RESUMO
Child undernutrition is a global health issue associated with a high burden of infectious disease. Undernourished children display an overabundance of intestinal pathogens and pathobionts, and these bacteria induce enteric dysfunction in undernourished mice; however, the cause of their overgrowth remains poorly defined. Here, we show that disease-inducing human isolates of Enterobacteriaceae and Bacteroidales spp. are capable of multi-species symbiotic cross-feeding, resulting in synergistic growth of a mixed community in vitro. Growth synergy occurs uniquely under malnourished conditions limited in protein and iron: in this context, Bacteroidales spp. liberate diet- and mucin-derived sugars and Enterobacteriaceae spp. enhance the bioavailability of iron. Analysis of human microbiota datasets reveals that Bacteroidaceae and Enterobacteriaceae are strongly correlated in undernourished children, but not in adequately nourished children, consistent with a diet-dependent growth synergy in the human gut. Together these data suggest that dietary cross-feeding fuels the overgrowth of pathobionts in undernutrition.
Assuntos
Microbioma Gastrointestinal/fisiologia , Desnutrição/microbiologia , Animais , Bacteroidetes/crescimento & desenvolvimento , Bacteroidetes/isolamento & purificação , Bacteroidetes/metabolismo , Criança , Técnicas de Cocultura , Dieta/efeitos adversos , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/metabolismo , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Escherichia coli/metabolismo , Fezes/microbiologia , Humanos , Intestinos/química , Intestinos/microbiologia , Camundongos , Nutrientes/análise , Nutrientes/metabolismo , SimbioseRESUMO
The gram-negative pathogen Shigella flexneri causes bacillary dysentery, an invasive disease of the human colonic mucosa. A major characteristic of the infectious process is the occurrence of an acute inflammatory reaction of mucosal tissues which is generally consequence of primary invasion and destruction of colonic epithelial cells by the pathogen. Confirming in vitro demonstration that S. flexneri is unable to invade the apical pole of colonic cells and that polymorphonuclear (PMN) cells may assist them in reaching the basal side of epithelial cells where they can invade, we have provided here in vivo evidence that S. flexneri enters the epithelial barrier essentially through the dome of lymphoid follicles at the early stage of infection and that subsequent invasion and destruction of the epithelium is primarily due to immigration of leukocytes, particularly PMN that destroy cohesion of the epithelial barrier. These conclusions are based on experiments carried out in infected rabbit ligated intestinal loops, with some animals treated by an anti-CD18 monoclonal antibody that blocked immigration of leukocytes into infected tissues.
Assuntos
Disenteria Bacilar/patologia , Inflamação/patologia , Mucosa Intestinal/patologia , Doença Aguda , Animais , Anticorpos Monoclonais/imunologia , Antígenos CD18/fisiologia , Epitélio/patologia , Mucosa Intestinal/ultraestrutura , Contagem de Leucócitos , Coelhos , Fator de Necrose Tumoral alfa/análiseRESUMO
To determine the role of humoral mucosal immune response in protection against shigellosis, we have obtained a monoclonal dimeric immunoglobulin A (IgA) antibody specific for Shigella flexneri serotype 5a lipopolysaccharide (mIgA) and used a murine pulmonary infection model that mimics the lesions occurring in natural intestinal infection. Adult BALB/c mice challenged with 10(7) S. flexneri organisms developed a rapid inflammatory response characterized by polymorphonuclear cell infiltration around and within the bronchi and strong systemic interleukin 6 response. Implantation of hybridoma cells in the back of mice, resulting in the development of a myeloma tumor producing mIgA in the serum and subsequently secretory mIgA in local secretions, or direct intranasal administration of these antibodies, protected the animals against subsequent intranasal challenge with S. flexneri serotype 5a. Absence of histopathological lesion and significant decrease in bacterial load of the lungs and of systemic interleukin 6 response were the three major criteria of protection. This protection was shown to be serotype-specific and dependent on local concentration of mIgA. These data demonstrate that mucosal antibodies directed against a single polysaccharidic surface epitope of Shigella can protect against the disease.
Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Disenteria Bacilar/prevenção & controle , Epitopos/imunologia , Imunização Passiva , Imunoglobulina A/imunologia , Lipopolissacarídeos/imunologia , Pneumonia Bacteriana/prevenção & controle , Shigella flexneri/imunologia , Administração Intranasal , Animais , Anticorpos Antibacterianos/administração & dosagem , Anticorpos Monoclonais/administração & dosagem , Modelos Animais de Doenças , Disenteria Bacilar/sangue , Disenteria Bacilar/microbiologia , Disenteria Bacilar/patologia , Feminino , Hibridomas/imunologia , Hibridomas/transplante , Imunoglobulina A/administração & dosagem , Interleucina-6/sangue , Pulmão/microbiologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Pneumonia Bacteriana/sangue , Pneumonia Bacteriana/microbiologia , Pneumonia Bacteriana/patologia , Sorotipagem , Shigella flexneri/classificação , Especificidade da EspécieRESUMO
Shigella flexneri is an enteroinvasive bacterium which causes bacillary dysentery in humans. A major feature of its pathogenic potential is the capacity to invade epithelial cells. Shigella entry into epithelial cells is considered a parasite-induced internalization process requiring polymerization of actin. Here we describe the cytoskeletal rearrangements during S. flexneri invasion of HeLa cells. After an initial contact of the bacterium with the cell surface, distinct nucleation zones of heavy chain actin polymerization appear in close proximity to the contact site underneath the parasite with long filaments being polymerized. These structures then push cellular protrusions that rise beside the entering bacterium, being sustained by tightly bundled long actin filaments organized in parallel orientation with their positive ends pointing to the cytoplasmic membrane. Finally, the cellular projections coalesce above the bacterial body, leading to its internalization. In addition, we found the actin-bundling protein plastin to be concentrated in these protrusions. Since plastin is known to bundle actin filaments in parallel orientation, colocalization of parallel actin filaments and plastin in the cellular protrusions strongly suggested a functional role of this protein in the architecture of parasite-induced cellular projections. Using transfection experiments, we show the differential recruitment of the two plastin isoforms (T- and L-) into Shigella entry zones. By transient expression of a truncated T-plastin which is deprived of one of its actin-binding sites, we also demonstrate the functional role of T-plastin in Shigella entry into HeLa cells.
Assuntos
Citoesqueleto/metabolismo , Proteínas de Neoplasias/fisiologia , Fosfoproteínas/metabolismo , Shigella flexneri/fisiologia , Actinas/metabolismo , Membrana Celular/microbiologia , Membrana Celular/ultraestrutura , Epitélio/microbiologia , Células HeLa , Humanos , Glicoproteínas de Membrana , Proteínas dos Microfilamentos , Proteínas de Neoplasias/genética , Fosfoproteínas/genética , Proteínas Recombinantes de Fusão/metabolismo , Shigella flexneri/ultraestruturaRESUMO
Bacterial type III secretion systems serve to translocate proteins into eukaryotic cells, requiring a secreton and a translocator for proteins to pass the bacterial and host membranes. We used the contact hemolytic activity of Shigella flexneri to investigate its putative translocator. Hemolysis was caused by formation of a 25-A pore within the red blood cell (RBC) membrane. Of the five proteins secreted by Shigella upon activation of its type III secretion system, only the hydrophobic IpaB and IpaC were tightly associated with RBC membranes isolated after hemolysis. Ipa protein secretion and hemolysis were kinetically coupled processes. However, Ipa protein secretion in the immediate vicinity of RBCs was not sufficient to cause hemolysis in the absence of centrifugation. Centrifugation reduced the distance between bacterial and RBC membranes beyond a critical threshold. Electron microscopy analysis indicated that secretons were constitutively assembled at 37 degrees C before any host contact. They were composed of three parts: (a) an external needle, (b) a neck domain, and (c) a large proximal bulb. Secreton morphology did not change upon activation of secretion. In mutants of some genes encoding the secretion machinery the organelle was absent, whereas ipaB and ipaC mutants displayed normal secretons.
