Immunophenotyping

免疫表型分型
  • 文章类型: Case Reports
    背景:脾B细胞淋巴瘤/具有突出核仁的白血病(SBLPN),即毛细胞白血病变体(HCL-v)是一种罕见的B细胞慢性淋巴增殖性疾病。主要的诊断挑战是区分SBLPN与经典毛细胞白血病(HCL-c),因为前者对治疗的反应较差,预后较差。
    目的:目的探讨3例SBLPN的临床血液学和免疫表型。
    方法:这是一项回顾性观察性研究。
    方法:从2011年到2021年,对所有诊断为HCL的病例进行了流式细胞术,收集3例CD25阴性或暗淡且血液学表现与SBLPN匹配的病例。
    方法:使用描述性统计。
    结果:所有病例均为男性。年龄从43岁到64岁不等。血红蛋白浓度中位数,白细胞总数,血小板计数为8.6g/dL,6.9×109/L,53×109/L,分别。非典型细胞为中等至大。所有三个都显示出突出的核仁。在所有病例中,骨髓活检均显示间质浸润。毛细胞CD20、CD11c、CD103CD25在一例中呈暗阳性。膜联蛋白A1在所有三个病例中均为阴性。在一个病例中进行了BRAFV600E突变分析,结果为突变阴性。
    结论:SBLPN是一种罕见的实体,通常在流式细胞术CD25阴性。然而,在昏暗的CD25阳性病例中,BRAFV600E突变分析有助于辨别SBLPN诊断并将其与HCL-c区分开。
    BACKGROUND: Splenic B-cell lymphoma/leukemia with prominent nucleoli (SBLPN) aka hairy cell leukemia variant (HCL-v) is a rare B-cell chronic lymphoproliferative disorder. The main diagnostic challenge is to differentiate SBLPN from Classical hairy cell leukemia (HCL-c), as the former faces inferior responses to therapies and a poor prognosis.
    OBJECTIVE: The aim is to discuss the clinic-hematological and immunophenotyping findings of three cases of SBLPN.
    METHODS: This is a retrospective observational study.
    METHODS: From the year 2011 to 2021, flow cytometry of all the cases with HCL diagnosis was reviewed, and three cases with negative or dim CD25 and hematological presentation matching with SBLPN were picked up.
    METHODS: Descriptive statistics is used.
    RESULTS: All the cases were male. The age ranges from 43 to 64 years. Median hemoglobin concentration, total leucocyte count, and platelet count were 8.6 g/dL, 6.9 × 109/L, and 53 × 109/L, respectively. The atypical cells were medium to large. All three showed prominent nucleoli. Bone marrow biopsies showed an interstitial pattern of infiltration in all the cases. The hairy cells were positive for CD20, CD11c, and CD103. CD25 was dim positive in one case. Annexin A1 was negative in all three cases. BRAF V600E mutation analysis was done in one case and turned out negative for the mutation.
    CONCLUSIONS: SBLPN is a rare entity, usually on-flow cytometry CD25 negative. However, in dim CD25-positive cases, BRAFV600E mutational analysis helps in discerning SBLPN diagnosis and differentiating it from HCL-c.
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  • 文章类型: Review
    微小残留病是指对化疗或放疗有抗性并导致疾病复发的白血病细胞群。MRD的评估对于做出疾病的准确预后和选择最佳治疗策略至关重要。这里,我们回顾了可用的遗传和表型方法的优缺点,并将重点放在多参数流式细胞术作为一种有前途的方法,具有更高的灵敏度,速度,标准化选项。此外,我们讨论了自动数据分析的应用如何胜过经典分析中窗户和大门的复杂组合的使用,从而消除了主观评价。
    Minimal residual disease refers to a leukemia cell population that is resistant to chemotherapy or radiotherapy and leads to disease relapse. The assessment of MRD is crucial for making an accurate prognosis of the disease and for the choice of optimal treatment strategy. Here, we review the advantages and disadvantages of the available genetic and phenotypic methods and focus on the multiparametric flow cytometry as a promising method with greater sensitivity, speed, and standardization options. In addition, we discuss how the application of automated data analysis outweighs the use of complex combinations of windows and gates in classical analysis, thus eliminating subjective evaluation.
