ΔN)” technique. When possible, FISH and SNP/CGH microarray studies were performed on sorted cell fractions.

Results

93% of BM specimens submitted post chemotherapy had a clearly identifiable CD34+CD56+ population present between 0.06% and 2.6% of total non-erythroid cells. An overlapping CD34+HLA-DRheterogeneous population was observed among 92% of patients at a lower frequency (0.04%–0.8% of total non-erythroid cells). In B-ALL-DS patients, the same CD34+CD56+ HLA-DRheterogeneous expression was observed. FACS-FISH/Array studies demonstrated no residual genetic clones in the DS-specific myeloid progenitor cells.

Conclusions

Non-malignant myeloid progenitors in the regenerating BM of patients who have undergone chemotherapy for either ML-DS or B-ALL-DS express an immunophenotype that is different from normal BM of non-DS patients. Awareness of this DS-specific non-malignant myeloid progenitor is essential to the interpretation of MRD by flow cytometry in patients with ML-DS.

背景:第一疗程化疗后,流式细胞术检测可测量的残留疾病检测(MRD)是衡量急性髓性白血病(AML)患者早期反应的标准指标。髓系白血病伴唐氏综合征(ML-DS)是AML的一种不同形式。有或没有退行性椎体滑移的患者在稳态和再生造血方面的差异尚不清楚。这一认识对于准确确定ML-DS患者是否存在残留白血病至关重要。方法采用“异于正常(ΔN)”技术,对45例ML-DS或DS前体b细胞急性淋巴细胞白血病(B-ALL-DS)化疗后115例随访骨髓(BM)标本中抗原的定量表达进行标准化抗体检测。在可能的情况下,对分选的细胞组分进行FISH和SNP/CGH微阵列研究。结果化疗后提交的BM标本中,93%的标本中CD34+CD56+细胞占非红系细胞总数的0.06% ~ 2.6%。在92%的患者中观察到重叠的CD34+ hla - dr异质群体,频率较低(占非红细胞总数的0.04%-0.8%)。在B-ALL-DS患者中,观察到相同的CD34+CD56+ hla - dr异质表达。FACS-FISH/Array研究表明,ds特异性髓系祖细胞中没有残留的基因克隆。结论在接受ML-DS或B-ALL-DS化疗的再生BM患者中,非恶性髓系祖细胞表达的免疫表型与非ds患者的正常BM不同。了解这种ds特异性的非恶性髓系祖细胞对于用流式细胞术解释ML-DS患者的MRD至关重要。
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引用次数: 2
Highly sensitive single tube B-lymphoblastic leukemia/lymphoma minimal/measurable residual disease test robust to surface antigen directed therapy 高度敏感的单管B淋巴细胞白血病/淋巴瘤最小/可测量的残留疾病测试,对表面抗原导向治疗具有稳健性。
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-03-30 DOI: 10.1002/cyto.b.22120
Qi Gao, Ying Liu, Umut Aypar, Jeeyeon Baik, Dory Londono, Xiaotian Sun, Jingping Zhang, Yanming Zhang, Mikhail Roshal

Background

Measurement of minimal/measurable residual disease (MRD) in B-lymphoblastic leukemia/lymphoma (B-ALL) has become a routine clinical evaluation tool and remains the strongest predictor of treatment outcome. In recent years, new targeted anti-CD19 and anti-CD22 antibody-based and cellular therapies have revolutionized the treatment of the high-risk B-ALL. The new treatments raise challenges for diagnostic flow cytometry, which relies on the presence of specific surface antigens to identify the population of interest. So far, reported flow cytometry-based assays are developed to either achieve a deeper MRD level or to accommodate the loss of surface antigens post-target therapies, but not both.

Methods

We developed a single tube flow cytometry assay (14-color-16-parameters). The method was validated using 94 clinical samples as well as spike-in and replicate experiments.

Results

The assay was well suited for monitoring response to targeted therapies and reached a sensitivity below 10−5 with acceptable precision (coefficient of variation < 20%), accuracy, and interobserver variability (κ = 1).

Conclusions

The assay allows for sensitive disease detection of B-ALL MRD independent of CD19 and CD22 expression and allows uniform analysis of samples regardless of anti-CD19 and CD22 therapy.

