Granular acute lymphoblastic leukaemia of childhood: a morphological phenomenon.

Three hundred and twenty consecutive children with lymphoblastic leukaemia (ALL), treated on the Medical Research Council UKALL VIII schedule, had their Romanowsky stained diagnostic marrows reviewed for the presence of azurophil granules in blast cell cytoplasm. Twenty patients (7%) had greater than 5% blasts showing this feature; 19 had the cell phenotype of "common ALL." Male children and those with French-American-British (FAB) L2 morphology predominantly showed this feature. There was also a strong correlation between granularity and non-diffuse acid phosphate positivity, but no obvious difference between the 20 patients in their response to treatment emerged during a minimum follow up of 15 months. The "granular" variant occurs in around 7% of children with ALL, but has no clear prognostic importance. Morphologists should be aware of its existence and incidence to avoid confusion with acute myeloid leukaemia.     

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.     

Translocation (2;8)(p12;q24) associated with a cryptic t(12;21)(p13;q22) TEL/AML1 gene rearrangement in a child with acute lymphoblastic leukemia

We report a case of childhood acute lymphoblastic leukemia with the simultaneous occurrence of a t(2;8)(p12;q24) typically associated with mature B cell or Burkitt leukemia, and a t(12;21)(p13;q22) exclusively associated with pre-B cell ALL. The lymphoblasts were characterized as L2 morphology by the French-American-British classification. However, there were atypical morphologic findings for L2 ALL, including vacuolization in some cells. The lymphoblasts were periodic acid-Schiff positive and myeloperoxidase negative. Immunophenotypic analysis revealed that the majority of lymphoblasts were TdT+, CD10+, CD19+, CD20-, and cytoplasmic mu+. These features were consistent with an immature pre-B cell leukemia phenotype with some characteristics of a mature B-cell leukemia. A t(2;8)(p12;q24)(p12;q24), characteristic of mature B-cell leukemia or Burkitt type leukemia, was detected by conventional cytogenetics with no other cytogenetic abnormalities. However, diagnostic peripheral blood and bone marrow specimens demonstrated simultaneous occurrence of a cryptic t(12;21)(p13;q22) by both FISH and RT-PCR. The simultaneous occurrence of these translocations in a pediatric patient have implications for the pathogenesis of leukemias with t(2;8)(p12;q24) as well as t(12;21)(p12;q22). Analysis of additional cases of leukemia with translocations involving the MYC locus on 8q24 will be required to determine the frequency of association with the cryptic t(12;21)(p13;22), and the prognostic significance of the simultaneous occurrence of the translocations.     

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.     

Symposium: classification of leukemia. 1. The classification of acute leukemia

Two main forms of acute leukemia have been recognized by the French-American-British (FAB) group: myeloid (AML) and lymphoblastic (ALL). Some types of AML can be diagnosed on well prepared bone marrow films stained with May-Grünwald-Giemsa. Poorly differentiated types, myeloblastic (M1) and monoblastic (M5PD), need confirmation by positive cytochemical reactions (Sudan Black B, myeloperoxidase and non-specific esterase). There are 2 sub-types of promyelocytic leukemia: M3 typical, hypergranular and M3 variant, microgranular. The M3 variant has a more acute course, higher WBC and may require cytochemistry to demonstrate promyelocytic differentiation. Electron microscopic cytochemistry can also help in the classification of difficult AML cases; the 'platelet-peroxidase' reaction, for example, is essential for the diagnosis of megakaryoblastic leukemia, a disorder often presenting as 'acute' myelosclerosis. Three morphological types are seen in ALL: L1, predominantly in children, L2, more frequently in adults, and the relatively rare L3 or Burkitt type. Immunological and enzyme markers (ALL and la antigens, terminal transferase, etc.) help define the cell phenotype: (1) non-B, non-T ALL with 3 forms (common, null and pre-B), (2) T-ALL, related to but distinct from T-lymphoblastic lymphoma, and (3) B-ALL, usually with L3 morphology, There is growing evidence that the FAB morphological types correlate with prognosis in ALL independently of other factors. The immunologically defined types also correlate with prognosis but not as an independent variable.     

