RESUMO
OBJECTIVE: To determine whether current retinopathy of prematurity (ROP) screening guidelines adequately identify treatable ROP in a contemporary cohort of extremely low gestation infants. STUDY DESIGN: Data from the Surfactant, Positive Pressure, and Pulse Oximetry Randomized Trial were used. Inborn infants of 24 (0)/7 to 27 (6)/7 weeks gestational age (GA) with consent before delivery were enrolled in 2005 to 2009. Severe ROP (type 1 ROP or treatment with laser, cryotherapy or bevacizumab) or death was the primary outcome for the randomized trial. Examinations followed the then current AAP (American Academy of Pediatrics) screening recommendations, beginning by 31 to 33 weeks postmenstrual age (PMA). RESULT: One thousand three hundred and sixteen infants were enrolled in the trial. Nine hundred and ninety-seven of the 1121 who survived to first eye exam had final ROP outcome determined. One hundred and thirty-seven (14% of 997) met criteria for severe ROP and 128 (93%) of those had sufficient data (without missing or delayed exams) to determine age of onset of severe ROP. PMA at onset was 32.1 to 53.1 weeks. In this referral center cohort, 1.4% (14/997) developed severe ROP after discharge. CONCLUSION: Our contemporary data support the 2013 AAP screening guidelines for ROP for infants of 24 (0)/7 to 27 (6)/7 weeks GA. Some infants do not meet treatment criteria until after discharge home. Post-discharge follow-up of infants who are still at risk for severe ROP is crucial for timely detection and treatment.
Assuntos
Guias de Prática Clínica como Assunto , Retinopatia da Prematuridade/diagnóstico , Feminino , Humanos , Recém-Nascido Prematuro , MasculinoRESUMO
OBJECTIVE: To describe the findings on fetal and postnatal magnetic resonance imaging (MRI) in premature infants with twin-twin transfusion syndrome (TTTS) and to determine whether currently used staging systems and other fetal and postnatal factors correlate with brain injury in this population. STUDY DESIGN: We performed a prospective study of 22 premature infants with TTTS whose mothers had fetal MRIs. Postnatal brain MRI was performed at term equivalent age (38 to 44 weeks) and medical records were reviewed. Brain injury was scored on fetal and postnatal MRIs using an injury scale incorporating hemorrhagic and nonhemorrhagic injury. RESULT: The median (range) gestational age (GA) was 31 weeks (26 to 35) and birth weight (BW) was 1296 g (762 to 2330). In all, 5/22 patients (23%) had brain injury seen on fetal MRI and 15/22 patients (68%) had brain injury seen on postnatal MRI. Quintero stage was the only predictor variable that was significantly correlated with the total brain injury score (P=0.05). CONCLUSION: Postnatal brain injury in premature infants with TTTS is correlated with Quintero stage. GA and BW are not predictive of brain injury in this cohort of infants.
Assuntos
Lesões Encefálicas/diagnóstico , Transfusão Feto-Fetal/diagnóstico , Recém-Nascido Prematuro , Imageamento por Ressonância Magnética/métodos , Diagnóstico Pré-Natal/métodos , Lesões Encefálicas/etiologia , Estudos de Coortes , Intervalos de Confiança , Feminino , Transfusão Feto-Fetal/complicações , Transfusão Feto-Fetal/mortalidade , Seguimentos , Idade Gestacional , Humanos , Recém-Nascido , Doenças do Prematuro/diagnóstico , Masculino , Idade Materna , Cuidado Pós-Natal/métodos , Valor Preditivo dos Testes , Gravidez , Resultado da Gravidez , Cuidado Pré-Natal/métodos , Estudos Prospectivos , Estatísticas não Paramétricas , Taxa de SobrevidaRESUMO
OBJECTIVE: To determine acceptance of donor human milk (DM) for feeding preterm infants and whether offering DM, alters mothers' milk (MM) feeding. STUDY DESIGN: Infant feeding data were collected from medical records of 650 very preterm infants enrolled between 2006-2011 in two hospital level III neonatal intensive care units (NICUs) in Cincinnati, Ohio. The study was conducted during the implementation of a program offering 14 days of DM. RESULT: From 2006-2011, any DM use increased from 8 to 77% of infants, largely replacing formula for the first 2 weeks of life; provision of MM did not change. DM was more likely to be given in the first 2 weeks of life, if infants never received MM or were >1000 g birth weight, but DM use did not differ by sociodemographic factors. CONCLUSION: Offering DM dramatically increased human milk feeding and decreased formula use, but did not alter MM feeding in hospital.
