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1.
Horm Res Paediatr ; 94(9-10): 390-398, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34673643

RESUMEN

Hypophosphatasia (HPP) is an inborn error of metabolism caused by loss-of-function mutations in the biomineralization-associated alkaline phosphatase gene, encoding tissue-nonspecific alkaline phosphatase (TNSALP). Symptoms include skeletal hypomineralization and extra-skeletal manifestations such as pyridoxine (B6)-responsive seizures due to impaired cerebral B6 passage. Since the introduction of enzyme replacement therapy (ERT), skeletal manifestations and B6-responsive seizures were reported to improve significantly. Nevertheless, there is an increasing evidence of B6-independent neurological manifestation of HPP including HPP-associated encephalopathy. Here, we present for the first time the brain alterations of an infant with neonatal HPP who died of neurological complications at the age of 5 months despite early initiation of ERT. CSF analysis showed normal concentrations of biogenic amines reflecting sufficient intracellular B6 availability. Postmortem histopathology revealed severe, localized affection of the cerebral cortex including cortical lesions in layers 2 and 3 in direct proximity to TNSALP-expressing neurons and hippocampal sclerosis. Our findings confirm that TNSALP deficiency may lead to a severe encephalopathy. We hypothesize that HPP-associated encephalopathy resistant to currently available ERT may develop in addition and probably independently of typical B6-responsive seizures in some patients. Prospective, controlled studies with close neurological follow-up including brain imaging are needed to identify patients at risk for severe neurological symptoms despite ERT.


Asunto(s)
Encefalopatías , Hipofosfatasia , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/metabolismo , Encefalopatías/tratamiento farmacológico , Encefalopatías/genética , Terapia de Reemplazo Enzimático/métodos , Humanos , Hipofosfatasia/diagnóstico , Hipofosfatasia/tratamiento farmacológico , Hipofosfatasia/genética , Lactante , Recién Nacido , Mutación , Estudios Prospectivos
2.
Artif Organs ; 45(12): 1582-1593, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34331775

RESUMEN

Hemadsorption via the cytokine-adsorber CytoSorb (CytoSorbents Europe, Berlin, Germany) has successfully been used as an adjunctive method in adults, mainly for the purpose of immunomodulation under acute inflammatory conditions such as sepsis and cardiac surgery. In recent years, there has been growing interest in its use in pediatric intensive care to improve outcomes in patients with multiple organ failure following an inflammatory illness. Literature on the application of CytoSorb in neonatal and pediatric patients is scarce, though the implication is that it could be an effective last-resort treatment option in critically ill pediatric patients. Herein we present the clinical cases of two pediatric patients successfully treated with a combination of the CytoSorb hemadsorber, continuous renal replacement therapy, and extracorporeal membrane oxygenation due to multiple organ failure following different underlying medical conditions. Patient 1 was a 7-month-old male child with Down's syndrome admitted to the Pediatric Intensive Care Unit (PICU) after congenital heart surgery, who developed antimicrobial-resistant septic shock and severe acute respiratory distress syndrome. Patient 2 was a 2-year-old male child admitted to the PICU with influenza A-associated acute liver failure resulting in hyperammonemia, lactate acidosis, hemodynamic instability, and acute kidney failure. In both patients, hemadsorption with CytoSorb was initiated as an adjunctive rescue therapy to treat refractory multisystem organ failure. Improvement of laboratory and clinical parameters was observed within hours of treatment initiation. The application of the hemadsorber-developed for use in adults-proved simple and safe for use in both of our low-weight pediatric patients.


Asunto(s)
Terapia de Reemplazo Renal Continuo/métodos , Oxigenación por Membrana Extracorpórea/métodos , Hemabsorción , Insuficiencia Multiorgánica/terapia , Síndrome de Dificultad Respiratoria/terapia , Preescolar , Síndrome de Down , Cardiopatías Congénitas/cirugía , Humanos , Lactante , Virus de la Influenza A , Gripe Humana/terapia , Fallo Hepático Agudo/terapia , Masculino , Choque Séptico/terapia , Resultado del Tratamiento
3.
Acta Paediatr ; 110(3): 805-810, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33074577

