RESUMEN
BACKGROUND AND OBJECTIVES: Transfusion-related acute lung injury (TRALI) is characterized by leukocyte transmigration and alveolar capillary leakage shortly after transfusion. TRALI pathogenesis has not been fully elucidated. In some cases, the infusion of alloantibodies (immune model), whereas in others the combination of neutrophil priming by proinflammatory molecules with the subsequent infusion of biological response modifiers (BRMs) in the hemocomponent (non-immune model) have been implicated. Our aim was to compare the pathological events involved in TRALI induced by antibodies or BRMs using murine models. MATERIALS AND METHODS: In the immune model, human HNA-2(+) neutrophils were incubated in vitro with a monoclonal antibody (anti-CD177, clone 7D8) directed against the HNA-2 antigen and injected i.v. in NOD/SCID mice. In the non-immune model, BALB/c mice were treated with low doses of lipopolysaccharide (LPS) followed by platelet-activating factor (PAF) infusion 2 h later. Forty minutes after PAF administration, or 6 h after neutrophil injection, lungs were isolated and histological analysis, determination of a variety of cytokines and chemokines including keratinocyte-derived chemokine (KC), MIP-2, the interleukins IL-1ß, IL-6, IL-8 as well as TNFα, cell influx and alveolar capillary leakage were performed. RESULTS: In both models, characteristic histological findings of TRALI and an increase in KC and MIP-2 levels were detected. In contrast to the immune model, in the non-immune model, there was a dramatic increase in IL-1ß and TNFα. However, capillary leakage was only detected if PAF was administrated. CONCLUSIONS: Regardless of the triggering event(s), KC, MIP-2 and integrins participate in TRALI pathogenesis, whereas PAF is essential for capillary leakage when two events are involved.
Asunto(s)
Lesión Pulmonar Aguda/inmunología , Lesión Pulmonar Aguda/patología , Reacción a la Transfusión , Lesión Pulmonar Aguda/etiología , Animales , Quimiocinas/inmunología , Modelos Animales de Enfermedad , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NOD , Ratones SCID , Neutrófilos/inmunología , Neutrófilos/patologíaRESUMEN
Regarded as an incidental finding, biliary sludge is often diagnosed in dogs on abdominal ultrasound. The aims of the present study were to assess the risk factors, biochemical markers and ultrasonographic findings and to estimate the prevalence and influence of different breeds, sexes, and ages on biliary sludge in dogs. Results demonstrate that the prevalence of biliary sludge is high, especially in senior dogs. The biochemical markers did not have a significant correlation with biliary sludge, and the type of diet was not considered to be the major risk factor. Hepatomegaly was frequently observed on the ultrasound scan of affected animals and of dogs on different systemic drugs and with cardiopathies, which have been referred to as risk groups for the development of inspissated bile.
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Bilis/diagnóstico por imagen , Enfermedades de los Perros/patología , Animales , Enfermedades de las Vías Biliares/diagnóstico por imagen , Enfermedades de las Vías Biliares/veterinaria , Enfermedades de los Perros/sangre , Enfermedades de los Perros/diagnóstico por imagen , Enfermedades de los Perros/metabolismo , Perros , Femenino , Masculino , Factores de Riesgo , UltrasonografíaRESUMEN
ABSTRACT The goal of this work was to perform a cytomorphologic study of rice plants upon salt stress (170 mM NaCl). The effects of salinity on cell structures were analyzed by optical and electron transmission microscopy. An early differentiation process could be observed in stressed plants, however the most striking deleterious effect was found in the thylacoid membranes of chloroplasts. These results indicate a correlation between the previously described accumulation of sodium and the cytomorphological changes in chloroplasts of mature tissues of rice plants.
RESUMO O objetivo deste trabalho foi realizar um estudo citomorfológico em plantas de arroz submetidas ao estresse salino (170 mM NaCl). Os efeitos da salinidade nas estruturas celulares foram analisados por meio de microscopia óptica e eletrônica de transmissão. Um processo de diferenciação precoce pôde ser observado na planta estressada, porém, o efeito deletério mais drástico foi encontrado nas membranas tilacóides dos cloroplastos. Estes resultados permitiram correlacionar o acúmulo de íons sódio, descritos na literatura, com os efeitos citomorfológicos do estresse salino em tecidos maduros de arroz.
