RESUMEN
We are entering the post-antibiotic era. Antimicrobial resistance (AMR) is a critical problem in chronic lung infections resulting in progressive respiratory failure and increased mortality. In the absence of emerging novel antibiotics to counter AMR infections, bacteriophages (phages), viruses that infect bacteria, have become a promising option for chronic respiratory infections. However, while personalized phage therapy is associated with improved outcomes in individual cases, clinical trials demonstrating treatment efficacy are lacking, limiting the therapeutic potential of this approach for respiratory infections. In this review, we address the current state of phage therapy for managing chronic respiratory diseases. We then discuss how phage therapy may address major microbiologic obstacles which hinder disease resolution of chronic lung infections with current antibiotic-based treatment practices. Finally, we highlight the challenges that must be addressed for successful phage therapy clinical trials. Through this discussion, we hope to expand on the potential of phages as an adjuvant therapy in chronic lung infections, as well as the microbiologic challenges that need to be addressed for phage therapy to expand beyond personalized salvage therapy.
Asunto(s)
Terapia de Fagos , Infecciones del Sistema Respiratorio , Humanos , Terapia de Fagos/métodos , Infecciones del Sistema Respiratorio/terapia , Infecciones del Sistema Respiratorio/microbiología , Infecciones del Sistema Respiratorio/virología , Bacteriófagos , Enfermedad Crónica , Antibacterianos/uso terapéuticoRESUMEN
Chronic wounds infected by Pseudomonas aeruginosa (Pa) are characterized by disease progression and increased mortality. We reveal Pf, a bacteriophage produced by Pa that delays healing of chronically infected wounds in human subjects and animal models of disease. Interestingly, impairment of wound closure by Pf is independent of its effects on Pa pathogenesis. Rather, Pf impedes keratinocyte migration, which is essential for wound healing, through direct inhibition of CXCL1 signaling. In support of these findings, a prospective cohort study of 36 human patients with chronic Pa wound infections reveals that wounds infected with Pf-positive strains of Pa are more likely to progress in size compared with wounds infected with Pf-negative strains. Together, these data implicate Pf phage in the delayed wound healing associated with Pa infection through direct manipulation of mammalian cells. These findings suggest Pf may have potential as a biomarker and therapeutic target in chronic wounds.
Asunto(s)
Inovirus , Infecciones por Pseudomonas , Infección de Heridas , Animales , Biopelículas , Humanos , Mamíferos , Estudios Prospectivos , Pseudomonas , Infecciones por Pseudomonas/terapia , Pseudomonas aeruginosa , Cicatrización de Heridas , Infección de Heridas/terapiaRESUMEN
Bacteriophages-viruses that infect bacteria-are abundant within our bodies, but their significance to human health is only beginning to be explored. Here, we synthesize what is currently known about our phageome and its interactions with the immune system. We first review how phages indirectly affect immunity via bacterial expression of phage-encoded proteins. We next review how phages directly influence innate immunity and bacterial clearance. Finally, we discuss adaptive immunity against phages and its implications for phage/bacterial interactions. In light of these data, we propose that our microbiome can be understood as an interconnected network of bacteria, bacteriophages, and human cells and that the stability of these tri-kingdom interactions may be important for maintaining our immunologic and metabolic health. Conversely, the disruption of this balance, through exposure to exogenous phages, microbial dysbiosis, or immune dysregulation, may contribute to disease.
Asunto(s)
Bacteriófagos , Microbiota , Virus , Bacterias , Bacteriófagos/genética , Humanos , Sistema InmunológicoRESUMEN
Bacteriophages have attracted extensive interest in vaccine design. This includes the use of phage display technology to select antigens, the use of engineered phages displaying target antigens in vaccine formulations, and phage DNA vaccines. However, the development of these approaches is limited in part by uncertainty regarding the underlying mechanisms by which phages elicit immunity. This has stymied the clinical development of this technology. Here we review the immunology of phage vaccines and highlight the gaps in our knowledge regarding the underlying mechanisms. First, we review the basic biology of phages and their use in vaccines. Next we discuss what is known about the mechanisms of immunity against engineered phages and phage DNA. Finally, we highlight the gaps in our understanding regarding the immunogenicity of these preparations. We argue that mechanistic insight into the immunology of phage vaccines is essential for the further development and clinical utility of these technologies.
