Perfil de parámetros hematológicos en el HIV pediátrico y los cambios asociados al tratamiento antirretroviral / Hematological parameters profile in pediatric HIV and changes associated with antiretroviral treatment / Perfil de parâmetros hematológicos no HIV pediátrico e as alterações associadas ao tratamento anti-retroviral

Resumen Las alteraciones hematológicas son comunes durante la infección por el virus de la inmunodeficiencia humana (HIV). El objetivo de este trabajo fue describir los perfiles hematológicos e inmunológicos de niños infectados, antes y después de 36 meses de implementado el tratamiento antirretroviral (TARV). Se revisaron historias clínicas de niños expuestos, atendidos en este hospital en el período 2008-2018, con edades entre 6 meses y 14 años. Fueron empleados un contador hematológico (ADVIA 2120), un citómetro de flujo (FACScalibur BD) y una PCR en tiempo real Nuclisens EasyQ (bioMérieux). En 486 historias clínicas se encontraron 58 pacientes sin TARV, 30 por diagnóstico reciente y 28 por adherencia incorrecta o abandono de tratamiento. En ambos grupos se observó disminución porcentual de hemoglobina (Hb) (53% y 43%), volumen corpuscular medio (VCM) (43% y 7%) y LTCD4+ (37% y 57%), respectivamente, sin alteraciones significativas en otros parámetros hematológicos. Veintidós niños con correcta adherencia al TARV incrementaron significativamente los niveles de LTCD4+ (t0:18,8±9%, t1:32,7±6%), Hb (t0:10,9±1,6 g/dL, t1:12,6±1,1g/dL) y VCM (t0:78,7±4,5 fL, t1:101,9±5,6 fL), con disminución simultánea de la carga viral (CV) (t0:4,4±0,75 log t1:<1,70 log) después del seguimiento. La disminución de Hb observada aproximadamente en el 50% de los pacientes sin TARV estaría asociada a la acción viral y al tiempo de evolución de la infección. El incremento en los niveles, asociados a macrocitosis, se relacionaría con el aumento de LTCD4+ y disminución de la CV.

Abstract Hematologic abnormalities are common during human immunodeficiency virus (HIV) infection. Our aim was to describe hematological and immunological profiles present in antiretroviral treatment (ART)-naïve infected children and the changes observed after 36 months of ART initiation. Medical records of exposed children attended at this hospital in the 2008-2018 period were reviewed. Children between 6 months and 14 years were included. An automated blood analyser ADVIA 2120, a FACScalibur BD flow cytometer, and a Nuclisens EasyQ bioMérieux real-time PCR were used to determine different parameters. In 486 medical records evaluated, 58 patients ART-naïve were found, 30 due to recent diagnosis and 28 for incorrect adherence or abandoned treatment. In both groups, a percentage decrease in hemoglobin (Hb) (53% and 43%), mean corpuscular volume (MCV) (43% and 7%) and LTCD4+ (37% and 57%) levels respectively, was observed, without significant alterations in other hematological parameters. Twenty-two children with ART correct adherence increased significantly CD4+T cells (t0:18.8±9%, t1:32.7±6%), Hb (t0:10.9±1.6 g/dL, t1:12.6±1.1 g/dL) and MCV (t0:78.7±4.5 fL, t1:101.9±5.6 fL) levels, with simultaneous decrease of viral load (VL), (t0:4.4±0.75 log, t1:<1.70 log), after 36 months of follow-up. The reduction in Hb levels observed in 50% approximately of patients without ART would be associated with viral action and time of evolution of the infection. The increase in Hb levels and an associated macrocytosis would be related to the CD4+ T cells increase and VL decrease.

Resumo Alterações hematológicas são comuns durante a infecção pelo vírus da imunodeficiência humana (HIV). Nosso objetivo foi descrever os perfis hematológicos e imunológicos em crianças infectadas, antes e após 36 meses de implementar o tratamento antirretroviral (TARV). Foram revisados os prontuários das crianças expostas atendidas neste hospital no período 2008-2018, com idade entre 6 meses e 14 anos. Um contador hematológico (ADVIA 2120), um citômetro de fluxo (FACScalibur BD) e um PCR em tempo real Nuclisens EasyQ (bioMérieux), foram usados. Em 486 prontuários foram encontrados 58 pacientes livres de TARV, 30 por diagnóstico recente e 28 por adesão incorreta ou abandono do tratamento. Em ambos os grupos, observou-se diminuição percentual de hemoglobina (Hb) (53% e 43%), volume corpuscular médio (VCM) (43% e 7%) e LTCD4+ (37% e 57%), respectivamente, sem alterações significativas nos demais parâmetros hematológicos. Vinte e duas crianças com adesão correta ao TARV aumentaram significativamente os níveis de LTCD4+ (t0:18,8±9%, t1:32,7±6%), Hb (t0:10,9±1,6 g/dL, t1:12,6±1,1 g/dL) e VCM (t0:78,7±4,5 fL, t1:101,9±5,6 fL), com diminuição simultânea da carga viral (CV) (t0:4,4±0,75 log, t1:<1,70 log), depois do seguimento. A diminuição dos níveis de Hb observada em aproximadamente 50% dos pacientes sem TARV estaria associada à ação viral e ao tempo de evolução da infecção. O aumento nos níveis, associados a macrocitose, estaria relacionado com o aumento de LTCD4+ e diminuição da CV.

