RESUMOOBJETIVO: Evidenciar a melhor estratégia de escolha de anticorpos para caracterizar a presença de clones de hemoglobinúria paroxística noturna (HPN) em pacientes submetidos à investigação diagnóstica de leucemias agudas ou em acompanhamento terapêutico. MATERIAIS E MÉTODOS: Foram analisadas 41 amostras de sangue periférico e medula óssea de pacientes portadores de leucemia aguda submetidos à investigação diagnóstica ou em acompanhamento terapêutico de dois hospitais oncológicos e públicos de Belém, de fevereiro a julho de 2015. RESULTADOS: Do total de amostras, 58,5% eram do gênero masculino, 41,5% estavam na faixa etária de 0-10 anos, 56,1% estavam em investigação diagnóstica e 43,9% em acompanhamento terapêutico. Dos casos em diagnóstico, 43,5% (10/23) eram de leucemia linfoblástica aguda de células B comum e 26,1% (6/23) de leucemia mieloide aguda. A presença de clones de HPN foi verificada em 9,8% (4/41) do total investigado, sendo 3/4 observados na investigação diagnóstica e 1/4 em paciente em acompanhamento terapêutico, sem recaída. A combinação dos anticorpos FLAER/CD59PE/CD45Per-Cy5 em blastos e linfócitos, FLAER/CD15PE/CD45Per-Cy5/CD24APC em granulócitos e FLAER/CD14PE/CD45Per-Cy5/ CD64APC em monócitos mostrou-se como a melhor estratégia para caracterizar a presença de clones de HPN nesses pacientes, independente do tipo de amostra. CONCLUSÃO: A presença de clones de HPN em portadores de leucemias agudas não dependeu da ontogenia celular ou do estágio do paciente (diagnóstico ou acompanhamento terapêutico). A melhor estratégia de anticorpos para a identificação de clones HPN em blastos leucêmicos, independente da sua ontogenia, foi por meio da combinação de FLAER com CD45.Palavras-chave: Hemoglobinúria Paroxística; Leucemia; Citometria de Fluxo.ABSTRACT OBJECTIVE: To highlight the best antibody selection strategy to characterize the presence of paroxysmal nocturnal hemoglobinuria (PNH) clones in patients undergoing diagnostic investigation of acute leukemia or in therapeutic follow-up. MATERIALS AND METHODS: Forty-one peripheral blood and bone marrow samples from patients with acute leukemia who underwent diagnostic investigation or therapeutic follow-up at two oncological and public hospitals in Belém, Pará State, Brazil, from February to July 2015, were analyzed. RESULTS: A total of 58.5% were male, 41.5% were 0-10 years old, 56.1% were under diagnostic investigation, and 43.9% were under therapeutic follow-up. Among cases under diagnostic investigation, 43.5% (10/23) were B-cell acute lymphoblastic leukemia and 26.1% (6/23) acute myeloid leukemia. The presence of PNH clones was observed in 9.8% (4/41) of cases, being 3/4 under diagnostic investigation and 1/4 under therapeutic follow-up, without relapse. The combination of FLAER/CD59PE/CD45Per-Cy5 antibodies in blasts and lymphocytes, FLAER/CD15PE/CD45Per-Cy5/CD24APC in granulocytes, and FLAER/CD14PE/CD45Per-Cy5/CD64APC in monocytes proved to be the best strategy to characterize the presence of PNH clones in these patients, regardless of sample ...

To better understand the relationship among cell adhesion molecules (CAM) and investigate the clinical diagnostic and prognostic application of ICAM-1 (ICAM1), LFA-1 (ITGAL), and L-selectin (SELL) proteins and mRNA corresponding expression in thyroid cancer. Gene expression was evaluated by RT–qPCR, and protein expression was evaluated by immunohistochemistry. We evaluated 275 patients (218 women, 57 men, 48.4 ± 14.5 years old), including 102 benign and 173 malignant nodules. The 143 papillary thyroid carcinoma (PTC) and 30 follicular thyroid carcinoma (FTC) patients were managed according to current guidelines and followed-up for 78.7 ± 54.2 months. Malignant and benign nodules differed concerning mRNA (p = 0.0027) and protein (p = 0.0020 for nuclear) expression of L-selectin and ICAM-1 (mRNA: p = 0.0001 and protein: p = 0.0014) and protein expression of LFA-1 (p = 0.0168), but not mRNA expression of LFA-1 (p = 0.2131). SELL expression was more intense in malignant tumors (p = 0.0027). ICAM1 (p = 0.0064) and ITGAL (p = 0.0244) mRNA expression was higher in tumors with lymphocyte infiltrate. ICAM-1 expression correlated with younger age at diagnosis (p = 0.0312) and smaller tumor size (p = 0.0443). Also, LFA-1 expression correlated with higher age at diagnosis (p = 0.0376) and was more intense at stage III and IV (p = 0.0077). In general, the protein expression of the 3 CAM decreased as the process of cellular dedifferentiation occurred. We suggest that the SELL and ICAM1 genes and L-selectin and LFA-1 protein expression may help confirm malignancy and assist in the histological characterization of follicular patterned lesions, but we were unable to correlate these CAMs with patient outcomes.

