Utility of white blood cell suspect flags and histogram pattern in the detection of acute leukemia by Advia-60 automated hematology analyzer

Blood samples from patients with acute leukemia, when analyzed with automated hematology counters, tend to introduce inaccuracies in the automated differential count and can cause diagnostic confusion without providing definite clues to the presence of abnormal cells. We designed this study to assess the utility of white blood cell (WBC) flags and histogram pattern generated by Advia-60 automated hematology analyzer in the recognition and categorization of acute leukemia. Data printouts of 31 newly diagnosed cases of acute leukemia, 22 with acute myeloid leukemia (AML) and 9 with acute lymphoblastic leukemia (ALL) were reviewed. All cases of AML and ALL generated the WBC suspect blastflag M2 associated with two of the non blast suspectflags G1 and G2. Among the cases of AML, 95.5% of the WBC histogram patterns were definitive of the presence of abnormal cells and were indicative of the myeloid nature of cells. Only 44.4% of the histograms in the cases of ALL could be definitive of the presence of abnormal cells and 33.3% were indicative of their lymphoid nature. Significantly, 55.5% of the histograms in ALL were normal. The false positives for both AML and ALL were 10.5% when only WBC flagging was considered and were reduced to 0.05% when the flags were combined with histogram patterns for interpretation. Combined flagging and histogram recognition can be of aid in identifying cases of acute leukemia and the morphologist can then assess these samples further. This ensures that cases of acute leukemia, especially in high output laboratories, are not inadvertently missed.     

Deficiency of complement decay-accelerating factor (DAF, CD55) in non-Hodgkin's lymphoma.

We have assessed levels of surface-expressed complement regulatory proteins, decay-accelerating factor (DAF) and membrane cofactor protein (MCP) on cells from patients with hematological malignancies. Neither malignant cells nor unaffected nucleated blood cells from the patients lacked MCP. On the other hand, complete deficiency of DAF was found in 2/10 of non-Hodgkin's lymphoma (NHL), while none of the 38 patients with acute nonlymphocytic leukemia (ANLL) (14 cases), chronic myelogenous leukemia (CML) (6 cases), acute lymphocytic leukemia (ALL) (12 cases) and chronic lymphocytic leukemia (CLL) (6 cases) lacked DAF. The two patients with DAF-negative NHL had no history of paroxysmal nocturnal hemoglobinuria (PNH), and their peripheral blood cells were DAF-positive. One DAF-negative NHL exhibited T cell markers and the other those of B cell. In both cases, treatment of the DAF-negative lymphoma cells with antibody against MCP (M177) followed by Mg(2+)-EGTA-serum resulted in efficient deposition of homologous C3. These results infer that some NHL specifically lack DAF and, through treatment with M177, are targeted by homologous C3.     

Chromosome studies in patients with acute nonlymphocytic or acute lymphocytic leukemia submitted to bone marrow transplantation--results of a European cooperative study

The chromosomal data of 58 acute nonlymphocytic leukemia (ANLL) patients and of 32 acute lymphocytic leukemia (ALL) patients submitted to bone marrow transplantation and collected from nine institutions are reported. Chromosomal studies were available at diagnosis in 19 cases with ANLL: seven had a partially or completely abnormal pattern. Forty-one patients had a chromosome study before bone marrow transplantation and all had a normal pattern. Thirteen patients with ALL were studied at diagnosis: five had a partially or completely abnormal karyotype. Of 20 cases analyzed before bone marrow transplantation, only one has maintained the abnormal pattern of diagnosis in part of the cells. Karyotypes were available in nine ANLL patients relapsed after bone marrow transplantations. Two showed the same clonal abnormalities seen at diagnosis; in three other cases the leukemic clone of relapse carried an additional chromosome abnormality with respect to the pattern at diagnosis and four more cases presented at relapse complex abnormalities; two of them had a cytogenetically normal pattern at diagnosis. In four of ten relapsed ALL cases chromosomes analyses were available. A relapse in donor cells and a hyperdiploid pattern were observed in two cases, respectively, while a normal, recipient pattern was documented in the other two cases. Serial chromosome studies performed in acute leukemia patients after bone marrow transplantation may allow the detection of different chromosomal patterns of relapse. In those cases who relapsed with a cell clone cytogenetically different from the pattern at diagnosis, a direct role played by the conditioning treatment in the pathogenesis of the relapsing disease may be hypothesized.     

