Diversidad de antígenos leucocitarios humanos A, B, DQB1 y DRB1 en células de sangre de cordón umbilical criopreservadas en el Hospital Universitario Dr. José Eleuterio González.

Antecedentes: el estudio del sistema HLA y la determinación de sus correspondientes antígenos son muy importantes en programas de trasplante de órganos (riñón y células hematoprogenitoras) para la selección adecuada de la pareja donador-receptor, y su uso es cada vez más frecuente en medicina por su relación entre algunas enfermedades y determinados antígenos del sistema. Objetivos: analizar los alelos y loci A, B de clase I y DQ, DR de clase II en las células de sangre de cordón umbilical, y determinar su diversidad en la población con base en las unidades de sangre de cordón criopreservadas. Material y métodos: análisis de la diversidad de los alelos HLA-A, B, DRB1 y DQB1, en 80 unidades de células de sangre de cordón criopreservadas en el servicio de hematología del Hospital Universitario, para tipificar los alelos mediante PCR-SSP, en laboratorio externo. Resultados: los resultados del análisis mostraron que la diversidad y distribución de alelos tienen frecuencia similar a la de otros estudios realizados en diversas poblaciones del país. Conclusiones: el Hospital Universitario cuenta con reserva de unidades de sangre de cordón con frecuencia de alelos similar a diversas poblaciones del país

Recolección de células hematopoyéticas periféricas para trasplante alogénico con una dosis intermedia de filgrastim

Antecedentes: las células hematopoyéticas de sangre periférica son un recurso para combatir diversas enfermedades que requieren un trasplante alogénico para su curación. El método internacionalmente aceptado para obtener estas células es la administración de 2 a 24 μg/kg de peso de un factor estimulante de colonias de granulocitos y realizar la aféresis al cuarto, quinto o sexto días de iniciada la estimulación del donador. Objetivo: determinar la dosis adecuada del factor estimulante de colonias de granulocitos y el día óptimo para realizar una sola aféresis. Pacientes y método: se incluyeron 11 donadores que recibieron factor estimulante de colonias de granulocitos en dosis diarias de 8 μg/kg de peso, durante cuatro días, y se realizaron dos aféresis: una en el cuarto y otra en el quinto día de estimulación. Se analizaron retrospectivamente los procedimientos de recolección de células hematopoyéticas de sangre periférica para trasplante alogénico realizados entre febrero del 2003 y mayo del 2005 en el Hospital Universitario Dr. José Eleuterio González de la UANL. Resultados: en 63% de los pacientes, la cantidad de células CD34+ recolectada fue significativamente mayor en el quinto día que en el cuarto (3.10 × 106 vs 2.9 × 106), ya que el volumen plasmático promedio procesado por aféresis fue menor en la primera ocasión que en la segunda (8,800 vs 14,080 mL). Conclusión: una dosis de 8 μg/kg de peso del factor estimulante de colonias de granulocitos es efectiva para favorecer la obtención de células hematopoyéticas periféricas. Una aféresis al quinto día de iniciada la estimulación es suficiente para obtener la cantidad necesaria de células CD34+ que aseguren la recuperación hematológica del paciente trasplantado. Este procedimiento reduce los efectos colaterales del factor y de la aféresis, así como el costo final del trasplante

Circulating microRNA expression profile in B-cell acute lymphoblastic leukemia

BACKGROUND: Acute lymphoblastic leukemia (ALL) is a highly diverse disease characterized by cytogenetic and molecularabnormalities, including altered microRNA (miRNA) expression signatures. AIM: We perform and validate a plasma miRNA expression profiling to identify potential miRNA involved in leukemogenesis METHODS: MiRNA expression profiling assay was realized in 39 B-ALL and 7 normal control plasma samples using TaqMan Low Density Array (TLDA) plates on Applied Biosystems 7900 HT Fast Real-Time PCR System. MiRNA validation was done for six miRNA differentially expressed by quantitative real-time PCR. RESULTS: Seventy-seven circulating miRNA differentially expressed: hsa-miR-511, -222, and -34a were overexpressed, whereas hsa-miR-199a-3p, -223, -221, and -26a were underexpressed (p values < 0.005 for both sets). According to operating characteristic curve analysis, hsa-miR-511 was the most valuable biomarker for distinguishing B-ALL from normal controls,with an area under curve value of 1 and 100% for sensitivity, and specificity respectively. CONCLUSIONS: Measuring circulating levels of specific miRNA implicated in regulation of cell differentiation and/or cell proliferation such as hsa-miRNA-511, offers high sensitivity and specificity in B-ALL detection and may be potentially useful for detection of disease progression, as indicator of therapeutic response, and in the assessment of biological and/or therapeutic targets for patients with B-ALL