ResumoO câncer de tireoide é a malignidade endócrina mais comum. A L-selectina é uma molécula de adesão celular que atua principalmente no momento de migração de leucócitos pelo endotélio vascular. Sua expressão gênica foi avaliada por PCR em tempo real e sua hiperexpressão correlacionada com tumores malignos da tireoide.

Objective: To better understand the relationship among cell adhesion molecules (CAM) and investigate the clinical diagnostic and prognostic application of ICAM-1 (ICAM1), LFA-1 (ITGAL), and L-selectin (SELL) proteins and mRNA corresponding expression in thyroid cancer. Design and Methods: Gene expression was evaluated by RT–qPCR, and protein expression was evaluated by immunohistochemistry. We evaluated 275 patients (218 women, 57 men, 48.4±14.5 years old), including 102 benign and 173 malignant nodules. The 143 papillary thyroid carcinoma (PTC) and 30 follicular thyroid carcinoma (FTC) patients were managed according to current guidelines and followed-up for 78.7±54.2 months. Results: Malignant and benign nodules differed concerning mRNA (p=0.0027) and protein (p=0.0020 for nuclear) expression of L-selectin and ICAM-1 (mRNA: p=0.0001 and protein: p=0.0014) and protein expression of LFA-1 (p=0.0168), but not mRNA expression of LFA-1 (p=0.2131). SELL expression was more intense in malignant tumors (p=0.0027). ICAM1 (p=0.0064) and ITGAL (p=0.0244) mRNA expression was higher in tumors with lymphocyte infiltrate. ICAM-1 expression correlated with younger age at diagnosis (p=0.0312) and smaller tumor size (p=0.0443). Also, LFA-1 expression correlated with higher age at diagnosis (p=0.0376) and was more intense at stage III and IV(p=0.0077). Conclusion: In general, the protein expression of the 3 CAM decreased as the process of cellular dedifferentiation occurred. We suggest that the SELL and ICAM1 genesand L-selectin and LFA-1 protein expression may help confirm malignancy and assist in the histological characterization of follicular patterned lesions, but we were unable to correlate these CAMs with patient outcomes.

Thyroid cancer biology is extremely diverse. While some cases never progress clinically or do so indolently, others evolve aggressively and may even lead to death. Cell adhesion molecules are glycoproteins present in the cell membrane and play an important role in inflammatory and neoplastic diseases by recruiting immune cells to these sites. The aim of the present study was to investigate the role of mRNA expression of SELL, ICAM1 and ITGAL in thyroid tumors and their relationship with lymphocyte infiltration. We evaluated by RT-qPCR technique 191 thyroid nodules including 97 benign (79 females, 17 males; 49.8±12.5 years old) and 94 malignant (71 females, 23 males; 48.3±15.5years old) cases. Clinical and pathology data were obtained from 47 goiters; 50 follicular adenomas (FA); 74 papillary thyroid carcinomas (PTC), including: 29 classic papillary thyroid carcinomas (CPTC), 21 follicular variant of PTC (FVPTC), 12oxifilic variant of PTC (OVPTC), 12 tall cell papillary thyroid carcinomas (TCPTC); and 20 follicular thyroid carcinomas (FTC). All patients were managed according to a standard protocol based on current guidelines and followed-up for 78.7±54.2 months. SELL was more expressed in malignant (0.85±1.54 UA) than in benign (0.54±0.71 UA, p=0.0027) nodules. The same occurred with ICAM1 (0.99±1.41 vs. 0.46±0.85, p=0.0001), but not with ITGAL gene expression (1.04±1.63 vs. 0.76±1.21, p=0.2131). In addition, the expression of SELL was different when we compared PTC with FA (0.94±1.62 UA vs. 0.47±0.72 UA, p=0.0018) and FTC with FA (0.82±2.38 UA vs. 0.47±0.72 UA, p=0.0078). ICAM1 expression was lower in goiters (0.46±0.90 UA) when compared with PTC (0.93±1.22 UA, p=0.0030) and FTC (1.03±3.30 UA, p=0.0207). Higher expression of ICAM1 (1.16±3.04 UA vs. 0.52±0.96 UA, p=0.0064) and ITGAL (1.17±1.54 UA vs. 0.49±1.39 UA, p=0.0244) was observed in tumors with lymphocyte infiltrate. Also, ITGAL gene expression was higher in tumors that had distant metastasis at diagnosis (1.53±2.18 UA vs. 0.57±1.10 UA, p=0.0217). We were not able to demonstrate any association between any of the investigated molecules and patients’ outcome. In conclusion, our data suggest that cell adhesion molecules may play an important role in neoplastic thyroid cells proliferation. In addition, our findings show that gene expression of SELL and ICAM1 may assist in the histological characterization of follicular patterned thyroid nodules.