Myeloperoxidase gene expression in acute leukemias

Since myeloperoxidase (MPO) is considered to be a critical marker of differentiating acute myelogenous leukemia (AML) from acute lymphocytic leukemia (ALL), the analysis of MPO gene expression may provide further insight into the leukemia classification and the lineage fidelity of leukemia cells. By Northern blot hybridization using full-length MPO cDNA as a probe, approximately 66% of AML cells (3/4 M1 cases, 2/4 M2 cases, 15/15 M3 cases, 11/15 M4 cases, and 2/12 M5 cases) were found to express MPO mRNAs, whereas none of 18 ALL cases did. MPO mRNA was detectable when AML cells contained at least 10% peroxidase-positive cells. APL (M3) cells expressed high levels of mRNA in accordance with heavy staining for peroxidase.     

Rapid immunocytochemical analysis of acute leukemias.

Immunophenotypic analysis of acute leukemias is time consuming and often requires flow cytometric analysis. A 1-hour alkaline phosphatase-labeled streptavidin-biotin immunocytochemical procedure was evaluated as an alternative. Seventeen cases of acute leukemia, including 10 acute lymphocytic (ALL) and 7 acute nonlymphocytic, were phenotyped by the rapid immunocytochemical procedure and the results were compared with standard analyses. In all 17 cases, the diagnoses made using standard cytochemical and immunologic methods were the same as obtained in blinded reviews by rapid immunocytochemical analysis. Nine cases of precursor B-cell ALL were positive for CD19 and/or CD22. Five CD19 + cases of ALL reacted with anti-myeloperoxidase, with one case also positive for CD15. CD15 positivity was confirmed on repeated study as well as with plastic section immunoperoxidase staining. Nine cases of ALL were positive for CD10 and eight were positive for terminal deoxynucleotidyl transferase. One case of ALL marked as T-cell ALL with CD1, CD2, CD3, and CD7. All cases of acute nonlymphocytic leukemia were positive for CD15, CD13, and/or CD33; anti-myeloperoxidase was positive in all but one case of monocytic leukemia. All cases of acute nonlymphocytic leukemia were negative for CD10 and one was positive for terminal deoxynucleotidyl transferase. Acute leukemias apparently may be phenotyped easily and accurately in 1 hour with this immunocytochemical technique, and slides may be stored permanently for review. There was in these 17 cases high correlation of the diagnoses with standard flow cytometric and cytochemical results. This rapid method allows a coordinated evaluation of morphologic features and immunophenotype; the latter features facilitated confirmation of unexpected reactivity of myeloid markers CD15 and MPO-7 in some cases of ALL.     

Interesting morphologic finding in T-cell acute lymphoblastic leukemia.

There is no specific morphologic finding that is representative of different phenotypic patterns of acute lymphoblastic leukemia (ALL). In 15 cases of T-cell ALL, there were ruptured white blood cells among the lymphoblasts, as seen in chronic lymphocytic leukemia. These 15 cases are presented in this report.     

Prognostic significance of morphologic and cytochemical markers in adult acute leukemia.

Survival times in 100 cases of acute leukemia (74 granulocytic, 14 lymphocytic, and 12 undifferentiated) were correlated with classic morphologic and cytochemical criteria. The 14 patients who had lymphocytic leukemia had significantly longer survival compared with the two other groups. Undifferentiated leukemias had a shorter survival time than granulocytic leukemias. Several subclasses of granulocytic leukemias were formed according to the presence or absence of Auer rods and the percentage of peroxidase-positive blasts. Neither of these two features significantly influenced the survival of these patients. In the lymphocytic leukemia group, PAS-positive and negative leukemias had similar survival expectancies. It is concluded that division into lymphocytic and nonlymphocytic leukemias is still helpful in predicting survival times of patients who have acute leukemia, but that further subclassification of these groups based on the presence or absence of Auer rods and the percentages of peroxidase-positive blasts is of no additional benefit.     