RESUMEN
We evaluated the expression of 10 adhesion molecules on peripheral blood tumor cells of 17 patients with chronic lymphocytic leukemia, 17 with mantle-cell lymphoma, and 13 with nodal or splenic marginal B-cell lymphoma, all in the leukemic phase and before the beginning of any therapy. The diagnosis of B-cell non-Hodgkin's lymphomas was based on cytological, histological, immunophenotypic, and molecular biology methods. The mean fluorescence intensity of the adhesion molecules in tumor cells was measured by flow cytometry of CD19-positive cells and differed amongst the types of lymphomas. Comparison of chronic lymphocytic leukemia and mantle-cell lymphoma showed that the former presented a higher expression of CD11c and CD49c, and a lower expression of CD11b and CD49d adhesion molecules. Comparison of chronic lymphocytic leukemia and marginal B-cell lymphoma showed that the former presented a higher expression of CD49c and a lower expression of CD11a, CD11b, CD18, CD49d, CD29, and CD54. Finally, comparison of mantle-cell lymphoma and marginal B-cell lymphoma showed that marginal B-cell lymphoma had a higher expression of CD11a, CD11c, CD18, CD29, and CD54. Thus, the CD49c/CD49d pair consistently demonstrated a distinct pattern of expression in chronic lymphocytic leukemia compared with mantle-cell lymphoma and marginal B-cell lymphoma, which could be helpful for the differential diagnosis. Moreover, the distinct profiles of adhesion molecules in these diseases may be responsible for their different capacities to invade the blood stream.
Asunto(s)
Moléculas de Adhesión Celular/biosíntesis , Leucocitos Mononucleares/metabolismo , Linfoma de Células B/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/metabolismo , Diagnóstico Diferencial , Femenino , Citometría de Flujo , Humanos , Leucemia Linfocítica Crónica de Células B/metabolismo , Linfoma de Células B de la Zona Marginal/metabolismo , Linfoma de Células del Manto/metabolismo , Masculino , Persona de Mediana EdadRESUMEN
We evaluated the expression of 10 adhesion molecules on peripheral blood tumor cells of 17 patients with chronic lymphocytic leukemia, 17 with mantle-cell lymphoma, and 13 with nodal or splenic marginal B-cell lymphoma, all in the leukemic phase and before the beginning of any therapy. The diagnosis of B-cell non-Hodgkin's lymphomas was based on cytological, histological, immunophenotypic, and molecular biology methods. The mean fluorescence intensity of the adhesion molecules in tumor cells was measured by flow cytometry of CD19-positive cells and differed amongst the types of lymphomas. Comparison of chronic lymphocytic leukemia and mantle-cell lymphoma showed that the former presented a higher expression of CD11c and CD49c, and a lower expression of CD11b and CD49d adhesion molecules. Comparison of chronic lymphocytic leukemia and marginal B-cell lymphoma showed that the former presented a higher expression of CD49c and a lower expression of CD11a, CD11b, CD18, CD49d, CD29, and CD54. Finally, comparison of mantle-cell lymphoma and marginal B-cell lymphoma showed that marginal B-cell lymphoma had a higher expression of CD11a, CD11c, CD18, CD29, and CD54. Thus, the CD49c/CD49d pair consistently demonstrated a distinct pattern of expression in chronic lymphocytic leukemia compared with mantle-cell lymphoma and marginal B-cell lymphoma, which could be helpful for the differential diagnosis. Moreover, the distinct profiles of adhesion molecules in these diseases may be responsible for their different capacities to invade the blood stream.
