Leukemia cell analysis in the diagnosis of ALL/AUL: clinical value of currently available methods

After morphology and cytochemistry, immunological leukemic cell diagnosis became a generally available method by the development of monoclonal antibodies. Besides an otherwise not possible classification of nonmyeloid leukemias according to T- or B-differentiation status, precision and standardization of leukemia diagnosis is accomplished by immunodiagnostic means in an experimental way, an important point for therapy studies. In children, the common ALL--a precursor B-ALL type--was prognostically more favorable than the T-ALL which differs also in clinical appearance and in prognostic factors. In adults, however, T-ALL had a better prognosis than common ALL in a prospective trial. The heterogeneous Null-ALL which makes up 23% of ALL in adults and 5% in children, was the prognostically most unfavorable form besides the rare B-ALL (1-2%), the immunophenotype being an independent prognostic variable. Immunodiagnostic means are therefore of utmost clinical importance compared to other methods such as cytogenetic and electron microscopy that are not performable in a corresponding extent.     

Ras activation in normal white blood cells and childhood acute lymphoblastic leukemia.

Ras is an important cellular switch, relaying growth-promoting signals from the plasma membrane to the nucleus. In cultured cells, Ras is activated by various hematopoietic cytokines and growth factors, but the activation state of Ras in peripheral WBCs and bone marrow cells has not been studied nor has Ras activation been assessed in cells from patients with acute lymphoblastic leukemia (ALL). Using an enzyme-based method, we assessed Ras activation in peripheral WBCs, lymphocytes, and bone marrow cells from normal subjects and from children with T-cell ALL (T-ALL) and B-lineage ALL (B-ALL). In normal subjects, we found mean Ras activations of 14.3, 12.5, and 17.2% for peripheral blood WBCs, lymphocytes, and bone marrow cells, respectively. All three of these values are higher than we have found in other normal human cells, compatible with constitutive activation of Ras by cytokines and growth factors present in serum and bone marrow. In 9 of 18 children with T-ALL, Ras activation exceeded two SDs above the mean of the corresponding cells from normal subjects, whereas in none of 11 patients with B-ALL did Ras show increased activation; activating genetic mutations in ras occur in less than 10% of ALL patients. Thus, Ras is relatively activated in peripheral blood WBCs, lymphocytes, and bone marrow cells compared with other normal human cells, and Ras is activated frequently in T-ALL but not in B-ALL. Increased Ras activation in T-ALL compared with B-ALL may contribute to the more aggressive nature of the former disease.     

Surface marker analysis of acute lymphocytic leukaemia in Taiwan, Republic of China

Acute lymphocytic leukaemia (ALL) is heterogeneous in clinical characteristics and immunological properties. Standard surface marker analysis has enabled us to subclassify 121 cases of ALL into four subtypes, i.e. T-ALL, common ALL, null ALL and B-ALL. We have also tried to correlate these subtypes with their clinical characteristics. Our patients had younger ages with a mean age of 13.75. A slight male predominance was observed. There were consistently higher incidences in northern Taiwan in each subtype, but no significant differences in incidences between rural and urban areas. Although there were high incidences of L1 type cell in each immunological subtype, there was no correlation between FAB classification and each subtype, nor did morphologic features relate to cellular origins. Clinical manifestations revealed significantly high incidence of CNS involvement and thymic mass in T-ALL. Hepatosplenomegaly was more common and complete remission rate was higher in children with ALL than in adults.     

急性淋巴细胞白血病免疫分型的特点及其临床意义(英文)