RESUMEN

AIM: Since therapeutic hypothermia (TH) is known for its inhibitory effects on leucocyte migration and cytokine synthesis, our aim was to underline the necessity of early monitoring for potential immunomodulatory risks. METHODS: Using a 13-year retrospective case-control study at the paediatric intensive care unit (PICU) of the Medical University in Vienna, all newborn infants and children receiving TH were screened and compared with a diagnosis-matched control group undergoing conventional normothermic treatment (NT). TH was accomplished by using a non-invasive cooling device. Target temperature was 32-34°C. Children with evident infections, a medical history of an immunodeficiency or undergoing immunosuppressive therapy, were excluded. RESULTS: During the observational period, 108 patients were screened, 27 of which underwent TH. Culture-proven infections occurred in 22% of the TH group compared with 4% of the normothermic controls (P = .1). From the second day following PICU admission, median C-reactive protein (CRP) values were higher in the TH group (day two P = .002, day three P = .0002, day six P = .008). CONCLUSION: Children undergoing TH showed earlier and higher increases in CRP levels when compared to normothermic controls. These data underline the necessity of early and continuous monitoring for possible infectious complications.


Asunto(s)
Hipotermia Inducida , Estudios de Casos y Controles , Niño , Frío , Humanos , Hipotermia Inducida/efectos adversos , Lactante , Recién Nacido , Unidades de Cuidado Intensivo Pediátrico , Estudios Retrospectivos
4.
Wien Med Wochenschr ; 171(1-2): 29-35, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33108806

RESUMEN

BACKGROUND: Sepsis is, worldwide, one of the leading causes of death among infants and children. Over the past two decades, mortality rates have declined due to advanced treatment options; however, the incidence of sepsis and septic shock is still on the rise in many hospital settings. The objective of this study was to evaluate the course of this disease in pediatric intensive care patients. METHODS: An evaluation of pediatric patients in the intensive care unit diagnosed with infections or sepsis between 2005 and 2015 was performed via a retrospective exploratory data analysis. RESULTS: During the observational period, 201 patients were diagnosed with infection or sepsis. The study population was divided into five age subgroups. The majority of patients were newborns, infants, and toddlers. Forty percent had sepsis; 6% had septic shock. Viral infection was the most prevalent (59%). The overall survival rate was 83%; newborns and adolescents had the lowest survival rates. CONCLUSION: With this registry, children divided into five age subgroups with infection or sepsis were evaluated and treatment strategies were examined. We have shown that our findings on children treated in our pediatric intensive care unit conform with current literature about pediatric sepsis. In addition to maintaining strict hygiene standards, optimal aspects of sepsis care should be stringently observed, such as the quick administration of empirical broad-spectrum antibiotics, initial adequate fluid resuscitation, and a reliable and frequent routine of source control.


Asunto(s)
Sepsis , Choque Séptico , Adolescente , Niño , Preescolar , Humanos , Lactante , Recién Nacido , Unidades de Cuidados Intensivos , Sistema de Registros , Estudios Retrospectivos
6.
J Leukoc Biol ; 106(5): 1177-1185, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31211458

RESUMEN

Plasmacytoid dendritic cells (pDCs) are key players in the antiviral immune response and type III IFNs such as IL-29 appear to play a pivotal role in pDC function. Pronounced susceptibility to viral infections in neonates is partly resulting from diminished antiviral immune mechanisms. Accordingly, the aim of the present study was to investigate the impact of IL-29 in the altered immune response of neonatal pDCs. PBMCs of adult and term newborns were stimulated with CpG-ODN2216 in the presence or absence of IL-29 and assessed for IFN-α production, downstream-signaling, and activation marker expression. A significantly lower IL-29 production after TLR9-specific stimulation was demonstrated in neonatal pDCs. IL-29 enhanced the IFN-α production of pDCs in adults compared to newborns. Newborn pDCs displayed a significantly lower surface expression of IL-10 and IL-28Rα receptor resulting in diminished STAT1 and IRF7 activation. Interestingly, concomitant stimulation with CpG-ODN2216/IL-29 had no impact on the expression of surface activation and maturation markers of pDCs in neither population. The diminished antiviral immune response of neonatal pDCs is associated with reduced production and cellular responses toward IL-29. Potential therapeutic agents enhancing the IL-29 response in neonatal pDCs possibly augment viral protection in newborns.


