Altered Distribution of Peripheral Blood Maturation-Associated B-Cell Subsets in Chronic Alcoholism.

BACKGROUND: Although decreased counts of peripheral blood (PB) B cells-associated with an apparently contradictory polyclonal hypergammaglobulinemia-have been reported in chronic alcoholism, no information exists about the specific subsets of circulating B cells altered and their relationship with antibody production. Here, we analyzed for the first time the distribution of multiple maturation-associated subpopulations of PB B cells in alcoholism and its potential relationship with the onset of liver disease. METHODS: PB samples from 35 male patients-20 had alcoholic hepatitis (AH) and 15 chronic alcoholism without liver disease (AWLD)-were studied, in parallel to 19 male healthy donors (controls). The distribution of PB B-cell subsets (immature/regulatory, naïve, CD27(-) and CD27(+) memory B lymphocytes, and circulating plasmablasts of distinct immunoglobulin-Ig-isotypes) was analyzed by flow cytometry. RESULTS: Patients with AH showed significantly decreased numbers of total PB B lymphocytes (vs. controls and AWLD), at the expense of immature, memory, and, to a lesser extent, also naïve B cells. AWLD showed reduced numbers of immature and naïve B cells (vs. controls), but higher PB counts of plasmablasts (vs. the other 2 groups). Although PB memory B cells were reduced among the patients, the percentage of surface (s)IgA(+) cells (particularly CD27(-) /sIgA(+) cells) was increased in AH, whereas both sIgG(+) and sIgA(+) memory B cells were significantly overrepresented in AWLD versus healthy donors. Regarding circulating plasmablasts, patients with AH only showed significantly reduced counts of sIgG(+) cells versus controls. In contrast, the proportion of both sIgA(+) and sIgG(+) plasmablasts-from all plasmablasts-was reduced in AH and increased in AWLD (vs. the other 2 groups). CONCLUSIONS: AH and AWLD patients display a significantly reduced PB B-cell count, at the expense of decreased numbers of recently produced immature/regulatory B cells and naïve B cells, together with an increase in Ig-switched memory B lymphocytes and plasmablasts, particularly of IgA(+) cells.

Decreased peripheral blood CD4+/CD25+ regulatory T cells in patients with alcoholic hepatitis.

BACKGROUND: Development of alcoholic hepatitis (AH) may be favored by the activation of the innate immune response. Recently, decreased numbers of circulating regulatory T cells (Tregs) have been reported in diseases associated with an immune activation status, but no studies have focused so far, in investigating the distribution of Tregs in chronic alcoholism and its potential association with liver disease. Here, we analyzed for the first time the frequency of peripheral blood (PB) Tregs and Treg subsets in AH and its relationship with the production of inflammatory cytokines by PB monocytes and dendritic cells (DCs). METHODS: PB samples from 25 male patients with AH were studied; in parallel, 15 male chronic alcoholic patients without liver disease (AWLD) and 17 male healthy donors were also studied, as controls. The distribution of CD4⁺CD25hiCD127-/lo Tregs and their maturation subsets (naïve, central memory, and peripheral memory Tregs) was analyzed by flow cytometry. Spontaneous and in vitro-stimulated production of inflammatory cytokines by PB monocytes and DCs was analyzed by flow cytometry at the cytoplasmic level. RESULTS: Patients with AH showed decreased (p < 0.05) numbers of PB CD4⁺CD25hiCD127-/lo Tregs at the expense of all maturation-associated subsets, while AWLD and healthy subjects showed a similar (p > 0.05) distribution of PB CD4⁺CD25hiCD127-/lo Tregs. Interestingly, significantly increased amounts of spontaneously produced inflammatory cytokines were found among circulating monocyte-derived DCs and monocytes from AH (and AWLD) patients in comparison with healthy donors. Conversely, the ability of these cell subsets to produce cytokines after in vitro stimulation was lower (p < 0.05) in AH versus the 2 control groups. CONCLUSIONS: PB CD4⁺CD25hiCD127-/lo Tregs are significantly decreased in patients with AH when compared to both healthy and AWLD; this may contribute to explain the more pronounced activation of the innate immune response observed in AH, as reflected by an increased secretion of inflammatory cytokines by PB DCs and monocytes, and could facilitate the development of liver disease.

Normal range for acid-labile subunit in paediatric patients in Spain and its association with age, sex, pubertal stage and other growth factors.

INTRODUCTION: The acid-labile subunit (ALS) plays an important role in the endocrine effects of insulin-like growth factors (IGFs) on target tissues. Historically, it has attracted limited attention. The aim of our study was to describe the normal range of ALS in healthy children and its association with other growth factors. PATIENTS AND METHODS: We designed a cross-sectional descriptive study. We collected data on age, height, body mass index, gestational age, anthropometry at birth and serum levels of ALS, IGF1 and IGFBP3 in healthy children aged 2-15 years with a normal height. The levels of ALS, IGF1 and IGFBP3 were measured by ELISA. We fitted GAMLSS normalization models to standardize the variables. RESULTS: Samples were collected from 446 children. In prepubertal children, the levels of ALS, IGF1 and IGFBP3 were positively correlated in both sexes and with age (P < .01). We found significant differences in the levels of ALS, IGF1 and IGFBP3 and the IGF1/IGFBP3 molar ratio between the sexes and higher levels in pubertal boys (P < .01). We generated normal probability plots for each sex for each of the components of the ternary complex and for the IGF1/IGFBP3 and IGFBP3/ALS molar ratios. In addition, we extracted equations from the models for the calculation of z-scores for age and sex. CONCLUSIONS: This study may contribute age- and sex-specific reference values for IGF1, IGFBP3 and ALS levels and IGF1/IGFBP3 and IGFBP3/ALS ratios in Spanish children and suggests an association between age, sex, and pubertal stage.

