脐血间充质干细胞体外培养方法的改良及其生物学特性研究

目的:对体外分离培养脐血间充质干细胞(UCB-MSCs)的传统方法进行改进,以提高培养成功率,同时观察其生物学特性.方法:无菌条件下取28份足月产新生儿脐血,以密度梯度离心法分离其中的单个核细胞,以含10% FBS的α-MEM培养基进行体外培养.原代培养5～7 d后半量换液,后每隔3～4 d全量换液1次.细胞贴壁之后按改良方法进行培养:方法一,当皿底圆形巨核细胞融合、梭形成纤维样细胞脱落时将细胞悬液移入新的皿中培养;方法二,待皿底圆形巨核细胞渐渐占取优势时,将培养基换为含15% FBS的α-MEM,当圆形巨核细胞大部脱落后换回含10?S的α-MEM培养基.显微镜下观察UCB-MSCs的形态,流式细胞仪测定细胞免疫表型,体外成骨及成脂诱导并进行细胞化学染色. 结果:28份脐血中20份培养出贴壁细胞,其中13份培养出能融合且可稳定传代的成纤维样细胞,成功率为46.4%,可传至22代而无形态上的变化,而且强烈表达CD105、CD29等MSCs表面标志,而CD34、CD45和CD106等表达呈阴性.在特定条件下,UCB-MSCs可分化为成骨细胞和脂肪细胞.结论:脐血中存在MSCs,培养方法经改良后可提高UCB-MSCs的培养成功率.UCB-MSCs具有与其他来源的MSCs类似的表型及分化潜能,且易于体外扩增、传代稳定,可望在骨组织工程学研究中有广阔的应用前景.     

126例慢性淋巴细胞白血病免疫表型分析

目的分析慢性淋巴细胞白血病(CLL)的免疫表型特征。方法应用系列相关单克隆抗体通过三色/四色流式细胞术对126例CLL进行免疫表型分析。结果126例CLL中117例为B系来源(92.9%),9例为T系来源(7.1%)。117例B-CLL均表达CD19,其他B系抗原CD20、CD22、CD23阳性率分别为88.6%、66%、74.2%。无一例表达FMC7和CD10。所有B-CLL中CD5 B-CLL占79.1%,CD5-B-CLL占20.9%。45例B-CLL进行CD38及ZAP-70的检测。共14例CD38 ZAP-70 ,4例CD38 ZAP-70-,3例CD38-ZAP-70 ,24例CD38-ZAP-70-。CD38与ZAP-70的表达具有相关性(P<0.05)。9例T-CLL患者均为成熟T细胞表型,仅表达T系抗原,不表达任何B系抗原。结论了解CLL的免疫表型特点对此类疾病的诊断及治疗后微小残余病灶(MRD)的检测具有重要意义。而免疫表型与预后的关系需作进一步研究。     

急性白血病免疫表型分析

目的探讨成人急性白血病(AL)的抗原表达及其临床意义。方法采用流式细胞仪检测分析45例AL的免疫表型。结果19例急性淋巴细胞白血病(ALL)髓系抗原阳性表达率为16.3%,其中CD13为14.6%,CD33为8.4%,ALL髓系抗原阳性表达率的CR率明显低于髓系抗原阴性表达的CR率(分别为35.2%和69.7%)。26例急性髓性白血病(AML)淋系抗原表达率为23.8%,其中CD7为12.4%,CD19为9.7%;AML淋系抗原阳性表达的CR率低于淋系抗原阴性表达的CR率(分别为41.6%和62.1%)。结论急性白血病免疫表型使诊断更精细、确切,对判断预后、鉴别诊断等方面意义重大。     

非霍奇金淋巴瘤细胞白血病的免疫分型与临床

采用ＡＢＣ酶抗体法对３０例非霍奇金淋巴瘤细胞白血病进行了免疫分型，结果Ｂ细胞型２０例，Ｔ细胞型１０例。病理分类以弥漫小裂细胞型、淋巴母细胞型及小淋巴细胞型易合并白血病。结合文献对Ｔ、Ｂ淋巴瘤细胞白血病的发生因素及临床特点等加以讨论。     

Expression of myeloid differentiation antigens in acute nonlymphocytic leukemia: Increased concentration of CD33 antigen predicts poor outcome—A report from the childrens cancer study group

Ninety-eight cryopreserved specimens of acute nonlymphocytic leukemia (ANLL) cells obtained at initial diagnosis of children enrolled on the Childrens Cancer Study Group 251 protocol (CCG 251) were examined by indirect immunofluorescence using four monoclonal antibodies to myeloid differentiation antigens. The relationship between the level of differentiation of ANLL cells as determined by their antigen phenotype and the clinical outcome of treatment, including complete remission (CR) rate, survival, and event-free survival, was evaluated. Most leukemic specimens were determined to express the CD33 antigen (L4F3), a 67-kD protein. Because the level of differentiation of normal myeloid cells is reflected by the concentration of the CD33 antigen expressed, samples were categorized as CD33-bright (immature) versus CD33-dull (mature). Patients with CD33-bright leukemic blasts had a marginally inferior CR rate to those with CD33-dull blasts (P = 0.08). With respect to survival and event-free survival, there was a significantly inferior outcome in the CD33-bright patients (P = 0.04 and P = 0.06, respectively). Reactions of ANLL with anti-CD15 antibody (1G10), anti-CD36 antibody (5F1), or anti-CD17 antibody (T5A7) did not predict clinical outcome. This study indicates that patients whose ANLL blasts displayed the CD33 antigen in an amount associated with immature myeloid cells experienced a worse outcome than patients with ANLL blasts that expressed a phenotype associated with more mature cells. © 1992 Wiley-Liss, Inc.     