Asunto(s)
Células Dendríticas/inmunología , Interferón-alfa/inmunología , Interferones/inmunología , Interleucinas/inmunología , Oligodesoxirribonucleótidos/farmacología , Adolescente , Adulto , Células Dendríticas/citología , Femenino , Humanos , Recién Nacido , Factor 7 Regulador del Interferón/inmunología , Interleucina-10/inmunología , Masculino , Receptores de Interferón/inmunología , Factor de Transcripción STAT1/inmunología , Receptor Toll-Like 9/agonistas , Receptor Toll-Like 9/inmunología
7.
Pediatr Res ; 83(1-1): 128-134, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-29278644

RESUMEN

BackgroundEndothelial cells (ECs) exert immunological functions such as production of proinflammatory cytokines/chemokines as well as facilitation of extravasation of immune cells into infected tissue. Limited data are available on the functionality of ECs from extremely preterm neonates during infection. Accordingly, the aim of our study was to investigate the immune response of premature ECs after proinflammatory stimulation.MethodsCell adhesion receptors' expression and function, nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NFκB) signaling, and chemokine production were analyzed in umbilical cord ECs from extremely preterm and term neonates after proinflammatory stimulation.ResultsP-selectin and E-selectin surface expression as well as NFκB signaling were lower after lipopolysaccharide (LPS) stimulation in premature ECs. Preterm ECs exhibited lower, but significant, cell-adhesive functions after LPS stimulation compared with term ECs. CCL2/CXCL8 chemokine secretion was significantly upregulated after proinflammatory stimulation in both groups. CXCL10 production was significantly increased in term but not in preterm ECs upon stimulation with tumor necrosis factor compared with unstimulated ECs.ConclusionExtremely premature ECs showed partly reduced expression levels and function of cell adhesion molecules. Both NFκB signaling and chemokine/cytokine production were reduced in premature ECs. The diminished endothelial proinflammatory immune response might result in impaired infection control of preterm newborns rendering them prone to severe infection.


Asunto(s)
Células Endoteliales/citología , Sistema Inmunológico/fisiología , FN-kappa B/metabolismo , Transducción de Señal , Adhesión Celular , Quimiocinas/inmunología , Citocinas/inmunología , Selectina E/metabolismo , Células Endoteliales/inmunología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Recien Nacido Extremadamente Prematuro , Recién Nacido , Inflamación , Lipopolisacáridos , Selectina-P/metabolismo , Fosforilación , Cordón Umbilical/metabolismo
8.
Pediatr Res ; 82(2): 215-225, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28288151

RESUMEN

BackgroundPentoxifylline (PTX), a methylxanthine derivate with immunomodulating properties, has been used as adjunctive treatment in severe neonatal sepsis. The aim of the study was to investigate the anti-inflammatory effects of PTX on Lipopolysaccharides (LPS)-stimulated monocytes of preterm neonates in vitro compared with monocytes of term infants and adult controls.MethodsWhole cord blood samples and control adult blood samples were incubated with LPS and PTX. The expression of surface markers, phagocytosis, cytokine secretion, and Toll-like receptor (TLR)4 signaling of monocytes were assessed by flow cytometry. Changes of TLR4-messenger RNA (mRNA) levels were confirmed by reverse-transcriptase PCR.ResultsThe expression of CD14, CD11b, CD64, CD71, and CD80 was downregulated by PTX in a dose-dependent manner; the greatest effect was observed on CD14 and CD11b in preterm infants. PTX markedly downregulated LPS-induced tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 levels in all age groups. Early IL-10 production was significantly downregulated by PTX in term and preterm neonates, while remaining unchanged in adults. Moreover, PTX downregulated TLR4 expression of monocytes on cellular and mRNA level, decreased signaling, and suppressed phagocytosis.ConclusionPTX downregulated TLR4 expression and signaling, thereby leading to strong anti-inflammatory properties in monocytes. Age-dependent differences were identified for CD14 and CD11b expression and IL-10 production.


Asunto(s)
Recien Nacido Prematuro , Inflamación/prevención & control , Lipopolisacáridos/efectos adversos , Monocitos/metabolismo , Pentoxifilina/farmacología , Adulto , Antígenos CD/metabolismo , Relación Dosis-Respuesta a Droga , Humanos , Técnicas In Vitro , Recién Nacido , Inflamación/inducido químicamente , Fagocitosis/efectos de los fármacos
9.
PLoS One ; 11(10): e0162667, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27695085