Assuntos
Aleitamento Materno , Hospitais Urbanos , Fórmulas Infantis/administração & dosagem , Doenças do Prematuro/terapia , Bancos de Leite Humano , Leite Humano , Peso ao Nascer , Aleitamento Materno/psicologia , Aleitamento Materno/estatística & dados numéricos , Extração de Leite/psicologia , Extração de Leite/estatística & dados numéricos , Feminino , Idade Gestacional , Hospitais Universitários , Humanos , Lactente , Recém-Nascido , Masculino , Bancos de Leite Humano/estatística & dados numéricos , Mães/psicologia , Motivação , Ohio , Melhoria de QualidadeRESUMO
OBJECTIVE: We sought to determine the incidence of necrotizing enterocolitis (NEC) and spontaneous intestinal perforation (SIP) in surviving extremely low-birth-weight (ELBW, <1000 g birth weight) infants and to establish the impact of NEC on outcomes by hospital discharge and at 18 to 22 months adjusted age in a large, contemporary, population-based practice. STUDY DESIGN: Hospital outcome data for all ELBW infants born in the greater Cincinnati region from 1998 to 2009 were extracted from the National Institute of Child Health Neonatal Research Network Database. Neurodevelopmental outcome at 18 to 22 months was assessed using Bayley Scales of Infant Development-II scores for Mental Developmental Index and Psychomotor Developmental Index. Multivariable logistic regression was used and adjusted odds ratios reported to control for confounders. RESULT: From 1998 to 2009, ELBW infants accounted for 0.5% of the 352 176 live-born infants in greater Cincinnati. The incidence of NEC was 12%, with a 50% case-fatality rate. Death before discharge, morbid complications of prematurity and neurodevelopmental impairment were all increased among infants diagnosed with NEC. Infants with surgical NEC and SIP had a higher incidence of death, but long-term neurodevelopmental outcomes were not different comparing surviving ELBW infants with medical NEC, surgical NEC and SIP. CONCLUSION: Although ELBW infants comprise a very small proportion of live-born infants, those who develop NEC and SIP are at an increased risk for death, morbid complications of prematurity and neurodevelopmental impairment. No significant differences in neurodevelopmental outcomes were observed between the medical and surgical NEC and SIP groups.
Assuntos
Desenvolvimento Infantil , Deficiências do Desenvolvimento/etiologia , Enterocolite Necrosante/complicações , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Doenças do Prematuro , Perfuração Intestinal/complicações , Enterocolite Necrosante/mortalidade , Enterocolite Necrosante/cirurgia , Humanos , Lactente , Recém-Nascido de Peso Extremamente Baixo ao Nascer/crescimento & desenvolvimento , Recém-Nascido , Recém-Nascido Prematuro , Doenças do Prematuro/cirurgia , Perfuração Intestinal/mortalidade , Perfuração Intestinal/cirurgiaRESUMO
AIM: To determine whether an imbalance of dendritic cell subsets might contribute to diminished adaptive host responses observed in newborn infants. It was hypothesized that the proportion of lymphoid dendritic cells would be greater than that of myeloid dendritic cells in cord blood. METHODS: To investigate this, dendritic cell subsets were evaluated in whole cord blood by flow cytometry. Circulating dendritic cells were also isolated from cord blood based on CD1c and BDCA-2 expression. Myeloid dendritic cells were also obtained by culturing cord and adult blood monocytes. Surface phenotypes of these cells were determined by flow cytometry using monoclonal antibodies directed against lineage, major histocompatibility, adhesion, co-stimulation and cytokine receptor molecules. Antigen-presenting functions of dendritic cell subsets were determined by mixed leukocyte reactions. RESULTS: Circulating myeloid dendritic cells were higher in cord blood than previously reported in adult blood, whereas lymphoid dendritic cell numbers were similar between cord and adult blood. Expression of CD11c, CD45RA and CD45RO did not accurately differentiate between dendritic cell subsets circulating in cord blood. Fresh and culture-derived cord blood myeloid dendritic cells stimulated adult allogeneic leukocyte proliferation, while lymphoid dendritic cells were less effective inducers of an adult allogeneic leukocyte response. Culture-derived dendritic cells induced modest autologous cord blood leukocyte proliferation, but freshly isolated myeloid and lymphoid dendritic cells did not stimulated autologous leukocytes. CONCLUSION: Contrary to the hypothesis, an imbalance in the ratio of circulating myeloid to lymphoid dendritic cell subsets does not exist and, therefore, does not contribute to diminished adaptive immune responses in newborn infants.
