ABSTRACT
El estudio de la megacariopoyesis humana se ha visto obstaculizado por la relativa escasez de megacariocitos en la médula ósea (0,05-0,2 % de las células medulares), lo que ha llevado a la optimización de protocolos de expansión in vitro a partir de precursores de diversos orígenes (cordón umbilical, médula ósea y sangre periférica con o sin movilización previa). Los cultivos celulares a partir de precursores han permitido la producción y el estudio tanto de megacariocitos así como de proplaquetas y plaquetas Sin embargo, la producción in vitro óptima de megacariocitos que culminen todos los estadios de diferenciación es un reto aún no resuelto. En este trabajo reportamos los hallazgos concernientes a la determinación de las condiciones y concentraciones de trombopoyetina para lograr una óptima relación entre la cantidad de trombopoyetina empleada y el porcentaje y grado de diferenciación megacariocítica en muestras obtenidas de cinco donantes alogénicos aceptados para trasplante de médula ósea.
The study of human megakaryocytopoiesis has been hampered by the relative scarcity of megakaryocytes in bone marrow (0.05-0.2 % of medullary cells), which has led to the optimization of protocols of in vitro expansion of precursors from diverse sources (umbilical cord, bone marrow and peripheral blood with or without previous mobilization). Cell cultures from different precursors have allowed the production and study of megakaryocytes as well as proplatelets and platelets. However, the in vitro production of megakaryocytes that culminate all stages of differentiation is a challenge that has not yet been resolved. In this work we report the findings related to the determination of thrombopoietin treatment conditions and concentrations to achieve an optimal relationship between the amount of thrombopoietin and the percentage and degree of megakaryocytic differentiation in five allogeneic donors that were accepted for bone marrow transplantation.
O estudo da megacariopoiese humana tem sido dificultado pela relativa escassez de megacariócitos na medula óssea (0,05-0,2 % das células medulares), o que levou à otimização dos protocolos de expansão in vitro a partir de precursores de diversas origens (cordão umbilical, medula óssea e sangue periférico com ou sem mobilização prévia). Culturas de células a partir de precursores permitiram a produção e o estudo tanto de megacariócitos e de proplaquetas e plaquetas. No entanto, a produção ótima in vitro de megacariócitos que culminam em todas as fases de diferenciação é um desafio ainda não resolvido. Neste trabalho, relatamos as descobertas relativas à determinação das condições e concentrações de trombopoietina para obter uma relação ótima entre a quantidade de trombopoietina usada e a taxa e o grau de diferenciação megacariocítica em amostras obtidas de cinco doadores alogênicos aceitos para transplante de medula óssea.
Subject(s)
Humans , Thrombopoietin/analysis , Megakaryocytes/cytology , Antigens, CD34/analysis , Cells, Cultured/cytology , Leukapheresis , Platelet Membrane Glycoprotein IIb/analysis , Integrin beta3/analysis , Culture Techniques/methodsABSTRACT
Introduction: CMV pp65-antigenemia (antigenemia) has been used for monitoring CMV viremia in allogeneic hematopoietic stem cell transplant (aHSCT) recipients. Recently, real time quantitative PCR (RT-qPCR ) has been used as a better approach than antigenemia for CMV diagnosis. The objective of this study was to assess the correlation of CMV viremia between RT-qPCR and antigenemia in aHSCT patients. Material and Methods: Observational prospective study of all aHSCT patients during 10 months in our center. CMV RT-qPCR in whole blood was performed weekly from day +7 to +100 after aHSCT. Simultaneous antigenemia was performed from engrafment to day +100. Concordance between both assays was evaluated. Results: Eighteen patients were included. In 120 simultaneous samples, 96 were concordant by both methods (80%). Kappa coefficient was 0.583. In 42% of cases without concordant results, patients were on antiviral therapy. Thirteen patients (72%) developed CMV infection (20 episodes). In 17 episodes, both the antigenemia and CMV RT-qPCR were positive. CMV RT-qPCR was detectable 1-2 weeks before antigenemia in 45% of the episodes. Conclusion: Both methods had a moderate concordance and CMV RT-qPCR detects CMV reactivations earlier than antigenemia, especially in neutropenic patients.
Introducción: La antigenemia pp65 (antigenemia) ha sido utilizada para monitorizar viremia de citomegalovirus (CMV) en pacientes sometidos a trasplantes alogeneicos de precursores hematopoiéticos (TPHa). Recientemente, la reacción de polimerasa en cadena cuantitativa en tiempo real (en inglés RT-qPCR) se ha usado como una mejor aproximación al diagnóstico de infección por CMV. El objetivo de este estudio fue evaluar la correlación de viremia por CMV a través de RT-qPCR con antigenemia, en pacientes que han recibido TPHa. Material y Métodos: Estudio prospectivo, observacional, de los pacientes sometidos a TPHa durante 10 meses. Se realizó RT-qPCR de CMV en sangre total semanalmente desde el día +7 al+100 después del trasplante y antigenemia en forma simultánea desde el prendimiento hasta el día +100. Se evaluó la concordancia entre ambos ensayos. Resultados: Dieciocho pacientes fueron incluidos. En 120 muestras simultáneas, 96 fueron concordantes por ambos métodos (80%). El coeficiente Kappa fue 0,583. En los casos no concordantes, el 42% se encontraba en terapia antiviral. Trece pacientes (72%) desarrollaron infección por CMV (20 episodios). En 17 episodios, ambos ensayos fueron positivos. La carga viral fue detectable 1-2 semanas antes que la antigenemia en 45% de los episodios. Conclusión: Existe una buena correlación entre ambas técnicas y la RT-qPCR detecta más precozmente que la antigenemia las reactivaciones de CMV, especialmente en pacientes neutropénicos.
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult , Cytomegalovirus Infections/diagnosis , Hematopoietic Stem Cell Transplantation/adverse effects , Antigens, Viral/blood , DNA, Viral/analysis , Early Diagnosis , Prospective Studies , Real-Time Polymerase Chain Reaction , Viral LoadABSTRACT
We present the neurological complications evaluated in a series of 1000 patients who underwent hematopoietic stem cell transplantation (HSCT). Central nervous system (CNS) neurological complications, particularly brain hemorrhages, were the most common, followed by seizures and CNS infections. An unusual neurological complication was Wernicke's encephalopathy. Less frequent neurological complications were metabolic encephalopathy, neuroleptic malignant syndrome, reversible posterior leukoencephalopathy syndrome, brain infarct and movement disorders. The most common neurological complication of the peripheral nervous system was herpes zoster radiculopathy, while peripheral neuropathies, inflammatory myopathy and myotonia were very rarely found.
Apresentamos as complicações neurológicas avaliadas em uma série de 1000 pacientes submetidos ao transplante de células tronco hematopoiéticas (TCTH). As complicações neurológicas do sistema nervoso central foram as mais encontradas, particularmente as hemorragias encefálicas, seguidas por crises convulsivas e por infecções. Uma complicação peculiar foi a encefalopatia de Wernicke. Menos freqüentemente foram encontrados casos de encefalopatia metabólica, síndrome maligna neuroléptica, leucoencefalopatia posterior reversível, infarto cerebral e os distúrbios do movimento. Entre as complicações neurológicas do sistema nervoso periférico a mais encontrada foi a radiculopatia pelo herpes zoster, enquanto que raramente se observaram casos de polineuropatias periféricas, miopatia inflamatória e de miotonia.