Normal variation of bone marrow B-cell precursors according to age - reference ranges for studies in myelodysplastic syndromes in Brazil.

BACKGROUND: Normal B lymphoid maturation occurs in bone marrow (BM) throughout life, but immature B-cell progenitors (BCPs) are more numerous in children than in adults. To assess the normal values according to age became important as BCPs are decreased in myelodysplastic syndromes and have been considered an important diagnostic and prognostic feature in these clonal disorders. METHODS: in a multicenter retrospective study from the Brazilian Group of Flow Cytometry we analyzed the variation of BCPs in normal BM according to age and technical peculiarities of each laboratory. We analysed of 45 BM donors and 89 cases examined for elucidation of transitory reactive cytopenias presenting a normal BM immunophenotyping. BCPs were enumerated as CD19+ /CD34+ /CD45dim /CD10+ cells (panel 1) or CD19+ /CD34+ /CD45dim cells (panel 2) among the total nucleated non-erythroid cells and as percentage of CD34+ cells. RESULTS: we included 134 cases. Panel 1 was applied in 88 cases and panel 2 was used in 46. Age range: 10 months to 89 years. In a multiple regression, % BCPs/total nucleated cells was an exponential function of age. Age explained alone 49.4% of the variance, while 'panel used' explained 1.8% and 'laboratory' explained 0.7%. Age explained only 24.9% of the variance of BCPs/CD34+ cells. CONCLUSIONS: in normal individuals, BM B-cell precursors varied mainly according to age, but were also dependent on technical peculiarities of operators and equipments. Analysis by phenotype and as percentage of total nucleated cells was more accurate and less susceptible to variation than evaluating % BCPs/total CD34+ cells. © 2017 International Clinical Cytometry Society.

Different expression patterns of LGALS1 and LGALS3 in polycythemia vera, essential thrombocythemia and primary myelofibrosis.

Despite all the knowledge, the cellular and molecular mechanisms involved in myeloproliferative neoplasm (MPN) pathophysiology remain unclear. Authors have shown galectin-1 (Gal-1) and 3 playing roles in tumour angiogenesis and fibrosis, which were correlated with poor prognosis in patients with MPN. In the present study LGALS1 and LGALS3 were differently expressed between polycythemia vera, essential thrombocythemia (ET) and primary myelofibrosis (PMF) diseases. Increased LGALS3 expression was associated with a negative JAK2 V617F status mutation in leucocytes from PMF but not in patients with ET without this mutation. However, a positive Janus kinase 2 (JAK2) V617F cell line established from patients with ET (SET-2 cells) when treated with JAK inhibitor presented high levels of LGALS3. Additionally, high LGALS1 expression was found in CD34(+) cells but not in leucocytes from patients with PMF, in absence of JAK2 V617F mutation, and also in SET-2 cells treated with JAK inhibitor. Thus, our findings indicate that differential expression of LGALS1 and/or LGALS3 in patients with MPN is linked with JAK2 V617F status mutation in these diseases and state of cell differentiation.

Application of five prognostic survival scores to primary myelofibrosis in 62 Brazilian patients.

Primary myelofibrosis (PM) is a Philadelphia-negative clonal hematopoietic stem cell disorder characterized by intense reactive changes of bone marrow stroma with collagen fibrosis, osteosclerosis and angiogenesis. PM usually affects elderly people, and approximately half of the patients present JAK2V617F mutation. PM clinical course varies from 1 to 30 years, evolving from asymptomatic into progressive bone marrow failure, symptomatic splenomegaly or acute leukemia in 10-20 % of cases. PM risk stratification is based on parameters predicting survival, and several attempts have been made to identify clinical and laboratory features that could predict PM patient survival. This study applied five prognostic scores: Dupriez, Cervantes, Mayo, IPSS and DIPSS-Plus in 62 Brazilians patients from three centers, and compared their relevance and clinical usefulness considering the scores' parameters, fibrosis, JAK2V617F mutation, splenomegaly, hepatomegaly and treatment. According to the Cervantes, Dupriez and Mayo scores, most patients were stratified into low-risk group. However, when IPSS and DIPSS-Plus were applied, most patients were classified into an intermediate range, being low risk in only 11 and 13 % of patients, respectively. Overall survival at 4 years was 84 %. The Cervantes score was the only one that remained significantly associated with survival in a multivariate analysis. In conclusion, the Cervantes score remains important to the prognostication of PM.

Myelodysplastic syndromes (MDS): prognostic factors and scoring systems.

OBJECTIVE: To evaluate the score systems of Cassano and Sanz and suggest a new one. DESIGN: Case series. LOCATION: Teaching hospitals: EPM UNIFESP and Faculdade de Medicina de Botucatu. PARTICIPANTS: 59 patients diagnosed from 1979 to 1992. INTERVENTION: Evaluation of clinical-laboratorial data. MEASUREMENT: Statistical comparison, uni and multivariate analysis and actuarial survival curves. RESULTS: Cassano's system divided the patients into high and low risk (p = 0.0966) while Sanz's gave high, intermediate and low risk (p = 0.0108). The univariate analysis showed hemoglobin, WBC count, E/M ratio, liver size and blast percentage in BM as statistically significant. The multivariate analysis showed blast percentage in BM (p = 0.004) and Hb (p = 0.050) as significant. Our system, considering the multivariate analysis data, divided the patients into high, intermediate and low risk (p = 0.0038). CONCLUSIONS: Sanz's system was more functional than Cassano's, while ours showed predictive survival value and ease of use in clinical practice.

