This second issue of 2021, in addition to the highlighted Special Section on Mycosis Fungoides and Sezary Syndrome (Craig, 2021), also include more papers than usual due to the abundance of accepted papers backlogged to publish. We expect the next few issues of 2021 to be similarly expanded until our abundance of riches are brought under better control. While all these contributions have been available on‐line, our publisher Wiley and the Editorial Office take very seriously our additional responsibility to bring accepted papers to their ultimate destination between two covers of the print or on‐line Journal. Thus, following find a description of these additional contributions.

Anti‐CD20 monoclonal antibody therapy, while initially envisioned as an anti‐neoplastic modality, has also found utility in targeting B‐cells involved in autoimmune disorders. The primary goal of such therapy is to eliminate the pool of pathogenic memory B‐cells that fuel the autoimmune process while simultaneously sparing the reservoir of those cells involved in the production of needed protective host antibodies. Effective therapy tends to profoundly deplete circulating B‐cells for up to a full year after administration. In this issue Gatti et al., 2021 describe a technical approach devised by the Italian Society for Clinical Cell Analysis [ISCCA] as a prospective national guideline. The study outlines an eight‐color 10‐marker high resolution flow cytometry protocol that can be applied to patients treated with anti‐CD20 monoclonal antibodies. The goal of the guidelines is to provide uniform support for needed clinical decision making in the safe usage of anti‐CD20 therapies. This current study underscores techniques and methodologies found in recent publications in this journal assessing the presence of lymphoid and plasma cell malignancies (Cherian et al., 2019; DiGiuseppe et al.,2019; Jevremovic et al., 2019; Seegmiller et al., 2019; Soh et al., 2020).

The RAM immunophenotype was described in 2016 from the Children's Oncology Group [COG] as a subtype of acute myelogenous leukemia (AML). This rare subtype of AML was characterized clinically by extremely poor prognosis (Eidenschink Broderson et al., 2016). This subtype of AML was immunophenotypically characterized with bright CD56, reduced expression of CD45 and CD38 and the absence of HLA‐DR expression. In this issue Panda et al., 2021 report on studying 1102 patients with a median age of 2 years, of which 11 were RAM‐AML patients. Additionally, CD36 expression was found to be consistently absent from the immunophenotypic signature. In addition to immunophenotyping, these authors included the morphological, cytochemical, cytogenetic and molecular characteristics comparing RAM‐AML to non‐RAM patients. Other recent studies related to the diagnosis of AML may be found in the journal in Rossi et al., 2020 and Zhou et al., 2019.

Chimeric antigen receptor (CAR) T‐cells provide a promising approach to the treatment of hematologic malignancies and solid tumors. Flow cytometry is a powerful analytical. modality, which plays an expanding role in all stages of CAR T therapy, from lymphocyte collection, to CAR T‐cell manufacturing, to in vivo monitoring of the infused cells and evaluation of their function in the tumor environment (Maryamchick et al., 2020). One of the most important factors that influences the outcome and durability of the response is the persistence of the CAR T‐cell infusion. In this issue Demaret et al., 2021 developed a flow cytometric test to monitor Yescarta™ (Kite, Gilead) CAR T‐cells in infusion bags and whole blood using the CAR T‐cells co‐expression of CD19+ and CD3+. This assay also provided the relative expression of CD4+ and CD8+ CAR T subsets and permits longitudinal studies of the CAR T‐cells persistence in treated patients.

As clinical labs expanded towards more comprehensive 10‐marker studies, it became important in many settings to ‘cocktail’ the antibodies and validate them into internal laboratory developed tests [LDT]. This saved tedious pipetting time as well as further guaranteed that single monoclonal antibodies would not be left out of a particular staining panel by incorrectly omitting them from the patient's staining sample. That is, it obviated the uncomfortable question of ‘…was a particular cell population lacking expression of a key marker, or was the antibody mistakenly omitted during the preparatory staining process?’ However, creating and storing such liquid monoclonal antibody cocktails requires routine validation as well as the capacity to reasonably store such refrigerated preparations. It was evident from such practices that a vendor's validated pre‐made monoclonal antibody cocktail would be extremely helpful in increased technical efficiency, standardization and patient safety. Furthermore, if such preparations were dried and stable for months and did not require refrigeration, additional obvious benefits were derived. However, the needed payment for such a preparation needed to be cost‐effective. An unsolved problem with such an approach is that ‘standardization’ and ultimate agreement of what combination of antibodies ‘the correct staining panel should consist of’ in our field, is to the moment, oxymoronic. This Editor recalls well an early Bethesda meeting to define ‘agreed upon’ staining panels; our passionate invited colleagues were admonished upon entering the auditorium that biting, spitting, kicking or any physical violence regarding such discussions would not be tolerated ☺!

Such disagreements aside, thus the attraction in selected laboratory settings for the dried ClearLLab 10C Panels which consist of a B‐ T‐ and two myeloid‐lineage staining tubes, each containing 10 monoclonal antibody conjugates to diagnose leukemia or lymphoma. Hedley et al., 2021 conducted a study of four clinical sites using 453 patient specimens to compare the dried preparations to the laboratories LDT and found a 98% agreement between the two methods in detecting the presence or absence of malignancy. The testing was performed only on the vendors Navios systems; other platforms were not part of the comparison.

Finally, included in this issue are two letters to the Editor (Hill et al., 2021; Liu et al., 2021) and two Brief Definitive Reports (Yu et al., 2021; Collins et al., 2021).