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Anciano de 80 o más Años , Moléculas de Adhesión Celular/biosíntesis , Leucocitos Mononucleares/metabolismo , Linfoma de Células B/metabolismo , Diagnóstico Diferencial , Citometría de Flujo , Leucemia Linfocítica Crónica de Células B/metabolismo , Linfoma de Células del Manto/metabolismo , Linfoma de Células B de la Zona Marginal/metabolismo , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/metabolismoRESUMEN
Acute promyelocytic leukemia (APL) is characterized by the expansion of blasts that resemble morphologically promyelocytes and harbor a chromosomal translocation involving the retinoic acid receptor alpha (RARalpha) and the promyelocytic leukemia (PML) genes on chromosomes 17 and 15, respectively. The expression of the PML/RARalpha fusion gene is essential for APL genesis. In fact, transgenic mice (TM) expressing PML/RARalpha develop a form of leukemia that mimics the hematological findings of human APL. Leukemia is diagnosed after a long latency (approximately 12 months) during which no hematological abnormality is detected in peripheral blood (pre-leukemic phase). In humans, immunophenotypic analysis of APL blasts revealed distinct features; however, the precise immunophenotype of leukemic cells in the TM model has not been established. Our aim was to characterize the expression of myeloid antigens by leukemic cells from hCG-PML/RARalpha TM. In this study, TM (N = 12) developed leukemia at the mean age of 13.1 months. Morphological analysis of bone marrow revealed an increase of the percentage of immature myeloid cells in leukemic TM compared to pre-leukemic TM and wild-type controls (48.63 +/- 16.68, 10.83 +/- 8.11, 7.4 +/- 5.46%, respectively; P < 0.05). Flow cytometry analysis of bone marrow and spleen from leukemic TM identified the asynchronous co-expression of CD34, CD117, and CD11b. This abnormal phenotype was rarely detected prior to the diagnosis of leukemia and was present at similar frequencies in hematologically normal TM and wild-type controls of different ages. The present results demonstrate that, similarly to human APL, leukemic cells from hCG-PML/RARalpha TM present a specific immunophenotype.
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Antígenos CD/inmunología , Leucemia Mieloide Aguda/inmunología , Leucemia Promielocítica Aguda/inmunología , Proteínas de Fusión Oncogénica/inmunología , Animales , Antígenos CD/genética , Médula Ósea/inmunología , Médula Ósea/patología , Catepsina G , Catepsinas , Citometría de Flujo , Genotipo , Inmunofenotipificación , Leucemia Mieloide Aguda/genética , Leucemia Promielocítica Aguda/genética , Ratones , Ratones Transgénicos , Proteínas de Fusión Oncogénica/genética , Serina Endopeptidasas , Bazo/inmunología , Bazo/patologíaRESUMEN
Acute promyelocytic leukemia (APL) is characterized by the expansion of blasts that resemble morphologically promyelocytes and harbor a chromosomal translocation involving the retinoic acid receptor a (RARa) and the promyelocytic leukemia (PML) genes on chromosomes 17 and 15, respectively. The expression of the PML/RARa fusion gene is essential for APL genesis. In fact, transgenic mice (TM) expressing PML/RARa develop a form of leukemia that mimics the hematological findings of human APL. Leukemia is diagnosed after a long latency (approximately 12 months) during which no hematological abnormality is detected in peripheral blood (pre-leukemic phase). In humans, immunophenotypic analysis of APL blasts revealed distinct features; however, the precise immunophenotype of leukemic cells in the TM model has not been established. Our aim was to characterize the expression of myeloid antigens by leukemic cells from hCG-PML/RARa TM. In this study, TM (N = 12) developed leukemia at the mean age of 13.1 months. Morphological analysis of bone marrow revealed an increase of the percentage of immature myeloid cells in leukemic TM compared to pre-leukemic TM and wild-type controls (48.63 ± 16.68, 10.83 ± 8.11, 7.4 ± 5.46 percent, respectively; P < 0.05). Flow cytometry analysis of bone marrow and spleen from leukemic TM identified the asynchronous co-expression of CD34, CD117, and CD11b. This abnormal phenotype was rarely detected prior to the diagnosis of leukemia and was present at similar frequencies in hematologically normal TM and wild-type controls of different ages. The present results demonstrate that, similarly to human APL, leukemic cells from hCG-PML/RARa TM present a specific immunophenotype.