Assuntos
Antígenos CD/imunologia , Células Dendríticas/imunologia , Antígenos HLA-DR/imunologia , Sistema Imunitário/embriologia , Imunidade Celular/fisiologia , Adaptação Fisiológica , Apresentação de Antígeno , Antígenos CD/análise , Medula Óssea/embriologia , Medula Óssea/fisiologia , Células Dendríticas/fisiologia , Biologia do Desenvolvimento , Feminino , Sangue Fetal/citologia , Citometria de Fluxo , Antígenos HLA-DR/análise , Humanos , Sistema Imunitário/fisiologia , Recém-Nascido , Ativação Linfocitária , Masculino , Sensibilidade e EspecificidadeRESUMO
Congenital bone marrow failure syndromes are associated with a number of congenital abnormalities affecting a wide range of organ systems. The underlying molecular abnormalities that cause these disorders affect normal embryonic development during the critical organogenesis phase (weeks 4 to 8). These syndromes predispose patients to leukemia and other malignancies, and these genetic disorders may represent the first hit of at least two hits necessary for malignant transformation. The molecular defects underlying these diseases are just beginning to be understood; mechanisms suggested by recent research include DNA repair (FA-A, FA-G); abnormalities of the ribosomes (DBA, DC); to disorders of electron transport (FA-C, Pearson's syndrome, Barth's syndrome). Understanding these molecular mechanisms provides the knowledge necessary to develop better therapy, possibly including gene therapy, offering for the first time the potential for curing the hematologic manifestations of these illnesses.
Assuntos
Doenças da Medula Óssea/congênito , Doenças da Medula Óssea/complicações , Deficiências do Desenvolvimento/etiologia , Leucemia/etiologia , Humanos , Lactente , SíndromeAssuntos
Doenças Hematológicas/diagnóstico , Doenças Hematológicas/terapia , Hematologia/métodos , Neonatologia/métodos , Transfusão de Eritrócitos , Eritropoetina/uso terapêutico , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Granulócitos/transplante , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Transfusão de Plaquetas , Medicina Preventiva/métodos , Proteínas Recombinantes/uso terapêuticoRESUMO
OBJECTIVE: To determine whether recombinant human granulocyte colony-stimulating factor (G-CSF) administration: 1) accelerates production of neutrophils; 2) increases bone marrow stored and precursor neutrophils; and 3) is safe in newborn infants with neutropenia and clinical signs of early-onset sepsis. STUDY DESIGN: We randomized 20 infants with neutropenia and clinical signs of early-onset sepsis in the first 3 days of life to receive G-CSF (10 microg/kg/d) or placebo for 3 days. Entry criteria included neutropenia as defined by Manroe criteria, an elevated immature to total neutrophil ratio [(I/T) >/=0.25], and a requirement for ventilatory support. Cultures were obtained and antibiotics initiated on all study infants. Circulating absolute neutrophil count (ANC), I/T ratio, bone marrow neutrophil storage pool (NSP) and neutrophil proliferative pool (NPP), and plasma G-CSF concentrations were evaluated. Also, severity of illness as determined using the Score for Neonatal Acute Physiology (SNAP), morbidity, and mortality were recorded. RESULTS: Circulating ANC increased in both G-CSF and placebo recipients by day 1. Also, the I/T neutrophil ratio decreased in both G-CSF and placebo recipients. There were no significant differences in the ANC or I/T ratio between the two groups during the study period. Similarly, bone marrow NSP and NPP did not differ between G-CSF and placebo recipients at study entry or day 2. No differences were observed in the secondary outcome measures including severity of illness, morbidity, and mortality. CONCLUSIONS: Administration of recombinant G-CSF to infants with neutropenia and clinical signs of early-onset sepsis did not increase circulating ANC, or bone marrow NSP and NPP compared with placebo. No differences were observed between G-CSF and placebo recipients in severity of illness, morbidity, or mortality. No adverse effects of G-CSF administrations were noted.
