RESUMEN
Pathogen immune responses are profoundly attenuated in fetuses and premature infants, yet the mechanisms underlying this developmental immaturity remain unclear. Here we show transcriptomic, metabolic and polysome profiling and find that monocytes isolated from infants born early in gestation display perturbations in PPAR-γ-regulated metabolic pathways, limited glycolytic capacity and reduced ribosomal activity. These metabolic changes are linked to a lack of translation of most cytokines and of MALT1 signalosome genes essential to respond to the neonatal pathogen Candida. In contrast, they have little impact on house-keeping phagocytosis functions. Transcriptome analyses further indicate a role for mTOR and its putative negative regulator DNA Damage Inducible Transcript 4-Like in regulating these metabolic constraints. Our results provide a molecular basis for the broad susceptibility to multiple pathogens in these infants, and suggest that the fetal immune system is metabolically programmed to avoid energetically costly, dispensable and potentially harmful immune responses during ontogeny.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Inmunidad Innata , Monocitos/inmunología , Proteína 1 de la Translocación del Linfoma del Tejido Linfático Asociado a Mucosas/inmunología , PPAR gamma/inmunología , Factores de Transcripción/inmunología , Adulto , Proteína 10 de la LLC-Linfoma de Células B/deficiencia , Proteína 10 de la LLC-Linfoma de Células B/genética , Proteína 10 de la LLC-Linfoma de Células B/inmunología , Proteínas Adaptadoras de Señalización CARD/deficiencia , Proteínas Adaptadoras de Señalización CARD/genética , Proteínas Adaptadoras de Señalización CARD/inmunología , Candida albicans/inmunología , Candida parapsilosis/inmunología , Humanos , Recién Nacido , Recien Nacido Prematuro , Interleucinas/deficiencia , Interleucinas/genética , Interleucinas/inmunología , Lectinas Tipo C/deficiencia , Lectinas Tipo C/genética , Lectinas Tipo C/inmunología , Lipopolisacáridos/farmacología , Análisis por Micromatrices , Monocitos/citología , Monocitos/efectos de los fármacos , Proteína 1 de la Translocación del Linfoma del Tejido Linfático Asociado a Mucosas/deficiencia , Proteína 1 de la Translocación del Linfoma del Tejido Linfático Asociado a Mucosas/genética , PPAR gamma/deficiencia , PPAR gamma/genética , Cultivo Primario de Células , Biosíntesis de Proteínas/inmunología , Serina-Treonina Quinasas TOR/deficiencia , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/inmunología , Factores de Transcripción/deficiencia , Factores de Transcripción/genética , Transcriptoma/inmunología , Factor de Necrosis Tumoral alfa/deficiencia , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Newborns are prone to fungal infections, largely due to Candida species. The immunological basis for this vulnerability is not yet fully understood. However, useful insights can be gained from the knowledge of the maturation of immune pathways during ontogeny, particularly when placed in context with how rare genetic mutations in humans predispose to fungal diseases. In this article, we review these most current data on immune functions in human newborns, highlighting pathways most relevant to the response to Candida. While discussing these data, we propose a framework of why deficiencies in these pathways make newborns particularly vulnerable to this opportunistic pathogen.
RESUMEN
Despite concerted international efforts, mortality from neonatal infections remains unacceptably high in some areas of the world, particularly for premature infants. Recent developments in flow cytometry and next-generation sequencing technologies have led to major discoveries over the past few years, providing a more integrated understanding of the developing human immune system in the context of its microbial environment. We review these recent findings, focusing on how in human newborns incomplete maturation of the immune system before a full term of gestation impacts on their vulnerability to infection. We also discuss some of the clinical implications of this research in guiding the design of more-accurate age-adapted diagnostic and preventive strategies for neonatal sepsis.