急性淋巴细胞白血病免疫表型与临床特征分析

目的探讨急性淋巴细胞白血病不同免疫表型的临床特点.方法使用8种抗CD单抗测定90例成人急性淋巴细胞白血病免疫表型,在此基础上对纯T,B系表型的病例进行临床特征分析.结果急性淋巴细胞免疫分型与FAB形态学分型基本符合,T系白细胞总数明显高于B系,在淋巴结肿大、肝脾肿大等体征上两者无显著性差异,两型的预后特点有待进一步观察.结论急性淋巴细胞白血病的免疫分型对其诊断和治疗有指导意义.     

人外周血淋巴细胞体外扩增培养前后19种细胞表型研究

目的:观察35例恶性肿瘤患者外周血淋巴细胞在体外经扩增培养后19种细胞表型的变化。方法:体外诱导扩增恶性肿瘤患者外周血淋巴细胞及单个核细胞,应用流式细胞术测定扩增前后CD3 等19种不同表型细胞百分比的变化。结果:经体外扩增培养后,CD3 、CD3 CD8 、CD3 CD16-CD56 、CD3 CD16 CD56-、CD3 CD16 CD56 ,CD45RO 细胞及其亚型CD8 CD45RO 、CD8 CD28-细胞比例较培养前明显增加(P<0.01);而CD19 、CD3 CD4 、CD3-CD16-CD56 、CD3-CD16 CD56-、CD3-CD16 CD56 ,CD45RA 细胞及其亚型CD4 CD45RA 、CD8 CD45RA ,CD4 CD45RO ,CD28 细胞及其亚型CD8 CD28 细胞比例则明显的降低(P<0.01)。结论:培养扩增后细胞具有直接杀伤肿瘤细胞的细胞毒活性。     

CD38‐negative relapse in multiple myeloma after daratumumab‐based chemotherapy

We present a case report of a patient relapsing after anti‐CD38 treatment (daratumumab). The phenotype of the disease changed during this treatment, and the myeloma clone became CD38 negative and daratumumab refractory. We expected clonal shift, however, based on immunophenotyping, cytogenetics and arrayCGH; the clone was identical as before daratumumab‐based treatment with the exception of CD38 negativity. We suggest that the downregulation or loss of CD38 might be an epigenetic “escape mechanism” of malignant plasma cells from antibody‐based treatment. The aim of our study was to point out the pitfalls of immunophenotyping and cytogenetics in both assessing the minimal residual disease and clone detection after monoclonal antibody‐based therapy.     

Relevance of immunophenotypes to prognostic subgroups of age,WBC, platelet count,and cytogenetics in de novo acute myeloid leukemia

Li X, Li J, Du W, Zhang J, Liu W, Chen X, Li H, Huang S, Li X. Relevance of immunophenotypes to prognostic subgroups of age, WBC, platelet count, and cytogenetics in de novo acute myeloid leukemia. APMIS 2010. Immunophenotyping is one of the independent prognostic factors in acute myeloid leukemia (AML). Relevance of immunophenotypes to prognostic subgroups of age, white blood cells (WBC), platelet count, and cytogenetics in de novo AML was comprehensively investigated in this study for the first time. Human leukocyte antigen (HLA)‐DR and CD14 expression associated with the elderly, highest WBC count, and unfavorable‐risk cytogenetics; CD4, CD7, and CD11b expression correlated with highest WBC count and unfavorable‐risk cytogenetics; CD64 expression was associated with higher WBC count while that of CD13 was associated with lower platelet count; CD22, CD34, CD123, and terminal deoxynucleotidyl transferase (TdT) expression correlated with unfavorable‐risk cytogenetics; CD5 expression was associated with normal platelet count while that of CD19 was associated with children and favorable‐risk cytogenetics; CD117 expression was associated with low WBC and lower platelet counts; myeloperoxidase (MPO) expression correlated with lower platelet count; and MPO and glycophorin A (Gly‐A) expression was associated with lower WBC count and favorable‐risk cytogenetics. The results of the relevance analysis revealed the distribution characteristics of antigen expression in different AML prognostic subgroups. The majority of antigens associated with good or poor prognostic subgroups were in accordance with the previous reports of correlation of expression of these antigens with prognosis. Antigens associated with good (or poor) prognostic subgroups were defined as good (or poor)‐risk antigens.