Nonrandom cytogenetic changes in human acute leukemia and their clinical implications

Sixty-three unselected Japanese cases of acute leukemia studied with chromosomal banding techniques and their hematologic and clinical evaluations were made. Salient features of this study were as follows: (1) Cases with t(8;21),t(15;17), and t(9;22) had characteristic clinical features, respectively; in particular, those with t(8;21) showed different hematologic findings than patients with other karyotypes with M2 type of leukemia. (2) Among cases of M0, M1, and M2 with normal karyotypes, those with initial white blood cell counts less than 2.0 X 10(4)/mm3 responded well to initial chemotherapy and had long remission durations, whereas most cases with white blood cell counts of more than 10.0 X 10(4)/mm3 failed to undergo first remissions and had a poor prognosis, (3) Acute nonlymphocytic leukemia patients with abnormal karyotypes, including t(8;21) and t(15;17), relapsed within 10 mo after the first complete remission, and no remarkable correlation between the specific chromosome changes and remission duration was observed.     

小儿急性白血病患者外周血T淋巴细胞亚群的研究

本文用单克隆抗体SACI-Ig花环法测定了小儿急性白血病及再生障碍性贫血患者外周血T淋巴细胞亚群的变化。结果显示,在急性淋巴细胞性白血病及再生障碍性贫血患儿OKT8~ 细胞均明显高于正常,OKT4~ /OKT8~ 比值显著降低,但在急性粒细胞性白血病患儿淋巴细胞亚群变化不明显。这提示,抑制性T细胞(OKT8~ )的增多可能与小儿急性淋巴细胞性白血病及再生障碍性贫血的发生或发展有关。     

分泌抗急性非淋巴细胞白血病M2型单克隆抗体细胞株的建立及其特异性

应用细胞杂交瘤技术建立了分泌抗急性非淋巴细胞白血病M_2型单克隆抗体1D_1、1C_2和1D_6细胞株,连续传代11个月,稳定地分泌单克隆抗体。用间接免疫荧光方法检测,这3株细胞分泌的单克隆抗体与急性非淋巴细胞白血病M_2型病人白血病细胞呈阳性反应;与M_1、M_3、M_4、M_5型病人白血病细胞呈阴性反应;与慢性粒细胞白血病、慢粒急变、急性淋巴细胞白血病及正常人白细胞也呈阴性反应,说明单克隆抗体有很好的特异性。     

Trisomy 4 in a case of acute lymphocytic leukemia (L1).

Trisomy 4 has been identified previously as a chromosome abnormality associated with acute nonlymphocytic leukemia (ANLL) with myelomonocytic lineage and in myelodysplastic syndromes (MDS). We report a case of acute lymphocytic leukemia (ALL) (French-American-British, FAB L1) in a 42-year-old Japanese man, with trisomy 4 as the sole chromosomal anomaly. Immunophenotypically, the leukemic blasts demonstrated reactivity with CD2, CD5, and CD7 and indicated on early stage of T cell.     

Significance of cytogenetic abnormalities in acute leukemias

Chromosome abnormalities of a clonal nature have been found in approximately 50 per cent of the patients with acute nonlymphocytic leukemia and acute lymphocytic leukemia. Patients with acute myeloblastic leukemia (M1 and M2) with only abnormal metaphases have the worst prognosis among patients with acute nonlymphocytic leukemia, whereas patients with acute myeloblastic leukemia (M1 and M2) with only normal metaphases have the best prognosis. Specific abnormalities include a t(8;21) in approximately 8 per cent of the patients with acute nonlymphocytic leukemia and a t(15;17) in approximately 50 per cent of the patients with either hypergranular (M3) or microgranular (M3 variant) promyelocytic leukemia. In patients who develop acute leukemia after cytotoxic or radiation therapy for a previous malignant disease, clonal chromosome abnormalities are almost always present.     

Granular acute lymphoblastic leukemia

Granules in blasts are most typical of acute myeloblastic leukemia. However, there have been scattered reports of patients with acute lymphoblastic leukemia (ALL) that have lymphoblasts with azurophilic cytoplasmic granules. These reports do not describe immunologic markers or cytogenetics. We report five additional cases with detailed cytologic, immunologic, and cytogenetic studies. At diagnosis one of these patients had central nervous system disease, while the others had no unusual features. Four of the five patients attained remission. The blasts of all five patients contained distinct cytoplasmic azurophilic granules. The granules were negative for peroxidase and chloroacetate esterase and positive for PAS, acid phosphatase, alpha-naphthyl acetate esterase incompletely fluoride inhibited, alpha-naphthyl butyrate esterase, and in one case, Sudan black B. In the one patient studied by electron microscopy, characteristic lymphoblasts contained membrane-bound electron lucent inclusions, which stained positively with non-specific esterase. Immunologic markers showed a common ALL phenotype. Different cytogenetic abnormalities were seen in all cases. It is important to recognize the characteristics of this morphologic subtype of ALL in order to avoid a misdiagnosis of acute nonlymphocytic leukemia.     

