髓系白血病转基因小鼠模型骨髓免疫表型分析

目的 分析髓系白血病转基因小鼠模型的免疫表型分型特点.方法 根据小鼠骨髓各系列分化抗原表达特性,结合多色流式细胞术分析5只髓系白血病转基因小鼠和10只BL6健康对照组小鼠的骨髓免疫表型分型特点,并通过细胞周期分析进一步确认细胞增殖情况.结果 白血病转基因小鼠的骨髓细胞中Mac-1+Gr-1+细胞、c-Kit+细胞的表达率分别为(72.6±6.5)%、(20.5±4.8)%,明显高于健康对照组的(52.8±4.8)%、(2.1±0.3)%,差异有统计学意义(t值分别为6.66、12.66,P均＜0.01);而B220+、CD3+、CD41+和Ter119+细胞的表达率分别为(2.7±1.1)%、(1.2±0.3)%、(1.2±0.6)%及(2.8±1.1)%,明显低于健康对照组的(20.2±2.1)%、(6.6±1.3)%、(4.7±1.1)%及(10.6±1.2)%,差异有统计学意义(t值分别为-17.63、-8.69、-6.30、-12.28,P均＜0.01).细胞周期中的S期和G2/M期的表达率分别为(25.7±4.2)%和(21.1±4.2)%,明显高于健康对照组的(11.8±2.1)%和(8.9±1.8)%,差异有统计学意义(t值分别为8.59、7.98,P均＜0.01).结论 髓系白血病小鼠模型的髓系标志Mac-1、Gr-1及造血干祖细胞标志c-Kit的表达明显增高,而B淋巴细胞系标志(B220)、T淋巴细胞系标志(CD3)、巨核细胞系标志(CD41)和红细胞系标志(Ter119)的表达明显受抑.     

利用多指标流式细胞术和外周血标本进行白血病免疫分型的研究

目的:利用CD45/SCC组合及设门技术的多参数流式细胞术,探讨采用外周血标本在白血病免疫分型中的技术要点和资料分析中的注意事项。方法:检测标本以CD45标记,构建CD45/SSC散点图,采用一组针对不同白细胞系列的单克隆抗体进行三色或双色荧光染色,用设门技术对白血病细胞群体加以限定,进行免疫表型分析。结果:113例急性白血病患者中106例在CD45/SSC散点图上出现可供分析白血病细胞独立群体,占分析标本的94%。描述了不同白血病免疫分型的特点。讨论了运用多参数FCM白血病免疫分型的技术要点和应注意的事项。结论:多数白血病标本在CD45/SCC散点图上可出现白血病细胞群体,对白血病细胞群体设门进行分析,可迅速而可靠地获取免疫分型结果。     

急性淋巴细胞白血病免疫分型研究

使用一组单克隆抗体及抗ＩｇＭ多抗对２４例急性淋巴细胞白血病（ＡＬＬ）细胞表面抗原标志进行分析。结果表明：急性非Ｔ细胞白血病（Ｎｏｎ－Ｔ－ＡＬＬ）为６２．５０％，急性Ｔ细胞白血病（Ｔ－ＡＬＬ）为２９．１７％，急性未分化型白血病（ＡＵＬ）为４．１７％，急性非淋巴细胞白血病（ＡＮＬＬ－Ⅱ）为４．１７％。Ｎｏｎ－Ｔ－ＡＬＬ－Ⅰ型以ＨＬＡ－ＤＰ＋，Ⅱ型以ＣＤ１９＋，Ⅲ型以ＣＤ１０＋，Ⅳ型以ＣＤ２０＋，Ⅵ型以ＳＩｇＭ＋为诊断条件。Ｔ－ＡＬＬ－Ⅰ期以ＣＤ７＋、Ⅲ期以ＣＤ３＋、ＣＤ４＋或ＣＤ８＋为诊断条件，其它表面抗原变化不定。Ｔ－ＡＬＬ中ＨＬＡ－ＤＰ表达率达４２．８６％。２４例ＡＬＬ中３例为Ｂ－Ｔ杂合型白血病，占１２．５０％，均分布在Ｎｏｎ－Ｔ－ＡＬＬ之中。ＦＡＢ形态学分型符合率为９５．６５％。非诊断性表面抗原标志的变化是进一步分亚型的依据，也可能与药物敏感性等治疗和预后相关，值得进一步探讨     

多形性横纹肌肉瘤临床病理特征分析

目的 探讨多形性横纹肌肉瘤（pleomorphic rhabdomyosarcoma, PRMS）的临床病理学特征及鉴别诊断要点。方法 回顾性收集2008年6月至2023年3月苏州大学附属第一医院收治的PRMS患者的临床表现、病理学特征、免疫表型、治疗经过,通过电话随访获取患者生存状态及有无复发和转移。结果 共纳入6例PRMS患者,其中男性5例,女性1例;年龄29～77岁,平均年龄54.17岁;发病部位分别为右上臂、右肾盂、左鼻窦/颈部/下颌、右肩背、右臀大肌和鼻咽部。4例临床资料完整的患者中,1例表现为涕中带血,喉部异物感;3例表现为肿块进行性增大,其中2例伴压痛,影像学均提示占位性病变。组织学上,3例（50%）有凝固性坏死,4例（66.7%）呈典型的多形性肉瘤形态,2例（33.3%）以异型的梭形细胞为主。免疫组化染色显示：6例（100%）弥漫表达desmin,5例（83.3%）灶性表达myogenin,4例（66.7%）灶性表达MyoD1,1例（16.7%）灶性表达SMA;Ki-67增殖指数30%～70%。6例患者均接受手术治疗;4例患者获得完整随访资料,均于术后进行放疗和（或）化...     

