Aberrant immunophenotypes detected by flow cytometry in acute lymphoblastic leukemia

The present study was designed to analyse the proportion of ALL patients in which the phenotypic detection of minimal residual disease (MRD) is feasible, based on the presence of aberrant phenotypes: lineage infidelity, asynchronous expression, overexpression and ectopic phenotype. For this purpose we have prospectively investigated the phenotype of blast cells from 25 patients at diagnosis using a large panel of monoclonal antibodies by multiparametric flow cytometry. The mean age was 23.3 +/- 17.3 with 10 children and 15 adults. 14 patients were classified as L1, 9 L2 and 2 L3 according to the FAB classification. 17 cases were B-lineage ALL and 8 T-ALL. 23 out of 25 cases (92%) included in this study displayed phenotypic aberrations at diagnosis (15 out of 17 cases of B-lineage ALL and all T-ALL patients). 76% of patients displayed two or more than two aberrancies. The phenotypic aberrations were lineage infidelity, found in 12 patients, asynchronous antigen expression detected in 17 patients, antigen overexpression in 4 patients and ectopic phenotype in 7 patients. In summary our results show that when a large panel of MoAbs is used for the immunophenotypical characterization of ALL, most patients display aberrant phenotypes, the coexistence of more than two aberrant antigen expressions being frequently detected. These results suggest that the use of immunological methods for the detection of MRD in ALL based on the existence of aberrant phenotypes could be of great help for the follow-up of patients in complete remission.     

Genuine CD7 expression in acute leukemia and lymphoblastic lymphoma.

CD7 has been used as a valuable marker for normal and malignant T cells and also for a proportion of acute nonlymphocytic leukemia (ANLL) cells. Difference in reactivity was noticed among CD7 antibodies, however, when tested against ANLL cells and myeloid/monocytoid cell lines; Tp40 antibody produced in our laboratories was not reactive with the HL-60 promyelocytic line, whereas 4A antibody was reactive, even though both detected a quite similar or an identical epitope on CD7 molecule. Preincubation of HL-60 cells with human immunoglobulin preparation clearly negated the reactivity by 4A, suggesting that 4A antibody is not reactive to CD7 itself, but it probably binds with immunoglobulin G Fc receptors expressed on HL-60 cells. Five cases of ANLL which were positive with 4A antibody were selected and tested with Tp40 antibody, and only two were found to be positive. Expression of CD7 mRNA in these two cases (but not in other cases) was also demonstrated by Northern blotting with a cDNA probe for CD7 recently cloned in our laboratories, indicating that CD7 is expressed on a certain fraction of ANLL, although the positive cases may be smaller than the reports so far appeared. A Northern blot study was also conducted with two acute lymphocytic leukemia cases and one lymphoblastic lymphoma case with CD7+, CD2-/+/-, CD5-/+/- phenotype and germline T cell receptor beta genes. CD7 mRNA is expressed in all three cases and CD3 mRNA is also observed in two cases, suggesting that these tumor cells are of T precursor origin.     

Childhood acute lymphoblastic leukemia: Integrating genomics into therapy

Acute lymphoblastic leukemia (ALL), the most common malignancy of childhood, is a genetically complex entity that remains a major cause of childhood cancer‐related mortality. Major advances in genomic and epigenomic profiling during the past decade have appreciably enhanced knowledge of the biology of de novo and relapsed ALL and have facilitated more precise risk stratification of patients. These achievements have also provided critical insights regarding potentially targetable lesions for the development of new therapeutic approaches in the era of precision medicine. In this review, the authors delineate the current genetic landscape of childhood ALL, emphasizing patient outcomes with contemporary treatment regimens as well as therapeutic implications of newly identified genomic alterations in specific subsets of ALL. Cancer 2015;121:3577–3590 . © 2015 American Cancer Society.     