Assuntos
Bacteriemia/terapia , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Doenças do Prematuro/terapia , Neutropenia/terapia , Bacteriemia/imunologia , Bacteriemia/mortalidade , Medula Óssea/efeitos dos fármacos , Medula Óssea/imunologia , Método Duplo-Cego , Fator Estimulador de Colônias de Granulócitos/sangue , Humanos , Recém-Nascido , Doenças do Prematuro/imunologia , Doenças do Prematuro/mortalidade , Contagem de Leucócitos/efeitos dos fármacos , Neutropenia/imunologia , Neutropenia/mortalidade , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Proteínas Recombinantes , Taxa de SobrevidaRESUMO
BACKGROUND: Clinical trials of erythropoietin (EPO) administration to preterm infants have not focused on infants weighing 750 gm or less, the population most likely to receive multiple transfusions because of large phlebotomy losses. It is unknown whether preterm infants weighing 750 gm or less will respond to EPO by accelerating erythropoiesis, or whether EPO administered to this population will decrease blood transfusions. METHODS: We randomly assigned 28 extremely low birth weight preterm infants (mean +/- SEM: 24.7 +/- 0.3 weeks' gestation, 662 +/- 14 gm birth weight), in the first 72 hours of life, to receive either EPO (200 U/kg/day) or placebo for 14 days and administered transfusions only according to protocol over a 21-day study period. All infants received 1 mg/kg/day iron dextran in their total parenteral nutrition solution during the 14-day treatment period. RESULTS: During the 21-day study period, a lower number and volume of transfusions were received by the EPO recipients (4.7 +/- 0.7 transfusions per patient and 70 +/- 11 ml/kg per patient) than by the placebo recipients (7.5 +/- 1.1 transfusions per patient and 112 +/- 17 ml/kg per patient; p < 0.05, EPO vs placebo), whereas hematocrits remained similar in the two groups. Reticulocyte counts were similar in both groups on day 1 but were greater in the EPO recipients on day 14 (EPO day 1, 351 +/- 53; EPO day 14, 359 +/- 40 x 10(3)/microl; placebo day 1, 334 +/- 64; placebo day 14, 120 +/- 10 x 10(3)/microl; p < 0.01, EPO vs placebo). Serum ferritin concentrations were similar in both groups at the beginning of the study but were greater in the placebo recipients by day 14 (EPO, 262 +/- 44 microg/L; placebo, 593 +/- 92 microg/L; p < 0.01). No adverse effects of EPO or iron were noted. CONCLUSION: The combination of EPO and parenteral iron stimulates erythropoiesis in preterm infants weighing 750 gm or less and results in fewer transfusions during their first 3 weeks of life.
Assuntos
Anemia Hemolítica/terapia , Transfusão de Sangue , Eritropoetina/uso terapêutico , Doenças do Prematuro/terapia , Recém-Nascido de muito Baixo Peso , Ferro/uso terapêutico , Anemia Hemolítica/sangue , Terapia Combinada , Método Duplo-Cego , Ferritinas/sangue , Idade Gestacional , Hematócrito/métodos , Humanos , Recém-Nascido , Doenças do Prematuro/sangue , Nutrição Parenteral Total , Proteínas Recombinantes , Contagem de ReticulócitosRESUMO
Interleukin-10 (IL-10) downmodulates phagocytic immune responses and accentuates humoral responses. Human neonates exhibit broad immune deficits that parallel actions of IL-10. We postulated that IL-10 production would be diminished in neonatal blood cells. We found that IL-10 production by lipopolysaccharide-stimulated peripheral blood mononuclear cells (PBMNCs) in vitro was greater by adult cells than by term cells and preterm cells. Additional studies were undertaken to identify mechanisms responsible for the developmental differences in IL-10 gene expression. IL-10 transcription was present in freshly isolated adult and neonatal cells in the absence of detectable levels of transcript. Transcription rates were not different between adult and neonatal cells. IL-10 transcripts were approximately 40% more abundant in adult cells than in term cells and were consistent with differences in secreted protein; however, no differences were noted in mRNA stability. IL-10 half-life was 60 minutes for both adult and term PBMNCs. We conclude that up-regulation of IL-10 gene expression in PBMNCs is modulated at the post-transcriptional level, that IL-10 protein production and mRNA content are greater in activated cells from adults compared with those from neonates, and that maturational differences in IL-10 expression are not due to differences in transcription rate or mRNA stability. Maturational differences in IL-10 expression might be due to differences in subpopulations of cytokine-producing cells or differences in nucleo-cytoplasmic transport.