Asunto(s)
Bacteriófagos , Vacunas , Bacteriófagos/genética , Técnicas de Visualización de Superficie CelularRESUMEN
Group B streptococci (GBSs) are the leading cause of neonatal meningitis. GBSs enter the CNS by penetrating the blood-brain barrier (BBB), which consists of specialized human brain microvascular endothelial cells (hBMECs). To identify GBS factors required for BBB penetration, we generated random mutant libraries of a virulent strain and screened for loss of hBMEC invasion in vitro. Two independent hypo-invasive mutants possessed disruptions in the same gene, invasion associated gene (iagA), which encodes a glycosyltransferase homolog. Allelic replacement of iagA in the GBS chromosome produced a 4-fold decrease in hBMEC invasiveness. Mice challenged with the GBS DeltaiagA mutant developed bacteremia comparably to WT mice, yet mortality was significantly lower (20% vs. 90%), as was the incidence of meningitis. The glycolipid diglucosyldiacylglycerol, a cell membrane anchor for lipoteichoic acid (LTA) and predicted product of the IagA glycosyltransferase, was absent in the DeltaiagA mutant, which consequently shed LTA into the media. Attenuation of virulence of the DeltaiagA mutant was found to be independent of TLR2-mediated signaling, but bacterial supernatants from the DeltaiagA mutant containing released LTA inhibited hBMEC invasion by WT GBS. Our data suggest that LTA expression on the GBS surface plays a role in bacterial interaction with BBB endothelium and the pathogenesis of neonatal meningitis.
Asunto(s)
Barrera Hematoencefálica/microbiología , Membrana Celular/metabolismo , Lipopolisacáridos/metabolismo , Streptococcus agalactiae/genética , Streptococcus agalactiae/patogenicidad , Ácidos Teicoicos/metabolismo , Animales , Transporte Biológico/fisiología , Barrera Hematoencefálica/metabolismo , Encéfalo/citología , Encéfalo/metabolismo , Encéfalo/microbiología , Encéfalo/patología , Células Endoteliales/citología , Células Endoteliales/microbiología , Endotelio Vascular/citología , Glucolípidos/metabolismo , Humanos , Recién Nacido , Meningitis Bacterianas , Ratones , Ratones Endogámicos C57BL , Streptococcus agalactiae/metabolismoRESUMEN
BACKGROUND: Modification of teichoic acids with D-alanine by the products of the dlt operon protects Gram-positive bacteria against major antimicrobial host defense molecules such as defensins, cathelicidins, myeloperoxidase or phospholipase. The graRS regulatory genes have recently been implicated in the control of D-alanylation in Staphylococcus aureus. RESULTS: To determine the impact of the GraRS regulatory system on resistance to antimicrobial host defense mechanisms and virulence of S. aureus, we compared inactivation of S. aureus SA113 wild type and its isogenic graRS deletion mutant by the human cathelicidin LL-37 or human neutrophil granulocytes in vitro, and the ability to cause infection in vivo. We show here that graRS deletion considerably alters bacterial surface charge, increases susceptibility to killing by human neutrophils or the defense peptide LL-37, and attenuates virulence of S. aureus in a mouse infection model. CONCLUSION: Our results indicate that S. aureus can regulate its surface properties in order to overcome innate host defenses.
Asunto(s)
Genes Reguladores , Infecciones Estafilocócicas/inmunología , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidad , Alanina/metabolismo , Animales , Péptidos Catiónicos Antimicrobianos/inmunología , Catelicidinas , Membrana Celular/metabolismo , Células Cultivadas , Citocromos c/metabolismo , Femenino , Eliminación de Gen , Genes Bacterianos , Humanos , Inmunidad Innata , Riñón/microbiología , Lisina/metabolismo , Ratones , Ratones Endogámicos BALB C , Neutrófilos/inmunología , Operón , Fosfatidilgliceroles/metabolismo , Ácidos Teicoicos/metabolismo , VirulenciaRESUMEN
Inflammatory bowel disease (IBD) is associated with risk variants in the human genome and dysbiosis of the gut microbiome, though unifying principles for these findings remain largely undescribed. The human commensal Bacteroides fragilis delivers immunomodulatory molecules to immune cells via secretion of outer membrane vesicles (OMVs). We reveal that OMVs require IBD-associated genes, ATG16L1 and NOD2, to activate a noncanonical autophagy pathway during protection from colitis. ATG16L1-deficient dendritic cells do not induce regulatory T cells (T(regs)) to suppress mucosal inflammation. Immune cells from human subjects with a major risk variant in ATG16L1 are defective in T(reg) responses to OMVs. We propose that polymorphisms in susceptibility genes promote disease through defects in "sensing" protective signals from the microbiome, defining a potentially critical gene-environment etiology for IBD.