目的为了探讨急性淋巴细胞白血病(ALL)各亚型免疫分型的特点及其临床意义。方法采用 CD45/SSC 双参数散点图设门,应用三色流式细胞术,对81例 ALL 的初诊患者骨髓标本进行免疫分型,并对其中45例进行核型分析。结果 (1)B-ALL 中 CD19表达最常见(阳性率为100%),而 T-ALL 中 CD5和 CD7表达阳性率最高,均为90%;B-ALL 和 T-ALL 都存在抗原交叉表达的现象;两组患者的完全缓解率(CR 率)并无显著差异(P>0.05)。(2)伴髓系抗原表达的急性淋巴细胞白血病(My~+ ALL)比较常见,本组达到39.5%,常累及 B 淋巴系统(占 My~+ ALL 的84.4%);各髓系抗原中以 CD13表达阳性率最高;此类患者的 CR 率较高,儿童 CR 率为72.2%,成人为78.6%。(3)急性杂合性白血病(HAL)的发病率为19.8%,以髓系、B 系共同表达者居多;并且 CD34表达阳性率较高(81.3%),该类患者 CR 率较低(儿童和成人分别为50%和40%)。(4)CD34在 B-ALL,My~+ALL 和 HAL 中表达阳性率较高,而 T-ALL 中少见(P<0.025)。结论免疫分型在诊断特殊类型的 ALL(如 HAL,My~+ ALL)中具有显著优势;CD19和 CD5诊断 B-ALL 和 T-ALL 的灵敏度较好,但特异性不高,存在抗原交叉表达;CD34和髓系抗原的表达与 CR 率无相关性,但在 HAL,CD34的表达与 CR 率成负相关。     

急性淋巴细胞白血病免疫分型的特点及其临床意义

目的为了探讨急性淋巴细胞白血病(ALL)各亚型免疫分型的特点及其临床意义。方法采用CD45/SSC双参数散点图设门,应用三色流式细胞术,对81例ALL的初诊患者骨髓标本进行免疫分型,并对其中45例进行核型分析。结果(1)B-ALL中CD19表达最常见(阳性率为100%),而T-ALL中CD5和CD7表达阳性率最高,均为90%;B-ALL和T-ALL都存在抗原交叉表达的现象;两组患者的完全缓解率(CR率)并无显著差异(P>0.05)。(2)伴髓系抗原表达的急性淋巴细胞白血病(My ALL)比较常见,本组达到39.5%,常累及B淋巴系统(占My ALL的84.4%);各髓系抗原中以CD13表达阳性率最高;此类患者的CR率较高,儿童CR率为72.2%,成人为78.6%。(3)急性杂合性白血病(HAL)的发病率为19.8%,以髓系、B系共同表达者居多;并且CD34表达阳性率较高(81.3%),该类患者CR率较低(儿童和成人分别为50%和40%)。(4)CD34在B-ALL,My ALL和HAL中表达阳性率较高,而T-ALL中少见(P<0.025)。结论免疫分型在诊断特殊类型的ALL(如HAL,My ALL)中具有显著优势;CD19和CD5诊断B-ALL和T-ALL的灵敏度较好,但特异性不高,存在抗原交叉表达;CD34和髓系抗原的表达与CR率无相关性,但在HAL,CD34的表达与CR率成负相关。     

Updates in the Pathology of Precursor Lymphoid Neoplasms in the Revised Fourth Edition of the WHO Classification of Tumors of Hematopoietic and Lymphoid Tissues

Purpose of Review

Acute lymphoblastic leukemias (ALL) are malignant disorders of immature B or T cells that occur characteristically in children, usually under the age of 6 (75%). Approximately 6000 new cases of ALL are diagnosed each year in the USA, 80–85% of which represent B-ALL forms. Most presentations of B-ALL are leukemic, whereas T-ALL presents with a mediastinal mass, with or without leukemic involvement. The revised fourth edition of the World Health Organization (WHO) classification (2017) has introduced some changes in both B and T-ALL. Here, we summarize the categories of lymphoblastic leukemia/lymphomas as defined by the WHO and recent developments in the understanding of this group of hematologic malignancy.

Recent Findings

Two provisional categories of B-ALL have now been identified including B-ALL, BCR-ABL1-like, and B-ALL with iAMP21. The Philadelphia chromosome-like B-ALL includes forms of the disease that shares the expression profiling of B-ALL with t(9;22) but lack such rearrangement. The second one shows amplification of part of the chromosome 21. Both entities are associated with worse prognosis. Within the T-ALL group, an early precursor T cell form has now been introduced as a provisional category. Such group demonstrates expression of stem cell and myeloid markers in conjunction with the T cell antigens.