The morphologic classification of acute lymphoblastic leukemia in childhood. Observations on concordance using a simple scoring system

The French-American-British (FAB) cooperative group proposed a simple scoring system in 1981 to improve concordance among researchers and clinicians for the subclassification of acute lymphoblastic leukemia (ALL) into subtypes L1 and L2. The authors subtyped 50 consecutively admitted patients with common ALL of childhood according to the FAB scoring system to assess concordance among a member of the FAB group (J.B.) and two pathologists (S.S. and C.C.) who are not FAB members. The initial agreement of 66% was unsatisfactory to the authors prompting the critical evaluation of the major sources of disagreement. Concordance was improved greatly by the use of two technics that are logical extensions of the scoring system: performing 100-200 differential cell counts to determine the correct percentages in borderline cases, and using reference drawings that illustrate blasts with cytoplasmic contents of 15%, 20%, and 25% in determining the cell's nuclear cytoplasmic ratio. The authors believe that the criteria explicitly given greater weight by the FAB scoring system (nuclear cytoplasmic ratio and nucleoli) are the most subjective. By following the criteria carefully and resolving differences together at a multiheaded microscope, a concordance of 92% was achieved.     

Terminal deoxynucleotidyl transferase-negative acute lymphoblastic leukemia

OBJECTIVE: Terminal deoxynucleotidyl transferase (TdT) is a useful marker in the diagnosis of acute lymphoblastic leukemia (ALL) (French-American-British [FAB] L1 and L2) and is most useful in distinguishing ALL from mature B-lymphoid neoplasms, such as Burkitt lymphoma (FAB L3) and other lymphoid malignancies. The frequency of TdT-negative ALL is not known. Here we report 3 TdT-negative ALL cases that met the criteria for T-cell ALL. DESIGN: We reviewed approximately 200 cases of ALL retrieved from the database at our institution. All cases were evaluated using Wright-Giemsa, myeloperoxidase, butyrate, and TdT staining; immunophenotyped using flow cytometry; and studied using Southern blot analyses for T-cell receptors and immunoglobulin gene rearrangement. RESULTS: All ALL cases (L1 and L2) were TdT-positive, except for 3 cases that were of early T-cell lineage. None of the 3 cases demonstrated positivity for TdT in immunofluorescence staining with polyclonal antibodies or flow cytometry with monoclonal antibodies. Flow cytometric analysis confirmed a pre-T-cell immunophenotype in all 3 cases. One of the cases showed rearrangement of a T-cell antigen receptor and immunoglobulin heavy chain (J(H)). A second case showed germline configuration of T-cell receptors, but also showed rearrangement of the J(H), despite the expression of T-cell markers only.     

A rare translocation (4;11)(q21;p14-15) in an acute lymphoblastic leukemia expressing T-cell and myeloid markers.

A 21-year-old male presented with a large mediastinal mass and a white cell count of 420 x 10(9)/L. A diagnosis of acute lymphoblastic leukemia (ALL) was made, with 90% of cells in the bone marrow (BM) and 99% in the peripheral blood (PB) being lymphoblasts (FAB L1). Cytogenetic analysis of these cells revealed a rare variant of the t(4;11) translocation involving chromosome arm 11p rather than 11q, namely t(4;11)(q21;p14-15). The standard form of the (4;11) translocation has been associated with leukemias with mixed-lineage phenotypes. Three cases of ALL with t(4q;11p) have previously been reported. One of these cases showed phenotypic heterogeneity involving myeloid and lymphoid lineages. The leukemia reported here also exhibits lymphoid/myeloid features. Immunophenotyping of the blasts showed that most of the cells were positive for CD2, CD5, CD7, CD10 (CALLA), CD34, and HLA-DR. A significant proportion of the cells expressed CD33. These results suggest a biphenotypic rather than a biclonal disease. Molecular analysis showed rearrangement of both immunoglobulin heavy-chain genes (JH) and of a single allele of the T-cell receptor (TCR) gamma 1 gene, while retaining germline TCR beta genes.     

Morphological metamorphosis in relapsing lymphoblastic leukemia.

Blast cell morphology was assessed at diagnosis and subsequent bone marrow relapse in 33 unselected patients with lymphoblastic leukemia (ALL). Each marrow was classified 'blind' according to the French-American-British (FAB) criteria, and it was found that 19 of 24 (79%) patients initially typed as FAB L1 changed to FAB L2 during the course of their disease, but no patient made the reverse morphological change (p is less than 0.001). Five patients retained FAB L1 appearances; these included three of the four who had T-cell markers. One patient typed as FAB L3 did so consistently. This study indicates that FAB L2 ALL frequently emerges as a treatment-resistant offshoot of FAB L1 and provides further evidence that this marks a more aggressive form of the disease.     