Rango normal de subunidad ácido-lábil (ALS) en niños españoles y su relación con la edad, el sexo, el estadio puberal y otros factores de crecimiento / Normal range for acid-labile subunit in paediatric patients in Spain and its association with age, sex, pubertal stage and other growth factors

Introducción: La subunidad ácido-lábil (ALS) tiene un papel importante en los efectos endocrinos de los factores de crecimiento similares a la insulina (IGF) en tejidos diana. Históricamente ha recibido una atención limitada. El objetivo de nuestro estudio fue describir el rango normal de ALS en niños sanos y su relación con otros factores de crecimiento. Pacientes y métodos: Se diseñó un estudio descriptivo transversal. Se recopilaron datos sobre edad, altura, índice de masa corporal, edad gestacional, antropometría al nacer y niveles séricos de ALS, IGF1 e IGFBP3 de niños sanos de 2 a 15años con estatura estándar. Los niveles de ALS, IGF1 e IGFBP3 se midieron mediante ELISA. Se utilizaron modelos de normalización GAMLSS para la estandarización de variables. Resultados: Se recogieron muestras de 446 niños. En niños prepúberes, los niveles de ALS, IGF1 e IGFBP3 se correlacionaron de manera positiva en ambos sexos y con la edad (p<0,01). Los niveles de ALS, IGF1 e IGFBP3 y la relación molar IGF1/IGFBP3 fueron significativamente diferentes entre ambos sexos y más altos en los niños puberales (p<0,01). Se realizaron gráficas de normalidad por género para cada uno de los componentes del complejo ternario y para las relaciones molares IGF1/IGFBP3 e IGFBP3/ALS. Además, se construyeron fórmulas modelo para calcular el Z Score según la edad y el sexo. ConclusionesEste estudio podría determinar valores de referencia específicos por edad y sexo para IGF1, IGFBP3, ALS, IGF1/IGFBP3 e IGFBP3/ALS en niños españoles y parece establecer la relación entre edad, sexo y estadio puberal. (AU)

Introduction: The acid-labile subunit (ALS) plays an important role in the endocrine effects of insulin-like growth factors (IGFs) on target tissues. Historically, it has attracted limited attention. The aim of our study was to describe the normal range of ALS in healthy children and its association with other growth factors. Patients and methods: We designed a cross-sectional descriptive study. We collected data on age, height, body mass index, gestational age, anthropometry at birth and serum levels of ALS, IGF1 and IGFBP3 in healthy children aged 2-15years with a normal height. The levels of ALS, IGF1 and IGFBP3 were measured by ELISA. We fitted GAMLSS normalization models to standardize the variables. Results: Samples were collected from 446 children. In prepubertal children, the levels of ALS, IGF1 and IGFBP3 were positively correlated in both sexes and with age (P<.01). We found significant differences in the levels of ALS, IGF1 and IGFBP3 and the IGF1/IGFBP3 molar ratio between the sexes and higher levels in pubertal boys (P<.01). We generated normal probability plots for each sex for each of the components of the ternary complex and for the IGF1/IGFBP3 and IGFBP3/ALS molar ratios. In addition, we extracted equations from the models for the calculation of z-scores for age and sex. Conclusions: This study may contribute to age- and sex-specific reference values for IGF1, IGFBP3 and ALS levels and IGF1/IGFBP3 and IGFBP3/ALS ratios in Spanish children and suggests an association between age, sex and pubertal stage. (AU)

Citoqueratinas (UBC y CYFRA 21-1) y proteínas de la matriz nuclear (NMP22) como marcadores tumorales en la orina en el diagnóstico del cáncer vesical / Cytokeratins (UBC and CYFRA 21-1) and molecular protein matrix as urinary tumor markers in the diagnosis of bladder cancer

Fundamento: Los marcadores tumorales en orina como UBC, CYFRA 21-1 y NMP22 son una alternativa no invasiva para el diagnóstico del cáncer vesical. Se comparó la sensibilidad individual y combinada de los marcadores urinarios en la detección del cáncer vesical con respecto a los métodos diagnósticos convencionales. Pacientes y métodos: Se recogieron consecutivamente las orinas precistoscopia de 237 individuos: 44 pacientes con sospecha de cáncer vesical primario y 193 pacientes en seguimiento de cáncer vesical. UBC y NMP22 se cuantificaron por enzimoinmunoanálisis, CYFRA 21-1 por electroquimioluminiscencia. Resultados: Tomando como puntos de corte 9,7 µg/l para UBC, 5,4 ng/ml para CYFRA 21-1 y 10,0 U/ml para NMP22 se encontraron unas sensibilidades del 70, del 69 y del 67 por ciento, para unas especificidades del 95, del 94 y del 80 por ciento, respectivamente. Todos los marcadores tumorales urinarios presentaron sensibilidades superiores a la de la citología urinaria (7 por ciento), la presencia de microhematuria (62 por ciento) y hematuria franca (10 por ciento), cuyas especificidades fueron del 99, del 78 y del 99 por ciento, respectivamente. La determinación conjunta CYFRA 21-1 y NMP22 fue la combinación que alcanzó la mayor sensibilidad (79 por ciento), ligeramente inferior a la determinación simultánea de los tres marcadores (80 por ciento). Conclusiones: La sensibilidad de los marcadores tumorales UBC, CYFRA 21-1 y NMP22 en orina para el diagnóstico de cáncer vesical puede justificar su determinación en sustitución de la citología urinaria. La similitud diagnóstica de las citoqueratinas individualmente y en cada tipo de pacientes en estudio desaconsejaría su determinación simultánea. La determinación conjunta de NMP22 y un marcador de citoqueratinas (CYFRA 21-1 o UBC) se presenta como la más aconsejable. (AU)