Alteraçöes citogenética como fatores prognósticos em leucemias mielóides agudas / Cytogenetic abnormalities as prognostic factors in acute myeloide leukemia

O estudo das alteraçoes cromossômica nas leucemias mielóides agudas (LMA) vem-se tornando importante no diagnóstico e na caracterizaçao de subtipos, pois associam-se a características clínicas, morfológicas e imunológicas definidas à resposta a tratamento e à sobrevida. OBJETIVO. O presente trabalho objetiva avaliar a importância relativa das alteraçoes citogenética em portadores de LMA. MATERIAL. Foram estudados, ao diagnóstico, 13 pacientes com LMA e com idade mediana igual a 38 anos. O estudo citogenético foi realizado em material medular.RESULTADOS. Os subtipos FAB M1 e M2 foram o mais freqüentes (61,6 por cento). A análise citogenética mostrou cariótipo anormal em 61,5 por cento dos casos e, dentre estes, apenas 15,3 por cento tinham alteraçoes indicadoras de bom prognóstico [t(l5;17) e t(8;21)]. Na data de avaliaçao do estudo havia três pacientes vivos, dois em remissao completa contínua e um em segunda remissao. A sobrevida mediana global foi de 7 meses. Os pacientes foram divididos em dois grupos: um intitulado "bom prognóstico", que englobou cinco indivíduos com cariótipo normal e dois com as translocaçoes t(l5;17) e t(8;21), e outro, "mau prognóstico", com oito pacientes com alteraçoes cromossômicas desfavoráveis. O grupo "bom prognóstico" teve sobrevida mediana de nove meses, enquanto outro, de 6,2 meses, mas sem diferença estatisticamente significante (p= 0,180084), provavelmente devido ao pequeno número de casos em cada grupo. Entretanto, ao se analisar os casos em separado nota-se que os pacientes com translocaçoes (8;21) e (15;17), tidas como de bom prognóstico, tiveram sobrevidas mais longas. CONCLUSAO. Concluímos que o trabalho evidenciou sobrevida desigual entre os dois grupos, ressaltando a importância da análise citogenética que permite distinguir o paciente que terá evoluçao favorável.

[Cytogenetic abnormalities as prognostic factors in acute myeloid leukemia]. / Alterações citogenéticas como fatores prognósticos em leucemias mielóides agudas.

UNLABELLED: The study of chromosomal abnormalities in AML has become very important in the diagnosis and in the characterization of subtypes since they are related to defined clinical, morphological and immunological features as well as treatment outcome and survival. PURPOSE: To evaluate the relative importance of cytogenetic abnormalities may have in AML patients. METHODS: 13 AML patients were studied during diagnosis. Cytogenetic study was performed on bone marrow aspirate material. RESULTS: M1 and M2 FAB subtypes were the most frequent (61.6%). The patients' median age was 38 years. Cytogenetic analysis showed abnormal karyotype in 61.5% of the cases and 15.3% of whom had abnormalities considered as good prognosis [t(15;17) and t(8;21)]. At the evaluation day there were 3 patients alive, two in continuous complete remission and 1 in a second remission. The median total survival time was 7 months. Patients were divided into two groups: a "good prognosis" one, that joined 5 patients with normal karyotype and 2 with the translocations t(15;17) and t(8;21) and another, the "bad prognosis" one, with 8 patients with unfavorable chromosomal abnormalities. The good prognosis group had a median survival time of 9 months versus 6.2 months in the other, but this was not statistically significant (p = 0.18), probably owing to the small number of cases in the groups. But when one observes the cases separately see that patients with translocations (8;21) and (15;17), known as good prognosis, had longer survivals. CONCLUSION: The different survival time between the two groups showed the importance of cytogenetic study to distinguish the patient who will have favorable evolution.

Chronic myeloid leukemia (CML): prognostic factors and survival analysis.

The prognostic value of different factors upon diagnosis of CML was analysed in 45 Philadelphia (Ph1)-positive patients. The median survival was 48 months. Univariate analysis showed 5 poor prognostic factors (male sex, under 45 years-old, bone marrow blasts greater than or equal to 10 percent, blood basophils greater than or equal to 6 percent and blood eosinophils greater than or equal to 6 percent) which provided for the development of a clinical staging system: Stage I with none or one factor and a two-year survival rate of 100 percent; Stage II with two or three factors and two-year survival of 72.2 percent; and Stage III with four or five factors and two-year survival of 0 percent (p = 0.00016). Multivariate survival analysis showed that combination of blood basophilia and bone marrow blasts had the strongest predictive relationship to survival time. We conclude that a combination of pretreatment factors identifies different risk subcategories in CML patients and is helpful in assessing the overall prognosis and the treatment approach.