RESUMEN
Septic diseases are characterized by an initial systemic, proinflammatory phase, followed by a period of anti-inflammation. In the context of the latter, monocytes have been described to display altered functions, including reduced TNF secretion and T cell-stimulating capacities in response to recall antigens. This hyporesponsiveness is supposed to be detrimental for coping with secondary infections. We here characterize bacterially reprogrammed PBMC-derived monocytes with special focus on their phagocytic activity. Hence, we have implemented a surrogate model of the early, postinflammatory period by exposing PBMCs to Escherichia coli on d0 and rechallenging them with bacteria on d2. This induced the emergence of a distinct monocytic phenotype with profound phagocytic impairments but a preserved ability for naïve T cell stimulation. The compromising effects on phagocytosis required the presence of bacteria and were not mimicked by TLR4 ligation or exposure to isolated cytokines alone. Moreover, the impairments were specific for the engulfment of bacteria and were coupled to a selective down-regulation of FcγR and SR expression. Intriguingly, this monocytic phenotype contributed to the stimulation of a T(H)17-polarized adaptive immune response in the context of secondary infection. Our findings extend the current knowledge of monocytic reprogramming and identify the phagocytic capacity of monocytes as a putative sepsis biomarker.
Asunto(s)
Infecciones por Escherichia coli/inmunología , Escherichia coli/inmunología , Activación de Linfocitos , Monocitos/inmunología , Fagocitosis/inmunología , Sepsis/inmunología , Células Th17/inmunología , Inmunidad Adaptativa , Adulto , Biomarcadores/metabolismo , Escherichia coli/metabolismo , Infecciones por Escherichia coli/metabolismo , Femenino , Humanos , Masculino , Monocitos/metabolismo , Receptores de IgG/inmunología , Receptores de IgG/metabolismo , Sepsis/metabolismo , Células Th17/metabolismo , Receptor Toll-Like 4/inmunología , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Trials of immune-modulating drugs in septic patients have mostly failed to demonstrate clinical efficacy. Thus, we sought to generate a surrogate model of myelomonocytic lineage differentiation that would potentially allow sepsis induction and preclinical testing of anti-inflammatory drugs. Comparing transplantation of cord blood-derived stem cells in neonatal NOD/SCID/IL2Rγ(null) (neonatal huNSG) mice with transplantation of adult peripheral mobilized stem cells into adult NSG (adult huNSG) recipients, we demonstrate that myelomonocytic lineage differentiation in neonatal huNSG mice is retarded and monocytes are phenotypically immature with respect to HLA-DR expression and the emergence of CD80(+)CD86(+) monocytes. Functionally, neonatal huNSG mice were less sensitive toward interferon-γ-induced upregulation of CD86 and exhibited a reduced T-cell stimulating capacity when compared with adult huNSG mice, whereas the phagocytic activity and the ability for cytokine secretion were mature. However, comparison of these data with data obtained from human neonates indicate that absence of the CD80(+)CD86(+) population and the reduced T-cell stimulating capacity of neonatal huNSG monocytes resemble functional immaturities observed in human neonatal monocytes. Thus, these two mouse models might well serve as 2 independent surrogate models for studying the neonatal myelomonocytic lineage differentiation or for testing the efficacy of immunomodulatory drugs on functionally mature monocytes.
Asunto(s)
Subunidad gamma Común de Receptores de Interleucina/genética , Monocitos/inmunología , Fenotipo , Animales , Diferenciación Celular/inmunología , Linaje de la Célula , Citocinas/metabolismo , Femenino , Humanos , Inmunofenotipificación , Mediadores de Inflamación/metabolismo , Subunidad gamma Común de Receptores de Interleucina/deficiencia , Receptores de Lipopolisacáridos/metabolismo , Activación de Linfocitos/inmunología , Masculino , Ratones , Ratones Endogámicos NOD , Ratones Noqueados , Ratones SCID , Monocitos/citología , Monocitos/metabolismo , Fagocitosis/inmunología , Linfocitos T/inmunologíaRESUMEN
Interactions between innate and adaptive immune functions in neonatal macrophages (MPhi) are still unclear. We therefore established a method to quantify bacterial phagocytosis and intracellular degradation, using green fluorescent protein (GFP)-labeled Escherichia coli in combination with phenotypic analysis. The kinetics of the proportion of phagocyting MPhi, phagocytosed bacteria per MPhi, and bacterial degradation were comparable for cord blood MPhi of term neonates and MPhi of healthy adults. Phenotyping revealed CD14 and CD16 to be down-modulated within minutes. GFP-labeled E. coli may be useful tools to further study MPhi subpopulations and determinants of phagocytosis in cord blood MPhi.