背景:测量B淋巴细胞白血病/淋巴瘤(B-ALL)的最小/可测量残留疾病(MRD)已成为常规临床评估工具,并且仍然是治疗结果的最强预测因素。近年来,新的靶向抗CD19和抗CD22抗体和细胞疗法彻底改变了高危B-ALL的治疗。新的治疗方法对诊断性流式细胞术提出了挑战,流式细胞仪依赖于特定表面抗原的存在来识别感兴趣的人群。到目前为止,已报道的基于流式细胞术的检测方法是为了达到更深的MRD水平或适应靶向治疗后表面抗原的损失,但不是两者兼而有之。方法:我们开发了一种单管流式细胞仪测定法(14个颜色16个参数)。该方法使用94个临床样本以及刺入和重复实验进行了验证。结果:该测定法非常适合监测对靶向治疗的反应,并达到10-5以下的灵敏度和可接受的精度(变异系数)。
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引用次数: 4
Correction to “Flow cytometry to detect bone marrow involvement by follicular lymphoma” 对“流式细胞术检测滤泡性淋巴瘤累及骨髓”的修正
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-03-16 DOI: 10.1002/cyto.b.22111

Aren, M., Marce, S., Jurado, R., Tapia, G., Puigdefabregues, L., Raya, M., Cortes, M., Garcia-Caro, M., Junca, J., Mozas, P., Viñets, E., Cabezon, M., Plensa, E., Miljkovic, M., Sancho, J.-M., Navarro, J.-T., Zamora, L., & Sorigue, M. (2022). Flow cytometry to detect bone marrow involvement by follicular lymphoma. Cytometry Part B: Clinical Cytometry, 102(6), 427–439. https://doi.org/10.1002/cyto.b.22098

Coauthor Esther Plensa's affiliation was incorrectly given as Department of Hematology, ICO-Mataro, Badalona, Spain. The correct affiliation is Department of Hematology and Hemotherapy, Consorci Sanitari del Maresme, Hospital de Mataró, Institut Català d'Oncologia, Mataró, Spain.

We apologize for this error.

Aren,M.,Marce,S.,Jurado,R.,Tapia,G.,Puigdefabregues,L.,Raya,M.,Cortes,M.,Garcia Caro,M.,Junca,J.,Mozas,P.,Viñets,E.,Cabezon,M.,Plensa,E.,Miljkovic,M.,Sancho,J.-M,Navarro,J.-T,Zamora,L;Sorigue,M.(2022)。流式细胞术检测滤泡性淋巴瘤累及骨髓。细胞测定学第B部分:临床细胞测定学,102(6),427–439。https://doi.org/10.1002/cyto.b.22098CoauthorEsther Plensa的隶属关系被错误地指定为西班牙巴达洛纳ICO Mataro血液科。正确的附属机构是西班牙马塔罗Catalàd‘Oncologia研究所马塔罗医院血液和血液治疗科。我们对此错误深表歉意。
{"title":"Correction to “Flow cytometry to detect bone marrow involvement by follicular lymphoma”","authors":"","doi":"10.1002/cyto.b.22111","DOIUrl":"10.1002/cyto.b.22111","url":null,"abstract":"<p>Aren, M., Marce, S., Jurado, R., Tapia, G., Puigdefabregues, L., Raya, M., Cortes, M., Garcia-Caro, M., Junca, J., Mozas, P., Viñets, E., Cabezon, M., Plensa, E., Miljkovic, M., Sancho, J.-M., Navarro, J.-T., Zamora, L., &amp; Sorigue, M. (2022). Flow cytometry to detect bone marrow involvement by follicular lymphoma. <i>Cytometry Part B: Clinical Cytometry</i>, 102(6), 427–439. https://doi.org/10.1002/cyto.b.22098</p><p>Coauthor Esther Plensa's affiliation was incorrectly given as Department of Hematology, ICO-Mataro, Badalona, Spain. The correct affiliation is Department of Hematology and Hemotherapy, Consorci Sanitari del Maresme, Hospital de Mataró, Institut Català d'Oncologia, Mataró, Spain.</p><p>We apologize for this error.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9193686","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}
引用次数: 1
Comparison of flowcytometry-based scoring system for the diagnosis of early T precursor-acute lymphoblastic leukemia 基于流式细胞术的评分系统在诊断早期T前体-急性淋巴细胞白血病方面的比较
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-03-09 DOI: 10.1002/cyto.b.22119
Deepak Marballi Basavaraju, Shruti Mishra, Gaurav Chhabra, Sudarshan Chougule

Background

Early T cell precursor-acute lymphoblastic leukemia (ETP-ALL) is a hematolymphoid malignancy where the blasts demonstrate T cell differentiation markers along with stem cell and myeloid antigen expression. The differential diagnosis of ETP-ALL from non-ETP ALL and mixed phenotype acute leukemia is often challenging due to its overlapping immunophenotypic picture with co-expression of myeloid antigens. In this study, we endeavored to describe the immune-phenotype profile of ETP-ALL in our patients and compared the utility of four different scoring systems for better discrimination of these entities.