Summary

The current review summarizes the recent updates to the WHO classification.

     

Peanut agglutinin, a marker for T-cell acute lymphoblastic leukemia with a good prognosis

Peanut agglutinin (PNA) binding was studied in cells from 74 children with acute lymphoblastic leukemia (ALL). PNA positivity occurred in 50% of T-ALL (12 of 24) and was rare in other types of ALL. There was no clear relationship between PNA and initial white blood cell count, French-American-British classification, stage of differentiation of leukemic cells, hand mirror cells, and organomegaly at diagnosis. Prognosis, however, was significantly better (continuous complete remission, death) in the PNA-positive T-ALLs. It seems that PNA is a useful marker for a subgroup of T-ALL with a better prognosis.     

Abnormalities of the short arm of chromosome 9 with partial loss of material in hematological disorders

Clinical, hematological, and cytogenetic data of 32 patients with loss of part of the short arm of chromosome 9 (9p-) are reviewed. There were 20 acute lymphoblastic leukemia (ALL), seven non-Hodgkin lymphoma (NHL), three acute myeloid leukemia, one refractory anemia with excess blasts in transformation, and one chronic myeloid leukemia (CML) in blast crisis. The cytogenetic findings were heterogeneous: 13 cases of del(9)(p21), among them four as sole karyotypic change; five cases of del(9)(p12), three of them as sole karyotypic change; four patients with i(9q), three with unbalanced translocations involving 9p12; and seven with unbalanced translocations involving 9p21. In addition, 10 patients showed known specific translocations for determined subgroups of ALL, NHL, and CML. The immunological phenotypes in the 20 ALL patients were common ALL (35%), pre-B-ALL (35%), B-ALL (5%), T-ALL (15%), and null ALL (10%). Three NHL were of T cell origin and the others of B cell origin. No specific association between the karyotypic change, immunophenotype, and clinical presentation could be ascertained for patients with ALL, acute myeloid leukemia, CML in blast crisis, and B-NHL. In T-NHL, three children with deletion of 9p, T immunoblastic lymphoma originating from common thymocyte and presenting with a mediastinal mass and pleural effusion may constitute a definite subgroup with good prognosis. All other cases had a poor outcome. Previously suggested association of 9p- with T-ALL and "lymphomatous features" was not confirmed.     

成人急性淋巴细胞白血病免疫表型特点分析

目的研究成人急性淋巴细胞白血病(ALL)患者中不同亚型的各种白血病细胞免疫表型分布特点。方法采用当前国际通用的四色流式细胞术图像分析系统检测并综合分析76例ALL患者的免疫表型及其发生规律与特点。结果①76例ALL免疫表型系列来源可分为三种不同亚型,其中T-ALL亚型5例(占6.57%)、B-ALL亚型68例(89.48%)、T和B细胞混合型(T/B-ALL)亚型3例(3.95%)②ALL早期抗原表达特点:在B-ALL亚型中CD38、CD34、HLA-DR呈高表达;在T-ALL亚型中,CD38和CD34呈高表达,但HLA-DR不表达;在T/B-ALL亚型中,HLA-DR表达,CD34和CD38不表达。③按免疫分型相关抗原的敏感性和特异性分析:在B-ALL中,特异性抗原cCD79a占91.18%,敏感性抗原CD19表达占97.06%;在T-ALL中,特异性抗原cCD3和敏感性抗原CD7均表达为100%;在T/B混合型ALL中,cCD3、cCD79a、CD19均表达,CD7表达2例;④伴髓系抗原交叉表达分析:在B-ALL中,伴髓系抗原表达占21例(30.88%);在T-ALL中,伴髓系抗原表达1例(20%);T/...     