Assuntos
Regulação da Expressão Gênica/imunologia , Interleucina-10/genética , Leucócitos Mononucleares/imunologia , Adulto , Células Cultivadas , Humanos , Interleucina-10/imunologia , RNA Mensageiro/análise , Transcrição GênicaRESUMO
Hematopoietic progenitors obtained from the bone marrow of healthy adults fail to undergo clonogenic maturation in vitro if a source of hematopoietic growth factors is not included in the culture dishes. In contrast, a fraction of similarly purified progenitors obtained from umbilical cord blood undergo clonogenic maturation even in the absence of added growth factors. We postulated that production of hematopoietic growth factors within the culture dishes containing the progenitors of umbilical cord blood origin might be responsible. We postulated further, that this production might be by non-progenitor cells co-plated along with the progenitors, or alternatively by CD34+ cells themselves, or by cells clonally derived from CD34+ cells. To test these possibilities we first assessed the effect of including in the cultures neutralizing antibody directed against various growth factors. Inclusion of anti-granulocyte macrophage colony-stimulating factor (GM-CSF) and anti-interleukin-3 (IL-3) (but not anti-IL-2) significantly reduced the growth factor independence of cord blood progenitors (P < .005 and P < .01). Inclusion of both anti-GM-CSF and anti-IL-3 almost completely ablated the spontaneous colony growth (P < .001). Inclusion of IL-10 also reduced, in a concentration-dependent fashion, the spontaneous generation of umbilical cord blood-derived colonies. Transcripts for GM-CSF and IL-3 were detected, by reverse transcriptase-polymerase chain reaction (RT-PCR), in the CD34+ cells from cord blood and from adult marrow. When plated without added growth factors, however, the CD34+ cells of adult marrow origin failed to produce colonies, whereas 6% of cord blood CD34+ cells similarly cultured did so. When these growth factor independent colonies were plucked from culture, transcripts for GM-CSF and IL-3 were identified in all. We conclude that production of GM-CSF and IL-3 occurs within culture dishes containing hematopoietic progenitors of umbilical cord origin, and that this explains some of their apparently unique features of in vitro growth.
Assuntos
Sangue Fetal/citologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/fisiologia , Células-Tronco Hematopoéticas/citologia , Interleucina-3/fisiologia , Adulto , Anticorpos/farmacologia , Antígenos CD , Antígenos CD34 , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Fator Estimulador de Colônias de Granulócitos e Macrófagos/antagonistas & inibidores , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Hematopoese/efeitos dos fármacos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Células-Tronco Hematopoéticas/metabolismo , Humanos , Recém-Nascido , Interleucina-10/farmacologia , Interleucina-3/antagonistas & inibidores , Interleucina-3/biossíntese , Interleucina-3/imunologia , Especificidade de Órgãos , RNA Mensageiro/biossínteseRESUMO
Studies were undertaken to delineate the actions of stem cell factor (SCF) on human fetal hematopoietic progenitors in vitro. Mononuclear cells from umbilical cord blood of term fetuses were "panned" immunologically, and the resulting hematopoietic progenitors were grown in methylcellulose culture containing various concentrations of SCF alone or in combination with other recombinant hematopoietic growth factors. Neutralizing antibodies to IL-3 and granulocyte-macrophage colony-stimulating factor were added to all plates to which recombinant IL-3 or granulocyte-macrophage colony-stimulating factor were not included to decrease any confounding effect resulting from production of small quantities of these factors within the culture plates. SCF, as a single agent, supported clonogenic maturation of fetal granulocyte-macrophage progenitors (granulocyte-macrophage colony-forming unit, p < 0.05), multipotent progenitors (CFU-MIX, p < 0.05), and erythroid progenitors (erythroid burst-forming unit, p < 0.05). When combined with subplateau concentrations (0.1 microgram/L) of IL-3 or granulocyte-macrophage colony-stimulating factor, SCF had an additive or synergistic effect on clonogenic maturation of granulocyte-macrophage colony-forming unit and CFU-MIX. When combined with higher concentrations (5.0 micrograms/L) of IL-3 or granulocyte-macrophage colony-stimulating factor, SCF generally did not enhance colony formation but did increase the number of cells per colony. Like other pleiotropic cytokines such as IL-6, IL-9, and IL-11, SCF had a broad spectrum of action of fetal hematopoietic progenitors.