2021年第二期，除了突出的《真菌病和塞萨里氏综合症的特殊章节》（克雷格，2021年）之外，由于积压的要发表的论文数量很多，因此收录的论文也比平时更多。我们预计2021年的下几个问题将以同样的方式扩大，直到我们的丰富财富得到更好的控制。尽管所有这些文稿都可以在线获得，但是我们的出版商Wiley和编辑部非常重视我们的额外责任，即将接受的论文带到印刷版或在线《 Journal》的两本书之间，并最终运送到它们的最终目的地。因此，下面找到这些额外贡献的描述。

抗CD20单克隆抗体疗法最初被设想为一种抗肿瘤方法，但也已发现可用于靶向涉及自身免疫性疾病的B细胞。这种治疗的主要目的是消除致病性记忆B细胞的池，这些池促进自身免疫过程，同时保留那些与产生所需保护性宿主抗体有关的细胞的池。有效治疗往往会在给药后长达整整一年的时间内大量消耗循环B细胞。在本期Gatti等人的文章（2021年）中描述由意大利临床细胞分析协会（ISCCA）设计的技术方法，作为前瞻性的国家指南。这项研究概述了可用于抗CD20单克隆抗体治疗的患者的八色10标记高分辨率流式细胞术方案。该指南的目标是为安全使用抗CD20疗法所需的临床决策提供统一支持。这项最新研究强调了该杂志最新出版物中发现的评估淋巴样和浆细胞恶性肿瘤存在的技术和方法（Cherian等人，2019 ; DiGiuseppe等人，2019 ; Jevremovic等人，2019 ; Seegmiller等人，2019 ; Soh等人，2020）。

2016年，儿童肿瘤学组[COG]将RAM免疫表型描述为急性骨髓性白血病（AML）的一种亚型。这种罕见的AML亚型在临床上以极差的预后为特征（Eidenschink Broderson et al。，2016）。这种AML亚型的免疫表型特征为明亮的CD56，减少的CD45和CD38表达以及不存在HLA-DR表达。在本期杂志中，Panda等人，2021年报告研究了1102名中位年龄为2岁的患者，其中11名为RAM‐AML患者。另外，发现免疫表型特征始终不存在CD36表达。除了免疫表型分析外，这些作者还包括将RAM‐AML与非RAM患者进行比较的形态，细胞化学，细胞遗传学和分子特征。与AML诊断有关的其他最新研究可在Rossi等人（2020）和Zhou等人（2019）的杂志中找到。

嵌合抗原受体（CAR）T细胞为血液系统恶性肿瘤和实体瘤的治疗提供了一种有前途的方法。流式细胞仪是一种功能强大的分析方法。这种方法在CAR T治疗的各个阶段都发挥着日益重要的作用，从淋巴细胞收集到CAR T细胞生产，再到体内监测注入的细胞并评估其在肿瘤环境中的功能（Maryamchick等人，2020年））。影响结果和反应持久性的最重要因素之一是CAR T细胞输注的持续性。在本期杂志中，Demaret等人，2021年开发了一种流式细胞术测试，以使用CD19 +和CD3 +的CAR T细胞共表达来监测输液袋和全血中的Yescarta™（Kite，Gilead）CAR T细胞。该测定法还提供了CD4 +和CD8 + CAR T亚型的相对表达，并允许对治疗患者的CAR T细胞持久性进行纵向研究。

随着临床实验室向更全面的10标记研究扩展，在许多情况下“结合”抗体并将其验证为内部实验室开发的测试[LDT]变得很重要。这节省了单调乏味的移液时间，并进一步确保了通过从患者的染色样品中不正确地省略单个单克隆抗体，不会将单个单克隆抗体排除在特定的染色面板之外。也就是说，它消除了“……特定细胞群是否缺乏关键标志物的表达，还是在准备性染色过程中错误地遗漏了抗体？”这一令人不安的问题。然而，创建和存储这种液体单克隆抗体混合物需要常规验证以及合理存储这种冷藏制剂的能力。从这种做法可以明显看出，卖方 经验证的预制单克隆抗体混合物将对提高技术效率，标准化和患者安全性非常有帮助。此外，如果将这些制剂干燥且稳定数月且不需要冷藏，则可获得其他明显的好处。但是，此类准备工作所需的付款必须具有成本效益。这种方法的一个尚未解决的问题是，“标准化”和对抗体组合的正确选择（在我们的领域中，“正确的染色小组应该组成”）的最终共识是矛盾的。这位编辑很好地回顾了贝塞斯达（Bethesda）早期的一次会议，该会议定义了“商定的”染色小组。我们热情洋溢的受邀同事在进入礼堂后被告诫，他咬人，随地吐痰，

除了这些分歧之外，干的ClearLLab 10C面板在选定的实验室环境中具有吸引力，该面板由BT和两个髓系谱管组成，每个都包含10个诊断白血病或淋巴瘤的单克隆抗体结合物。Hedley等人（2021年）使用453个患者标本对四个临床部位进行了研究，以将干燥的制剂与实验室LDT进行比较，发现这两种方法在检测恶性肿瘤存在与否方面有98％的一致性。该测试仅在供应商Navios系统上执行；其他平台不属于比较范围。

最后，在本期中包括给编辑的两封信（Hill等人，2021； Liu等人，2021）和两份简短的权威性报告（Yu等人，2021； Collins等人，2021）。