RESUMEN

Toll-like receptors (TLR) are crucial sensors of microbial agents such as bacterial or viral compounds. These receptors constitute key players in the induction of inflammation, e.g. in septic or chronic inflammatory diseases. Colony-stimulating factors (CSFs) such as granulocyte-macrophage-CSF (GM-CSF) or granulocyte-CSF (G-CSF) have been extensively investigated in their capacity to promote myelopoiesis in febrile neutropenia or to overcome immunosuppression in patients suffering from sepsis-associated neutropenia or from monocytic immunoincompetence. We report here that GM-CSF, downregulates TLR1, TLR2 and TLR4 in a time- and dose-dependent fashion in human monocytes. Diminished pathogen recognition receptor expression was accompanied by reduced downstream p38 and extracellular-signal-regulated kinase (ERK) signaling upon lipoteichoic acid (LTA) and lipopolysaccharide (LPS) binding-and accordingly led to impaired proinflammatory cytokine production. Knockdown experiments of the transcription factors PU.1 and VentX showed that GM-CSF driven effects on TLR regulation is entirely PU.1 but not VentX dependent. We further analysed monocyte TLR and CD14 expression upon exposure to the IMID® immunomodulatory drug Pomalidomide (CC-4047), a Thalidomide analogue known to downregulate PU.1. Indeed, Pomalidomide in part reversed the GM-CSF-mediated effects. Our data indicate a critical role of PU.1 in the regulation of TLR1, 2, 4 and of CD14, thus targeting PU.1 ultimately results in TLR modulation. The PU.1 mediated immunomodulatory properties of GM-CSF should be taken into consideration upon usage of GM-CSF in inflammatory or infection-related conditions.


Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/fisiología , Monocitos/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Receptores Toll-Like/biosíntesis , Transactivadores/metabolismo , Citocinas/biosíntesis , Regulación hacia Abajo , Citometría de Flujo , Humanos , Receptores de Lipopolisacáridos/biosíntesis , Monocitos/fisiología , Proteínas Proto-Oncogénicas/fisiología , Interferencia de ARN , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptor Toll-Like 1/biosíntesis , Receptor Toll-Like 2/biosíntesis , Receptor Toll-Like 4/biosíntesis , Transactivadores/fisiología
10.
Pediatr Res ; 80(2): 311-8, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-27057737

RESUMEN

BACKGROUND: Preterm neonates display an impaired vaccine response. Neonatal antigen-presenting cells (APCs) are less effective to induce an adaptive immune response and to promote the development of immunological memory. Efficient adjuvantal toll-like receptor (TLR)-triggering may overcome the neonatal immunological impairment. Accordingly, the aim of this study was to investigate the immunostimulatory action of R-848 and CpG-B on neonatal APCs. METHODS: Surface marker and cytokine secretion of APCs were evaluated after incubation of cord blood and peripheral blood mononuclear cells with the indicated adjuvants and were analyzed using flow cytometry. RESULTS: TLR-specific stimulation resulted in a significant induction of costimulatory molecules on neonatal APCs. Stimulation with R-848 resulted in significant higher secretion of TNFα, IL-6, IL-10, IL-12/IL-23p40, IL-12p70, and IFN-γ. Interestingly, CpG-B resulted in significant higher secretion of TNFα and IL-6. CONCLUSION: In summary, the incubation of TLR-agonists induced activation and maturation of neonatal APCs. These data show that modern TLR-specific adjuvants achieve a direct effect and potent upregulation of activation and maturation markers and cytokines in preterm neonates. We thus conclude that agents triggering TLRs might possibly overcome neonatal lack of vaccine responses.


Asunto(s)
Células Presentadoras de Antígenos/inmunología , Imidazoles/farmacología , Oligodesoxirribonucleótidos/farmacología , Receptores Toll-Like/metabolismo , Adyuvantes Inmunológicos/química , Células Presentadoras de Antígenos/citología , Citocinas/metabolismo , Femenino , Sangre Fetal/citología , Sangre Fetal/efectos de los fármacos , Citometría de Flujo , Voluntarios Sanos , Humanos , Sistema Inmunológico , Recién Nacido , Recien Nacido Prematuro , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Masculino , Regulación hacia Arriba
11.
J Leukoc Biol ; 100(3): 607-12, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-26965638