急性淋巴细胞白血病患者 ASB2和 Jak3基因mRNA 表达的研究

目的：研究急性淋巴细胞白血病（ALL）患者骨髓中锚蛋白重复序列和抑制细胞因子信号盒蛋白2（ASB2）和 Janus 激酶3（Jak3）mRNA 的表达及其两者的相关性。方法收集初诊的48例 ALL患者（37例 B 细胞 ALL,11例 T 细胞 ALL）和34例非白血病患者（对照组）骨髓,采用实时荧光定量 PCR检测骨髓中 ASB2和 Jak3 mRNA 表达情况。结果B 细胞 ALL 和 T 细胞 ALL 患者骨髓中 ASB2 mRNA表达量相对于对照组分别升高了32．7倍和68．5倍,差异均有统计学意义（t ＝20．1,P ＜0．01;t ＝23．1, P ＜0．01）,Jak3 mRNA 表达量较对照组分别升高了2336．3和7131．5倍（t ＝70．2,P ＜0．01;t ＝90．4, P ＜0．01）。ASB2和 Jak3 mRNA 表达量具有相关性（r ＝0．523,P ＜0．001）。结论ASB2和 Jak3在 ALL患者骨髓中异常表达,且具有正相关性,两者可能共同参与白血病细胞的恶性增殖和异常分化。     

Purine metabolism in childhood acute lymphoblastic leukemia: biochemical markers for diagnosis and chemotherapy

Adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), 5'nucleotidase (5'NT), ecto-5'NT, hypoxanthine-guanine phosphoribosyltransferase(HGPRT), adenine phosphoribosyltransferase(APRT), adenosine kinase(AK), AMP deaminase (AMPD) and adenylate kinase(AdKin) activities were assayed in leukemic cells from bone marrow and/or peripheral blood of 43 newly diagnosed children with acute lymphoblastic leukemia(ALL). These enzyme activities have been investigated in relation to some immunological markers. ADA activity was higher in E-rosette positive leukemia(E+ ALL), while HGPRT, APRT, PNP, 5'NT, ecto-5'NT and AdKin activities were found to be lower in E+ ALL as compared to E- ALL. In common ALL (cALL) antigen positive leukemia, mean ADA activity was significantly lower as compared to cALL- leukemia, whereas PNP, 5'NT, ecto-5'NT and AdKin activities were significantly higher. cALL cells with cytoplasmic immunoglobulin M(IgM) heavy chains were found to have mean 5'NT activities twice as high as cALL cells lacking cytoplasmic IgM heavy chains. In two patients who had surface immunoglobulins on their cell membranes, low 5'NT activities were found. When measuring enzyme activities after 2-4 days of prednisone monotherapy, only mean ADA and HGPRT activities decreased in non-B, non-T ALL. These decreases were not significant in T-ALL patients. Mean enzyme activities in the leukemic cells of five patients with relapse were comparable to those in newly diagnosed patients, except for 5'NT, which was found to be within the activity range of control peripheral blood lymphocytes. It is concluded that ADA and AdKin activities are suitable as markers for E+ ALL and cALL+ leukemias respectively. 5'NT might help to distinguish between cALL cells having and lacking pre-B characteristics. Since 5'NT activity may also be decreased in B-ALL, it is not suitable as a T-ALL marker. Enzymes of purine metabolism in leukemic relapse need further investigation.     

ASPP1, a common activator of TP53, is inactivated by aberrant methylation of its promoter in acute lymphoblastic leukemia

We have analyzed the regulation and expression of ASPP members, genes implicated in the regulation of the apoptotic function of the TP53 tumor-suppressor gene, in acute lymphoblastic leukemia (ALL). Expression of ASPP1 was significantly reduced in ALL and was dependent on hypermethylation of the ASPP1 gene promoter. Abnormal ASPP1 expression was associated with normal function of the tumor-suppressor gene TP53 in ALL. The analyses of 180 patients with ALL at diagnosis showed that the ASPP1 promoter was hypermethylated in 25% of cases with decreased mRNA expression. Methylation was significantly higher in adult ALL vs childhood ALL (32 vs 17%, P = 0.03) and T-ALL vs B-ALL (50 vs 9%, P = 0.001). Relapse rate (62 vs 44%, P = 0.05) and mortality (59 vs 43%, P = 0.05) were significantly higher in patients with methylated ASPP1. DFS and OS were 32.8 and 33.7% for patients with unmethylated ASPP1 and 6.1 and 9.9% for methylated patients (P < 0.001 y P < 0.02, respectively). On the multivariate analysis, methylation of the ASPP1 gene promoter was an independent poor prognosis factor in ALL patients. Our results demonstrate that decreased expression of ASPP1 in patients with ALL is due to an abnormal methylation of its promoter and is associated with a poor prognosis.     