Flow cytometry data analysis of CD34+/CD133+ stem cells in bone marrow and peripheral blood and T, B, and NK cells after hematopoietic grafting

This article provides flow cytometry information regarding levels of expression for hematopoietic stem cell markers CD34 and CD133 obtained simultaneously of the bone marrow and peripheral blood from recipients of allogeneic and autologous transplants of PB hematoprogenitors for treating hematological malignancies and who were clinically healthy after ≥100 days following the procedure. CD34 and CD133 expression is compared regarding type of transplant (autologous vs. allogeneic) and sample cell source (bone marrow vs. peripheral blood). Patients were conditioned with a reduced-intensity conditioning regimen. Also shown is the flow cytometry analysis of mononuclear cell and lymphocyte populations in the peripheral blood of both types of recipients, as well as the characterization of immune cells, including T lymphocyte antigenic make up markers CD3, CD4 and CD8, B lymphocytes and NK cells, including total NK, bright and dim subtypes in the peripheral blood of both types of recipients. For further information and discussion regarding interpretation and meaning of post-transplant flow cytometry analysis, please refer to the article “Assessment of immune reconstitution status in recipients of a successful hematopoietic stem cell transplant from peripheral blood after reduced intensity conditioning” [1]. Keywords: Bone Marrow, CD34+ cells, CD133+ hematoprogenitors, Flow cytometry, Hematopoietic transplant, Peripheral blood, T, B and NK cell

Comparison of CD4+/CD8+ Lymphocytic Subpopulations Pre- and Post-Antituberculosis Treatment in Patients with Diabetes and Tuberculosis

Is there a CD4+ and CD8+ immunity alteration in patients with pulmonary tuberculosis (TB) and diabetes (DM) that does not recover after antituberculosis treatment? This prospective comparative study evaluated CD4+ and CD8+ lymphocytic subpopulations and antituberculosis antibodies in patients with diabetes and tuberculosis (TB-DM), before and after antituberculosis treatment. CD4+ T cell counts were lower in patients with TB-DM compared to those with only TB or only DM, and these levels remained low even after two months of anti-TB treatment. Regarding the CD8+ T cell analysis, we identified higher blood values in the DM-only group, which may be explained by the high prevalence of latent tuberculosis (LTBI) in patients with DM. IgM antituberculosis antibodies levels were elevated in patients with only TB at baseline, and 2 months post-anti-TB treatment, IgG did not express any relevant alterations. Our results suggest an alteration in CD4+ immunity in patients with TB-DM that did not normalize after antituberculosis treatment

Identification of Differentially Expressed Genes Associated with Prognosis of B Acute Lymphoblastic Leukemia

Background. Acute lymphoblastic leukemia type B (B-ALL) is a neoplastic disorder with high mortality rates. The aim of this study was to validate the expression profile of 45 genes associated with signaling pathways involved in leukemia and to evaluate their association with the prognosis of B-ALL. Methods. 219 samples of peripheral blood mononuclear cells obtained from 73 B-ALL patients were studied at diagnosis, four, and eight weeks after starting treatment. Gene expression was analyzed by quantitative real-time polymerase chain reaction. Results. Normalized delta Cq values of 23 genes showed differences between B-ALL and controls at diagnosis time (P values < 0.05). There were significant associations between B-ALL patients relapse/death and the expression levels of IL2RA, SORT1, DEFA1, and FLT3 genes at least in one of the times evaluated (P values < 0.05 and odds ratio ranges: 3.73–27). The association between FLT3 deregulation and relapse/death was a constant in the times studied and their overexpression significantly increased the odds of relapse/death in a range of 3.73 and 6.05 among study population (P values < 0.05). Conclusions. Overexpression of FLT3 and DEFA1 genes retained independent prognostic significance for B-ALL outcome, reflected as increased risks of relapse/death among the study population