RESUMEN
The influence of non-opsonized and opsonized S. aureus 2879M and E. coli 321 strains on the total strength of interaction between the endothelial cell and neutrophil during the docking process was studied using in vitro model of experimental septicemia. We observed a decrease in the force and work of adhesion between receptors of neutrophils and endothelial cells under the influence of non-opsonized strains and further decrease in the affinity of single interactions between cells under the influence of opsonized S. aureus, which was compensated by an increase in the number of contacts, as well as an increase in the force of adhesion under the influence of opsonized E. coli compared to non-opsonized bacteria, which remained below the control level, while adhesion work reaches the control level. Thus, opsonization of S. aureus aggravates the "immunological uncoupling" between neutrophils and endothelial cells, while opsonization of E. coli reduces the pathological effect compared to non-opsonized bacteria.
Asunto(s)
Células Endoteliales , Escherichia coli , Neutrófilos , Sepsis , Staphylococcus aureus , Neutrófilos/inmunología , Neutrófilos/metabolismo , Escherichia coli/inmunología , Staphylococcus aureus/inmunología , Staphylococcus aureus/patogenicidad , Sepsis/inmunología , Sepsis/microbiología , Sepsis/metabolismo , Sepsis/patología , Células Endoteliales/inmunología , Células Endoteliales/metabolismo , Células Endoteliales/microbiología , Humanos , Fagocitosis , Adhesión Celular/inmunología , Proteínas Opsoninas/metabolismo , Proteínas Opsoninas/inmunología , Adhesión Bacteriana , AnimalesRESUMEN
The bacterial pathogen Staphylococcus aureus responds to the host environment by increasing the thickness of its cell wall. However, the impact of cell wall thickening on susceptibility to host defenses is unclear. Using bacteria incubated in human serum, we show that host-induced increases in cell wall thickness led to a reduction in the exposure of bound antibody and complement and a corresponding reduction in phagocytosis and killing by neutrophils. The exposure of opsonins bound to protein antigens or lipoteichoic acid (LTA) was most significantly reduced, while opsonization by IgG against wall teichoic acid or peptidoglycan was largely unaffected. Partial digestion of accumulated cell wall using the enzyme lysostaphin restored opsonin exposure and promoted phagocytosis and killing. Concordantly, the antibiotic fosfomycin inhibited cell wall remodeling and maintained the full susceptibility of S. aureus to opsonophagocytic killing by neutrophils. These findings reveal that host-induced changes to the S. aureus cell wall reduce the ability of the immune system to detect and kill this pathogen through reduced exposure of protein- and LTA-bound opsonins. IMPORTANCE: Understanding how bacteria adapt to the host environment is critical in determining fundamental mechanisms of immune evasion, pathogenesis, and the identification of targets for new therapeutic approaches. Previous work demonstrated that Staphylococcus aureus remodels its cell envelope in response to host factors and we hypothesized that this may affect recognition by antibodies and thus killing by immune cells. As expected, incubation of S. aureus in human serum resulted in rapid binding of antibodies. However, as bacteria adapted to the serum, the increase in cell wall thickness resulted in a significant reduction in exposure of bound antibodies. This reduced antibody exposure, in turn, led to reduced killing by human neutrophils. Importantly, while antibodies bound to some cell surface structures became obscured, this was not the case for those bound to wall teichoic acid, which may have important implications for vaccine design.
Asunto(s)
Pared Celular , Neutrófilos , Proteínas Opsoninas , Fagocitosis , Staphylococcus aureus , Pared Celular/inmunología , Pared Celular/metabolismo , Humanos , Neutrófilos/inmunología , Neutrófilos/microbiología , Staphylococcus aureus/inmunología , Proteínas Opsoninas/metabolismo , Proteínas Opsoninas/inmunología , Opsonización/inmunología , Anticuerpos Antibacterianos/inmunología , Anticuerpos Antibacterianos/sangre , Ácidos Teicoicos/metabolismo , Ácidos Teicoicos/inmunología , Evasión Inmune , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/microbiología , Interacciones Huésped-Patógeno/inmunologíaRESUMEN
Introduction: Innate immunity is crucial to reducing parasite burden and contributing to survival in severe malaria. Monocytes are key actors in the innate response and, like macrophages, are plastic cells whose function and phenotype are regulated by the signals from the microenvironment. In the context of cerebral malaria (CM), monocyte response constitutes an important issue to understand. We previously demonstrated that decreased percentages of nonclassical monocytes were associated with death outcomes in CM children. In the current study, we postulated that monocyte phagocytosis function is impacted by the severity of malaria infection. Methods: To study this hypothesis, we compared the opsonic and nonopsonic phagocytosis capacity of circulant monocytes from Beninese children with uncomplicated malaria (UM) and CM. For the CM group, samples were obtained at inclusion (D0) and 3 and 30 days after treatment (D3, D30). The phagocytosis capacity of monocytes and their subsets was characterized by flow cytometry and transcriptional profiling by studying genes known for their functional implication in infected-red blood cell (iRBC) elimination or immune escape. Results: Our results confirm our hypothesis and highlight the higher capacity of nonclassical monocytes to phagocyte iRBC. We also confirm that a low number of nonclassical monocytes is associated with CM outcome when compared to UM, suggesting a mobilization of this subpopulation to the cerebral inflammatory site. Finally, our results suggest the implication of the inhibitory receptors LILRB1, LILRB2, and Tim3 in phagocytosis control. Discussion: Taken together, these data provide a better understanding of the interplay between monocytes and malaria infection in the pathogenicity of CM.