Defects of natural killer cell activity in children with untreated acute lymphocytic leukemia

Summary Natural killer (NK) activity against cells of the K-562 line was significantly depressed in 12 of 18 children (66%) with untreated acute lymphocytic leukemia (ALL). No suppression of allogeneic NK activity was observed with sera of the patients, regardless of the level of NK depression. The ability of peripheral blood lymphocytes (PBLs) to suppress allogeneic NK activity was tested in two ALL patients — one with no detectable NK activity, and one with high NK activity. No NK-suppressive activity was found with PBLs of the areactive patient; PBLs of the reactive patients exhibited some suppressive activity, but only at a particular suppressor-to-effector cell ratio. Leukemic blasts were resistant to killing by autologous NK cells stimulated by IFN, as well as to killing by allogeneic, IFN-stimulated PBLs. Leukemic blasts of an ALL patient inhibited lysis of K-562 cells in an 18-h, but not in a 4-h NK assay. The inhibition could partly be reversed by pretreatment of ALL cells with alpha interferon, suggesting that the blasts might inhibit the lysis of K-562 targets in a competitive manner. Disturbed function and/or regulation of NK cells may influence attempts at NK cell activation by lymphokines.Abbreviations ALL acute lymphocytic leukemia - AML acute myeloid leukemia - CLL chronic lymphocytic leukemia - CML chronic myeloid leukemia - IFN interferon - IL-2 interleukin-2 - LGL large granular lymphocyte - NK cell natural killer cell - PBL peripheral blood lymphocyte - Poly I:C polyinosinic-polycytidylic acid     

Chromosome patterns in 26 South African children with acute nonlymphocytic leukemia (ANLL)

Of 46 black leukemic children 52% had acute nonlymphocytic leukemia (ANLL), whereas only 11% of 62 white leukemic children had the disease. An abnormal karyotype was found in 73% of the 26 children with ANLL, and the majority of abnormal karyotypes were pseudodiploid. "Balanced" translocations were noted in 10 children, of whom four had t(8;21) associated with M2 ANLL, two had t(15;17) and M3 ANLL, two had a t(9;22), one child with M5 ANLL had t(10p;11q), and an infant with congenital M5 ANLL had t(8;16). Monosomy #7 was detected in two preleukemic children who subsequently developed M4 ANLL. Hyperdiploidy was present in only three cases. These patterns were compared with those of other published series, confirming the increased frequency of chromosome abnormalities in children with ANLL. The differing ratio of ANLL:ALL, some of the distinctive clinical features, and the high frequency of detectable chromosome abnormalities in black children may be reflections of a particular oncogenic agent(s) within their environmental background that could be responsible for the initiation of the leukemic process.     

Prognostic implications of cytology in acute leukemia in the adult: The case for subacute leukemia

One hundred sixty-nine cases of acute leukemia in adults have been prospectively classified on the basis of the original peripheral blood smear and bone marrow into two subclasses of acute lymphatic leukemia (acute lymphocytic leukemia and subacute lymphocytic leukemia) and five subclasses of acute nonlymphocytic leukemia (acute myelogenous leukemia, subacute myelogenous leukemia, acute myelomonocytic leukemia, erythroleukemia, and acute promyelocytic leukemia). A number od differences in presenting symptoms and physical findings were found among the various subclasses of acute leukemia. Most significant, however, was the marked difference in median survival between patients with acute lymphocytic leukemia (10 months) and subacute lymphocytic leukemia (16.5 months; p=0.01) and acute myelogenous leukemia (2 months) and subacute myelogenous leukemia (6 months; p0.01). The division on the basis of morphology of the acute leukemias into acute and subacute forms has important prognostic significance in terms of survival and perhaps also in terms of response to therapy.     