Assuntos
Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Membrana Eritrocítica/metabolismo , Eritrócitos/microbiologia , Hemólise , Shigella flexneri/metabolismo , Animais , Antígenos de Bactérias/genética , Azidas/farmacologia , Proteínas de Bactérias/genética , Centrifugação , Vermelho Congo/farmacologia , Endopeptidase K/metabolismo , Membrana Eritrocítica/microbiologia , Membrana Eritrocítica/ultraestrutura , Eritrócitos/citologia , Eritrócitos/metabolismo , Eritrócitos/ultraestrutura , Genes Bacterianos , Hemólise/efeitos dos fármacos , Humanos , Microscopia Eletrônica , Estrutura Molecular , Mutação , Concentração Osmolar , Ovinos , Shigella flexneri/química , Shigella flexneri/patogenicidade , Shigella flexneri/ultraestrutura , TemperaturaRESUMO
To propel itself in infected cells, the pathogen Shigella flexneri subverts the Cdc42-controlled machinery responsible for actin assembly during filopodia formation. Using a combination of bacterial motility assays in platelet extracts with Escherichia coli expressing the Shigella IcsA protein and in vitro analysis of reconstituted systems from purified proteins, we show here that the bacterial protein IcsA binds N-WASP and activates it in a Cdc42-like fashion. Dramatic stimulation of actin assembly is linked to the formation of a ternary IcsA-N-WASP-Arp2/3 complex, which nucleates actin polymerization. The Arp2/3 complex is essential in initiation of actin assembly and Shigella movement, as previously observed for Listeria monocytogenes. Activation of N-WASP by IcsA unmasks two domains acting together in insertional actin polymerization. The isolated COOH-terminal domain of N-WASP containing a verprolin-homology region, a cofilin-homology sequence, and an acidic terminal segment (VCA) interacts with G-actin in a unique profilin-like functional fashion. Hence, when N-WASP is activated, its COOH-terminal domain feeds barbed end growth of filaments and lowers the critical concentration at the bacterial surface. On the other hand, the NH(2)-terminal domain of N-WASP interacts with F-actin, mediating the attachment of the actin tail to the bacterium surface. VASP is not involved in Shigella movement, and the function of profilin does not require its binding to proline-rich regions.
Assuntos
Actinas/metabolismo , Proteínas de Ciclo Celular/metabolismo , Proteínas Contráteis , Proteínas do Citoesqueleto , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação ao GTP/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Fatores de Transcrição/metabolismo , Proteína 2 Relacionada a Actina , Proteína 3 Relacionada a Actina , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Plaquetas/citologia , Plaquetas/metabolismo , Encéfalo/citologia , Encéfalo/metabolismo , Bovinos , Moléculas de Adesão Celular/metabolismo , Proteínas de Ligação a DNA/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Escherichia coli/fisiologia , Células HeLa , Humanos , Listeria/fisiologia , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Movimento , Mutação , Proteínas do Tecido Nervoso/química , Fosfoproteínas/metabolismo , Polímeros , Profilinas , Prolina/metabolismo , Shigella flexneri/genética , Shigella flexneri/fisiologia , Fatores de Transcrição/genética , Proteína Neuronal da Síndrome de Wiskott-Aldrich , Proteína cdc42 de Saccharomyces cerevisiae de Ligação ao GTPRESUMO
Bacterial lipopolysaccharide (LPS) at the apical surface of polarized intestinal epithelial cells was previously shown to be transported from the apical to the basolateral pole of the epithelium (Beatty, W.L., and P.J. Sansonetti. 1997. Infect. Immun. 65:4395-4404). The present study was designed to elucidate the transcytotic pathway of LPS and to characterize the endocytic compartments involved in this process. Confocal and electron microscopic analyses revealed that LPS internalized at the apical surface became rapidly distributed within endosomal compartments accessible to basolaterally internalized transferrin. This compartment largely excluded fluid-phase markers added at either pole. Access to the basolateral side of the epithelium subsequent to trafficking to basolateral endosomes occurred via exocytosis into the paracellular space beneath the intercellular tight junctions. LPS appeared to exploit other endocytic routes with much of the internalized LPS recycled to the original apical membrane. In addition, analysis of LPS in association with markers of the endocytic network revealed that some LPS was sent to late endosomal and lysosomal compartments.