Acute lymphoblastic leukemia. Survey of immunophenotype, French-American-British classification, frequency of myeloid antigen expression, and karyotypic abnormalities in 210 pediatric and adult cases

Immunophenotypic studies are essential to distinguish acute lymphoblastic leukemia (ALL) from minimally differentiated acute myeloid leukemia (AMLM0) and to classify ALL into immunologic subtypes. Frequently, immunophenotyping identifies myeloid antigen expression in ALL, causing a potential diagnostic problem. To evaluate the immunophenotype of ALL, we studied 210 cases of pediatric and adult ALL by flow cytometry and compared the results with the French-American-British (FAB) Cooperative Group classification and the karyotypic findings. Myeloid-associated antigens were expressed in 78 (45.6%) of precursor B-cell ALL cases. Pediatric precursor B ALLs had a higher frequency of myeloid antigen expression than did adult cases. All mature B-cell ALL cases were negative for TdT and myeloid antigens. Myeloid antigen expression was less frequent in T-cell ALL cases compared with precursor B-cell ALL cases. Of the 192 cases submitted for cytogenetic analysis, 147 were abnormal. The most common chromosomal translocation was the Philadelphia chromosome, which was more likely to have L2 blast morphology and a precursor B immunophenotype. Myeloid antigen expression was present in 70.8% of Ph-positive cases (P = .008). Chromosome rearrangements involving 11q23 also showed an increased frequency of myeloid antigen expression. Chromosome translocations involving regions of T-cell receptor genes were present in 24% of T-cell ALL cases. A high percentage of ALL cases, however, had various other cytogenetic abnormalities, many of which involved less well-studied chromosomal regions.     

Immunophenotypic analysis of acute lymphoblastic leukemia using routinely processed bone marrow specimens

Monoclonal antibodies have been recently developed that react with antigens expressed on T and B lymphocytes in routinely processed, paraffin-embedded lymphoid tissues. In this study, we assessed bone marrow clot and/or core biopsy sections of 19 cases of acute lymphoblastic leukemia (ALL) using routinely decalcified, B5- or formalin-fixed, paraffin-embedded sections and a panel of monoclonal antibodies, including LN1, LN2, L26, Leu-22, UCHL-1, and LCA. Each case had been previously phenotyped using freshly obtained aspirate material and a standard immunophenotypic protocol. Our results demonstrate the utility of the LN2 antibody in differentiating between precursor B-cell (pre-B) and precursor T-cell ALL. The LN2 antibody stained 11 of 12 cases of pre-B ALL and did not react with any of the seven T-cell ALLs. The other antibodies tested were less helpful. The Leu-22 antibody stained both pre-B and T-cell ALLs, while the results with UCHL-1 revealed peculiar nuclear staining of pre-B and T-cell ALLs; this we attributed to processing artifact. The L26 antibody reacted with only one case of pre-B ALL (also CD20 antigen positive), while the LN1 antibody did not react with any pre-B ALLs. Neither L26 nor LN1 stained any cases of T-cell ALL. The LCA antibody stained in only four (21%) of 19 cases, two pre-B and two T-cell ALLs. The results also suggest that this panel of antibodies may be useful in differentiating ALL from mature B-cell and T-cell lymphomas involving the bone marrow.     

Acute lymphoblastic leukaemia of the L3 subtype in adults in the Northern health region of England 1983-99

AIM: Acute lymphoblastic leukaemia (ALL) with an L3 morphological FAB type is regarded by some as being synonymous with B cell ALL or ALL with a Burkitt-type chromosomal translocation-t(8;14), t(2;8), t(8;22). This paper describes a series from a population based study of 24 patients with L3 ALL presenting over 17 years. METHODS: Clinical data were collected prospectively from all adult patients presenting with acute leukaemia in the Northern region since 1982. Data from all patients diagnosed with FAB type L3 ALL were analysed. RESULTS: Overall, L3 ALL accounts for 8.6% of all adult ALL and it is more common in the elderly than has hitherto been recognised. In addition to classic Burkitt-type translocations (11 of 24 cases), the t(14;18) translocation, which is characteristically found in lower grade lymphomas such as follicular lymphoma, is frequently present (five of 24 cases). CONCLUSION: The presence of L3 ALL is often associated with non-Burkitt-type translocations and the presence of a t(14;18) translocation may indicate that in some cases a clinically non-apparent lymphoproliferative disorder, such as a low grade follicular lymphoma, has transformed to a more aggressive form and, thus, presents as a de novo acute leukaemia.     