Asunto(s)
Interacciones Huésped-Patógeno/inmunología , Inmunidad , Sepsis Neonatal/etiología , Animales , Modelos Animales de Enfermedad , Desarrollo Fetal/inmunología , Hematopoyesis , Humanos , Inmunidad Mucosa , Lactante , Recién Nacido , Ratones , Membrana Mucosa/inmunología , Membrana Mucosa/metabolismo , Membrana Mucosa/microbiología , Sepsis Neonatal/diagnóstico , Sepsis Neonatal/epidemiología , Evaluación del Resultado de la Atención al PacienteRESUMEN
BACKGROUND: Antimicrobial responses have been shown to be profoundly attenuated in very preterm neonates when examined on cord blood. However, we lack data on these responses at the time these neonates are most vulnerable to infections. METHODS: Multiple cytokine responses to two prototypic Toll-like receptor (TLR) agonists: lipopolysaccharide (LPS) (TLR4) and R848 (TLR7/8) were prospectively measured in preterm neonates born ≤30 wk of gestation (n = 50) during the first 28 d of age using whole blood and single-cell multiparameter flow cytometry assays. Results were compared to term neonates (n = 30) and adult controls (n = 25). RESULTS: In preterm neonates, LPS and R848 responses remained attenuated in both cord blood and in the first 28 d of age. These responses showed significant maturation over time after adjusting for gestational age and were confirmed in monocytes and dendritic cells on a per-cell basis. We detected no major contribution of chorioamnionitis, maternal antenatal corticosteroids or magnesium sulfate treatment, labor, or mode of delivery to the maturation of cytokine responses. CONCLUSION: Innate immune antimicrobial defenses are profoundly attenuated developmentally in very preterm neonates during the neonatal period, suggesting that exogenous factors drive the sustained systemic inflammation that has been linked to increased morbidities in these infants.
Asunto(s)
Inmunidad Innata , Recien Nacido Prematuro/inmunología , Adulto , Estudios de Casos y Controles , Citocinas/sangre , Citocinas/inmunología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Femenino , Sangre Fetal/inmunología , Citometría de Flujo , Edad Gestacional , Humanos , Imidazoles/farmacología , Inmunidad Innata/efectos de los fármacos , Recién Nacido , Recien Nacido Prematuro/sangre , Lipopolisacáridos/farmacología , Masculino , Monocitos/efectos de los fármacos , Monocitos/inmunología , Estudios Prospectivos , Factores de Tiempo , Receptor Toll-Like 4/agonistas , Receptor Toll-Like 4/sangre , Receptor Toll-Like 4/inmunología , Receptor Toll-Like 7/agonistas , Receptor Toll-Like 7/sangre , Receptor Toll-Like 7/inmunología , Receptor Toll-Like 8/agonistas , Receptor Toll-Like 8/sangre , Receptor Toll-Like 8/inmunologíaRESUMEN
Interleukin-1ß (IL-1ß) production is impaired in cord blood monocytes. However, the mechanism underlying this developmental attenuation remains unclear. Here, we analyzed the extent of variability within the Toll-like receptor (TLR)/NLRP3 inflammasome pathways in human neonates. We show that immature low CD14 expressing/CD16(pos) monocytes predominate before 33 weeks of gestation, and that these cells lack production of the pro-IL-1ß precursor protein upon LPS stimulation. In contrast, high levels of pro-IL-1ß are produced within high CD14 expressing monocytes, although these cells are unable to secrete mature IL-1ß. The lack of secreted IL-1ß in these monocytes parallels a reduction of NLRP3 induction following TLR stimulation resulting in a lack of caspase-1 activity before 29 weeks of gestation, whereas expression of the apoptosis-associated speck-like protein containing a CARD and function of the P2×7 receptor are preserved. Our analyses also reveal a strong inhibitory effect of placental infection on LPS/ATP-induced caspase-1 activity in cord blood monocytes. Lastly, secretion of IL-1ß in preterm neonates is restored to adult levels during the neonatal period, indicating rapid maturation of these responses after birth. Collectively, our data highlight important developmental mechanisms regulating IL-1ß responses early in gestation, in part due to a downregulation of TLR-mediated NLRP3 expression. Such mechanisms may serve to limit potentially damaging inflammatory responses in a developing fetus.