Assuntos
Mucosa Intestinal/metabolismo , Lipopolissacarídeos/metabolismo , Shigella flexneri/metabolismo , Biomarcadores , Polaridade Celular , Endossomos/metabolismo , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/microbiologia , Lipopolissacarídeos/farmacologia , Lisossomos/metabolismoRESUMO
Shigella flexneri, the causative agent of bacillary dysentery, has the ability to enter nonphagocytic cells. The interferon (IFN) family of cytokines was found to inhibit Shigella invasion of cultured epithelial cells. We show here that IFN-alpha inhibits a Src-dependent signaling cascade triggered by Shigella that leads to the reorganization of the host cell cytoskeleton. Immunofluorescence studies showed that IFN-alpha inhibits Shigella-induced actin polymerization required for bacterial entry into cells. Phosphorylation of cortactin, a Src-substrate specifically tyrosyl-phosphorylated during Shigella entry, was inhibited by IFN-alpha. Overexpression of a dominant interfering form of pp60c-src led to inhibition of Shigella-induced cytoskeletal rearrangements and decreased cortactin phosphorylation indicating a role for Src in Shigella entry. Also, Shigella uptake in cells that expressed constitutively active Src was unaffected by IFN-alpha treatment. We conclude that Src kinase activity is necessary for Shigella invasion of epithelial cells and that IFN-alpha inhibits this Src-dependent signaling pathway.
Assuntos
Citoesqueleto/metabolismo , Células Epiteliais/microbiologia , Interferon-alfa/metabolismo , Proteínas Proto-Oncogênicas pp60(c-src)/genética , Shigella flexneri/metabolismo , Actinas/metabolismo , Cortactina , Células Epiteliais/metabolismo , Células HeLa , Humanos , Proteínas dos Microfilamentos/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas pp60(c-src)/metabolismo , Transdução de Sinais/fisiologia , Tirosina/metabolismoRESUMO
In vivo and in vitro, Shigella flexneri, an invasive pathogen of the human colon, cannot invade epithelial cells through their apical pole. To identify ways by which it may reach the cellular basolateral domain in order to invade, we have established an assay using the human colonic T-84 cell line grown on permeable filters. Human PMN were added to the basal pole of the cells, and invasive shigellae to their apical pole. Apical addition of bacteria induced strong transmigration of PMN, reaching a maximum after 1 h of incubation. Transmigration depended on a receptor-specific interaction since it was inhibited by an anti-CD18 monoclonal antibody that antagonizes binding of MAC1 on its putative epithelial cell receptor. After 1 h of PMN transmigration, shigellae started to invade the monolayer in areas of intense PMN infiltration. Invasion was clearly dependant on PMN transmigration since it was also inhibited by addition of an anti-CD18 monoclonal antibody. This in vitro assay is consistent with in vivo observations showing early PMN efflux within colonic crypts in the course of shigellosis. PMN transmigration may therefore allow invasion in the colon by opening the paracellular pathway to invasive microorganisms.
Assuntos
Colo/fisiologia , Neutrófilos/fisiologia , Shigella flexneri/fisiologia , Shigella flexneri/patogenicidade , Anticorpos Monoclonais/farmacologia , Antígenos CD/imunologia , Antígenos CD/fisiologia , Antígenos CD18 , Linhagem Celular , Movimento Celular , Colo/microbiologia , Colo/ultraestrutura , Epitélio/microbiologia , Epitélio/fisiologia , Epitélio/ultraestrutura , Feminino , Humanos , Masculino , Microscopia Eletrônica , Modelos Biológicos , Shigella flexneri/ultraestruturaRESUMO
Peritoneal macrophages undergoing apoptosis induced by Shigella flexneri infection release the inflammatory cytokine interleukin 1 (IL-1), but not IL-6 or tumor necrosis factor alpha (TNF alpha). Wild type shigella causes a very fast and significant release of IL-1 from prestimulated peritoneal macrophages, before the cell's integrity is compromised. Both IL-1 alpha and IL-1 beta are released, IL-1 beta in its mature processed form. IL-1 is released from presynthesized cytoplasmic pools. These results demonstrate that bacteria-induced apoptosis of macrophages may play an active role in vivo by releasing IL-1, which in turn mediates an early inflammatory response in epithelial tissues.