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Animales , Ratones , Antígenos CD/inmunología , Leucemia Mieloide Aguda/inmunología , Leucemia Promielocítica Aguda/inmunología , Proteínas de Fusión Oncogénica/inmunología , Antígenos CD/genética , Médula Ósea/inmunología , Médula Ósea/patología , Catepsinas , Citometría de Flujo , Genotipo , Inmunofenotipificación , Leucemia Mieloide Aguda/genética , Leucemia Promielocítica Aguda/genética , Ratones Transgénicos , Proteínas de Fusión Oncogénica/genética , Serina Endopeptidasas , Bazo/inmunología , Bazo/patologíaRESUMEN
The multidrug resistance P-glycoprotein is a transmembrane efflux pump expressed by lymphocytes and is involved in their cytolytic activity. In the present study, we investigated the age-related changes of P-glycoprotein function in normal peripheral blood lymphocytes. Blood samples from 90 normal volunteers (age range, 0 to 86 years) were analyzed. P-glycoprotein function was assessed by the flow cytometric rhodamine 123 assay. P-glycoprotein function was highest in cord blood and progressively declined with age in peripheral blood T CD4+ and CD8+ cells. In contrast, P-glycoprotein function did not vary with age in CD19+ B or CD16+CD56+ natural killer cells. These data suggest that the decline in P-glycoprotein function in T CD4+ and CD8+ lymphocytes as a function of age may contribute to the decrease in T cell cytolytic activity with aging.
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Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/fisiología , Resistencia a Múltiples Medicamentos/fisiología , Linfocitos T/fisiología , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Envejecimiento/fisiología , Niño , Preescolar , Citometría de Flujo , Colorantes Fluorescentes , Humanos , Lactante , Recién Nacido , Células Asesinas Naturales/fisiología , Persona de Mediana Edad , Rodamina 123 , Células Madre/fisiologíaRESUMEN
The multidrug resistance P-glycoprotein is a transmembrane efflux pump expressed by lymphocytes and is involved in their cytolytic activity. In the present study, we investigated the age-related changes of P-glycoprotein function in normal peripheral blood lymphocytes. Blood samples from 90 normal volunteers (age range, 0 to 86 years) were analyzed. P-glycoprotein function was assessed by the flow cytometric rhodamine 123 assay. P-glycoprotein function was highest in cord blood and progressively declined with age in peripheral blood T CD4+ and CD8+ cells. In contrast, P-glycoprotein function did not vary with age in CD19+ B or CD16+CD56+ natural killer cells. These data suggest that the decline in P-glycoprotein function in T CD4+ and CD8+ lymphocytes as a function of age may contribute to the decrease in T cell cytolytic activity with aging.
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Adolescente , Recién Nacido , Lactante , Preescolar , Niño , Adulto , Persona de Mediana Edad , Humanos , Resistencia a Múltiples Medicamentos , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP , Linfocitos T , Factores de Edad , Anciano de 80 o más Años , Citometría de Flujo , Colorantes Fluorescentes , Rodamina 123RESUMEN
The distinction between normal and leukemic bone marrow (BM) B-precursors is essential for the diagnosis and treatment monitoring of acute lymphoblastic leukemia (ALL). In order to evaluate the potential use of quantitative fluorescence cytometry (QFC) for this distinction, we studied 21 normal individuals and 40 patients with CD10+ ALL. We characterized the age-related changes of the CD10, CD19, TdT, CD34 and CD79a densities in normal and leukemic BM. Compared to normal adults, the B-precursors from normal children expressed significantly lower values of CD34-specific antibody binding capacity (SABC) (median value of 86.6 vs 160.2 arbitrary units (a.u.) in children and adults, respectively). No significant age-related difference was observed in the expression of the other markers in the normal BM, or in any of the markers in the leukemic BM. Based on the literature, we set the cut-off value for the normal CD10 expression at 45 x 10(3) a.u. for both age groups. For the remaining markers we established the cut-off values based on the minimum-maximum values in the normal BM in each age group. The expression of CD10 was higher than the cut-off in 30 ALL cases and in 18 of them there was a concomitant aberrant expression of other markers. In 9 of the 10 CD10+ ALL with normal CD10 SABC values, the expression of at least one other marker was aberrant. In conclusion, the distinction between normal and leukemic cells by QFC was possible in 38/40 CD10+ ALL cases.