Asunto(s)
Bacteroides fragilis/inmunología , Proteínas Portadoras/genética , Microbioma Gastrointestinal/inmunología , Interacción Gen-Ambiente , Enfermedades Inflamatorias del Intestino/genética , Enfermedades Inflamatorias del Intestino/microbiología , Proteína Adaptadora de Señalización NOD2/genética , Adulto , Anciano , Animales , Autofagia/inmunología , Proteínas Relacionadas con la Autofagia , Células Dendríticas/inmunología , Vesículas Extracelulares/inmunología , Femenino , Predisposición Genética a la Enfermedad , Genoma Humano , Humanos , Enfermedades Inflamatorias del Intestino/inmunología , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Polimorfismo Genético , Linfocitos T Reguladores/inmunologíaRESUMEN
The commensal microbiota impacts specific immune cell populations and their functions at peripheral sites, such as gut mucosal tissues. However, it remains unknown whether gut microbiota control immunity through regulation of hematopoiesis at primary immune sites. We reveal that germ-free mice display reduced proportions and differentiation potential of specific myeloid cell progenitors of both yolk sac and bone marrow origin. Homeostatic innate immune defects may lead to impaired early responses to pathogens. Indeed, following systemic infection with Listeria monocytogenes, germ-free and oral-antibiotic-treated mice display increased pathogen burden and acute death. Recolonization of germ-free mice with a complex microbiota restores defects in myelopoiesis and resistance to Listeria. These findings reveal that gut bacteria direct innate immune cell development via promoting hematopoiesis, contributing to our appreciation of the deep evolutionary connection between mammals and their microbiota.
Asunto(s)
Tracto Gastrointestinal/microbiología , Hematopoyesis , Listeriosis/inmunología , Microbiota/inmunología , Animales , Carga Bacteriana , Vida Libre de Gérmenes , Ratones , Análisis de SupervivenciaRESUMEN
The discovery that microorganisms can be etiologic agents of disease has driven clinical, research and public health efforts to reduce exposure to bacteria. However, despite extensive campaigns to eradicate pathogens (via antibiotics, vaccinations, hygiene, sanitation, etc.), the incidence and/or severity of multiple immune-mediated diseases including, paradoxically, infectious disease have increased in recent decades. We now appreciate that most microbes in our environment are not pathogenic, and that many human-associated bacteria are symbiotic or beneficial. Notably, recent examples have emerged revealing that the microbiome augments immune system function. This review will focus on how commensal-derived signals enhance various aspects of the host response against pathogens. We suggest that modern lifestyle advances may be depleting specific microbes that enhance immunity against pathogens. Validation of the notion that absence of beneficial microbes is a risk factor for infectious disease may have broad implications for future medical practices.
Asunto(s)
Biota , Enfermedades Gastrointestinales/microbiología , Tracto Gastrointestinal/microbiología , Metagenoma/efectos de los fármacos , Interacciones Huésped-Patógeno , Humanos , Modelos Biológicos , Factores de Riesgo , SimbiosisRESUMEN
All animals live in symbiosis. Shaped by eons of co-evolution, host-bacterial associations have developed into prosperous relationships creating mechanisms for mutual benefits to both microbe and host. No better example exists in biology than the astounding numbers of bacteria harbored by the lower gastrointestinal tract of mammals. The mammalian gut represents a complex ecosystem consisting of an extraordinary number of resident commensal bacteria existing in homeostasis with the host's immune system. Most impressive about this relationship may be the concept that the host not only tolerates, but has evolved to require colonization by beneficial microorganisms, known as commensals, for various aspects of immune development and function. The microbiota provides critical signals that promote maturation of immune cells and tissues, leading to protection from infections by pathogens. Gut bacteria also appear to contribute to non-infectious immune disorders such as inflammatory bowel disease and autoimmunity. How the microbiota influences host immune responses is an active area of research with important implications for human health. This review synthesizes emerging findings and concepts that describe the mutualism between the microbiota and mammals, specifically emphasizing the role of gut bacteria in shaping an immune response that mediates the balance between health and disease. Unlocking how beneficial bacteria affect the development of the immune system may lead to novel and natural therapies based on harnessing the immunomodulatory properties of the microbiota.