RESUMEN

Premature infants are highly susceptible to severe bacterial infections. The impaired infection control related to the functional immaturity of the neonatal innate immune system is an important cause of infection. Different monocyte subpopulations have been described and functionally characterized. However, data from preterm infants are scarce. We analyzed constitutive monocyte TLR2, TLR4, CD163, and HLA-DR expression in preterm cord blood. We further investigated activation of the signaling proteins ERK1/2 and NF-κB in monocyte subpopulations after ex vivo stimulation with the bacterial TLR agonists LPS and lipoteichoic acid. The functional outcome of the stimulation was determined by the intracellular production of TNF. Furthermore, the phagocytic activity was measured via flow cytometry. TLR4 and HLA-DR showed a gestational age-dependent increase. However, activation of ERK1/2 and NF-κB was impaired in neonatal monocyte subpopulations after stimulation with TLR agonists. Accordingly, intracellular TNF was diminished in preterm monocytes, especially in nonclassic monocytes. Premature monocytes showed high phagocytic activity, with significantly lower acidification of the phagosome. The reduced functional response of nonclassic monocytes of preterm neonates appears to be part of the diminished early immune response to bacterial cell wall components and is likely to contribute to their susceptibility to bacterial infection.


Asunto(s)
Sangre Fetal/inmunología , Recien Nacido Prematuro/inmunología , Monocitos/inmunología , Transducción de Señal/efectos de los fármacos , Factor de Necrosis Tumoral alfa/farmacología , Adolescente , Adulto , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Estudios de Casos y Controles , Femenino , Sangre Fetal/efectos de los fármacos , Sangre Fetal/metabolismo , Edad Gestacional , Antígenos HLA-DR/metabolismo , Humanos , Lactante , Recién Nacido , Recien Nacido Prematuro/metabolismo , Masculino , Monocitos/citología , Monocitos/metabolismo , Receptores de Superficie Celular/metabolismo , Receptor Toll-Like 3/metabolismo , Receptor Toll-Like 4/metabolismo , Adulto Joven
12.
J Med Genet ; 51(4): 254-63, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24431329

RESUMEN

BACKGROUND: Subtelomeric deletions and duplications may cause syndromic disorders that include features of immunodeficiency. To date, no phenotype of immunological pathology has been linked to partial trisomy 19. We report here on two unrelated male patients showing clinical and laboratory signs of immunodeficiency exhibiting a duplication involving Chromosome 19p13. METHODS: Both patients underwent a detailed clinical examination. Extended laboratory investigations for immune function, FISH and array comparative genome hybridization (CGH) analyses were performed. RESULTS: The reported patients were born prematurely with intrauterine growth retardation and share clinical features including neurological impairment, facial dysmorphy and urogenital malformations. Array CGH analyses of both patients showed a largely overlapping terminal duplication affecting Chromosome 19p13. In both affected individuals, the clinical course was marked by recurrent severe infections. Signs of humoral immunodeficiency were detected, including selective antibody deficiency against polysaccharide antigens in patient 1 and reduced IgG1, IgG3 subclass levels and IgM deficiency in patient 2. Class-switched B memory cells were almost absent in both patients. Normal numbers of T cells, B cells and natural killer cells were observed in both boys. Lymphocytic proliferation showed no consistent functional pathology, however, function of granulocytes and monocytes as assessed by oxidative burst test was moderately reduced. Moreover, natural killer cytotoxicity was reduced in both patients. Immunoglobulin substitution resulted in a decreased number and severity of infections and improved thriving in both patients. CONCLUSIONS: Partial trisomy 19p13 represents a syndromic disorder associating organ malformation and hitherto unrecognised immunodeficiency.


Asunto(s)
Cromosomas Humanos Par 19/genética , Síndromes de Inmunodeficiencia/genética , Trisomía/genética , Autoanticuerpos/inmunología , Niño , Preescolar , Femenino , Humanos , Inmunidad Celular/genética , Inmunidad Humoral/genética , Síndromes de Inmunodeficiencia/inmunología , Lactante , Recién Nacido , Masculino , Fenotipo , Embarazo
13.
Neonatology ; 99(1): 51-6, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-20616571

RESUMEN

BACKGROUND: Concise regulation of the Toll signaling pathway is mandatory in neonatal innate immunity. The microRNA-146 family (miR-146a/b) was recently reported to be a regulator of Toll-like receptor 4 (TLR4) through a negative feedback loop mechanism. Acting as a potent regulator, miRNA helps to protect the organism from developing overwhelming proinflammatory immune responses leading to septic shock or chronic inflammatory diseases. OBJECTIVE: We investigated for the first time whether miRNA-146a/b plays a regulatory role in human monocytes derived from infant cord or adult blood, and whether differences in miRNA-146 expression exist. METHODS: Expression profiles of miR-146a/b and TLR4 were studied by real-time PCR upon stimulation with lipopolysaccharide. RESULTS: Both members of the miRNA-146 family showed a time-dependent upregulation. For miR-146a, a statistically higher significant increase was found after 24 h of stimulation in monocytes from cord blood compared to those derived from adults. In contrast, no differences were found for miR-146b and TLR4, respectively. CONCLUSION: We conclude that differences between the negative regulatory role for miR-146a obviously exist in neonatal and adult TLR4 signaling, and suggest that more intense research in the involvement of miRNA in immune regulation will facilitate the understanding of the development and function of the innate immune system of neonates.