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  • 文章类型: Case Reports
    肾脏受累的青少年原发性干燥综合征(pSS)极为罕见,报告了大约50名儿童,主要是女孩。这里,我们介绍了第一例报道的男性儿童患有青少年pSS眼表疾病(以前是干燥性角膜结膜炎),颌下腺受累,和肾小管间质性肾炎。首先,两种症状在临床上表现明显.我们在这里说明应该积极寻找肾脏参与pSS,因为青少年pSS可能与无症状的肾脏受累有关。使用多色流式细胞术对外周血细胞进行免疫表型分型,在诊断时显示了适应性(T记忆细胞和B记忆细胞)的变化,和先天免疫(自然杀伤细胞的激活增加,以及单核细胞和中性粒细胞,和中间单核细胞的代表增加)。我们的病例报告指出了肾脏检查的重要性,青少年pSS的早期诊断和治疗,以及强调国际合作以获得这种罕见疾病的更多数据。
    Juvenile primary Sjögren syndrome (pSS) with renal involvement is extremely rare, reported approximately in 50 children, predominantly girls. Here, we present the first reported case of a male child with juvenile pSS with ocular surface disease (previously keratoconjunctivitis sicca), submandibular salivary gland involvement, and tubulointerstitial nephritis. First, two symptoms were clinically apparent at presentation. We illustrate here that kidney involvement in pSS should be actively looked for, as juvenile pSS may be associated with asymptomatic renal involvement. Immunophenotyping of peripheral blood cells using multicolor flow cytometry revealed at the time of diagnosis changes in both adaptive (T memory cells and B memory cells), and innate immunity (an increased activation of natural killer cells, as well as monocytes and neutrophils, and an increased representation of intermediate monocytes). Our case report points to the importance of kidney examination, early diagnosis and therapy in juvenile pSS, as well as highlights international collaboration to obtain more data for this rare disease.
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  • 文章类型: Review
    To improve the diagnosis of atypical lymphocytes and reduce the misdiagnosis rate,we analyzed the medical records of 2 cases with cell morphology suggestive of atypical lymphocytes.One case was diagnosed with infectious mononucleosis and the other with aggressive NK cell leukemia.The purpose of this paper is to emphasize that the diagnosis of atypical lymphocytes based only on morphological interpretation of cells may be incorrect,which should be combined with clinical symptoms,signs,imaging examination,cell immunophenotype,and disease outcome.
    为探讨异型淋巴细胞的鉴别诊断,减少误诊率,分析2例细胞形态提示异型淋巴细胞的病历资料,其中1例诊断传染性单核细胞增多症,1例诊断侵袭性NK细胞白血病。本文旨在强调仅凭细胞形态学判读的异型淋巴细胞可能有误,需结合患者的临床症状、体征、影像学检查、细胞免疫表型、疾病转归等综合诊断。.
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  • 文章类型: Journal Article
    骨髓增生异常综合征(MDS)的诊断很复杂。骨髓单核细胞区室的流式细胞术分析可能是有帮助的,但它是高度主观的,非专门组的可重复性尚不清楚。通过流式细胞术对红系谱系的分析正在出现,因为它可能更具可重复性和更容易进行。同时保持高诊断性能。
    我们回顾了这方面的证据,包括1)使用公认的标志物-CD71和CD36-和其他不太公认的标志物和参数;2)使用红细胞谱系流式细胞术评分;3)其他方面,包括计算工具的出现和流式细胞术在诊断之外的作用.最后,我们讨论目前证据的局限性,包括1)样品处理方案和试剂对结果的影响,2)缺乏标准的门控策略,3)现有出版物中的概念化和设计问题。
    最后,我们通过流式细胞术对红细胞谱系分析提供了当前使用的建议-以及我们个人对价值的看法。
    The diagnosis of myelodysplastic syndrome (MDS) is complex. Flow cytometric analysis of the myelomonocytic compartment can be helpful, but it is highly subjective and reproducibility by non-specialized groups is unclear. Analysis of the erythroid lineage by flow cytometry is emerging as potentially more reproducible and easier to conduct, while keeping a high diagnostic performance.
    We review the evidence in this area, including 1) the use of well-established markers - CD71 and CD36 - and other less well-established markers and parameters; 2) the use of flow cytometric scores for the erythroid lineage; and 3) additional aspects, including the emergence of computational tools and the roles of flow cytometry beyond diagnosis. Finally, we discuss the limitations with the current evidence, including 1) the impact of the sample processing protocol and reagents on the results, 2) the lack of a standard gating strategy, and 3) conceptualization and design issues in the available publications.
    We end by offering our recommendations for the current use - and our personal take on the value - of the analysis of erythroid lineage by flow cytometry.
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  • 文章类型: Journal Article
    经典霍奇金淋巴瘤,结节性淋巴细胞为主的霍奇金淋巴瘤,和富含T细胞/组织细胞的大B细胞淋巴瘤形成一组独特的淋巴瘤,具有相似的形态学生长模式(在突出的细胞背景中偶有肿瘤细胞),在病理生物学上相关。通过流式细胞术区分这些实体历来是困难的;然而,我们实验室开发了能够对这些淋巴瘤进行免疫分型的抗体-荧光染料组合.此外,背景反应性淋巴细胞的表征可以帮助缩小鉴别诊断范围。这篇综述总结了在这一独特的淋巴瘤组中发现的肿瘤和反应性人群的免疫表型特征和见解。
    Classic Hodgkin lymphoma, nodular lymphocyte predominant Hodgkin lymphoma, and T cell/histiocyte-rich large B cell lymphoma form a unique set of lymphomas with similar morphologic growth patterns (occasional neoplastic cells within a prominent cellular cell background) that are pathobiologically related. Distinguishing these entities has been historically difficult by flow cytometry; however, our laboratory has developed antibody-fluorochrome combinations capable of immunophenotyping these lymphomas. Additionally, characterization of the background reactive lymphocytes can aid in narrowing the differential diagnosis. This review summarizes the immunophenotypic features and insights of the neoplastic and reactive populations found in this unique group of lymphomas.