Mixed T Helper1/T Helper2/T Cytotoxic Profile in Subjects with Chronic Chagas Disease with Hypersensitivity Reactions to Benznidazole.

Dermatitis is the most common adverse event during treatment with benznidazole in chronic Chagas disease and is probably mediated by T cells. A set of molecules representative of the different type IV hypersensitivity reactions was evaluated in the circulation and skin biopsies of Trypanosoma cruzi-infected subjects presenting dermatitis during benznidazole administration. Through cytometric bead assays and enzyme-linked immunosorbent assay capture techniques, the serum levels of cytokines, chemokines, proapoptotic molecules, and mediators of the activation and migration of eosinophils and T cells were measured in subjects infected with Trypanosoma cruzi who exhibited skin adverse events (n = 22) and compared with those without adverse events (n = 37) during benznidazole therapy. Serum levels of interleukin- 5 (IL-5), soluble Fas cell surface death receptor ligand (FAS-L), and interferon γ-induced protein (IP-10) significantly increased at 7 to 30 days posttreatment with benznidazole and decreased thereafter in subjects with dermatitis but not in those without dermatitis. Circulating eotaxin levels were lower in subjects with dermatitis than in those without. Two patterns emerged in the skin biopsies: a T helper 1/T cytotoxic profile and a T helper 2/T cytotoxic profile with the presence of CD4+ and CD8+ T cells. Increased low-density lipoprotein (LDL), glutamic-oxaloacetic transaminase (GOT), uremia, and T cell activation emerged as risk factors for the development of dermatitis during benznidazole administration. These results support a delayed-type hypersensitivity reaction to benznidazole, involving CD4+ and CD8+ T cells and eosinophils, and a mixed cytokine profile. This study provides new insights for better management of adverse drug reactions to benznidazole. IMPORTANCE This study identified the risk factors for the development of adverse reactions to benznidazole and identified a set molecule to monitor the appearance of these reactions. This knowledge might improve the safety of benznidazole administration.

Erythropoietin in children with hemolytic uremic syndrome: a pilot randomized controlled trial.

BACKGROUND: The efficacy of recombinant human erythropoietin (rHuEPO) in sparing red blood cell (RBC) transfusions in children with hemolytic uremic syndrome related to Shiga toxin-producing Escherichia coli (STEC-HUS) is uncertain. METHODS: We conducted a pilot randomized controlled open trial between December 2018 and January 2021. Children were randomized to the intervention (subcutaneous rHuEPO 50 U/kg three times weekly until discharge + RBC transfusion if hemoglobin ≤ 7 g/dL and/or hemodynamic instability) or to the control arm (RBC transfusion if hemoglobin ≤ 7 g/dL and/or hemodynamic instability). Primary outcome was the number of RBC transfusions received during hospitalization. Secondary outcomes were to explore whether baseline EPO levels were adequate to the degree of anemia, to correlate selected acute phase parameters with the number of RBC transfusions, and to assess possible adverse events. RESULTS: Twelve patients per arm were included; they were comparable at recruitment and throughout the disease course. Median number of RBC transfusions was similar between groups (1.5, p = 0.76). Most patients had baseline EPO levels adequate to the degree of anemia, which did not correlate with the number of transfusions (r = 0.19, p = 0.44). Conversely, baseline (r = 0.73, p = 0.032) and maximum lactic dehydrogenase levels (r = 0.78, p = 0.003), creatinine peak (r = 0.71, p = 0.03) and dialysis duration (r = 0.7, p = 0.04) correlated significantly with RBC requirements. No side effects were recorded. CONCLUSION: In children with STEC-HUS, the administration of rHuEPO did not reduce the number of RBC transfusions. Larger studies addressing higher doses and similar severity of kidney failure at rHuEPO initiation (e.g. at start of dialysis) are warranted. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT03776851. A higher resolution version of the Graphical abstract is available as Supplementary information.

Respuesta humoral y persistencia de anticuerpos anti-SARS-CoV-2 en pacientes hospitalizados / SARS-CoV-2 antibodies kinetics in hospitalized patients