Assuntos
Apoptose , Interleucina-1/biossíntese , Macrófagos Peritoneais/imunologia , Shigella flexneri/patogenicidade , Animais , Bioensaio , Células Cultivadas , Escherichia coli , Interleucina-1/análise , Interleucina-1/metabolismo , Interleucina-6/análise , Interleucina-6/biossíntese , Interleucina-6/metabolismo , Lipopolissacarídeos/toxicidade , Macrófagos Peritoneais/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos C3HRESUMO
The effect of human recombinant interleukin-1 receptor antagonist on intestinal inflammation, tissue destruction, and bacterial invasion during experimental shigellosis caused by Shigella flexneri was studied in the rabbit-ligated loop infection model. Intravenous infusion of the inhibitor at a dose of 2 mg/kg per h, was initiated 30 min before intestinal loops were ligated and infected, and continued during the 8-h period of infection. The animals treated with IL-1 receptor antagonist showed a striking decrease in inflammation, destruction, and bacterial invasion of their tissues, both at the level of the villous intestine and Peyer's patches. This is conclusive evidence that interleukin-1 plays a critical role in the pathogenesis of shigellosis. This proinflammatory cytokine is here proposed as a major trigger of the inflammatory reaction which is characteristic of this invasive disease of the intestine, due to the particular interaction existing between S. flexneri and macrophages.
Assuntos
Disenteria Bacilar/etiologia , Interleucina-1/fisiologia , Shigella flexneri , Animais , Atrofia , Disenteria Bacilar/imunologia , Disenteria Bacilar/patologia , Exsudatos e Transudatos , Humanos , Inflamação , Proteína Antagonista do Receptor de Interleucina 1 , Interleucina-1/antagonistas & inibidores , Mucosa Intestinal/patologia , Contagem de Leucócitos , Macrófagos/fisiologia , Neutrófilos , Perfusão , Nódulos Linfáticos Agregados/imunologia , Coelhos , Sialoglicoproteínas/farmacologia , Fator de Necrose Tumoral alfa/análise , Úlcera/patologiaRESUMO
The ability to discriminate between pathogenic and non-pathogenic bacteria is extremely important for epithelial cells lining mucosal surfaces and is particularly so in colonic epithelial cells. Accumulating evidence suggests that bacterial recognition systems used by epithelial cells are very different from those in cells of the myeloid lineage and are likely to have developed to maintain mucosal surfaces in a state of homeostasis with the normal microbial flora. Bacterial invasion of epithelial cells or breach of the epithelial barrier provides a signal to epithelial cells to initiate inflammatory responses, which are key events for the clearance of the infecting microbe. Therefore, elucidation of the mechanisms by which epithelial cells recognize bacteria and bacterial products, and of the nature of the innate immune responses that are triggered by these factors are important for our understanding of both the immunology of mucosal surfaces and bacterial pathogenesis.
Assuntos
Infecções Bacterianas/imunologia , Animais , Apoptose , Bactérias/imunologia , Bactérias/patogenicidade , Infecções Bacterianas/genética , Infecções Bacterianas/microbiologia , Infecções Bacterianas/patologia , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Expressão Gênica , Humanos , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Modelos Biológicos , Transdução de SinaisRESUMO
Several changes in cell morphology take place during the capping of surface receptors in Entamoeba histolytica. The amoebae develop the uroid, an appendage formed by membrane invaginations, which accumulates ligand-receptor complexes resulting from the capping process. Membrane shedding is particularly active in the uroid region and leads to the elimination of accumulated ligands. This appendage has been postulated to participate in parasitic defense mechanisms against the host immune response, because it eliminates complement and specific antibodies bound to the amoeba surface. The involvement of myosin II in the capping process of surface receptors has been suggested by experiments showing that drugs that affect myosin II heavy-chain phosphorylation prevent this activity. To understand the role of this mechanoenzyme in surface receptor capping, a myosin II dominant negative strain was constructed. This mutant is the first genetically engineered cytoskeleton-deficient strain of E. histolytica. It was obtained by overexpressing the light meromyosin domain, which is essential for myosin II filament formation. E. histolytica overexpressing light meromyosin domain displayed a myosin II null phenotype characterized by abnormal movement, failure to form the uroid, and failure to undergo the capping process after treatment with concanavalin A. In addition, the amoebic cytotoxic capacities of the transfectants on human colon cells was dramatically reduced, indicating a role for cytoskeleton in parasite pathogenicity.