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Antígenos CD/análisis , Células de la Médula Ósea/inmunología , Leucemia-Linfoma Linfoblástico de Células Precursoras/inmunología , Adolescente , Adulto , Biomarcadores , Células de la Médula Ósea/citología , Células de la Médula Ósea/patología , Estudios de Casos y Controles , Niño , Preescolar , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Inmunoelectroforesis Bidimensional , Inmunofenotipificación , Lactante , Modelos Lineales , Persona de Mediana Edad , Neoplasia Residual/diagnóstico , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Estadísticas no Paramétricas , EsternónRESUMEN
The distinction between normal and leukemic bone marrow (BM) B-precursors is essential for the diagnosis and treatment monitoring of acute lymphoblastic leukemia (ALL). In order to evaluate the potential use of quantitative fluorescence cytometry (QFC) for this distinction, we studied 21 normal individuals and 40 patients with CD10+ ALL. We characterized the age-related changes of the CD10, CD19, TdT, CD34 and CD79a densities in normal and leukemic BM. Compared to normal adults, the B-precursors from normal children expressed significantly lower values of CD34-specific antibody binding capacity (SABC) (median value of 86.6 vs 160.2 arbitrary units (a.u.) in children and adults, respectively). No significant age-related difference was observed in the expression of the other markers in the normal BM, or in any of the markers in the leukemic BM. Based on the literature, we set the cut-off value for the normal CD10 expression at 45 x 10Ý a.u. for both age groups. For the remaining markers we established the cut-off values based on the minimum-maximum values in the normal BM in each age group. The expression of CD10 was higher than the cut-off in 30 ALL cases and in 18 of them there was a concomitant aberrant expression of other markers. In 9 of the 10 CD10+ ALL with normal CD10 SABC values, the expression of at least one other marker was aberrant. In conclusion, the distinction between normal and leukemic cells by QFC was possible in 38/40 CD10+ ALL cases
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Humanos , Lactante , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Antígenos CD/análisis , Células de la Médula Ósea , Citometría de Flujo , Inmunofenotipificación , Leucemia-Linfoma Linfoblástico de Células Precursoras , Biomarcadores , Estudios de Casos y Controles , Técnica del Anticuerpo Fluorescente , Técnica del Anticuerpo Fluorescente Directa , Inmunoelectroforesis Bidimensional , Modelos Lineales , Neoplasia Residual/diagnóstico , Estadísticas no Paramétricas , Esternón/citologíaRESUMEN
A 46-year-old woman with a previous diagnosis of sarcoidosis presented with morphologically typical large granular lymphocyte (LGL) leukemia/lymphoma with an aggressive clinical course. Epstein-Barr virus DNA was detected in peripheral blood mononuclear cells by PCR. The phenotype was typical of the T cell lineage (CD2+ CD3+ CD5+ CD7+ CD8+ TCRalphabeta+) but with the absence of the CD16, CD56, CD57 NK cell markers. In addition, the LGLs expressed CD122 (p75) in the absence of CD25 which is characteristic of LGLs. These leukemic LGLs did not exhibit NK activity. The clonal nature of this proliferation was demonstrated by the rearrangement of the TCRgamma gene. This phenotypically unusual but morphologically typical LGL leukemia/lymphoma may represent the clonal expansion of a minor normal subset of T-LGLs which do not express any NK cell markers, probably corresponding to in vivo activated T cells.