Resumen Luego de la infección por SARS-CoV-2 se producen anticuerpos específicos y continúa siendo objeto de estudio su cinética, cuantificación y umbral protector. Nuestro objetivo fue estudiar la cinética de aparición de anticuerpos IgG/IgM anti SARS-CoV-2, magnitud de respuesta y duración en el tiempo, en 55 pacientes hospitalizados por COVID-19, y relacionar el patrón de respuesta con diferentes parámetros demográ ficos y clínicos. Medidas con un ensayo cualitativo automatizado, VIDAS® SARS-Cov-2 (Biomerieux - ELFA) se evaluaron las variaciones en la concentración de anticuerpos a lo largo del tiempo con un modelo generalizado de efectos fijos. Todos los pacientes seroconvirtieron IgM e IgG, al día 10 y 10.5 respectivamente, patrón sincrónico mayoritario; no siendo de utilidad la IgM aislada como indicador de respuesta aguda. La sensibilidad clínica fue: semana 1, 30%, semanas 2 y 3, 72%, 4: 91% y 8: 96%. IgG permaneció detectable hasta los 6 meses (período de seguimiento) con gran heterogeneidad de magnitud; IgM negativizó en el 90.9% de los pacientes. Observamos un nivel mayor de IgM en los pacientes > 56 años, y en hombres respecto a mujeres. En pacientes con enfer medad pulmonar obstructiva crónica la respuesta de IgM está aumentada; los inmunocomprometidos y aquellos con enfermedad pulmonar intersticial difusa tuvieron menor IgM e IgG respectivamente. De nuestro grupo de pacientes, aquellos que requirieron cuidados críticos, asistencia respiratoria mecánica y los que fallecieron no presentaron diferencias significativas en magnitud de respuesta humoral respecto de quienes tuvieron un curso menos grave. La metodología utilizada refleja adecuadamente la cinética de anticuerpos.

Abstract Specific antibodies are produced after infection by SARS-CoV2. Currently, the understanding of antibody responses following infection with SARS-CoV-2 is limited including the magnitude, duration of responses and correlates of protective immunity following infection. Here we intended to characterize humoral immune response in a cohort of 55 hospitalized patients for COVID-19 and its relationship with different demographic and clinical parameters. The ELFA assay VIDAS® SARS-Cov-2 (Biomerieux) measured IgG/IgM antibodies. Their concentration over time was evaluated with a fixed effects generalized linear model. All patients seroconverted IgM and IgG, at day 10 and 10.5 respectively, showing a majority synchronous pattern; IgM alone would not be useful as a marker of acute response. Clini cal sensitivity was: week 1, 30%, weeks 2 and 3, 72%, 4: 91% and 8: 96%. IgG seropositivity was sustained in patients up to 180 days (last time point measured), in contrast, IgM response was short-lived (91days) in 90.9% of patients. Longer term follow-up is needed to determine the duration of IgG responses. We observed a higher level of IgM in patients > 56 years, and in men compared to women. In chronic obstructive pulmonary disease patients, the IgM response is increased, while in immunocompromised and interstitial lung disease patients, IgM and IgG were lower, respectively. Those patients who required critical care, mechanical ventilation and those who died did not present significant differences in the magnitude of humoral response compared to those who had a less severe course. The methodology used adequately reflects the kinetics of antibodies.

[SARS-CoV-2 antibodies kinetics in hospitalized patients]. / Respuesta humoral y persistencia de anticuerpos anti-SARS-CoV-2 en pacientes hospitalizados.

Specific antibodies are produced after infection by SARS-CoV2. Currently, the understanding of antibody responses following infection with SARS-CoV-2 is limited including the magnitude, duration of responses and correlates of protective immunity following infection. Here we intended to characterize humoral immune response in a cohort of 55 hospitalized patients for COVID-19 and its relationship with different demographic and clinical parameters. The ELFA assay VIDAS® SARS-Cov-2 (Biomerieux) measured IgG/IgM antibodies. Their concentration over time was evaluated with a fixed effects generalized linear model. All patients seroconverted IgM and IgG, at day 10 and 10.5 respectively, showing a majority synchronous pattern; IgM alone would not be useful as a marker of acute response. Clinical sensitivity was: week 1, 30%, weeks 2 and 3, 72%, 4: 91% and 8: 96%. IgG seropositivity was sustained in patients up to 180 days (last time point measured), in contrast, IgM response was short-lived (91days) in 90.9% of patients. Longer term follow-up is needed to determine the duration of IgG responses. We observed a higher level of IgM in patients > 56 years, and in men compared to women. In chronic obstructive pulmonary disease patients, the IgM response is increased, while in immunocompromised and interstitial lung disease patients, IgM and IgG were lower, respectively. Those patients who required critical care, mechanical ventilation and those who died did not present significant differences in the magnitude of humoral response compared to those who had a less severe course. The methodology used adequately reflects the kinetics of antibodies.