Assuntos
Sangue Fetal/citologia , Fatores de Crescimento de Células Hematopoéticas/farmacologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Sinergismo Farmacológico , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/administração & dosagem , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Hematopoese/efeitos dos fármacos , Hematopoese/fisiologia , Fatores de Crescimento de Células Hematopoéticas/administração & dosagem , Fatores de Crescimento de Células Hematopoéticas/fisiologia , Células-Tronco Hematopoéticas/citologia , Humanos , Técnicas In Vitro , Interleucina-3/administração & dosagem , Interleucina-3/farmacologia , Gravidez , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Fator de Células-TroncoRESUMO
We postulated that defective generation of granulocyte colony-stimulating factor (G-CSF) by cells of newborn infants might underlie their deficiencies in upregulating neutrophil production and function during bacterial infection. To test this, we isolated monocytes from the blood of preterm neonates, term neonates, and adults and, after stimulation with various concentrations of interleukin-1 alpha (IL-1 alpha) or lipopolysaccharide (LPS), quantified G-CSF concentrations in cell supernatants and G-CSF mRNA in cell lysates. When stimulated with plateau concentrations of IL-1 alpha for 24 hours, G-CSF concentrations were higher in supernatants of adult cells (8,699 +/- 5,529 pg/10(6) monocytes) than in those from term infants (2,557 +/- 442 pg, P < .05) or from preterm infants (879 +/- 348 pg, P < .05 v adults). When stimulated with plateau concentrations of LPS, supernatants of monocytes from preterm neonates had less G-CSF than did those from term neonates or adults. G-CSF mRNA content was low in cells from preterm infants, higher in those from term infants, and highest in those from adults. On the basis of the in vitro studies, we speculated that serum G-CSF concentrations might be less elevated in neutropenic neonates than in neutropenic adults. Indeed, serum concentrations were relatively low in all nonneutropenic subjects; 92 +/- 34 pg/mL (mean +/- SEM) in 10 preterm neonates, 114 +/- 21 pg/mL in 16 term neonates, and 45 +/- 13 pg/mL in 11 healthy adults. Serum concentrations were not elevated in 7 neutropenic neonates (39 +/- 17 pg/mL) but were in 8 neutropenic adults (2101 +/- 942 pg/mL, P < .05 v healthy adults). Other studies suggested that the lower G-CSF production in neonates is not counterbalanced by a heightened sensitivity of G-CSF--responsive progenitors to G-CSF. Therefore, we speculate that newborn infants, particularly those delivered prematurely, generate comparatively low quantities of G-CSF after inflammatory stimulation, and that this might constitute part of the explanation for their defective upregulation of neutrophil production and function during infection.
Assuntos
Sangue Fetal/metabolismo , Fator Estimulador de Colônias de Granulócitos/biossíntese , Recém-Nascido/sangue , Recém-Nascido Prematuro/sangue , Monócitos/metabolismo , Adulto , Sequência de Bases , Células Cultivadas , Fator Estimulador de Colônias de Granulócitos/genética , Fator Estimulador de Colônias de Granulócitos/farmacologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Humanos , Dados de Sequência Molecular , RNA Mensageiro/análiseRESUMO
Developmental delays, which impair antibacterial host defense, are present in the neutrophil system of human preterm neonates. We hypothesized that diminished production of interleukin-8 (IL-8), a neutrophil chemotactic peptide, might in part be responsible for these defects. To test this, monocytes from the blood of preterm neonates, term neonates, and adults were isolated immunologically by "negative panning" and subsequently stimulated with interleukin-1 alpha (IL-1 alpha), tumor necrosis factor alpha (TNF-alpha), or lipopolysaccharide (LPS), after which IL-8 levels in the supernatants were measured by ELISA. Total cellular RNA was extracted and IL-8 mRNA was assessed by Northern blotting and by competitive polymerase chain reaction (PCR) analyses. After stimulation with IL-1 alpha, IL-8 accumulation was lowest in supernatants of monocytes from preterm neonates, intermediate in supernatants of monocytes from term neonates and greatest from monocytes of adults. Similarly, when stimulated with TNF-alpha or LPS, monocytes from preterm neonates produced less IL-8 than cells from term neonates and adults. These differences in IL-8 concentrations paralleled differences in IL-8 mRNA expression.
Assuntos
Sangue Fetal/imunologia , Recém-Nascido/imunologia , Recém-Nascido Prematuro/imunologia , Interleucina-8/genética , Monócitos/imunologia , Transcrição Gênica , Adulto , Sequência de Bases , Northern Blotting , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Feminino , Humanos , Recém-Nascido/sangue , Recém-Nascido Prematuro/sangue , Interleucina-1/farmacologia , Interleucina-8/análise , Interleucina-8/biossíntese , Lipopolissacarídeos/farmacologia , Dados de Sequência Molecular , Monócitos/efeitos dos fármacos , Oligodesoxirribonucleotídeos , Placenta , Reação em Cadeia da Polimerase/métodos , Gravidez , RNA Mensageiro/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Valores de Referência , Transcrição Gênica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
We quantified circulating and storage neutrophils, their precursors and progenitors, and mRNA for some of the cytokines involved in granulocytopoiesis, in newborn and adult mice following intrapulmonary inoculation of Escherichia coli. Four hours following inoculation of adult and newborn mice with a quantity of organisms 2 logs below the LD100, all animals were neutropenic. After 24 h, adults had recovered from the neutropenia but neonates had not (p < 0.001). Accelerated neutrophil production was evident in the infected adults, and correlated with the appearance of granulocyte colony-stimulating factor (G-CSF) transcripts in the liver, spleen, and lung, and interleukin-6 (IL-6) transcripts in the spleen and lung. An increase in neutrophil production was not observed in the neonates, and none of their organs tested had transcripts for either G-CSF or IL-6, but they did have transcripts for cytokines not involved in granulocytopoiesis; macrophage colony-stimulating factor and its receptor (c-fms). We speculate that the failure to increase neutrophil production in infected neonatal mice is the result of failure to increase production of relevant cytokines.