Assuntos
Entamoeba histolytica/metabolismo , Entamoeba histolytica/patogenicidade , Subfragmentos de Miosina/metabolismo , Miosinas/fisiologia , Animais , Antígenos Glicosídicos Associados a Tumores/metabolismo , Células CACO-2 , Sobrevivência Celular , Entamoeba histolytica/genética , Humanos , Mutagênese , Miosinas/genética , Fenótipo , Receptores de Superfície Celular/metabolismo , Transfecção , VirulênciaAssuntos
Vacinas Bacterianas , Diarreia/prevenção & controle , Vacinas Protozoárias , Vacinas Virais , Adulto , Animais , Criança , Pré-Escolar , Cólera/prevenção & controle , Países em Desenvolvimento , Diarreia/etiologia , Entamoeba histolytica/imunologia , Entamebíase/prevenção & controle , Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/prevenção & controle , Saúde Global , Humanos , Lactente , Rotavirus/imunologia , Infecções por Rotavirus/prevenção & controle , Vibrio cholerae/imunologiaRESUMO
Shigella, the causative agent of bacillary dysentery, invades epithelial cells by reorganizing the cell cytoskeleton during bacterial entry. This entry process requires the Shigella Ipa proteins that are secreted by a type III secretion apparatus and that act in concert to fine tune cell responses. Actin polymerization at the site of entry is dependent on the IpaB and IpaC proteins, whereas IpaA further modulates cytoskeletal rearrangements by binding to vinculin.
Assuntos
Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Células Epiteliais/microbiologia , Shigella/fisiologia , Animais , Apoptose , Células CHO , Cricetinae , Citoesqueleto/fisiologia , Células Epiteliais/citologia , GTP Fosfo-Hidrolases/metabolismo , Humanos , Receptores de Hialuronatos/metabolismo , Integrinas/metabolismo , Shigella/patogenicidade , Vinculina/metabolismo , VirulênciaRESUMO
Infection of cells by some pathogenic bacteria triggers host cell apoptosis. Bacteria-induced apoptosis appears to promote an inflammatory response that causes tissue damage and further bacterial colonization. Shigella pathogenesis offers a paradigm for the role of apoptosis in bacterial infections.
Assuntos
Apoptose/imunologia , Disenteria Bacilar/imunologia , Inflamação/fisiopatologia , Infecções Bacterianas/imunologia , Disenteria Bacilar/epidemiologia , Humanos , MacrófagosRESUMO
Shigella flexneri is a model for the entry of bacterial pathogens into nonphagocytic epithelial cells. On contact with the epithelial cell surface, the Ipa proteins are secreted from the bacterium. The Ipa complex then triggers a reorganization of the host-cell cytoskeleton leading to the formation of membrane ruffles, which engulf the bacterium.