Luego de la infección por SARS-CoV-2 se producen anticuerpos específicos y continúa siendo objeto de estudio su cinética, cuantificación y umbral protector. Nuestro objetivo fue estudiar la cinética de aparición de anticuerpos IgG/IgM anti SARS-CoV-2, magnitud de respuesta y duración en el tiempo, en 55 pacientes hospitalizados por COVID-19, y relacionar el patrón de respuesta con diferentes parámetros demográficos y clínicos. Medidas con un ensayo cualitativo automatizado, VIDAS® SARS-Cov-2 (Biomerieux - ELFA) se evaluaron las variaciones en la concentración de anticuerpos a lo largo del tiempo con un modelo generalizado de efectos fijos. Todos los pacientes seroconvirtieron IgM e IgG, al día 10 y 10.5 respectivamente, patrón sincrónico mayoritario; no siendo de utilidad la IgM aislada como indicador de respuesta aguda. La sensibilidad clínica fue: semana 1, 30%, semanas 2 y 3, 72%, 4: 91% y 8: 96%. IgG permaneció detectable hasta los 6 meses (período de seguimiento) con gran heterogeneidad de magnitud; IgM negativizó en el 90.9% de los pacientes. Observamos un nivel mayor de IgM en los pacientes > 56 años, y en hombres respecto a mujeres. En pacientes con enfermedad pulmonar obstructiva crónica la respuesta de IgM está aumentada; los inmunocomprometidos y aquellos con enfermedad pulmonar intersticial difusa tuvieron menor IgM e IgG respectivamente. De nuestro grupo de pacientes, aquellos que requirieron cuidados críticos, asistencia respiratoria mecánica y los que fallecieron no presentaron diferencias significativas en magnitud de respuesta humoral respecto de quienes tuvieron un curso menos grave. La metodología utilizada refleja adecuadamente la cinética de anticuerpos.

Armonización de la determinación de ANCA por inmunofluorescencia indirecta: primera reunión argentina / Harmonization of the determination of ANCA by indirect immunofluorescence: first argentine meeting / Harmonização da determinação de ANCA por imunofluorescência indireta: primeira reunião argentina

Resumen La determinación de anticuerpos anti-citoplasma de neutrófilos (ANCA) es utilizada en la clínica diaria como una herramienta diagnóstica en distintas formas de vasculitis de pequeños vasos (vasculitis asociadas a ANCA), incluyendo la granulomatosis con poliangeítis (antes: granulomatosis de Wegener), poliangeítis microscópica y granulomatosis eosinofílica con poliangeítis (antes: síndrome de Churg-Strauss), y como apoyo diagnóstico de colitis ulcerosa, colangitis esclerosante primaria y enfermedad de Crohn. Estos anticuerpos están dirigidos contra distintos epítopos antigénicos de diferentes proteínas presentes en el citoplasma del neutrófilo. La determinación de ANCA, habitualmente realizada por la técnica de inmunofluorescencia indirecta (IFI), presenta cierto grado de complejidad en la definición de las imágenes de los diferentes patrones, variabilidad en la forma de trabajo y expresión de resultados. El 24 de octubre de 2018 en el marco del X Congreso Argentino de la Calidad en el Laboratorio Clínico y VIII Jornadas Latinoamericanas de la Calidad en el Laboratorio Clínico, en la Ciudad de Buenos Aires, se realizó una reunión de Armonización de la Determinación de ANCA por IFI con el objeto de presentar, discutir y consensuar los distintos aspectos que se presentan en esta técnica. Las propuestas iniciales fueron discutidas arribándose a recomendaciones generales para proporcionar estándares de trabajo e interpretación de imágenes con el objeto de disminuir la variabilidad de resultados entre los laboratorios clínico-inmunológicos.

Abstract Anti-neutrophil cytoplasmic antibodies (ANCA) tests are widely used in daily clinical practice as a useful tool for the diagnosis of pathologies such as granulomatosis with polyangiitis, microscopic polyangiitis, pauci-immune necrotizing segmental glomerulonephritis and eosinophilic granulomatosis with polyangiitis, ulcerative colitis, primary sclerosing cholangitis, Crohn's disease, etc. These antibodies are directed against different antigenic epitopes of various proteins which are present in the neutrophil cytoplasm. ANCA testing is usually carried out by using indirect immunofluorescence (IIF) method. The determination of ANCA presents some difficulties in the definition of the images of the different patterns, work protocols and result reports uniformity. The "Harmonization Conference on the Determination of ANCA by IIF" was held within the framework of the X Argentine Congress on Quality in the Clinical Laboratory and VIII Latin American Conference on Quality in the Clinical Laboratory, in Buenos Aires, Argentina, on October 24, 2018, in order to present, discuss and agree on different aspects of this method. Some initial proposals were discussed, arriving at general recommendations to provide standards of work and an image interpretation, with the aim of reducing the variability of results among the clinical-immunological laboratories.

Resumo A determinação de anticorpos anti-citoplasma de neutrófilos (ANCA) é utilizada na prática clínica diária como um suporte diagnóstico para várias patologias, como granulomatose com poliangiite, poliangiite microscópica, glomerulonefrite segmentar necrotizante pauci-imune e granulomatose eosinofílica com poliangiite, poliarterite nodosa, colite ulcerosa, colangite esclerosante primária, doença de Crohn. Esses anticorpos são dirigidos contra diferentes epítopos antigênicos de diferentes proteínas presentes no citoplasma do neutrófilo. A determinação de ANCA, normalmente realizada pela técnica de imunofluorescência indireta IFI, apresenta certo grau de complexidade na definição das imagens dos diferentes padrões, variabilidade na forma de trabalho e expressão de resultados. Em 24 de outubro de 2018, no âmbito do X Congreso Argentino de la Calidad en el Laboratorio Clínico (X Congresso Argentino da Qualidade no Laboratório Clínico) e VIII Jornadas Latinoamericanas de la Calidad en el Laboratorio Clínico (VIII Jornadas Latino-americanas da Qualidade no Laboratório), na cidade de Buenos Aires, foi realizada uma Jornada de Harmonização da Determinação de ANCA pelo IFI a fim de apresentar, discutir e concordar sobre os diferentes aspectos que são apresentados nesta técnica. As propostas iniciais foram discutidas, chegando a recomendações gerais para fornecer padrões de trabalho e interpretação de imagens visando reduzir a variabilidade de resultados entre os laboratórios clínico-imunológicos.