急性淋巴细胞白血病伴CD13表达免疫表型分析

目的探讨急性淋巴细胞白血病(acutelymphocytic leukemia,ALL)伴CD13表达的免疫表型特点,对包括CD33在内的其他抗原进行相关性分析.方法对85例初诊ALL患者进行免疫学分型,以是否表达My把B-ALL和T-ALL分为My+ALL和My-ALL,进行组间分析.结果全部85例B-ALL和T-ALL患者均高表达B系和T系相关抗原(100%,100%).所有的B-ALL均不表达T-ALL相关抗原,所有T-ALL均不表达B-ALL相关抗原.CD13的表达率为31%(B-ALL31.4%,T-ALL 28.6%).My+B-ALL和My+T-ALL患者的CD13阳性细胞的中位数高于My-B-ALL和My-T-ALL患者,P值分别为0.013和0.04.My+B-ALL患者的CD15阳性细胞的中位数高于My-B-ALL患者,P=0.000 1.结论白血病免疫分型对于白血病的诊断、治疗和预后判断均有很大帮助,其在临床诊断中的推广将有助于患者的诊断和治疗.CD13在My+ALL中表达较高(31%),白血病细胞在恶性演变过程中不同时期表现出特征反映,其临床意义有待于重新评价.     

急性淋巴细胞白血病伴CD13表达免疫表型分析

目的：探讨急性淋巴细胞白血病（acutelymphocytic leukemia,ALL）伴CD13表达的免疫表型特点,对包括CD33在内的其他抗原进行相关性分析.方法：对85例初诊ALL患者进行免疫学分型,以是否表达My把B-ALL和T-ALL分为My＋ALL和My-ALL,进行组间分析.结果：全部85例B-ALL和T-ALL患者均高表达B系和T系相关抗原（100%,100%）.所有的B-ALL均不表达T-ALL相关抗原,所有T-ALL均不表达B-ALL相关抗原.CD13的表达率为31%（B-ALL31.4%,T-ALL 28.6%）.My＋B-ALL和My＋T-ALL患者的CD13阳性细胞的中位数高于My-B-ALL和My-T-ALL患者,P值分别为0.013和0.04.My＋B-ALL患者的CD15阳性细胞的中位数高于My-B-ALL患者,P=0.000 1.结论：白血病免疫分型对于白血病的诊断、治疗和预后判断均有很大帮助,其在临床诊断中的推广将有助于患者的诊断和治疗.CD13在My＋ALL中表达较高（31%）,白血病细胞在恶性演变过程中不同时期表现出特征反映,其临床意义有待于重新评价.     

伴髓系抗原CD13高表达的急性B细胞白血病形态学及流式细胞学特点分析

目的 分析伴异常表达CD13急性B细胞白血病形态学及流式细胞学特点,提高诊断此类疾病的诊断水平.方法 对确诊的4例患者的形态学及流式细胞术散点图进行分析,研究并复习相关文献,总结特点.结果 急性B细胞白血病伴异常表达CD13的患者形态学上多数具有高度异质性与分化发育的不协调性,且此类细胞均高表达CD34.结论 急性淋巴细胞白血病伴异常表达髓系抗原的确诊必须依靠免疫学分型,同时该类原始细胞既具有典型B细胞的免疫标志又有髓系细胞的特有标志,具有髓系原始细胞及原始淋巴细胞的双重特点,是形态学误诊的最主要原因.     

多参数流式细胞术观察251例白血病细胞CD14表达的意义

目的　探讨 CD14在白血病细胞的表达及其在白血病诊断和鉴别诊断中的意义。方法　采用 CD45 /SSC参数设门多色流式细胞术分析 2 5 1例白血病细胞 CD14及其它白血病相关抗原的表达情况。结果　 2 5 1例中有 19例 ( 7.5 % )白血病细胞表达 CD14,95例急性髓系白血病 ( AML )中 16例 ( 16 .8% )白血病细胞表达 CD14,130例淋系白血病未见 CD14阳性的病例。 M4和 M5b CD14的阳性率 ( 9/ 14;6 / 6 )明显高于 M2 ( 1/ 30 ) ( P=0 .0 0 1;P=3.6× 10 - 6 )及 M5a( 0 / 5 ) ( P=0 .0 3;P=2 .2× 10 - 3 )。标准 CD14单抗均不与 5例 M5a细胞反应 ,而自制 CD14新克隆 2 F9单抗则全部阳性。5例 M5a中 1例 CD15和 2 9例 M2 中 13例 CD11b的阳性率明显低于 M5b( 6 / 6 ,P=0 .0 2 ;6 / 6 ,P=0 .0 2 )。在 AML 组中 ,CD14的表达与 CD117呈负相关 ( r=- 0 .2 ,n=81,P=0 .0 3) ,与 CD11b、CD15及HL A- DR呈正相关 ( r=0 .5 ,n=93,P=0 .0 0 1;r=0 .4,n=95 ,P=0 .0 0 1;r=0 .2 ,n=95 ,P=0 .0 2 )。结论　 CD14有助于淋巴系白血病、髓系白血病及其单核细胞相关性白血病的诊断与鉴别 ,其对单核细胞相关性白血病免疫分型诊断中的特异性为 96 .7% ,敏感性为 6 0 .0 %。     

CD34 antigen expression in children with Philadelphia chromosome-positive acute lymphoblastic leukemia.