先天性白血病临床病理学分析（附6例尸检报告）

目的　探讨先天性白血病的病理临床特点 .方法　收集 6例死亡病例 ,进行尸检及临床分析 .结果　 6例检查中 ,肿瘤细胞广泛侵犯内脏器官 ,心、肝、脾、肺、肾和消化道的有 6例 ;骨髓、淋巴结、胸腺、胰、肾上腺有 5例 ;脑和甲状腺 4例 ;膀胱 3例 ;脑膜和皮肤 2例 ;舌咽、睾丸、子宫各 1例 .6例中急淋占 4例 ,急非淋 (急单 ) 2例 ;免疫分型 4例急淋全为T细胞性 ,表现为CD4 5RO( )、CD3( )表达 ;2例急非淋 ;1例CD6 8( )、Lyso( )表达 ,诊断为急单可能性大 (本例生后 1小时死亡 ,临床未及血液学检查、FAB分类 ) ;另 1例为早期病例 ,骨髓涂片FAB分型M5 ,未进行免疫检测分型 .结论　先天性白血病在尸检材料中见肿瘤细胞呈广泛性内脏器官浸润 ,并且血管腔内瘤栓形成 ,沿血道传播全身 .本病有发病早、病情急而凶险、治疗困难预后差、早期死亡的特点 .     

High resolution chromosomes of the t(9;22) positive leukemias

A combined bromodeoxyuridine (BrdU) and actinomycin D (AMD) treatment of methotrexate (MTX) synchronized bone marrow cells from four patients with chronic myelogenous leukemia (CML), and two each with acute lymphocytic (ALL) and acute nonlymphocytic (ANLL) leukemia were used to define the breakpoints involved in the t(9;22) found in subgroups of these three disorders. An additional 20 patients with CML were studied with the MTX technique alone. All cases showed the same breakpoints at subbands 9q34.1 and 22q11.2. In CML, it was possible to further define the breakpoint of chromosome 22 to subband q11.21.     

白血病髓外浸润的针吸细胞病理学诊断

目的 探讨对白血病髓外浸润行针吸细胞病理学(FNAc)诊断和鉴别诊断的要点。方法 对65例白血病髓外浸润的临床及FNAc诊断结果进行了分析总结。为确诊,对疑为白血病者,用油镜观察细胞形态并做外周血涂片镜检以助确诊,部分病例做了髓过氧化物酶(MPO)、氯醋酸AS-D奈酚酯酶(CE)、酸性非特异性酯酶(ANAE)及糖原(PAS)染色。部分病例做了组织病理学、免疫细胞化学及流式细胞仪检查。结果 65例髓外浸润中急性淋巴细胞性白血病(ALL)24例、急性髓细胞性白血病(AML)25例、慢性淋巴细胞性白血病(CLL)6例、慢性髓细胞性白血病(CML)10例。各部位中以淋巴结浸润最多,占73．8％。结论 白血病髓外浸润的FNAC诊断应紧密结合临床及有关实验室检查,特别是以局部肿块为首发症状而就诊的初诊白血病。各类白血病髓外浸润以原始细胞增生为主者,需与非霍奇金淋巴瘤(NHL)鉴别。FNAC拟诊为NHL者均应及时用油镜观察细胞形态并做外周血涂片镜检。     

伴7号染色体异常的急性白血病32例分析

目的探讨7号染色体异常在急性白血病中的发生率及预后意义。方法采用R带常规显带技术进行染色体检查,对410例急性白血病患者的核型进行分析。结果410例急性白血病患者中检出7号染色体异常患者32例,占7.8%;其中-7/7q-19例(59.4%),t(7;11)3例(9.4%),其他为少见的7号染色体改变der(7), 7,t(2;7),t(5;7),t(7;9),t(7;8),dic(1;7)。-7/7q-中,急性髓细胞白血病12例,其中M0、M1、M2型的-7/7q-发生率高于其他类型急性髓细胞白血病。32例患者中20例同时有其他染色体异常,最常见的是t(9;22)伴-7, 8,-5。30例进行正规化疗的患者,11例缓解,缓解率低于同期急性白血病的总缓解率(36.7%vs65.8%);伴-7/7q-的急性髓细胞白血病的缓解率低于染色体正常的急性髓细胞白血病患者(25%vs55.6%);伴-7/7q-的急性淋巴细胞白血病的缓解率与染色体正常的急性淋巴细胞白血病患者无差异(57.1%vs77.8%),但缓解的4例急性淋巴细胞白血病患者均于短期内复发。伴其他7号染色体异常的11例患者仅4例缓解。结论-7/7q-是7号染色体异常中最为常见的核型改变,且多见于急性髓细胞白血病的M0、M1、M2型,并可能与急性白血病的发病有关;伴7号染色体异常的急性白血病患者预后较差。