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Biomarcadores de Tumor/análisis , Células Asesinas Naturales/patología , Leucemia de Células T/patología , Linfoma de Células T/patología , Subgrupos de Linfocitos T/patología , Linaje de la Célula/inmunología , ADN Viral/análisis , Femenino , Herpesvirus Humano 4/genética , Humanos , Inmunofenotipificación , Células Asesinas Naturales/inmunología , Leucemia de Células T/inmunología , Leucemia de Células T/virología , Linfoma de Células T/inmunología , Linfoma de Células T/virología , Persona de Mediana Edad , Subgrupos de Linfocitos T/inmunologíaRESUMEN
BACKGROUND: The immunomodulatory effects of allogeneic blood transfusions have been attributed to the white cells (WBCs) present in the cellular blood components transfused to patients. STUDY DESIGN AND METHODS: The effect of the transfusion of allogeneic red cells (RBCs) or allogeneic prestorage WBC-reduced RBCs (WBC-reduced RBCs) on host immune responsiveness was evaluated by measuring the lymphocyte subsets and the in-vitro cytokine production in response to phytohemagglutinin stimulation of WBCs of orthopedic surgery patients. Forty-seven patients undergoing hip replacement surgery were randomly assigned to receive allogeneic RBCs (n = 17) or WBC-reduced RBCs (n = 14; 99.95% WBC removal). Sixteen patients were not transfused. Patient blood samples taken before surgery and on Days 1 and 4 after surgery were tested for complete blood count, lymphocyte subset analysis, and measurement of cytokine levels. RESULTS: After surgery, the lymphocyte count was significantly decreased in patients transfused with > or = 3 units of allogeneic RBCs (2.0 +/- 0.5 vs. 1.3 +/- 0.3 x 10(9)/L; p = 0.017), but not in patients transfused with > or = 3 units of WBC-reduced RBCs (2.0 +/- 0.9 vs. 1.7 +/- 0.8 x 10(9)/L). Compared with preoperative levels, on Day 4 after surgery, patients transfused with > or = 3 units of allogeneic RBCs also had a decrease in the number of natural killer cells (0.07 +/- 0.05 vs. 0.04 +/- 0.03 x 10(9)/L; p = 0.018). Postoperatively, interleukin-2 was decreased in one patient who received WBC-reduced RBCs compared with that in four patients transfused with allogeneic RBCs (p = 0.32), and eight untransfused patients (p = 0.01). On Day 4, about 70 percent of patients transfused with allogeneic RBCs showed a 20-percent decrease in the interferon gamma level. CONCLUSION: Taken together, these data support the hypothesis that transfusion of > or = 3 units of allogeneic RBCs is associated with early postoperative lymphopenia in otherwise healthy individuals undergoing surgery. These findings were not observed in those individuals transfused with RBCs that had undergone prestorage WBC reduction.
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Artroplastia de Reemplazo de Cadera , Transfusión de Eritrocitos/métodos , Adulto , Anciano , Anciano de 80 o más Años , Formación de Anticuerpos/fisiología , Recuento de Células , Citocinas/biosíntesis , Femenino , Humanos , Interleucina-6/metabolismo , Leucaféresis , Subgrupos Linfocitarios/citología , Masculino , Persona de Mediana Edad , Atención Perioperativa , Estudios Prospectivos , Trasplante Homólogo/inmunologíaRESUMEN
The presence of phenotypically immature lymphocytes in umbilical cord blood has been a controversial topic. Moreover, their changes with age have not been systematically evaluated. In the present study, relative and absolute numbers of CD34+, CD10+CD19+, and CD4+CD8+ cell subsets were determined in umbilical cord blood from 12 full-term normal newborns, 43 children aged 1 month to 6 years, and 10 young adults. The samples were processed by whole-blood lysis and monoclonal antibody staining, and cells were analyzed by flow cytometry. Immature cells were present in cord blood and progressively declined in both absolute and percentage numbers with age, each according to a particular curve, reaching youth values roughly at age 2-4 years. These results demonstrate that phenotypically immature cells normally circulate at low levels in peripheral blood, mostly at birth and during infancy, but also during youth.