[Seroprevalence of anti-Mycoplasma pneumoniae antibodies in otherwise healthy children]. / Seroprevalencia de anticuerpos anti-Mycoplasma pneumoniae en niños menores de 12 años.

INTRODUCTION: Mycoplasma pneumoniae (Mypn) infection could be occurring at an earlier age due to social pheno mena such as attending daycare centers more frequently and earlier than decades ago. OBJECTIVE: to estimate the prevalence of anti-Mypn antibodies in children aged 0-12 years, and to explore whether age, attendance to daycare center/school, overcrowding or the presence of children aged below 12 years in the households increase the risk of seropositivity. PATIENTS AND METHOD: Cross-sectional stu dy including healthy children aged 0-12 years which required blood draws for routine laboratory tests. In all cases, the aforementioned variables were recorded and anti-Mypn IgG was determined by enzyme immunoassay. The association between predictors and seropositivity was assessed in a logistic regression model. RESULTS: We included 232 patients (average age 56.4 ± 40.0 months). 56.9% attended a daycare center/school, 63.8% co-habited with children under 12 years old, and 15.9% lived in overcrowded households. The prevalence of anti-Mypn antibodies was 14.6%. There were no significant differences between seropositive and seronegative children regarding age (63.1 ± 40.7 vs. 55.4 ± 41.3 months), school/day-care attendance (64.7% vs. 55.5%), overcrowding (14.7% vs. 14.9%), or co-habiting with children (64.7% vs. 63.6%). Age was not an independent predictor of seropositivity in the multivariate model. CONCLUSION: The prevalence of anti-Mypn antibodies in children was 14.6% and age was not a predictor of seropositivity.

Seroprevalencia de anticuerpos anti-Mycoplasma pneumoniae en niños menores de 12 años / Seroprevalence of anti-Mycoplasma pneumoniae antibodies in otherwise healthy children

Resumen: Introducción: La infección por Mycoplasma pneumoniae (Mypn) podría estar ocurriendo a edades más tempranas, debido a fenómenos sociales como concurrencia a centros de cuidado diurno en forma más frecuente y precoz. Objetivo: estimar la prevalencia de anticuerpos anti-Mypn en niños de 0-12 años, y explorar si la edad, asistencia a centro de cuidados diurnos/escuela, hacinamiento o convivencia con niños incrementan el riesgo de seropositividad. Pacientes y Método: Estudio transversal incluyendo niños de 0-12 años de edad que requirieron extracciones de sangre para control, por lo demás sanos. En todos los casos se consignaron las variables mencionadas y se determinó IgG anti-Mypn mediante enzimoinmunoanálisis. Se evaluó la asociación entre predictores y seropositividad en un modelo de regresión logística. Resultados: Se incluyeron 232 pacientes (edad promedio 56,4 ± 40,0 meses). El 56,9% concurría a centro de cuidado diurno/escuela, 63,8% convivían con menores de 12 años y 15,9% presentaban hacinamiento. El 14,6% presentaba anticuerpos anti-Mypn. Los niños seroposi- tivos no mostraron diferencias significativas con aquellos seronegativos en relación a edad (63,1 ± 40,7 vs. 55,4 ± 41,3 meses), escolaridad (64,7% vs 55,5%), hacinamiento (14,7% vs 14,9%), ni con vivencia con menores (64,7% vs 63,6%). La edad tampoco se mostró como predictor independiente de seropositividad en el modelo multivariado. Conclusión: La prevalencia de anticuerpos anti-Mypn fue 14,6%. La edad no fue predictor de seropositividad.

Abstract: Introduction: Mycoplasma pneumoniae (Mypn) infection could be occurring at an earlier age due to social pheno mena such as attending daycare centers more frequently and earlier than decades ago. Objective: to estimate the prevalence of anti-Mypn antibodies in children aged 0-12 years, and to explore whether age, attendance to daycare center/school, overcrowding or the presence of children aged below 12 years in the households increase the risk of seropositivity. Patients and Method: Cross-sectional stu dy including healthy children aged 0-12 years which required blood draws for routine laboratory tests. In all cases, the aforementioned variables were recorded and anti-Mypn IgG was determined by enzyme immunoassay. The association between predictors and seropositivity was assessed in a logistic regression model. Results: We included 232 patients (average age 56.4 ± 40.0 months). 56.9% attended a daycare center/school, 63.8% co-habited with children under 12 years old, and 15.9% lived in overcrowded households. The prevalence of anti-Mypn antibodies was 14.6%. There were no significant differences between seropositive and seronegative children regarding age (63.1 ± 40.7 vs. 55.4 ± 41.3 months), school/day-care attendance (64.7% vs. 55.5%), overcrowding (14.7% vs. 14.9%), or co-habiting with children (64.7% vs. 63.6%). Age was not an independent predictor of seropositivity in the multivariate model. Conclusion: The prevalence of anti-Mypn antibodies in children was 14.6% and age was not a predictor of seropositivity.