Assuntos
Infecções por Escherichia coli/fisiopatologia , Fator Estimulador de Colônias de Granulócitos/genética , Interleucina-6/genética , Neutrófilos/patologia , RNA Mensageiro/metabolismo , Animais , Animais Recém-Nascidos , Células Sanguíneas/patologia , Medula Óssea/efeitos dos fármacos , Medula Óssea/patologia , Divisão Celular , Sobrevivência Celular/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/patologia , Leucócitos/patologia , Fator Estimulador de Colônias de Macrófagos/genética , Camundongos , Camundongos Endogâmicos C57BL , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Timidina/farmacologia , Fatores de TempoRESUMO
A recently cloned human cytokine, interleukin-11 (IL-11), has functional similarities to IL-6. We tested the hypothesis that the hematopoietic actions of IL-11 in vitro also resemble those of IL-6. The effect of IL-11 on the cell cycle status of fetal and adult hematopoietic progenitors was assessed using serum-free incubations followed by tritiated thymidine suicide studies. Its effect on clonogenic maturation was assessed by including IL-11, either as a single agent or with subplateau or plateau concentrations of other recombinant cytokines, in cultures that contained neutralizing monoclonal antibodies directed against relevant growth factors. Similar to IL-6, IL-11 resulted in accelerated cycling of fetal colony-forming units-mixed (CFU-MIX), CFU-granulocyte macrophage (CFU-GM), and erythroid burst-forming units (BFU-E). This effect was additive to that of submaximal, but not to plateau, concentrations of IL-6. However, no effect of IL-11 was observed on cycling status of adult progenitors. As a single agent, IL-11 failed to support clonal maturation of either fetal or adult progenitors. IL-11 was additive to GM-CSF in supporting clonal maturation of CFU-GM from adult marrow but not from fetal blood. We conclude that the in vitro hematopoietic actions of IL-11 on cell cycle status of hematopoietic progenitors resemble those of IL-6. However, unlike IL-6, IL-11 as a single agent failed to support clonal maturation of fetal CFU-GM, BFU-E, and CFU-MIX.
Assuntos
Células da Medula Óssea , Ciclo Celular , Sangue Fetal/citologia , Células-Tronco Hematopoéticas/citologia , Interleucinas/farmacologia , Adulto , Células Cultivadas , Células Clonais/citologia , Sinergismo Farmacológico , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Granulócitos/citologia , Humanos , Interleucina-11 , Interleucina-6/farmacologia , Macrófagos/citologia , Proteínas Recombinantes/farmacologiaRESUMO
Several deficiencies in antibacterial defense have been described in neonates. Among those best characterized are delayed maturation of B cells into antibody producing cells, deficient T-cell maturation, and delayed cycling of hematopoietic progenitor cells after an infectious challenge. No unifying theory has been forwarded, however, to explain the concomitance of these three developmental deficiencies. IL-6, a cytokine produced primarily by monocytes and macrophages in response to stimulation by IL-1, is involved in the regulation of these three processes. Thus, we postulated that defective production of IL-6 could be a mechanism underlying these immune deficiencies of neonates. Indeed, we observed that a peak production, cells of five term neonates produced only one half as much IL-6 (14 120 +/- 2590 pg IL-6/10(6) monocytes) as those of five adults (28 940 +/- 1680 pg, p less than 0.001). Peak production was lower still by monocytes of six preterm neonates (7190 +/- 1400 pg, p less than 0.001 versus term). Production of IL-6 protein was inhibited by actinomycin D and the IL-6 mRNA content of monocytes from neonates, as assessed by competitive polymerase chain reaction, was less than that of adult monocytes. We speculate that defective IL-6 transcription might underlie some of the defects in immune regulation observed in neonates.