One characteristic of Philadelphia chromosome (Ph')-positive acute leukemia is the occasional presence of both lymphoid and myeloid features in the same leukemia. This phenomenon supports the theory that this subtype of acute leukemia arises from lymphoid-myeloid stem cell, pluripotent progenitors. Very few reports, however, describe the immunophenotype, especially CD34 antigen, of Ph'-positive acute lymphoblastic leukemia (ALL). It has been shown that CD34, the human progenitor cell antigen, is found on 1% or less of normal human bone marrow cells, approximately 30% of acute leukemias, and multipotent progenitor cells; CD34 is not found on normal peripheral blood cells. A high frequency of CD34 expression was found in children with Ph'-positive ALL: CD34 was positive for all six patients tested, and one had an acute mixed-lineage leukemia. These findings suggest the involvement of a pluripotent stem cell in Ph'-positive ALL.     

Significance of CD66c expression in childhood acute lymphoblastic leukemia

Upon analyzing 696 childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cases, we identified the characteristics of CD66c expression. In addition to the confirmation of strong correlation with BCR-ABL positivity and hyperdiploid, we further observed that CD66c is frequently expressed in CRLF2-positive (11/15, p < 0.01 against chimeric gene-negative) as well as hypodiploid cases (3/4), whereas it is never expressed in ETV6-RUNX1, MLL-AF4, MLL-AF9, MLL-ENL, and E2A-PBX1-positive cases. Although the expression of CD66c itself is not directly linked to the prognosis, the accompanying genetic abnormalities are important prognostic factors for BCP-ALL, indicating the importance of CD66c expression in the initial diagnosis of BCP-ALL.     

Detection of aneuploid cells in acute lymphoblastic leukemia with flow cytometry before and after therapy

We have evaluated the proliferative activity and DNA content of immunophenotyped hematopoietic cells applying flow cytometry. After indirect immunofluorescence the cell membrane was permeabilized for propidium iodide staining of DNA. Compared with single parameter detection of DNA content this method has more certainty in the determination of aneuploidies in lymphatic leukemia cells. Immunophenotyped residual normal hematopoietic cells were used as an internal standard. If this method was tested for evaluation of therapeutic effects after chemotherapy greater sensitivity in detection of minimal residual disease was observed than when using microscopic evaluation or single parameter DNA analysis in cases of aneuploid lymphoblastic leukemias.     

Pattern of subtypes of acute lymphoblastic leukemia in India

Leukemic cells from 124 acute lymphoblastic leukemia (ALL) and 31 chronic lymphatic leukemia (CLL) were examined for sheep erythrocyte receptor (E), surface immunoglobulin (SIg) and their reactivity with a panel of monoclonal antibodies recognizing specific surface antigens including pan-T, Common ALL and Ia antigens. In acute lymphatic leukemia, 33% of patients reveal T-cell receptor associated with higher age group, mediastinal mass and high WBC count. Common ALL was predominant between 2 and 9-yr age group. Among chronic lymphatic leukemia, 2 patients were found to be T-CLL while 29 revealed presence of SIg. Ia antigen was detected in 44.4% of ALL and 64% fo CLL patients. The pattern of surface marker observed in our series may be related to our life style, socio-economic and environmental factors.     

Immunological subtypes of acute lymphoblastic leukemia in north India

Pretreatment immunologic marker analysis in 152 adult and childhood patients of ALL and ALL/lymphoma employing multiple monoclonal antibodies and hetero-antisera revealed three major subgroups, i.e. T-ALL (37.7%), N-ALL (33.1%) and C-ALL (21.5%). The early age peak was absent, males predominated in all the subgroups and T-ALL had increased incidence of thymic mass. Leucocyte counts of 50,000 X 10(6)/l were equally frequent in the three groups. T-ALL showed marked heterogeneity by showing a variety of markers such as T-helper/inducer, T-suppressor/cytotoxic, p-24, Ia and CALLA. These results show a high prevalence of unfavourable prognostic factors in ALL in our geographic region which might be related to socioeconomic and/or environmental factors.     