Restoration of recent thymic emigrant CD4+ T cells is associated with sustained adherence to antiretroviral treatment in HIV-infected children.

To evaluate the levels of recent thymic emigrant (RTE) CD4+ T cells in HIV-infected children and to explore the associations among their frequency, antiretroviral treatment (ART) adherence, and CD4+ T cell restoration. The group evaluated comprised 85 HIV-infected patients classified as subjects with moderate or severe immunosuppression or as those with no evidence of immunosuppression. To evaluate the association between the frequency of RTE CD4+ T cells and ART adherence, 23 of the 85 patients were evaluated at two different time points during a one-year follow-up period. Children with severe immunosuppression had lower frequencies of RTE CD4+ T cells compared with children without evidence of immunosuppression (P < .001). The frequency of RTE CD4+ T cells in children with a high rate of adherence was significantly higher (P < .05) than that observed among those with suboptimal adherence. The latter group presented with infectious intercurrences on admission that decreased after initiation of treatment along with improved CD4+ and RTE naïve CD4+ T cells counts. The adequate ART adherence is essential for immune reconstitution, which might be reflected by the levels of RTE CD4+ T cells.

Inmunodeficiencia combinada con compromiso cutáneo asociada a mutación en DOCK8 / Combined immunodeficiency with cutaneous manifestations associated with DOCK8 mutation

Diferentes inmunodeficiencias primarias se caracterizan por niveles elevados de IgE e infecciones cutáneas de origen viral. Describimos el caso de un niño de 2 años y 8 meses de edad, con inmunodeficiencia combinada, dermatitis y molusco contagioso diseminado. El paciente presentaba niveles aumentados de IgE, eosinofilia y marcada linfopenia a predominio de TCD8. Se encontraron alteraciones en los ensayos funcionales por cultivo y en la respuesta a la vacunación. Resultados normales de la proteína ZAP-70, funcionalidad NK y niveles de HLA I, tendientes a verificar alteraciones cuantitativas y funcionales de las células citotóxicas, llevaron a la sospecha de deficiencia en el gen DOCK8. El resultado positivo del estudio molecular, junto con las características clínicas e inmunológicas del paciente, confirmaron el diagnóstico de esta nueva inmunodeficiencia, que, de acuerdo con nuestro conocimiento, sería el primer caso diagnosticado en un hospital pediátrico en nuestro país.

Different primary immunodeficiencies present increased levels of IgE and cutaneous infections of viral etiology. We report a case of a 2 y, 8 m old boy with combined immunodeficiency, dermatitis and disseminated molluscum contagiosum. The patient presented high titers of IgE, eosinophilia and pronounced TCD8 lymphopenia. Impaired proliferation assays and abnormal antibody response to vaccination were found. Normal results of ZAP-70 protein, NK function, and HLA I levels, to test quantitatives and functional defects of cytotoxic cells, lead us to suspect a mutation in DOCK8 gene. Positive result in molecular study together with clinical and immunology features in the patient confirmed the diagnosis of this new immunodeficiency, being to the authors ́ knowledge the first case recorded in a paediatric hospital in our country.

[Combined immunodeficiency with cutaneous manifestations associated with DOCK8 mutation]. / Inmunodeficiencia combinada con compromiso cutáneo asociada a mutación en DOCK8.

Different primary immunodeficiencies present increased levels of IgE and cutaneous infections of viral etiology. We report a case of a 2 y, 8 m old boy with combined immunodeficiency, dermatitis and disseminated molluscum contagiosum. The patient presented high titers of IgE, eosinophilia and pronounced TCD8 lymphopenia. Impaired proliferation assays and abnormal antibody response to vaccination were found. Normal results of ZAP-70 protein, NK function, and HLA I levels, to test quantitatives and functional defects of cytotoxic cells, lead us to suspect a mutation in DOCK8 gene. Positive result in molecular study together with clinical and immunology features in the patient confirmed the diagnosis of this new immunodeficiency, being to the authors' knowledge the first case recorded in a paediatric hospital in our country.

Inmunodeficiencia combinada con compromiso cutáneo asociada a mutación en DOCK8. / [Combined immunodeficiency with cutaneous manifestations associated with DOCK8 mutation].