Asunto(s)
Inmunidad Innata/fisiología , MicroARNs/fisiología , Monocitos/inmunología , Adulto , Células Cultivadas , Sangre Fetal/citología , Perfilación de la Expresión Génica , Humanos , Recién Nacido , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Lipopolisacáridos/farmacología , Monocitos/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/inmunología
14.
Vaccine ; 28(40): 6621-6, 2010 Sep 14.
Artículo en Inglés | MEDLINE | ID: mdl-20656033

RESUMEN

To assess B-cell function in patients under immunoglobulin (IgG)-replacement therapy, the non-licensed artificial bacteriophage (ΦX174)-neo-antigen may be used despite limited availability and experience. Active immunization against tick-borne encephalitis (TBE) is performed in few European countries. To test the feasibility of using licensed TBE vaccination as (neo-)antigen to determine residual or restored B-cell function in patients under regular IgG substitution, TBE-IgG levels were analyzed in 18 patients with ≥ 1-2 years of regular intravenous or subcutaneous IgG substitution and in pharmaceutical IgG-preparations (n=21 batches, 10 products). Six individuals were boosted against TBE. Although TBE-specific IgG was detectable in concentrates (281-57,100 VieU/0.5 µL), levels were only borderline in patient sera (n=31, 18 individuals; median 132 VieU; positive >155). Thus, TBE vaccination may be used to test B-cell function under IgG replacement therapy because IgG substitution appears insufficient to yield protective TBE-specific antibody levels in children.


Asunto(s)
Anticuerpos Antivirales/biosíntesis , Encefalitis Transmitida por Garrapatas/prevención & control , Inmunización Pasiva , Inmunoglobulina G/biosíntesis , Síndromes de Inmunodeficiencia/inmunología , Vacunas Virales/inmunología , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Linfocitos B/inmunología , Niño , Preescolar , Virus de la Encefalitis Transmitidos por Garrapatas/inmunología , Encefalitis Transmitida por Garrapatas/inmunología , Estudios de Factibilidad , Femenino , Humanos , Inmunoglobulina G/sangre , Masculino , Adulto Joven
15.
Int J Mol Med ; 23(2): 253-9, 2009 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19148550

RESUMEN

The purpose of this study was to investigate the expression of different CD44 and hyaluronan synthase isoforms in cartilage, their alterations during the chondrocyte dedifferentiation process in monolayer culture and during the redifferentiation process on 3D scaffolds. Chondrocytes isolated from human articular cartilage were cultured as a monolayer for up to 36 days and were seeded on two different 3D scaffolds (HYAFF 11 and Bio-Gide). Expression levels of CD44s, CD44-lt, CD44-st, HAS1, HAS2, HAS3 and UDPGD were determined by real-time RT-PCR at different time points. At the protein level CD44 and CD90 were analyzed by flow cytometry. HAS2 was found to be the predominantly expressed hyaluronan synthase in chondrocytes and was not subjected to any regulation during the dedifferentiation process. CD44s, CD44-lt, CD44-st and UDPGD, however, were upregulated immediately after cell isolation. In addition, a high cell density was found to significantly increase CD44-st and CD44-lt expression. Redifferentiation on 3D scaffolds reversed the increase of the CD44 expression. Our data point out that CD44 expression does not correlate with matrix assembly in chondrocytes and that CD44 has a regulatory function in chondrocytes, not necessarily on differentiation, but probably on proliferation.