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Linfocitos B/clasificación , Células Madre Hematopoyéticas/clasificación , Linfocitos T/clasificación , Adulto , Antígenos CD/análisis , Antígenos CD34/análisis , Linfocitos B/inmunología , Niño , Preescolar , Células Madre Hematopoyéticas/inmunología , Humanos , Inmunofenotipificación , Lactante , Recién Nacido , Linfocitos T/inmunologíaRESUMEN
In order to assess the age-related changes in CD10 and CD19 fluorescence intensity (FI) the present study analyzed by flow cytometry 56 sternal biopsies from 'normal' infants, children and adults undergoing cardiac surgery. The CD10(+weak) subset was predominant in all age groups, representing approximately 50% of the bone marrow (BM) lymphoid cells in children younger than 4 years. Both CD10+ subsets significantly decreased with age but their ratio did not differ significantly. Moreover, the intensity of CD10 and CD19 fluorescence in the strong and weak subsets did not vary with age. The CD19 intensity was significantly higher in CD10(+weak) than in CD10(+strong) cells. In addition, we classified as CD10(+weak) or CD10(+strong) the leukemic cells from BM aspirates of 117 patients with common acute lymphoblastic leukemia (cALL) (78 children and 39 adults). A higher frequency of cases expressing the CD19+ CD10(+strong) phenotype was observed both in children and adults. Children of the CD10(+weak) group tended to be older than those of the CD10(+strong) group (median = 7 vs. 4 years, P = 0.07), and presented a significantly higher frequency of splenomegaly (93.7 vs. 55%, P = 0.04), which was massive in about 60% of these cases. Among adults, a significantly higher frequency of cases expressing the CD10(+weak) phenotype was observed in females. No other clinical or biological difference was detected between the two groups either for children or adults. Concerning the treatment outcome, we did not observe significant differences in complete remission rate (CRR) or in disease free survival (DFS) among the 32 children and 28 adults analyzed. Finally, we compared the CD10 and CD19 intensity in normal and leukemic BM. Overexpression of either or both antigens in leukemic cells was observed in 42.4% of the cALL cases. In these cases, using cut off values of 110 afu for the CD10 FI and of 100 afu for the CD19 FI, the detection of leukemic cells was possible at levels of 0.2% based on CD10 analysis, of 0.6% based on CD19, and 0.02% when both antigens were overexpressed. In conclusion, we demonstrated that the heterogeneity of CD10 and CD19 fluorescence intensity is of no clinical relevance in cALL, although its study may be helpful for the diagnosis and the detection of minimal residual disease.
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Antígenos CD19/química , Linfocitos B/inmunología , Neprilisina/química , Leucemia-Linfoma Linfoblástico de Células Precursoras/inmunología , Células Madre/inmunología , Adolescente , Adulto , Análisis de Varianza , Células de la Médula Ósea/inmunología , Niño , Femenino , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Valores de Referencia , Resultado del TratamientoRESUMEN
To address a possible impairment of multidrug resistance mechanisms in acquired aplastic anaemia (AA), the functions of P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP) were respectively assessed by rhodamine 123 (Rh123) and daunorubicin (DNR) efflux in peripheral blood lymphocytes from AA patients. The proportion of Rh123-effluxing T cells was significantly decreased in AA, relative to controls. Interestingly, these changes were also present in patients with AA in remission. Conversely, Rh123 efflux in B and natural killer (NK) cells and DNR efflux in peripheral blood lymphocytes were unchanged. These data indicated that P-gp activity was decreased in AA not only during the development of the disease, but also after remission, introducing a new concept on the pathophysiology of AA by suggesting that it may contribute to drug-induced injury to haemopoietic cells in some cases of AA, by increasing the proportion of susceptible cells.
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Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Anemia Aplásica/metabolismo , Adolescente , Adulto , Anemia Aplásica/inducido químicamente , Niño , Preescolar , Daunorrubicina/metabolismo , Resistencia a Múltiples Medicamentos , Femenino , Colorantes Fluorescentes/metabolismo , Humanos , Masculino , Rodamina 123/metabolismoRESUMEN
There is a paucity of information in the literature concerning the age-related changes of the lymphocyte subsets in bone marrow (BM), and the available reports disagree about the characteristics of the population studied and the methods for obtaining, handling, and analyzing the samples. The purpose of the present study was to determine the distribution of lymphoid subsets in the BM from infants, children, and adults by analyzing fragments of sternum obtained during cardiovascular surgery. The samples were studied by flow cytometry employing the whole blood lysis method and excluding from the analysis the contamination of the lymphoid window by erythroid precursors. We observed that in the first 4 years of life the B subset represented more than 65% of all cells in the lymphoid window, most of them (80%) exhibiting the immature phenotype CD19+CD100+. Conversely, the T subset was composed of mature CD4+ or CD8+ cells, with the CD4/CD8 ratio being less than 1 in all age groups. With age there was a progressive decrease in the percentage of B cells and an increase of T cells, reaching similar proportions in the BM from adults (33.6% and 34.8%, respectively). Furthermore, the percentage of CD10+ cells in the B subset decreased independently, whereas the CD20 expression increased. The percentage of NK cells did not change with age.