Asunto(s)
Malaria Cerebral , Monocitos , Fagocitosis , Humanos , Malaria Cerebral/inmunología , Malaria Cerebral/parasitología , Monocitos/inmunología , Masculino , Preescolar , Femenino , Niño , Lactante , Plasmodium falciparum/inmunología , Proteínas Opsoninas/metabolismo , Proteínas Opsoninas/inmunología , Eritrocitos/parasitología , Eritrocitos/inmunología , Inmunidad InnataRESUMEN
Introduction: The California purple sea urchin, Strongylocentrotus purpuratus, relies solely on an innate immune system to combat the many pathogens in the marine environment. One aspect of their molecular defenses is the SpTransformer (SpTrf) gene family that is upregulated in response to immune challenge. The gene sequences are highly variable both within and among animals and likely encode thousands of SpTrf isoforms within the sea urchin population. The native SpTrf proteins bind foreign targets and augment phagocytosis of a marine Vibrio. A recombinant (r)SpTrf-E1-Ec protein produced by E. coli also binds Vibrio but does not augment phagocytosis. Methods: To address the question of whether other rSpTrf isoforms function as opsonins and augment phagocytosis, six rSpTrf proteins were expressed in insect cells. Results: The rSpTrf proteins are larger than expected, are glycosylated, and one dimerized irreversibly. Each rSpTrf protein cross-linked to inert magnetic beads (rSpTrf::beads) results in different levels of surface binding and phagocytosis by phagocytes. Initial analysis shows that significantly more rSpTrf::beads associate with cells compared to control BSA::beads. Binding specificity was verified by pre-incubating the rSpTrf::beads with antibodies, which reduces the association with phagocytes. The different rSpTrf::beads show significant differences for cell surface binding and phagocytosis by phagocytes. Furthermore, there are differences among the three distinct types of phagocytes that show specific vs. constitutive binding and phagocytosis. Conclusion: These findings illustrate the complexity and effectiveness of the sea urchin innate immune system driven by the natSpTrf proteins and the phagocyte cell populations that act to neutralize a wide range of foreign pathogens.
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Fagocitos , Fagocitosis , Proteínas Recombinantes , Animales , Fagocitosis/inmunología , Fagocitos/inmunología , Fagocitos/metabolismo , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/genética , Unión Proteica , Strongylocentrotus purpuratus/inmunología , Strongylocentrotus purpuratus/genética , Inmunidad Innata , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Erizos de Mar/inmunología , Vibrio/inmunología , Proteínas Opsoninas/metabolismo , Proteínas Opsoninas/inmunologíaRESUMEN
In normal cells, binding of the transmembrane protein CD47 to signal regulatory protein-α (SIRPα) on macrophages induces an antiphagocytic signal. Tumor cells hijack this pathway and overexpress CD47 to evade immune destruction. Macrophage antitumor activity can be restored by simultaneously blocking the CD47-SIRPα signaling axis and inducing a prophagocytic signal via tumor-opsonizing antibodies. We identified a novel, fully human mAb (BMS-986351) that binds SIRPα with high affinity. BMS-986351 demonstrated broad binding coverage across SIRPα polymorphisms and potently blocked CD47-SIRPα binding at the CD47 binding site in a dose-dependent manner. In vitro, BMS-986351 increased phagocytic activity against cell lines from solid tumors and hematologic malignancies, and this effect was markedly enhanced when BMS-986351 was combined with the opsonizing antibodies cetuximab and rituximab. A phase I dose-escalation/-expansion study of BMS-986351 for the treatment of advanced solid and hematologic malignancies is underway (NCT03783403). SIGNIFICANCE: Increasing the phagocytotic capabilities of tumor-associated macrophages by modulating macrophage-tumor cell surface signaling via the CD47-SIRPα axis is a novel strategy. Molecules targeting CD47 have potential but its ubiquitous expression necessitates higher therapeutic doses to overcome potential antigen sink effects. The restricted expression pattern of SIRPα may limit toxicities and lower doses of the SIRPα antibody BMS-986351 may overcome target mediated drug disposition while maintaining the desired pharmacology.
Asunto(s)
Neoplasias Hematológicas , Neoplasias , Humanos , Antígeno CD47/genética , Receptores Inmunológicos/genética , Fagocitosis , Macrófagos , Neoplasias/tratamiento farmacológico , Anticuerpos Antineoplásicos/metabolismo , Proteínas Opsoninas/metabolismo , Neoplasias Hematológicas/metabolismoRESUMEN
Se comunican los resultados de un trabajo realizado para determinar el efecto de la tuftsina sintética en la capacidad oxidativa de células polimorfonucleares de niños recién nacidos pequeños para su edad gestacional, ya que se ha demostrado que está disminuida la actividad fagocítica de macrófagos y polimorfonucleares (PMNs) comparada con las células de niños con peso adecuado para su edad gestacional. En los resultados se observó un efecto de estimulación de la capacidad oxidativa de los PMNs con tuftsina sintética a través de incrementar la reducción de nitroazul de tetrazolio, lo que sugiere que la baja capacidad oxidativa de las células de recién nacidos pequeños para su edad gestacional no se relaciona con un defecto intrínseco celular