CD1d expression on B-precursor acute lymphoblastic leukemia subsets with poor prognosis.

Acute lymphoblastic leukemia (ALL) is the most frequent malignancy of childhood. Although therapeutical advances have been achieved, some ALL subgroups still fare poorly. CD1d is a monomorphic molecule that provides a suitable target for immunotherapy in view of the characterization of a glycolipid, alpha-galactosylceramide (alpha-GalCer), capable of being presented to CD1d-restricted T cells with cytotoxic potential. We investigated CD1d expression in 80 pediatric B-cell precursor (BCP) ALL cases defined according to immunophenotype, cytogenetic features and age at onset. CD1d was detected on ALL cells in 15% of the patients. CD1d+ ALLs were significantly associated with infant leukemia, pro-B phenotype and mixed-lineage leukemia (MLL)/AF4 gene rearrangement. Accordingly, overall survival of patients with CD1d+ ALL was significantly shorter. CD1d+ leukemic blasts were able to present alpha-GalCer via CD1d to cytotoxic CD1d-restricted T cells, which induced apoptosis of ALL cells that was inhibited by mAb to CD1d. CD1d+ blasts loaded with alpha-GalCer elicited cytokine secretion by CD1d-restricted T cells. Analysis of bone marrow (BM) cells derived from normal donors revealed that CD19+/CD1d+ cells were mostly mature B lymphocytes. However, a minority of BCPs expressed CD1d. Thus, expression of CD1d in ALL cases heralds an adverse prognosis but may provide a therapeutic tool.     

CD56 expression predicts occurrence of CNS disease in acute lymphoblastic leukemia

We examined the pre-treatment bone marrow samples from 200 consecutive adult patients with acute lymphoblastic leukemia (ALL) treated on various protocols at the University of Texas, M.D. Anderson Cancer Center between 1986 and 1998. Standard MFC techniques were used to determine CD56 expression on the leukemia blasts cells. The expression of CD56 was correlated with clinical characteristics at diagnosis, response to therapy, survival and disease-free survival.Blast expression of CD56 (> or = 20% of leukemic blasts) was seen in 16 (8%) of patients, with a median expression of 67% (range 20-99%). CD56 expression was associated with a higher incidence of central nervous system (CNS) disease at diagnosis (19% versus 4%; P=0.016). Incidence of CNS disease at any time was higher in patients with CD56+ disease (31% versus 14%; P=0.057). Among the 109 patients uniformly treated with the hyperCVAD regimen, CD56 expression was associated with a statistically significant higher incidence of CNS disease (33% versus 9%; P=0.026). CD56 expression in ALL is uncommon but may predict a higher risk for CNS disease. If these results are confirmed, CD56 expression could be used in combination with other high-risk features (e.g. lactate dehydrogenase (LDH), S-phase fraction, mature B-cell phenotype) to design a risk-oriented approach to CNS prophylaxis.     

Monitoring of acute myeloid leukemia by flow cytometry

Monitoring of minimal residual disease (MRD) becomes increasingly important in the risk-adapted management of patients with acute myeloid leukemia (AML). In selected patients with AML, multiparameter flow cytometry has shown accuracy and sensitivity in the quantification of MRD levels with independent prognostic impact. The applicability of this approach is superior to that of other methods such as quantitative polymerase chain reaction: Up to 80% of all patients can be monitored by flow cytometry. Nonetheless, significant technical advances are anticipated to extend the applicability of flow cytometry to 100% and to improve its sensitivity. Large clinical trials will determine the role of immunologic monitoring in the prognostic stratification of patients with AML.     

双克隆型多发性骨髓瘤五例

 目的　研究双克隆型多发性骨髓瘤（MM）患者的临床特点及治疗。方法　分析该科自1996年1月至2010年6月收治的5例双克隆型MM患者,男性2例,女性3例,年龄59～76岁,中位年龄72岁。其中IgG+IgA型3例,IgM+IgG／A型2例。结果　本组患者中诊断时均为DS分期Ⅲ期,治疗效果为2／4达到部分缓解（PR）,1／4为稳定（SD）,1／4为进展（PD）,无达到完全缓解（CR）的病例。结论　双克隆型MM较为罕见,目前对此型MM的诊断和治疗尚缺乏更深刻的认识,需要更多病例的分析。