Asunto(s)
Inmunidad Mucosa/inmunología , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Simbiosis/inmunología , Animales , Humanos , Tolerancia InmunológicaRESUMEN
In humans, Streptococcus pneumoniae (SPN) is the leading cause of bacterial meningitis, a disease with high attributable mortality and frequent permanent neurological sequelae. The molecular mechanisms underlying the central nervous system tropism of SPN are incompletely understood, but include a primary interaction of the pathogen with the blood-brain barrier (BBB) endothelium. All SPN strains possess a gene encoding the surface-anchored sialidase (neuraminidase) NanA, which cleaves sialic acid on host cells and proteins. Here, we use an isogenic SPN NanA-deficient mutant and heterologous expression of the protein to show that NanA is both necessary and sufficient to promote SPN adherence to and invasion of human brain microvascular endothelial cells (hBMECs). NanA-mediated hBMEC invasion depends only partially on sialidase activity, whereas the N-terminal lectinlike domain of the protein plays a critical role. NanA promotes SPN-BBB interaction in a murine infection model, identifying the protein as proximal mediator of CNS entry by the pathogen.
Asunto(s)
Barrera Hematoencefálica/microbiología , Células Endoteliales/metabolismo , Meningitis Bacterianas/metabolismo , Neuraminidasa/metabolismo , Streptococcus pneumoniae/metabolismo , Animales , Adhesión Celular/efectos de los fármacos , Adhesión Celular/fisiología , Cartilla de ADN/genética , Humanos , Masculino , Meningitis Bacterianas/genética , Meningitis Bacterianas/microbiología , Ratones , Ratones Endogámicos BALB C , Ácido N-Acetilneuramínico/análogos & derivados , Ácido N-Acetilneuramínico/farmacología , Neuraminidasa/antagonistas & inhibidores , Neuraminidasa/genética , Plásmidos/genética , Streptococcus pneumoniae/genéticaRESUMEN
Group A Streptococcus (GAS) is a leading human pathogen associated with a wide spectrum of mucosal and invasive infections. GAS expresses a large number of virulence determinants whose expression is under the control of several transcriptional regulatory networks. Here we performed the first mutational analysis of a genetic locus immediately upstream of the streptolysin S biosynthetic operon in several GAS genome sequences, including that of the M1T1 serotype, the leading isolates associated with serious invasive disease. The locus consists of a predicted RofA-like stand-alone transcriptional regulator (RALP3) and the largest open reading frame in the GAS genome, encoding a predicted LPXSG motif cell wall-anchored protein we have named LSA (for "large surface-anchored" protein). Comparative reverse transcription-PCR analysis of wild-type M1T1 GAS and an isogenic RALP3-deficient mutant identifies RALP3 as a global transcriptional regulator affecting expression of numerous virulence factor genes, including those for strong repression of the hyaluronic acid capsule and cysteine protease production. RALP3 contributed to GAS epithelial cell invasion and bloodstream survival. LSA was found to be under negative regulation by RALP3 and to influence GAS-epithelial cell interactions and GAS antimicrobial peptide sensitivity. Isogenic M1T1 GAS mutants lacking either RALP3 or LSA were attenuated in a murine model of systemic infection, indicating that this locus plays a role in the virulence potential of the organism.
Asunto(s)
Proteínas Bacterianas/metabolismo , Operón/genética , Streptococcus pyogenes/genética , Streptococcus pyogenes/patogenicidad , Estreptolisinas/metabolismo , Animales , Cápsulas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Línea Celular , Supervivencia Celular , Medios de Cultivo , Cisteína Endopeptidasas/metabolismo , Femenino , Humanos , Ácido Hialurónico/metabolismo , Ratones , Estreptolisinas/genética , VirulenciaRESUMEN
Responsible for many childhood diseases, group A Streptococcus (GAS) is a rare cause of central nervous system infections. We report the case of a previously healthy boy with brain abscesses caused by M/emm type 12 GAS and review the case in the context of the published literature and recent epidemiological data.