Asunto(s)
Cartílago Articular/citología , Condrocitos/citología , Glucuronosiltransferasa/genética , Receptores de Hialuranos/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Cartílago Articular/metabolismo , Recuento de Células , Diferenciación Celular , Condrocitos/metabolismo , Expresión Génica , Glucuronosiltransferasa/metabolismo , Humanos , Receptores de Hialuranos/metabolismo , Hialuronano Sintasas , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , ARN Mensajero/metabolismo
16.
Diagn Microbiol Infect Dis ; 63(1): 10-5, 2009 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-18990529

RESUMEN

Infection with Toxoplasma gondii during pregnancy is often asymptomatic and may cause severe fetal damage. A quantitative TaqMan minor groove binder real-time polymerase chain reaction (PCR) assay was developed for the specific and sensitive detection of the previously described 529-bp repeat element occurring up to 200 to 300 times in T. gondii genome. The qualitative and quantitative detection limits determined were 6 and 20 marker copies (1/30 to 1/50 of 1 parasite) per PCR, respectively. In addition to standard PCR cycling conditions, 3 different fast PCR protocols were evaluated to minimize run time. A higher variability but no loss of specificity was observed. For the evaluation of clinical applicability, a total of 135 amniotic fluid samples were analyzed targeting both 529-bp and B1 gene. The sensitivity and specificity were 88.0% and 100.0% for B1, and 100.0% and 98.2% for 529-bp PCR assay (positive predictive value and negative predictive value: 100.0% and 97.4%, and 92.6% and 100.0%, respectively). Our results demonstrated an increased sensitivity of the 529-bp PCR assay even in a faster protocol.


Asunto(s)
Líquido Amniótico/microbiología , Reacción en Cadena de la Polimerasa/métodos , Toxoplasma/aislamiento & purificación , Toxoplasmosis Congénita/microbiología , Análisis de Varianza , Animales , Líquido Ascítico/microbiología , ADN Protozoario/genética , Femenino , Genes Protozoarios/genética , Humanos , Ratones , Lavado Peritoneal/veterinaria , Valor Predictivo de las Pruebas , Embarazo , Secuencias Repetitivas de Ácidos Nucleicos/genética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Toxoplasma/genética
17.
J Infect Dis ; 195(2): 296-302, 2007 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-17191175

RESUMEN

The impaired infection control related to the functional immaturity of the neonatal immune system is an important cause of infection in preterm newborns. We previously reported that constitutive Toll-like receptor (TLR) 4 expression and cytokine secretion on lipopolysaccharide (LPS) stimulation increases with gestational age. Here, we analyzed constitutive monocyte TLR2 expression and evaluated the expression profiles of the proximal downstream adapter molecule myeloid differentiation factor 88 (MyD88). We further investigated activation of protein kinases p38 and extracellular regulated kinsase (ERK) 1/2 in CD14 monocytes after ex vivo stimulation with bacterial TLR ligands (LPS and lipoteichoic acid [LTA]). The functional outcome of the stimulation was determined by cytokine secretion. Monocytes from 31 preterm newborns (<30 weeks of gestation, n=16; 30-37 weeks of gestation, n=15), 10 term newborns, and 12 adults were investigated. In contrast to TLR4 expression, TLR2 levels did not differ between age groups. However, MyD88 levels were significantly lower in preterm newborns. Activation of p38 and ERK1/2 was impaired in all newborn age groups after stimulation with TLR-specific ligands. Accordingly, after LTA stimulation, the levels of interleukin (IL)-1 beta , IL-6, and IL-8 cytokine production were substantially lower (P<.001) in preterm newborns than in adults. The reduced functional response to bacterial cell wall components appears to be part of the functional immaturity of the neonatal immune system and might predispose premature newborns to bacterial infection.


Asunto(s)
Envejecimiento/inmunología , Inmunidad Innata/fisiología , Recién Nacido/inmunología , Recien Nacido Prematuro/inmunología , Monocitos/inmunología , Transducción de Señal/inmunología , Receptores Toll-Like/metabolismo , Adulto , Femenino , Sangre Fetal/citología , Sangre Fetal/inmunología , Regulación del Desarrollo de la Expresión Génica , Edad Gestacional , Humanos , Inmunidad Innata/inmunología , Factor 88 de Diferenciación Mieloide/metabolismo , Embarazo , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/metabolismo
18.
Eur J Immunol ; 36(2): 361-70, 2006 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-16402404