尖锐湿疣病损组织粗提蛋白对树突状细胞免疫表型的影响

目的 研究尖锐湿疣病损组织粗提蛋白对树突状细胞免疫表型的影响。方法 贴壁法分离脐血和外周血中的单核细胞,流式细胞仪检测树突状细胞特异性表面分子的表达。结果 rhGM-CSF、rhIL-4和LPS可诱导脐血和外周血来源的单核细胞分化为成熟的树突状细胞。尖锐湿疣病损粗提蛋白刺激后的树突状细胞高表达CD86穴脐血91.7%熏外周血88.6%雪和HLA-DR穴脐血88.4%熏外周血85.0%雪熏而包皮刺激组的CD86和HLA-DR的表达较低穴CD86押脐血87.1%熏外周血77.2%;HLA-DR押脐血68.0%熏外周血59.7%雪,空白对照组的CD86表达更低穴脐血83.5%熏外周血62.4%雪,HLA-DR的表达介于前两者之间(脐血72.7%熏外周血68.3%)。结论 尖锐湿疣病损粗提蛋白使树突状细胞部分抗原递呈相关分子表达呈上调趋势,提示尖锐湿疣粗提蛋白可能增强树突状细胞的抗原递呈功能并进而促进T细胞的激活。     

A Rare Case of Adult T Cell Leukemia/Lymphoma: Clinicopathological Correlation with Autopsy Confirmation

Adult T cell leukaemia/lymphoma (ATLL) is a rare T lymphoproliferative disorder which is etiologically linked with human T cell lymphotropic virus type-1 (HTLV-1). HTLV-1 is endemic in Japan, Caribbean and Africa. The highest incidence of ATLL is in Japan although sporadic cases have been reported elsewhere in the world. We describe a case of ATLL with an unusual presentation with clinic-pathological correlation and autopsy confirmation. A 56 year old male was referred to Command Hospital (Southern Command) for an incidental finding of lymphocytosis on a routine Hemogram. Clinical examination did not reveal hepatosplenomegaly, lymphadenopathy, jaundice or skin lesions. Laboratory investigations showed lymphocytosis with predominance of atypical lymphomonocytoid cells. Immunophenotyping of the bone marrow mononuclear cells showed positivity for CD45, CD2, CD3, CD4, CD5 and negative for CD7, CD8, CD13, CD33, CD19, which is characteristic of ATLL phenotype. Clonality was confirmed by PCR for TCR gene rearrangement on post mortem tissue. He succumbed to his illness after 40 days of initial presentation and 16 days of being diagnosed as ATLL. Here, we discuss the pathogenesis and characteristics of ATLL with clinico-pathological correlation and autopsy confirmation.     

Diagnostic application of monoclonal antibody KB90 (CD11c) in acute myeloid leukaemia

Summary The expression of membrane CD11c by leukaemic blast cells was examined (indirect immunorosetting) in 75 cases of acute leukaemia (myeloid, n=60; lymphoid, n=15) and evaluated as a potential marker for the diagnostic discrimination between monocytic (AMML-M4 and AMoL-M5) and non-monocytic (M1, M2 and M3) AML subtypes. Preliminary studies of normal bone marrow cells indicated that CD11c expression was not restricted to cells of monocytic lineage but was also present, with apparent lower density, on significant proportions of mature and immature granulocytes. Examination of acute myeloid leukaemia (AML) subtypes revealed that the non-monocytic leukaemias (n=33) were CD11c^–, defined as <30% positive cells, whereas all but one of the AMML-M4 (n=13) and AMoL-M5 (n=14) cases were CD11c⁺. All 15 cases of lymphoblastic leukaemia (ALL) showed <5% CD11c⁺ blasts. Membrane CD11c expression was also compared to the more widely used markers of monocytic differentiation; cytoplasmic alpha-naphthyl acetate esterase (ANAE) and membrane CD14 expression. This analysis showed that all 13 AMML-M4 leukaemias studied, including seven cases that were CD14^– and eight that were ANAE^–, were CD11c⁺. In addition, the AMoL-M5 cases (all of which were ANAE⁺) could be phenotypically subdivided into CD11c⁺ CD14⁺ (n=9), CD11c⁺ CD14^– (n=4) and CD11c^– CD14^– (n=1) subgroups. The study also confirmed that the discriminitive ability and sensitivity of the immunorosetting procedure for the detection of membrane CD11c compared favourably to immunofluorescent staining intensities as measured by flow cytometry.     