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  • 文章类型: Journal Article
    流式细胞术的自动化最近从部分实验室自动化和机器人胰岛发展起来,更全面的集成系统。本文回顾了三家制造商的最新样品制备系统:BeckmanCellMek,SysmexPS-10和BDFACSDuet。这三种仪器能够执行流式细胞术样品处理中的许多手动步骤(移液,染色,lysing,washing,fixing).一般说明,能力,优势,并对每个系统的缺点进行了比较。总的来说,这些系统有可能成为今天繁忙的临床流式细胞术实验室的主要项目,并为实验室工作人员节省大量的动手时间。
    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.
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  • 文章类型: Journal Article
    单克隆B细胞淋巴细胞增多症(MBL)在2008年WHO分类中作为新实体引入,定义为循环B细胞克隆<5.109/L,没有器官肿大和先前和/或同时发生的淋巴增殖性疾病。MBL被细分为MBLCLL类型(最常见),MBL非典型CLL型和MBL非CLL型(文献报道较少)。这里是诊所,细胞学,从34例病例中描述了MBL非CLL型的免疫学和遗传学特征。正如以前报道的那样,本病例与MZL具有免疫学和遗传相似性,可能与新提出的实体CBL-MZ(边缘区起源的克隆B细胞淋巴细胞增多症)相关.此外,少数病例与脾弥漫性红髓淋巴瘤(SDRPL)相似。总之,根据文献,具有非CLL型(与CBL-MZ同化)的MBL可能是MZL和/或SDRPL的恶化前状态。
    The monoclonal B-cell lymphocytosis (MBL) introduced as new entities in the 2008 WHO classification, are defined by circulating B-cell clone < 5.109/L without organomegaly and previous and/or simultaneous lymphoproliferative disorders. The MBL were subclassified in MBL CLL type (the most frequent), MBL atypical CLL type and MBL non-CLL type (rarely reported in literature). Here the clinic, cytologic, immunologic and genetic features of MBL non-CLL type were described from a series of 34 cases. As previously reported, present cases presented immunologic and genetic similarities to MZL and could be associated to the new proposed entity CBL-MZ (clonal B-cell lymphocytosis of marginal zone origin). In addition, few cases presented similarities to splenic diffuse red pulp lymphoma (SDRPL). In conclusion, according to the literature, MBL with non-CLL type (assimilated to CBL-MZ) may be a premalignant state of MZL and/or SDRPL.
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  • 文章类型: Review
    目的:在癌症患者的血液中发现循环肿瘤细胞(CTCs),包括头颈部鳞状细胞癌(HNSCC)。目的是回顾CTC在HNSCC患者中的应用的最新状态。
    方法:PubMed中的英文文章。
    方法:综述了文献中关于CTCs在HNSCCs中的所有研究。
    结论:关于CTC在HNSCC中的诊断和预后价值有新的信息。证据还强调了各种下游分析方法相对于循环肿瘤DNA(ctDNA)的优势,例如单CTC分析,离体,和CTC的体内扩增。多种表型表面标记(细胞角蛋白,EpCAM,波形蛋白,等。),用于使用不同的免疫测定法表征CTC,可以预测疾病进展以及患者对治疗效果的反应。CTC中的免疫检查点抑制剂状态也可以更好地了解治疗。CTC的克隆扩增和CTC的单细胞分析是当今最新兴的领域,可以提供对肿瘤进化机制以及治疗功效的理解。虽然几项临床试验正在进行中,在CTC的检测和表征方面仍然存在局限性。由于缺乏黄金标准协议,CTC计数方法的敏感性和特异性各不相同。
    结论:在将CTC用作HNSCC患者的临床治疗中的诊断和预后标志物之前,仍需要进行前瞻性临床试验。
    Circulating tumor cells (CTCs) are found in the blood of patients with cancer, including head and neck squamous cell carcinomas (HNSCCs). The aim is to review the most up-to-date status of CTCs for applications in patients with HNSCC.
    English articles in PubMed.
    All the studies on CTCs in HNSCCs in the literature were reviewed.
    There is emerging information on the diagnostic and prognostic value of CTCs in HNSCCs. Evidence also highlights the advantages of various downstream analysis approaches over circulating tumor DNA (ctDNA), such as single-CTC analysis, ex vivo, and in vivo expansion of CTCs. Multiple phenotypic surface markers (cytokeratins, EpCAM, vimentin, etc.), used for CTCs characterization using different immunoassays, could predict disease progression as well as patients\' response to treatment efficacy. Immune checkpoint inhibitors\' status in CTCs could also provide better insight into treatment. Clonal expansion of CTCs and single-cell analysis of CTCs are the most emerging fields nowadays which may offer an understanding of the mechanism of tumor evolution as well as therapeutic efficacy. Although several clinical trials are ongoing, limitations still exist in the detection and characterization of CTCs. Due to the lack of a gold standard protocol, the sensitivity and specificity of CTC enumeration methods vary.
    Prospective clinical trials are still needed before CTCs can be employed as diagnostic and prognostic markers in the clinical management of patients with HNSCC.
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