Immunological subtypes of childhood T-cell acute lymphoblastic leukemia and their prognostic significance

The data on examination and treatment of 39 children with T-cell acute lymphoblastic leukemia (T-ALL) were assessed; all patients received ALL BFM-90-M treatment. The fraction of children with T-ALL among ALL patients in Belarus was 12.2% (pre-T-ALL--15, cortical T-ALL--46 and mature T-ALL--23%). Also, a subtype of T-ALL with atypical expression of markers was identified (13%). Overall 7-year survival in T-ALL patients was 47(20%). The worst prognosis was recorded in the T-ALL subgroup with atypical expression of markers (p < 0.001 as compared with the other subgroups). As for outcome--from best to worst, T-ALL subtypes ranged as follows: cortical T-ALL, mature T-ALL, pre-T-ALL and T-ALL with atypical expression of markers.     

miR-128在急性淋巴细胞白血病中的表达

目的 探讨miR-128在急性淋巴细胞白血病(ALL)中的表达及其意义.方法 采用实时荧光定量聚合酶链反应法对62例ALL患者和20例骨髓正常患者骨髓单个核细胞miR-128的相对表达量进行检测,并与患者的临床资料进行相关性分析.结果 miR-128在ALL患者中的相对表达量高于健康对照组(P＜0.05),但其在急性B淋巴细胞白血病(B-ALL)及急性T淋巴细胞白血病(T-ALL)中的表达水平差异无统计学意义(P＞0.05),miR-128的表达与是否存在bcr-abl融合基因及其mRNA的表达水平均无相关性(均P＞ 0.05).结论 miR-128在ALL患者中表达上调,可作为临床诊断ALL的潜在分子标志物,bcr-abl融合基因的表达对miR-128在ALL中的表达无明显影响.     

Cell markers and acute leukaemia subtypes in Chile

A panel of monoclonal antibodies were used to define acute myeloblastic leukaemia (AML) and acute lymphoblastic leukaemia (ALL) with its different subgroups. Thirty-three patients were studied in a period of one year. ALL was diagnosed in 80% of the children and 20% of the adults. AML was present in 22% of the children and 78% of adults. In children, only one of seven was common ALL, three of seven were T-ALL and three of seven were B-ALL. Five of 12 were unclassifiable (U-ALL). In adults the 4 ALL were U-ALL.     

Granulocyte-Colony Stimulating Factor, Granulocyte-Macrophage Colony Stimulating Factor, PIXY-321, Stem Cell Factor, Interleukin-3, and Interleukin-7: Receptor Binding and Effects on Clonogenic Proliferation in Acute Lymphoblastic Leukemia

Cytokines are frequently used after chemotherapy of leukemias and solid tumors to augment recovery of normal hematopoiesis. While the regulation of normal and leukemic myelopoiesis is well investigated, little is known about effects of cytokines on growth and differentiation of lymphoblastic leukemia. In this study, we investigated the expression of receptors for G-CSF, GM-CSF, SCF, IL-3, and IL-7 on acute lymphoblastic leukemia (ALL) blasts and the effects of these growth factors (GF) on ALL blast colony formation. The binding of fluorescence-tagged cytokines to receptors on ALL blasts was studied by flow-cytometry in 27 cases of ALL (24 precursor B-ALL, 3 T-ALL). Receptor-binding for myeloid-associated GF was observed in the majority of precursor B-ALL (G-CSF = 100%, GM-CSF = 65%, IL-3 = 83%, SCF = 74%), but not in T-ALL. Binding of labelled IL-7 was detected in both precursor B- (92%) and T-ALL (100%). The presence of receptors for SCF in ALL was confirmed by polymerase chain reaction for c-kit mRNA in 19/21 cases tested. Expression of receptors for G-CSF, GM-CSF, IL-3, and SCF was not associated with expression of myeloid antigens, or with specific cytogenetic abnormalities. The effects of these GF on clonogenic cells were tested in the ALL blast colony assay and varied between samples, but all cytokines were able to increase clonogenic growth. The GM-CSF/IL-3 fusion molecule PIXY-321 was most effective in promoting colony growth. In some cases inhibition of colony formation was found. We conclude that ALL blast cells have receptors not only for IL-7, but also for G-CSF, GM-CSF, SCF, and IL-3. ALL precursors can respond to these GF with changes in their clonogenic growth indicating the presence of functional receptors. Results may have implications, for therapeutic approaches combining cytokines and chemotherapy.     