Methods

This retrospective analysis included 31 ETP-ALL out of 860 acute leukemia cases consecutively diagnosed at the two tertiary care centers. Flowcytometry-based immunophenotype was reviewed for all the cases, and the utility of four flow-based objective scorings was assessed for the diagnosis of ETP-ALL. Receiver operating curves were drawn to compare the different flow-based scoring systems.

Results

The prevalence of ETP-ALL was 40% (n = 31/77 T-ALL) in our study group, comprised mainly of adults with a median age of 20 years. The five-marker scoring system had the maximum area under the curve, followed by the seven-marker scoring system. A cut-off of ≥2.5 was more specific (sensitivity: 91%; specificity: 100%), while a score of ≥1.5 was more sensitive but slightly less specific (sensitivity: 94%, specificity: 96%).

Conclusion

The WHO criteria for the diagnosis of ETP-ALL should be followed across all laboratories to avoid confusion and for better treatment stratification. Flow-based scoring systems can be objectively employed for better detection of cases.

背景 早期 T 细胞前体-急性淋巴细胞白血病(ETP-ALL)是一种血淋巴细胞恶性肿瘤,其胚胎表现出 T 细胞分化标记以及干细胞和髓系抗原表达。ETP-ALL与非ETP ALL和混合表型急性白血病的鉴别诊断往往具有挑战性,因为其免疫表型与髓抗原的共同表达存在重叠。在本研究中,我们试图描述 ETP-ALL 患者的免疫表型特征,并比较四种不同评分系统的效用,以更好地区分这些实体。 方法 这项回顾性分析包括两个三级医疗中心连续诊断的 860 例急性白血病病例中的 31 例 ETP-ALL。对所有病例进行了基于流式细胞术的免疫表型审查,并评估了四种基于流式细胞术的客观评分对诊断 ETP-ALL 的实用性。绘制了接收者操作曲线,以比较不同的流式评分系统。 结果 在我们的研究小组中,ETP-ALL的发病率为40%(n = 31/77 T-ALL),主要由中位年龄为20岁的成年人组成。五指标评分系统的曲线下面积最大,其次是七指标评分系统。≥2.5分界点的特异性更高(灵敏度:91%;特异性:100%),而≥1.5分界点的灵敏度更高,但特异性稍低(灵敏度:94%;特异性:96%)。 结论 所有实验室都应遵循世界卫生组织的 ETP-ALL 诊断标准,以避免混淆并更好地进行治疗分层。基于流程的评分系统可客观地用于更好地检测病例。
{"title":"Comparison of flowcytometry-based scoring system for the diagnosis of early T precursor-acute lymphoblastic leukemia","authors":"Deepak Marballi Basavaraju,&nbsp;Shruti Mishra,&nbsp;Gaurav Chhabra,&nbsp;Sudarshan Chougule","doi":"10.1002/cyto.b.22119","DOIUrl":"10.1002/cyto.b.22119","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Early T cell precursor-acute lymphoblastic leukemia (ETP-ALL) is a hematolymphoid malignancy where the blasts demonstrate T cell differentiation markers along with stem cell and myeloid antigen expression. The differential diagnosis of ETP-ALL from non-ETP ALL and mixed phenotype acute leukemia is often challenging due to its overlapping immunophenotypic picture with co-expression of myeloid antigens. In this study, we endeavored to describe the immune-phenotype profile of ETP-ALL in our patients and compared the utility of four different scoring systems for better discrimination of these entities.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>This retrospective analysis included 31 ETP-ALL out of 860 acute leukemia cases consecutively diagnosed at the two tertiary care centers. Flowcytometry-based immunophenotype was reviewed for all the cases, and the utility of four flow-based objective scorings was assessed for the diagnosis of ETP-ALL. Receiver operating curves were drawn to compare the different flow-based scoring systems.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The prevalence of ETP-ALL was 40% (<i>n</i> = 31/77 T-ALL) in our study group, comprised mainly of adults with a median age of 20 years. The five-marker scoring system had the maximum area under the curve, followed by the seven-marker scoring system. A cut-off of ≥2.5 was more specific (sensitivity: 91%; specificity: 100%), while a score of ≥1.5 was more sensitive but slightly less specific (sensitivity: 94%, specificity: 96%).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The WHO criteria for the diagnosis of ETP-ALL should be followed across all laboratories to avoid confusion and for better treatment stratification. Flow-based scoring systems can be objectively employed for better detection of cases.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9076006","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}
引用次数: 0
Addition of formaldehyde releaser imidazolidinyl urea and MOPS buffer to urine samples enables delayed processing for flow cytometric analysis of urinary cells: A simple, two step conservation method of urinary cells for flow cytometry 在尿液样本中加入甲醛释放剂咪唑烷基脲和 MOPS 缓冲液,可延迟尿液细胞流式细胞术分析的处理时间:用于流式细胞仪的尿液细胞两步简单保存法
IF 3.4 3区 医学 Q1 Medicine Pub Date : 2023-03-07 DOI: 10.1002/cyto.b.22117
Paul Freund, Christopher M. Skopnik, Diana Metzke, Nina Goerlich, Jan Klocke, Emil Grothgar, Luka Prskalo, Falk Hiepe, Philipp Enghard