RESUMEN

Toll-like receptors (TLR) represent an ancient front-line defence system that enables the host organism to sense the presence of microbial components within minutes. As inducers of inflammation, TLR act as important triggers of distinct entities such as sepsis or autoimmune disease exacerbation. We report here that vitamin D3 [1alpha,25-dihydroxycholecalciferol, 1,25(OH)(2)D3] suppresses the expression of TLR2 and TLR4 protein and mRNA in human monocytes in a time- and dose-dependent fashion. Despite 1,25(OH)(2)D3-induced up-regulation of CD14, challenge of human monocytes with either LPS or lipoteichoic acid resulted in impaired TNF-alpha and procoagulatory tissue factor (CD142) production, emphasizing the critical role of TLR in the induction of inflammation. Moreover, reduced TLR levels in 1,25(OH)(2)D3-treated phagocytes were accompanied by impaired NF-kappaB/RelA translocation to the nucleus and by reduced p38 and p42/44 (extracellular signal-regulated kinase 1/2) phosphorylation upon TLR-ligand engagement. Both TLR down-regulation and CD14 up-regulation were substantially inhibited by the vitamin D receptor (VDR) antagonist ZK 159222, indicating that the immunomodulatory effect of 1,25(OH)(2)D3 on innate immunity receptors requires VDR transcription factor activation. Our data provide strong evidence that 1,25(OH)(2)D3 primes monocytes to respond less effectively to bacterial cell wall components in a VDR-dependent mechanism, most likely due to decreased levels of TLR2 and TLR4.


Asunto(s)
Colecalciferol/farmacología , Regulación hacia Abajo/efectos de los fármacos , Activación de Macrófagos/efectos de los fármacos , Monocitos/inmunología , Receptor Toll-Like 2/biosíntesis , Receptor Toll-Like 4/biosíntesis , Transporte Activo de Núcleo Celular/efectos de los fármacos , Transporte Activo de Núcleo Celular/inmunología , Calcitriol/análogos & derivados , Calcitriol/farmacología , Células Cultivadas , Colecalciferol/inmunología , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/inmunología , Humanos , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Receptores de Lipopolisacáridos/biosíntesis , Receptores de Lipopolisacáridos/inmunología , Lipopolisacáridos/inmunología , Lipopolisacáridos/farmacología , Activación de Macrófagos/inmunología , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Monocitos/metabolismo , Monocitos/patología , Fosforilación/efectos de los fármacos , Receptores de Calcitriol/antagonistas & inhibidores , Receptores de Calcitriol/inmunología , Receptores de Calcitriol/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunología , Ácidos Teicoicos/inmunología , Ácidos Teicoicos/farmacología , Receptor Toll-Like 2/inmunología , Receptor Toll-Like 4/inmunología , Factor de Transcripción ReIA/inmunología , Factor de Transcripción ReIA/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/inmunología , Proteínas Quinasas p38 Activadas por Mitógenos/inmunología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
19.
Pediatr Res ; 58(1): 121-4, 2005 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-15879290

RESUMEN

Premature newborns are highly susceptible to severe bacterial infections. This is partially due to their immature innate immune system, characterized by decreased neutrophil and monocyte activity as well as by reduced concentrations of complement factors. However, additional mechanisms might be important for innate immunity and are still the subject of considerable debate. The importance of pattern recognition domains such as Toll-like receptors (TLR) has been fully acknowledged within the last few years. Therefore, we investigated age-related monocyte TLR4 expression and lipopolysaccharide-induced cytokine secretion from very low birth weight infants (VLBWI) and from newborns after wk 30 of gestation in comparison to healthy adults. In VLBWI, expression of TLR4 surface protein, detected by flow cytometry, and TLR4-specific mRNA, quantified by real time-PCR, were significantly reduced in comparison to mature infants and to adults. Reduced TLR4 expression was paralleled by significantly diminished ex vivo LPS stimulated IL-1beta, IL-6, and tumor necrosis factor-alpha secretion into whole blood. We conclude that, in VLBWI, the minimized expression of TLR4 contributes to the susceptibility of VLBWI to infections with Gram-negative bacteria due to the lack of cytokines to boost initial immune response.


Asunto(s)
Sangre Fetal/metabolismo , Regulación del Desarrollo de la Expresión Génica , Lipopolisacáridos/metabolismo , Glicoproteínas de Membrana/biosíntesis , Monocitos/metabolismo , Receptores de Superficie Celular/biosíntesis , Citocinas/metabolismo , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Edad Gestacional , Humanos , Recién Nacido , Recién Nacido de muy Bajo Peso , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Receptores de Lipopolisacáridos/biosíntesis , Microscopía Fluorescente , Neutrófilos/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Receptor Toll-Like 4 , Receptores Toll-Like , Factor de Necrosis Tumoral alfa/metabolismo
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