Avaliação do perfil de infiltração e relevância do uso de estabilizantes celulares nas amostras de líquido cefalorraquidiano recebidas no Laboratório de Marcadores Celulares do Centro de Hematologia e Hemoterapia de Santa Catarina / Infiltration profile evaluation and relevance of the use of cellular stabilizers in cerebrospinal fluid samples received by Cell Labelling Laboratory in Hematology and Hemotherapy Center of Santa Catarina

Objetivo: O presente estudo pretende analisar o perfil e as condições das amostras de líquido cefalorraquidiano (LCR) recebidas no setor de Marcadores Celulares do Centro de Hematologia e Hemoterapia de Santa Catarina (HEMOSC). Métodos: Os dados foram obtidos retrospectivamente através do sistema HemoSis e dos planos de trabalho contidos no laboratório num intervalo de 12 meses. Resultados: Das 117 amostras avaliadas, observa-se que 54% das provenientes do interior do estado de Santa Catarina são pouco viáveis, enquanto que apenas 9% das amostras da grande Florianópolis apresentaram este problema. No total de amostras avaliadas, quatro laudos foram inconclusivos devido à baixa viabilidade. Em relação ao perfil das amostras, somam-se LCR referentes a 65 pacientes sem predomínio de sexo, majoritariamente em idade adulta e se sobressaem as investigações de Linfoma Não-Hodgkin B e leucemias agudas. Dos pacientes, nove revelaram amostras infiltradas. Entre as neoplasias que acometem o sistema nervoso central, há um predomínio de infiltração por doenças linfoides nas amostras recebidas pelo laboratório. Além disso, a baixa viabilidade celular em algumas amostras pode estar associada a resultados inconclusivos ou de baixa confiabilidade. Conclusão: É preciso adequar as coletas do LCR e inserir o uso dos estabilizantes na rotina para evitar resultados inconclusivos e possíveis recoletas de amostra.

Evaluation of CD307a expression patterns during normal B-cell maturation and in B-cell malignancies by flow cytometry.

BACKGROUND: Flow cytometric immunophenotyping is deemed a fundamental tool for the diagnosis of B-cell neoplasms. Currently, the investigation of novel immunophenotypic markers has gained importance, as they can assist in the precise subclassification of B-cell malignancies by flow cytometry. Therefore, the purpose of the present study was to evaluate the expression of CD307a during normal B-cell maturation and in B-cell malignancies as well as to investigate its potential role in the differential diagnosis of these entities. METHODS: CD307a expression was assessed by flow cytometry in normal precursor and mature B cells and in 115 samples collected from patients diagnosed with precursor and mature B-cell neoplasms. CD307a expression was compared between neoplastic and normal B cells. RESULTS: B-acute lymphoblastic leukemia cases exhibited minimal expression of CD307a, displaying a similar expression pattern to that of normal B-cell precursors. Mantle cell lymphoma (MCL) cases showed the lowest levels of CD307a among mature B-cell neoplasms. CD307a expression was statistically lower in MCL cases than in chronic B lymphocytic leukemia (CLL) and marginal zone lymphoma (MZL) cases. No statistical differences were observed between CD307a expression in neoplastic and normal plasma cells. CONCLUSION: These results indicate that the assessment of CD307a expression by flow cytometry could be helpful to distinguish CLL from MCL, and the latter from MZL. Although these results are not entirely conclusive, they provide a basis for further studies in a larger cohort of patients. © 2018 International Clinical Cytometry Society.

Determination of normal expression patterns of CD86, CD210a, CD261, CD262, CD264, CD358, and CD361 in peripheral blood and bone marrow cells by flow cytometry.

In 2010, new monoclonal antibodies were submitted to the 9th International Workshop on Human Leukocyte Differentiation Antigens, and there are few studies demonstrating normal expression patterns of these markers. Thus, the objective of this study was to determine the normal patterns of cell expression of CD86, CD210a, CD261, CD262, CD264, CD358, and CD361 in peripheral blood (PB) and bone marrow (BM) samples by flow cytometry. In the present study, CD86 was expressed only in monocytes and B lymphocytes in PB and in monocytes and plasma cells in BM. Regarding CD210a expression, in PB samples, monocytes and NK cells showed weak expression, while neutrophils, B and T lymphocytes, and basophils showed weak and partial expression. In BM samples, expression of CD210a was observed in eosinophils, monocytes, and B and T/NK lymphocytes. Weak expression of CD210a was also observed in neutrophilic cells and plasma cells. All B cell maturation stages had weak expression of CD210a except for immature B cells, which did not express this marker. In the present study, no cell type in PB samples showed positivity for CD261 and, in BM samples, there was very weak expression in neutrophilic series, monocytes, and B lymphocytes. Conversely, plasma cells showed positivity for CD261 with a homogeneous expression. For CD262, there was weak expression in monocytes, neutrophils, and B lymphocytes in PB samples and weak expression in monocytes, B lymphocytes, and plasma cells in BM samples. The evaluation of CD264 showed very weak expression in B cells in PB samples and no expression in BM cells. Very weak expression of CD358 was observed in neutrophils, monocytes, and B lymphocytes in PB and BM samples. In addition, in BM samples, plasma cells and T lymphocytes showed weak expression of CD358. In relation to the maturation stages of B cells, there was weak expression in pro-B cel, pre-B cell, and mature B cell. In the present study, it was possible to observe expression of CD361 in all cell types analyzed in PB and BM samples. The analyzed markers presented varied profiles of expression and, in some cases, these profiles were different from those observed in other studies. Further studies are needed to evaluate these molecules, mainly in relation to a possible application in the diagnosis of hematological malignancies or as new therapeutic targets for the treatment of hematological neoplasms or autoimmune diseases.