Introduction

Kidney diseases are a major health concern worldwide. Currently there is a large unmet need for novel biomarkers to non-invasively diagnose and monitor kidney diseases. Urinary cells are promising biomarkers and their analysis by flow cytometry has demonstrated its utility in diverse clinical settings. However, up to date this methodology depends on fresh samples, as cellular event counts and the signal-to-noise-ratio deter over time. Here we developed an easy-to-use two-step preservation method for conservation of urine samples for subsequent flow cytometry.

Methods

The protocol utilizes a combination of the formaldehyde releasing agent imidazolidinyl urea (IU) and MOPS buffer, leading to gentle fixation of urinary cells.

Results

The preservation method increases acceptable storing time of urine samples from several hours to up to 6 days. Cellular event counts and staining properties of cells remain comparable to fresh untreated samples.

Outlook

The hereby presented preservation method facilitates future investigations on flow cytometry of urinary cells as potential biomarkers and may enable broad implementation in clinical practice.

导言 肾脏疾病是全球关注的主要健康问题。目前,对新型生物标记物的需求仍未得到满足,而这些标记物可用于无创诊断和监测肾脏疾病。尿液细胞是一种很有前景的生物标记物,流式细胞仪对其进行的分析已在不同的临床环境中证明了其实用性。然而,迄今为止,这种方法依赖于新鲜样本,因为细胞事件计数和信噪比会随着时间的推移而降低。在此,我们开发了一种简单易用的两步保存法,用于保存尿液样本,以便随后进行流式细胞术分析。 方法 该方法结合使用甲醛释放剂咪唑烷基脲(IU)和 MOPS 缓冲液,温和地固定尿液细胞。 结果 该保存方法可将尿液样本的可接受保存时间从数小时延长至 6 天。细胞事件计数和细胞染色特性仍与未经处理的新鲜样本相当。 展望 本文介绍的保存方法有助于今后对尿液细胞作为潜在生物标记物进行流式细胞术研究,并可在临床实践中广泛应用。
{"title":"Addition of formaldehyde releaser imidazolidinyl urea and MOPS buffer to urine samples enables delayed processing for flow cytometric analysis of urinary cells: A simple, two step conservation method of urinary cells for flow cytometry","authors":"Paul Freund,&nbsp;Christopher M. Skopnik,&nbsp;Diana Metzke,&nbsp;Nina Goerlich,&nbsp;Jan Klocke,&nbsp;Emil Grothgar,&nbsp;Luka Prskalo,&nbsp;Falk Hiepe,&nbsp;Philipp Enghard","doi":"10.1002/cyto.b.22117","DOIUrl":"10.1002/cyto.b.22117","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>Kidney diseases are a major health concern worldwide. Currently there is a large unmet need for novel biomarkers to non-invasively diagnose and monitor kidney diseases. Urinary cells are promising biomarkers and their analysis by flow cytometry has demonstrated its utility in diverse clinical settings. However, up to date this methodology depends on fresh samples, as cellular event counts and the signal-to-noise-ratio deter over time. Here we developed an easy-to-use two-step preservation method for conservation of urine samples for subsequent flow cytometry.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The protocol utilizes a combination of the formaldehyde releasing agent imidazolidinyl urea (IU) and MOPS buffer, leading to gentle fixation of urinary cells.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The preservation method increases acceptable storing time of urine samples from several hours to up to 6 days. Cellular event counts and staining properties of cells remain comparable to fresh untreated samples.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Outlook</h3>\u0000 \u0000 <p>The hereby presented preservation method facilitates future investigations on flow cytometry of urinary cells as potential biomarkers and may enable broad implementation in clinical practice.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22117","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9413362","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}
引用次数: 0
期刊
Cytometry Part B: Clinical Cytometry
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