Asunto(s)
Proteínas Portadoras/inmunología , Desarrollo Fetal/inmunología , Inflamasomas/inmunología , Interleucina-1beta/inmunología , Macrófagos/inmunología , Monocitos/inmunología , Adenosina Trifosfato/farmacología , Adulto , Proteínas Adaptadoras de Señalización CARD , Proteínas Portadoras/genética , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/inmunología , Sangre Fetal/citología , Sangre Fetal/efectos de los fármacos , Sangre Fetal/inmunología , Feto , Regulación del Desarrollo de la Expresión Génica , Edad Gestacional , Humanos , Recién Nacido , Recien Nacido Prematuro , Inflamasomas/genética , Interleucina-1beta/genética , Lipopolisacáridos/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Monocitos/citología , Monocitos/efectos de los fármacos , Proteína con Dominio Pirina 3 de la Familia NLR , Receptores Purinérgicos P2X7/genética , Receptores Purinérgicos P2X7/inmunología , Transducción de Señal , Receptores Toll-Like/genética , Receptores Toll-Like/inmunologíaRESUMEN
The appetite suppressing hormone, leptin is now established as an important component of the immune response to pathogens partly via the induction of brain IL-1beta. We have previously demonstrated that this hormone acts on microglia to induce the release of IL-1beta through actions on its functional receptors. In the present study, we extended these findings by demonstrating that leptin's action on microglia is that of a modulator rather than a direct trigger of inflammation. Using primary microglia cultures prepared from rat brain we show that pre-incubation of these cells with leptin for 24h prior to treatment with LPS increased the IL-1beta output 2-fold. This effect was not limited to IL-1beta but was also true for another cytokine, TNF-alpha and chemokines such as CINC-1 and MIP-2. The role of leptin in potentiating the microglial response to LPS appeared to be linked to morphological changes rendering the microglia more reactive. These results suggest that leptin has an important role in microglial function in inflammation and given that its circulating levels fluctuate across a number of conditions, these findings can have important implications for an individual's ability to mount an efficient and complete response to invading pathogens.
Asunto(s)
Citocinas/metabolismo , Leptina/farmacología , Microglía/metabolismo , Microglía/ultraestructura , Animales , Encéfalo/citología , Química Encefálica/efectos de los fármacos , Química Encefálica/fisiología , Células Cultivadas , Quimiocinas/metabolismo , Relación Dosis-Respuesta a Droga , Inmunoensayo , Inmunohistoquímica , Inflamación/inducido químicamente , Inflamación/patología , Interleucina-1beta/biosíntesis , Lipopolisacáridos/farmacología , Microglía/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Cardiovascular dysfunction as a result of sepsis is the leading cause of death in the critically ill. Cardiomyocytes respond to infectious pathogens with a Toll-like receptor-initiated proinflammatory response in conjunction with a decrease in contractility, although the downstream events linking Toll-like receptor activation and reduced cardiac contractility remain to be elucidated. Using microarray analysis of cardiac tissue exposed to systemic lipopolysaccharide (LPS), we discovered that 2 small calcium-regulating proteins (S100A8 and S100A9) are highly upregulated. HL-1 cardiomyocytes, isolated primary cardiomyocytes, and live mice were exposed to LPS, whereas beating HL-1 cells had S100A8 and S100A9 overexpressed and their calcium flux quantified. Using in vivo microbubble technology, we delivered S100A8 and S100A9 to normal mouse hearts; using the same technology, we inhibited S100A9 production in mouse hearts and subsequently exposed them to LPS. Coimmunoprecipitation of S100A8 and S100A9 identified interaction with RAGE (the receptor for advanced glycation end products), the cardiac function and postreceptor signaling of which were investigated. HL-1 cardiomyocytes, isolated primary cardiomyocytes, and whole hearts exposed to LPS have large increases in S100A8 and S100A9. Cardiac overexpression of S100A8 and S100A9 led to a RAGE-dependent decrease in calcium flux and, in the intact mouse, to a decreased cardiac ejection fraction, whereas knockdown of S100A9 attenuated LPS-induced cardiac dysfunction. Cardiomyocytes exposed to LPS express S100A8 and S100A9, leading to a RAGE-mediated decrease in cardiomyocyte contractility. This finding provides a novel mechanistic link between circulating pathogen-associated molecular products and subsequent cardiac dysfunction.