Assuntos
Recém-Nascido/imunologia , Interleucina-6/biossíntese , Monócitos/imunologia , Adulto , Humanos , Técnicas In Vitro , Recém-Nascido Prematuro , Monócitos/metabolismo , RNA Mensageiro/metabolismoRESUMO
Very high concentrations of erythropoietin (epo), in clonogenic cultures, result in reduced production of neutrophils, and fetal progenitors are more sensitive to this effect of epo than are those of adults. However, the significance of this observation is unclear because no evidence of reduced neutrophil production has been presented following administration of recombinant epo to human or animal subjects. In the present study we injected newborn rats, beginning on the first day of life, with 20, 200, or 2,000 U epo/kg body weight, and measured serum epo concentrations after 2, 8, 24, or 48 hours. After selecting a dose that resulted in serum concentrations greater than 1,000 mU/mL (a concentration that resulted in down-modulation of neutrophil production from neonatal rat progenitors in vitro) other newborn rats were treated for 3 days with that dose (1,000 U epo/kg) or a vehicle control. Administration of epo resulted in increased hematocrits (P less than .001), reticulocyte counts (P less than .001), normoblasts/femur (P less than .05), and normoblasts/spleen (P less than .001). Recipients of epo also had more erythroid colony-forming units (CFU-E) (P less than .001) and higher CFU-E tritiated thymidine suicide rates (P less than .01) than did controls. However, femurs and spleens of epo recipients contained fewer postmitotic neutrophils (femur, P less than .01; spleen, P less than .01), proliferative neutrophils (femur, P less than .01; spleen, P less than .02), granulocyte-macrophage colony-forming units (CFU-GM) (P less than .005), and lower CFU-GM tritiated thymidine suicide rates (P less than .01). Seven and nine days after twice-daily administration of 2,000 U epo/kg, blood neutrophil concentrations had diminished (P less than .05). Thus, administration of high doses of recombinant epo to newborn rats resulted in diminished neutrophil production accompanying accelerated erythropoiesis.
Assuntos
Animais Recém-Nascidos , Eritropoetina/farmacologia , Neutrófilos/citologia , Animais , Peso Corporal , Ciclo Celular/efeitos dos fármacos , Eritropoese/efeitos dos fármacos , Eritropoetina/administração & dosagem , Eritropoetina/sangue , Fêmur/citologia , Células-Tronco Hematopoéticas/citologia , Contagem de Leucócitos/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Baço/citologiaRESUMO
We assessed the effect of interleukin-9 (IL-9) on clonogenic maturation and cell-cycle status of hematopoietic progenitors of fetal (umbilical cord blood) and adult (bone marrow) origin. As a single agent IL-9 supported, in a concentration-dependent fashion, maturation of burst-forming units-erythroid (BFU-E) of adult and fetal origin. However, only 1/3 the number of adult BFU-E colonies developed, as did in response to granulocyte-macrophage colony-stimulating factor (GM-CSF), and only 1/6 the number developed as did in response to IL-3. In contrast, the effect of IL-9 on fetal BFU-E colonies was equal to that of GM-CSF and IL-3. Synergistic effects of IL-9 with low concentrations (0.1 ng/mL) of GM-CSF and IL-3 were seen on adult BFU-E colony formation, but no effect was apparent at higher concentrations (1.0 ng/mL). In contrast, using fetal cells, synergistic effects of IL-9 with low and high concentrations of GM-CSF and IL-3 were apparent. Addition of IL-9 to plates containing fetal cells plus GM-CSF and IL-3 not only resulted in more BFU-E colonies, but also in more multicentered (greater than or equal to 10 individual centers) colonies, and more cells per colony. IL-9 had a wider spectrum of action on progenitors of fetal origin than on progenitors of adult origin, supporting the generation of fetal multipotent colony-forming unit (CFU)-Mix and CFU-GM colonies. Incubation with IL-9 did not accelerate cycling of adult or fetal BFU-E, CFU-Mix, or CFU-GM to the extent observed after incubation with IL-6. Thus, IL-9 primarily supported maturation of erythroid progenitors of adult origin, and its addition to plates containing GM-CSF and IL-3 (1.0 ng/mL) did not result in maturation of additional clones. In contrast, IL-9 had a wider spectrum of action on fetal progenitors and, when combined with IL-3 and GM-CSF, resulted in clonogenic maturation of progenitors that did not undergo maturation after stimulation with IL-3 and GM-CSF.