Different primary immunodeficiencies present increased levels of IgE and cutaneous infections of viral etiology. We report a case of a 2 y, 8 m old boy with combined immunodeficiency, dermatitis and disseminated molluscum contagiosum. The patient presented high titers of IgE, eosinophilia and pronounced TCD8 lymphopenia. Impaired proliferation assays and abnormal antibody response to vaccination were found. Normal results of ZAP-70 protein, NK function, and HLA I levels, to test quantitatives and functional defects of cytotoxic cells, lead us to suspect a mutation in DOCK8 gene. Positive result in molecular study together with clinical and immunology features in the patient confirmed the diagnosis of this new immunodeficiency, being to the authors knowledge the first case recorded in a paediatric hospital in our country.

[Intestinal cryptosporidiosis in HIV infected children]. / Criptosporidiosis intestinal en niños con HIV/Sida.

Cryptosporydium parvum is an intracellular parasite that infects gastrointestinal epithelium and produces diarrhea that is self-limited in immunocompetent persons but potentially life-threatening in immunocompromised, especially those with the acquired immunodeficiency syndrome (AIDS). C. parvum enteric infection's incidence in a pediatric HIV/AIDS cohort, during a 6 years period, was studied. Clinical and immunologic characteristics of the dual infection were also recorded. Highly active antiretroviral therapy (HAART) was started or continued by all the patients during follow-up. Azithromicyn was used as antiparasitic drug. Cryptosporidiosis incidence was 13.7%; 33 out 240 children showed chronic diarrhea lasting 14 days at least, or recurrent, without dehydration and electrolytic disturbance. Peripheral blood T CD4+ percentage levels of the patients were variable and without relationship with C. parvum presence. Viral load levels in 31 out 33 patients were over cut-off at the enteric episode time. Mild or moderate eosinophilia were recorded in 23% of the patients and other intestinal parasites were present in 11 children. When the number of enteric episodes were compared with the clinical and immunological patient's status, not significant differences were recorded. HAART is the best treatment to improve immune function in HIV patients avoiding potentially fatal complications that accompany acute diarrhea during concomitant infection with C. parvum.

Criptosporidiosis intestinal en niños con HIV/SIDA / Intestinal cryptosporidiosis in HIV infected children

El Cryptosporidium parvum, protozoo parásito intracelular, infecta el epitelio gastrointestinal produciendo diarrea autolimitada en individuos inmunocompetentes pero potencialmente grave en pacientes inmunocomprometidos, especialmente en aquellos con Sida. En este trabajo se evaluó, durante un lap-so de 6 años, la incidencia de infección intestinal por C. parvum en una población pediátrica con HIV/Sida analizando las características clínicas e inmunológicas de la coinfección. Todos los pacientes iniciaron o continuaron el tratamiento antirretroviral de alta eficacia HAART durante el período de estudio, mientras que la infección intestinal fue tratada con azitromicina. La incidencia de criptosporidiosis fue de 13.7%. 33 de los 240 niños en seguimiento presentaron diarrea crónica de más de 14 días de evolución o recurrente, sin complicaciones hidroelectrolíticas. Los pacientes evaluados presentaron niveles porcentuales variables de células T CD4+ en sangre periférica, y la presencia del parásito no estuvo en relación con el compromiso inmunitario. Al momento del cuadro entérico 31 de los 33 pacientes tuvieron niveles plasmáticos de carga viral que superaban el límite de detección. Se observó eosinofilia leve o moderada en el 23% de los pacientes y la coinfección con otros parásitos fue detectada en 11 niños. No se obtuvieron diferencias significativas al relacionar el número de episodios intestinales con los estadios clínico-inmunológicos de los pacientes. La correcta implementación del HAART con la subsecuente restauración de la función inmune se relacionaría con la ausencia de cuadros diarreicos agudos y de las complicaciones hidroelectrolíticas derivadas de la coinfección con C. parvum.

Cryptosporydium parvum is an intracellular parasite that infects gastrointestinal epithelium and produces diarrhea that is self-limited in immunocompetent persons but potentially life-threatening in immunocompromised, especially those with the acquired immunodeficiency syndrome (AIDS). C. parvum enteric infection's incidence in a pediatric HIV/AIDS cohort, during a 6 years period, was studied. Clinical and immunologic characteristics of the dual infection were also recorded. Highly active antiretroviral therapy (HAART) was started or continued by all the patients during follow-up. Azithromicyn was used as antiparasitic drug. Cryptosporidiosis incidence was 13.7%; 33 out 240 children showed chronic diarrhea lasting 14 days at least, or recurrent, without dehydration and electrolytic disturbance. Peripheral blood T CD4+ percentage levels of the patients were variable and without relationship with C. parvum presence. Viral load levels in 31 out 33 patients were over cut-off at the enteric episode time. Mild or moderate eosinophilia were recorded in 23% of the patients and other intestinal parasites were present in 11 children. When the number of enteric episodes were compared with the clinical and immunological patient's status, not significant differences were recorded. HAART is the best treatment to improve immune function in HIV patients avoiding potentially fatal complications that accompany acute diarrhea during concomitant infection with C. parvum.