Morphological and immunological classification and response to chemotherapy in adult patients with acute leukemias

Three hundred forty four adult patients with acute leukemia were classified according to the FAB classification, and revealed 109 patients were ALL and 235 were AML. Surface marker analysis were done in 52 cases with ALL and 51 with AML. Patients with ALL were treated with VP or LVP therapy and those with AML were treated with DC(M)P, BH-AC DMP, DCMP-85 or DBMP-85. L 1 was 56% of ALL patients and L 2 was 44%. Complete remission (CR) rate of L 1 was 78.8% and that of L 2 was 62.5%, however 5 year survival rate was less than 20% in both groups. c-ALL was 62% of ALL, T-ALL and N-ALL were 15% and 13% respectively. L + M ALL was 8% and B + T ALL was 2%. CR rate was almost same among each groups, however disease free survival (DFS) was better in T-ALL and worse in c-ALL. M 1 was 23.4% of AML, M 2, M 3, M 4, M 5 and M 6 were 42.6%, 15.7%, 11.9%, 5.1% and 1.3% respectively. CR rate of all AML was 71.9% and that of M 3, M 5 and M 6 was worse than the average. Five years continued complete remission (CCR) rate of M 2, M 3 and M 4 were 20-25%, while that of M 1 and M 5 were worse than the former. Myeloid-AML was 67% of AML, L + M-AML and lymphoid-AML were 27% and 6% respectively. CR and CCR rate of L + M-AML and lymphoid-AML were seemed to be better than those of myeloid-AML. Thus, the patients with AML, even though they have the blasts which express lymphoid markers, can be treated with AML directed therapy.     

French American British (FAB) morphological classification of childhood lymphoblastic leukaemia and its clinical importance

As part of the Medical Research Council Leukaemia Trial UKALL VIII, 738 unselected children with acute lymphoblastic leukaemia (ALL) had the morphology of their marrow blast cells reviewed by a panel of three haematologists. Ninety four (13%) showed appearances classifiable as type L2 by the French American and British (FAB) cooperative group's criteria, five (0.7%) were typed L3, and the remaining 639 (86%) as L1. Disregarding the patients classified as L3, those with the L2 variant showed an inferior disease free survival to that of the remainder (p less than 0.01), and more of them failed to remit after receiving "standard" remission induction treatment (p less than 0.01). They included an excess of older children (p less than 0.01) with less profound marrow failure at diagnosis, and fewer of them expressed the common ALL antigen (p = 0.05). There was no association between L2 morphology and the diagnostic white cell count, sex, or the presence of a mediastinal mass. These findings confirm earlier reports that FAB L2 ALL is associated with a poor prognosis and that it occurs more commonly in older children. The high remission failure rate is a recent observation and indicates that alternative early treatment may be appropriate for such patients.     

Terminal deoxynucleotidyl transferase activity in childhood and adult acute lymphoblastic leukaemia.

The measurement of terminal deoxynucleotidyl transferase (TDT) activity in leukaemic blasts of 26 cases of acute lymphoblastic leukaemia (ALL) (13 children less than 14 years, 13 adults greater than 14 years) demonstrated significantly greater activities of the enzyme on a proportion of the adults. The predominant cytological sub-type in the adult patients was L2 (FAB classification) whereas L1 cytological sub-type dominated in the childhood group. There was no relation between TDT values and FAB sub-type but the highest activities in the childhood group were seen in patients assessed at the time of relapse. We conclude that continued use of quantitative TDT estimations may provide useful information in further characterising the currently recognised cytological and immunological sub-types in ALL.     