Association of two ARID5B gene variant single nucleotide polymorphisms with acute lymphoblastic leukemia in the Egyptian population

Background: ARID5B SNPs have been linked to ALL in many research studies in which it was identified as a risk factor. From this context, we had great interest to investigate the relationship between ARID5B rs4948488 and ARID5B rs2893881 genotypes and ALL susceptibility and relapse in this study. Materials and Methods: Peripheral blood mononuclear cells were analyzed for ARID5B rs4948488 and rs2893881 gene polymorphisms by real-time quantitative polymerase chain reaction in 80 ALL patients and 80 controls. Results: Our results showed that the C/C genotype of ARID5B rs4948488 and A/G genotype and G-allele of rs2893881 were linked to higher ALL incidence. Regarding the relapse of ALL, rs4948488 C/C genotype and C-alleles were significantly associated with relapse of ALL. Meanwhile, rs4948488 C/C genotype and rs2893881 A/A genotype and A-allele are associated with T-ALL, while rs2893881 A/G genotype and G-allele are associated with B-ALL. Conclusion: The results of our study suggested that ARID5B rs4948488 and rs2893881 SNPs might be used risk factors for genetic susceptibility for B-ALL and T-ALL, and that ARID5B s4948488 is related to relapse in ALL patients.     

Childhood T-cell acute lymphoblastic leukemia: the Dana-Farber Cancer Institute acute lymphoblastic leukemia consortium experience.

PURPOSE: T-cell acute lymphoblastic leukemia (T-ALL) accounts for 10% to 15% of newly diagnosed cases of childhood acute lymphoblastic leukemia (ALL). Historically, T-ALL patients have had a worse prognosis than other ALL patients. PATIENTS AND METHODS: We reviewed the outcomes of 125 patients with T-ALL treated on Dana-Farber Cancer Institute (DFCI) ALL Consortium trials between 1981 and 1995. Therapy included four- or five-agent remission induction; consolidation therapy with doxorubicin, vincristine, corticosteroid, mercaptopurine, and weekly high-dose asparaginase; and cranial radiation. T-ALL patients were treated the same as high-risk B-progenitor ALL patients. Fifteen patients with T-cell lymphoblastic lymphoma were also treated with the same high-risk regimen between 1981 and 2000. RESULTS: The 5-year event-free survival (EFS) rate for T-ALL patients was 75% +/- 4%. Fourteen of 15 patients with T-cell lymphoblastic lymphoma were long-term survivors. There was no significant difference in EFS comparing patients with T-ALL and B-progenitor ALL (P =.56), although T-ALL patients had significantly higher rates of induction failure (P <.0001), and central nervous system (CNS) relapse (P =.02). The median time to relapse in T-ALL patients was 1.2 years versus 2.5 years in B-progenitor ALL patients (P =.001). There were no pretreatment characteristics associated with worse prognosis in patients with T-ALL. CONCLUSION: T-ALL patients fared as well as B-progenitor patients on DFCI ALL Consortium protocols. Patients with T-ALL remain at increased risk for induction failure, early relapse, and isolated CNS relapse. Future studies should focus on the identification of and treatment for T-ALL patients at high risk for treatment failure.