Routine immunophenotyping of acute leukaemias

Summary Recent progress in immunophenotyping includes the availability of monoclonal antibodies (MAbs), knowledge of specificity and reactivity patterns of these reagents, and the technical improvements and standardization of immunofluorescence and immunocytology staining procedures, including flow cytometry. These advances have contributed significantly to the establishment of immunophenotyping as an essential diagnostic tool in the differential diagnosis of types of acute leukaemia. Immunophenotyping allows for the objective and reproducible distinction of acute lymphoblastic leukaemia (ALL) from acute myeloblastic leukaemia (AML) and of T-lineage from B-lineage ALL. Immunologically defined ALL and AML subtypes have been found to convey prognostic significance. Using cell lineage-specific and differentiation stage-specific MAbs, cases of T- and B-lineage ALL and of AML can be further classified into a number of different subtypes. Routine immunophenotyping concentrates on the diagnostic enquiry into a few major, clinically relevant subtypes; only a limited number of crucial reagents are employed that are commercially available. The simplification and standardization of discriminatory immunomarker panels make immunophenotyping a reliable diagnostic instrument for the provision of critical data to make a differential diagnosis. An effort to identify the nature and origin of the blast cells precisely, immunological typing definitely plays an important part in the multiple-marker analysis of acute leukaemia (morphology, cytochemistry, karyotying, genotyping) for applied diagnostic and fundamental research purposes.     

弥漫性大B细胞淋巴瘤免疫分型与分子遗传学异常的研究

目的 分析弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)的免疫表型分型和分子遗传学异常,并探讨二者之间的相关性及其对于预后的意义.方法 用免疫组织化学链霉亲和素-过氧化物酶法检测59例DLBCL患者中CD10、BCL6、MUM1的表达,采用Hans免疫分型法将DLBCL分为生发中心B细胞(germinal center B-cell,GCB)型和非GCB型;分别应用BCL6双色断裂点分离探针、IgH/BCL2双色双融合探针及MYC双色断裂点分离探针在石蜡包埋的淋巴瘤组织切片上进行荧光原位杂交分析,检测BCL6、BCL2和MYC基因的易位和扩增情况.结果 在59例DLBCL患者中,GCB型占28.8％(17/59),非GCB型占71.2％ (42/59).BCL6、BCL2和MYC基因易位的发生率分别为24.1％(14/58)、1.7％(1/59)和5.3％(3/57),BCL6、BCL2和MYC基因扩增的发生率分别为17.2％ (10/58)、22.0％(13/59)和21.1％(12/57).BCL6扩增与BCL6易位无相关性(P=0.424),但与BCL2及MYC扩增呈正相关(列联系数C值分别为0.405和0.403,P值均＜0.01).在GCB型中BCL6易位的发生率高于非GCB型,而BCL6、BCL2或MYC扩增更常见于非GCB型,差异接近但均无统计学意义(P值分别为0.089和0.106).在单因素分析中,免疫分型和国际预后指数积分对总生存率(overall survival,OS)均有显著影响(P值分别为0.047和0.001),而BCL6易位和3基因扩增对OS率均无明显影响(P值分别为0.150和0.444);在多因素分析中,国际预后指数积分是影响OS唯一的独立预后因素(RR=3.843,P=0.017).结论 本组DLBCL患者中GCB亚型较少见,常见的遗传学异常为BCL6易位和BCL6、BCL2及MYC扩增,且3基因扩增之间密切相关,而BCL2易位的发生率低.免疫表型分型对预后的预测意义较小,遗传学异常不能用于预测DLBCL患者的预后.     

Allogeneic Hematopoietic Cell Transplantation May Alleviate the Negative Prognostic Impact of Monosomal and Complex Karyotypes on Patients with Acute Myeloid Leukemia

Monosomal karyotype (MK) and complex karyotype (CK) are well known to be associated with a very poor clinical outcome in patients with acute myeloid leukemia (AML). However, whether or not the prognostic impact of MK and CK remains relevant for patients who have undergone allogeneic hematopoietic cell transplantation (allo-HCT) is still unclear. We retrospectively analyzed the status of MK and CK, as well as other clinical laboratory features, in 148 allo-HCT AML patients at our institution and correlated with their event-free survival (EFS) and overall survival (OS) after transplantation. MK and CK were identified in 14 (9%) and 19 (13%) cases, respectively. On univariate analysis, only age (≥60 years) and WBC count (≥15 × 10⁹/L) were significant adverse predictors for EFS (P < .001 and P = .017, respectively) and OS (P = .002 and P = .021, respectively). MK, CK, and other relevant parameters analyzed did not affect the clinical outcome. Multivariable analysis confirmed that both older age and high WBC count were independent prognostic factors for a shorter OS (P = .001 and P = .003, respectively) and a shorter EFS (P < .001 and P = .001, respectively). Our results indicate that neither MK nor CK are high-risk factors in AML patients undergoing allo-HCT.