[L-selectin expression on T lymphocytes and neutrophils in HIV infected children]. / Expresión de L-selectina en linfocitos T y neutrófilos de niños infectados con HIV.

The ability of leukocytes to leave the circulation and migrate into tissues is a critical feature of the immune response. L-selectin (CD62L), the leukocyte selectin, mediates the binding of lymphocytes to high endothelial venules of peripheral lymph nodes and is also involved in lymphocyte, neutrophil and monocyte attachment to vascular endothelium at sites of inflammation. In this study L-selectin expression on peripheral T cells and neutrophils was evaluated in 25 HIV infected children, who had not received antiretroviral therapy, and 25 healthy controls. The expression level of L-selectin on T cells was also evaluated in 10 out 25 patients after 6 months of antiretroviral therapy. L-selectin expression on CD3+, CD4+ and CD8+ T cells were significantly lower in HIV infected children than in the control group. The percentage of neutrophils expressing CD62L was significantly reduced in patients with severe immunologic suppression. A positive correlation between the number of CD4+ T cells and the percentage of neutrophils CD62L+ was found. L-selectin expression on both CD4+ and CD8+ T cells did not significantly vary after 6 months of treatment. Altered leukocyte functions such as migration and homing resulting from reduced expression of CD62L may be an important contributor of the progressive dysfunction of the immune system in HIV infected children.

Expresión de L-selectina en linfocitos T y neutrófilos de niños infectados con HIV. / [L-selectin expression on T lymphocytes and neutrophils in HIV infected children]

The ability of leukocytes to leave the circulation and migrate into tissues is a critical feature of the immune response. L-selectin (CD62L), the leukocyte selectin, mediates the binding of lymphocytes to high endothelial venules of peripheral lymph nodes and is also involved in lymphocyte, neutrophil and monocyte attachment to vascular endothelium at sites of inflammation. In this study L-selectin expression on peripheral T cells and neutrophils was evaluated in 25 HIV infected children, who had not received antiretroviral therapy, and 25 healthy controls. The expression level of L-selectin on T cells was also evaluated in 10 out 25 patients after 6 months of antiretroviral therapy. L-selectin expression on CD3+, CD4+ and CD8+ T cells were significantly lower in HIV infected children than in the control group. The percentage of neutrophils expressing CD62L was significantly reduced in patients with severe immunologic suppression. A positive correlation between the number of CD4+ T cells and the percentage of neutrophils CD62L+ was found. L-selectin expression on both CD4+ and CD8+ T cells did not significantly vary after 6 months of treatment. Altered leukocyte functions such as migration and homing resulting from reduced expression of CD62L may be an important contributor of the progressive dysfunction of the immune system in HIV infected children.

Comportamiento de la forma soluble de L-selectina en niños infectados con HIV / Behavior of soluble L-selectin in HIV infected children

La L-selectina es una molécula de adhesión responsable de la adhesión inicial de los leucocitos al endotelio. Esta molécula es liberada desde la superficie celular por clivaje proteolítico después de la activación leucocitaria. Se midieron los niveles séricos de la forma soluble de la L-selectina (sL-selectina) en 51 niños HIV(+) y en 15 controles sanos HIV(-). Estos valores se compararon con parámetros de activación inmune y de progresión de enfermedad. La concentración de sL-selectina se encontró significativamente aumentada en el grupo de pacientes HIV(+). Dichos niveles eran más altos en los pacientes con mayor inmunosupresión. Se encontró una correlación positiva entre los niveles de sL-selectina y el número relativo de LTCD8, y entre sL-selectina y los niveles de la forma soluble de la molécula de adhesión intercelular-1(sICAM-1) y la inmunoglobulina A(IgA). No se obtuvo correlación significativa entre sL-selectina y los valores de la carga viral (CV) plasmática. El aumento en los niveles de sL-selectina sería otro componente entre las múltiples alteraciones inmunológicas producidas por el HIV

Comportamiento de la forma soluble de L-selectina en niños infectados con HIV / Behavior of soluble L-selectin in HIV infected children

La L-selectina es una molécula de adhesión responsable de la adhesión inicial de los leucocitos al endotelio. Esta molécula es liberada desde la superficie celular por clivaje proteolítico después de la activación leucocitaria. Se midieron los niveles séricos de la forma soluble de la L-selectina (sL-selectina) en 51 niños HIV(+) y en 15 controles sanos HIV(-). Estos valores se compararon con parámetros de activación inmune y de progresión de enfermedad. La concentración de sL-selectina se encontró significativamente aumentada en el grupo de pacientes HIV(+). Dichos niveles eran más altos en los pacientes con mayor inmunosupresión. Se encontró una correlación positiva entre los niveles de sL-selectina y el número relativo de LTCD8, y entre sL-selectina y los niveles de la forma soluble de la molécula de adhesión intercelular-1(sICAM-1) y la inmunoglobulina A(IgA). No se obtuvo correlación significativa entre sL-selectina y los valores de la carga viral (CV) plasmática. El aumento en los niveles de sL-selectina sería otro componente entre las múltiples alteraciones inmunológicas producidas por el HIV