Assuntos
Shigella flexneri/patogenicidade , Aderência Bacteriana/genética , Aderência Bacteriana/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Citoesqueleto/microbiologia , Disenteria Bacilar/etiologia , Disenteria Bacilar/microbiologia , Epitélio/microbiologia , Genes Bacterianos , Células HeLa , Humanos , Chaperonas Moleculares/genética , Chaperonas Moleculares/fisiologia , Plasmídeos/genética , Shigella flexneri/genética , Shigella flexneri/fisiologia , Transdução de Sinais , Virulência/genéticaRESUMO
Acute infectious colitis remains a major pediatric issue of worldwide impact because it still represents a significant public health burden among the larger group of diarrheal diseases with the highest mortality rate. It is also a relevant model of inflammatory bowel diseases (IBD), such as Crohn's disease and ulcerative colitis. Among cases of acute colitis of infectious origin, shigellosis is certainly the one that has benefited the most from a significant research effort. Shigella, the causative agent, is a Gram-negative bacterium that has the capacity to invade, disrupt, and cause inflammatory destruction of the intestinal epithelial barrier. The molecular and cellular bases of this invasive phenotype essentially encompass crossing of the epithelial lining, apoptotic killing of macrophages, entry into epithelial cells, and escape into the cytoplasm, followed by cell-to-cell spread. Intracellular colonization is likely to protect the micro-organisms from killing by humoral and cellular effectors of the innate immune response. Concurrently, the capacity of Shigella to reprogram invaded epithelial cells to produce proinflammatory mediators plays a major role in the strong inflammatory profile of the disease. This profile is likely to impact on the nature and quality of the adaptive response, which is dominated by humoral protection at the mucosal level.
Assuntos
Disenteria Bacilar/imunologia , Vacinas contra Shigella/isolamento & purificação , Shigella/imunologia , Shigella/patogenicidade , Adaptação Fisiológica , Animais , Proteínas de Bactérias/imunologia , Células Dendríticas/microbiologia , Disenteria Bacilar/etiologia , Disenteria Bacilar/patologia , Disenteria Bacilar/prevenção & controle , Células Epiteliais/imunologia , Células Epiteliais/patologia , Humanos , Imunidade Inata , Imunidade nas Mucosas , Técnicas In Vitro , Intestinos/imunologia , Intestinos/patologia , Macrófagos/microbiologiaRESUMO
A randomized study of 12 treated patients and seven controls was conducted in order to evaluate HPA-23 anti-HIV activity in HIV-infected patients. The antiviral activity was assessed by determining HIV p24 antigenemia. A persistence or even increase in antigenemia was shown in treated patients and thrombocytopenia was observed in nine out of the 12 patients. This suggests that HPA-23 should not be used in anti-HIV therapy.
Assuntos
Antimônio/uso terapêutico , Antivirais/uso terapêutico , Produtos do Gene gag/sangue , Antígenos HIV/sangue , Infecções por HIV/tratamento farmacológico , Compostos de Tungstênio , Tungstênio/uso terapêutico , Proteínas do Core Viral/sangue , Antimônio/efeitos adversos , Antimônio/farmacologia , Antivirais/efeitos adversos , Antivirais/farmacologia , Avaliação de Medicamentos , Seguimentos , Proteína do Núcleo p24 do HIV , Infecções por HIV/sangue , Infecções por HIV/imunologia , Humanos , Contagem de Leucócitos , Contagem de Plaquetas , Distribuição Aleatória , Inibidores da Transcriptase Reversa , Trombocitopenia/induzido quimicamente , Tungstênio/efeitos adversos , Tungstênio/farmacologiaRESUMO
Shigella infections lead to severe inflammation associated with destruction of colonic mucosa. We assessed the effect of in vivo blockade of CD14 on the outcome of experimental Shigella infection in rabbits. A total of 17 rabbits were divided into two groups: 8 received a single i.v. dose of anti-rabbit CD14 monoclonal antibody prior to infection with an invasive Shigella flexneri strain; the remainder served as controls. The anti-CD14-treated rabbits exhibited more severe tissue destruction and a 50-fold increase in bacterial invasion of the intestinal mucosa when compared to controls. Similar numbers of polymorphonuclear leukocytes were recruited to the intestinal mucosa in both groups despite the massive bacterial invasion seen in the CD14-blocked group. No statistically significant differences were seen in levels of IL-1beta nor in the ratio of IL-1RA/IL-1beta for either group. In contrast, higher quantities of TNF-alpha were observed in the CD14-blocked group. To conclude, anti-CD14 treatment had a detrimental effect on the capacity of Shigella-infected animals to clear the infection.