免疫表型与急性白血病的分型诊断

对426例急性白血病采用APAAP法23种单抗进行免疫分型研究.结果发现:(1)426例中,71.6%可确定为淋巴系或髓系或髓/淋混合型,28.4%不能确定细胞系列.FAB结果仅62.75%得到免疫表型证实,29.5%不能判断细胞起源,7.75%两者分型结果不一致;(2)ANLL140例中,44.29%免疫表型证实为髓细胞型,17.14%仅表达淋巴系标志,38.57%不能判断细胞起源,各髓系标志无明显FAB亚型特异性;(3)ALL260例中,72.67%进一步证实为淋巴细胞型,2. 69%单纯表达髓系标志或混合表达髓系和淋巴系抗原,另外24.62%不能确定细胞起源;(4)FAB不能分型26例中,23例可确定为ALL或ANLL,另外3例仍不能诊断.研究认为,免疫分型是FAB分型的必要补充.     

Terminal deoxynucleotidyl transferase (TdT) in acute nonlymphocytic leukemia. A clinical, morphologic, cytochemical, immunologic, and ultrastructural study

The classification of acute leukemia is essential for proper therapy and may be based on morphologic, cytochemical, immunologic, or even ultrastructural studies. Terminal deoxynucleotidyl transferase (TdT) is expressed in most patients with acute lymphocytic leukemia (ALL) and a minority of patients with acute nonlymphocytic leukemia (ANLL). Thirteen patients with ANLL and greater than 30% blasts positive for TdT were studied to establish the clinical, light microscopic, cytochemical, immunologic, and ultrastructural correlates of this phenomenon. Most patients demonstrated some morphologic and cytochemical features of monocytic differentiation. On cytochemical stains, nine had greater than 3% Sudan black-positive blasts. Diffuse alpha naphthyl acetate esterase (ANAE) staining of leukemic cells was present in nine cases, though extremely weak in seven. Blasts in ten patients did not express any other markers of lymphoid differentiation except TdT. However, two patient's immature cells bore CD10 common ALL antigen (CALLA) and CD19 (B4). Ultrastructural studies confirmed nonlymphoid differentiation in all ten patients studied, with a prominent monocytic component present in nine. In no case was a second population of lymphoblasts identified to account for TdT positivity. These patients responded poorly to conventional therapy for ANLL, with complete remissions in 3 of 13 (23%). With conventional therapy for ALL, complete remission was achieved in only two of nine (22%) patients. However, four of seven (57%) patients had a complete remission with high-dose cytosine arabinoside regimens. The authors' studies suggest that patients with TdT-positive ANLL represent a distinct subset that usually displays ultrastructural evidence for monocytic differentiation and is clinically significant in that these patients respond poorly to conventional therapy for both ALL and ANLL. Recognition of the monocytic lineage of these cases by light microscopic examination is difficult because they are often poorly differentiated morphologically and express only weak nonspecific esterase positivity.     

Chloroacetate esterase positivity in acute lymphoblastic leukemia

Chloroacetate esterase (CAE) reactivity is associated with acute nonlymphocytic leukemia (ANLL) and, to the author's knowledge, has not been reported in acute lymphoblastic leukemia (ALL). The authors have identified a single patient whose blasts displayed CAE reactivity from a review of 400 newly diagnosed patients with ALL. The diagnosis of ALL in this case was based upon morphologic evaluation (FAB:L1), presence of terminal deoxynucleotidyl transferase (TdT), the absence of reactivity with myeloperoxidase, and the failure of leukemic blasts to mark with a panel of anti-myeloid monoclonal antibodies. Remission was induced promptly with standard ALL therapy. This case demonstrates that punctate CAE positivity rarely can occur in ALL, and, therefore, CAE is not entirely specific for ANLL. The authors conclude that CAE positivity should not be used as a sole criterion to diagnose ANLL in the absence of supporting morphologic, cytochemical, immunocytologic, or clinical information.     

t(2;14)(p13;q32): a recurring abnormality in lymphocytic leukemia. A Pediatric Oncology Group study.

Chromosome banding studies of 1,411 children with newly diagnosed acute lymphocytic leukemia (ALL) identified two patients with the t(2;14)(p13;q32) chromosome abnormality and a third patient with a complex three-way translocation involving the same breakpoints on chromosomes 2 and 14 but also involving chromosome 12 at band q11. The three cases demonstrated variability of immunophenotypes: one was a T-cell ALL, and two were early pre-B ALLs. All three patients achieved complete remissions and have remained in remission for 14-19 months.