Asunto(s)
Envejecimiento/fisiología , Subgrupos de Linfocitos B/fisiología , Células de la Médula Ósea/fisiología , Subgrupos de Linfocitos T/fisiología , Adolescente , Adulto , Anticuerpos Monoclonales , Antígenos CD , Biopsia con Aguja , Niño , Preescolar , Femenino , Citometría de Flujo , Humanos , Lactante , Recién Nacido , Células Asesinas Naturales/fisiología , Subgrupos Linfocitarios , Masculino , Persona de Mediana Edad , Esternón/citologíaRESUMEN
In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anaemia (AA) we measured the expression of the Fas receptor (membrane protein that triggers apoptosis), Fas ligand (FasL), bcl-2 (cytoplasmatic protein that blocks apoptosis) and p53 (nuclear protein that induces apoptosis) in CD3 and CD19 lymphocytes from the peripheral blood or bone marrow of controls, patients with AA, aplastic anaemia in complete remission (AA-CR) and multiply transfused patients without aplastic anaemia. The Fas receptor was overexpressed in both T and B lymphocytes from the peripheral blood and bone marrow from patients with AA. These abnormalities were not detected in AA-CR or multiply transfused patients. CD3/FasL cells were not increased and no FasL expression was detected in B lymphocytes. Bcl-2 was highly expressed in lymphocytes from controls, AA, AA-CR and multiply transfused patients (> 99% of positive cells) whereas p53 was not detected in any group. To further characterize the functional activity of the Fas receptor we performed a Fas-induced apoptosis assay in peripheral blood lymphocytes using an anti-Fas monoclonal antibody. The crosslinking of the Fas receptor transduced an increased apoptotic signal in lymphocytes from AA patients, but not in lymphocytes from controls, AA-CR patients or multiply transfused patients. Taken together, these data suggest that a Fas-based mediated apoptosis without the apparent participation of bcl-2 or p53 is a possible mechanism of lymphocyte depletion in patients with AA. In addition, these findings suggest that Fas expression is a continuous event occurring from progenitor bone marrow cells to mature cells.
Asunto(s)
Anemia Aplásica/patología , Genes bcl-2 , Genes p53 , Glicoproteínas de Membrana/metabolismo , Adolescente , Adulto , Anemia Aplásica/genética , Apoptosis , Niño , Proteína Ligando Fas , Femenino , Citometría de Flujo , Humanos , Subgrupos Linfocitarios/patología , Linfocitos/patología , Masculino , Persona de Mediana EdadRESUMEN
To address the effect of extremely low frequency electromagnetic fields on programmed cell death we assessed both the spontaneous and dexamethasone (Dex)-induced apoptosis of thymocytes and spleen cells from mice submitted to a long-term continuous exposure of a 0.4-1.0 microT 60 Hz magnetic field or an 8-20 microT direct current (DC) magnetic field. Dex-induced apoptosis but not spontaneous apoptosis was substantially increased in thymocytes from 0.4 to 1.0 microT 60 Hz field-exposed animals. Spontaneous apoptosis and Dex-induced apoptosis of spleen cells were not affected by the 0.4-1.0 microT 60 Hz field exposure. In addition, spontaneous apoptosis and Dex-induced apoptosis of thymocytes and spleen cells from mice exposed to an 8-20 microT DC field were similar to the controls. These findings represent the first demonstration that thymocytes from mice exposed to a long-term 0.4-1.0 microT 60 Hz field may show abnormal response to Dex apoptotic stimuli.