RESUMO
CDC has updated the interim guidance for U.S. health care providers caring for pregnant women with possible Zika virus exposure in response to 1) declining prevalence of Zika virus disease in the World Health Organization's Region of the Americas (Americas) and 2) emerging evidence indicating prolonged detection of Zika virus immunoglobulin M (IgM) antibodies. Zika virus cases were first reported in the Americas during 2015-2016; however, the incidence of Zika virus disease has since declined. As the prevalence of Zika virus disease declines, the likelihood of false-positive test results increases. In addition, emerging epidemiologic and laboratory data indicate that, as is the case with other flaviviruses, Zika virus IgM antibodies can persist beyond 12 weeks after infection. Therefore, IgM test results cannot always reliably distinguish between an infection that occurred during the current pregnancy and one that occurred before the current pregnancy, particularly for women with possible Zika virus exposure before the current pregnancy. These limitations should be considered when counseling pregnant women about the risks and benefits of testing for Zika virus infection during pregnancy. This updated guidance emphasizes a shared decision-making model for testing and screening pregnant women, one in which patients and providers work together to make decisions about testing and care plans based on patient preferences and values, clinical judgment, and a balanced assessment of risks and expected outcomes.
Assuntos
Pessoal de Saúde , Guias de Prática Clínica como Assunto , Complicações Infecciosas na Gravidez/prevenção & controle , Infecção por Zika virus/prevenção & controle , Centers for Disease Control and Prevention, U.S. , Feminino , Humanos , Gravidez , Estados UnidosRESUMO
INTRODUCTION: Antimicrobial resistance threatens to undermine treatment of severe infection; new therapeutic strategies are urgently needed. Preclinical work shows that augmented passive immunotherapy with P4 peptide increases phagocytic activity and shows promise as a novel therapeutic strategy. Our aim was to determine ex vivo P4 activity in a target population of patients admitted to critical care with severe infection. METHODS: We prospectively recruited UK critical care unit patients with severe sepsis and observed clinical course (≥3 months postdischarge). Blood samples were taken in early (≤48âh postdiagnosis, nâ=â54), latent (7 days postdiagnosis, nâ=â39), and convalescent (3-6 months postdiagnosis, nâ=â18) phases of disease. The primary outcome measure was killing of opsonized Streptococcus pneumoniae by neutrophils with and without P4 peptide stimulation. We also used a flow cytometric whole blood phagocytosis assay to determine phagocyte association and oxidation of intraphagosomal reporter beads. RESULTS: P4 peptide increased neutrophil killing of opsonized pneumococci by 8.6% (confidence interval 6.35-10.76, Pâ<â0.001) in all phases of sepsis, independent of infection source and microbiological status. This represented a 54.9% increase in bacterial killing compared with unstimulated neutrophils (15.6%) in early phase samples. Similarly, P4 peptide treatment significantly increased neutrophil and monocyte intraphagosomal reporter bead association and oxidation, independent of infection source. CONCLUSIONS: We have extended preclinical work to demonstrate that P4 peptide significantly increases phagocytosis and bacterial killing in samples from a target patient population with severe sepsis. This study supports the rationale for augmented passive immunotherapy as a therapeutic strategy in severe sepsis.
Assuntos
Imunização Passiva/métodos , Oligopeptídeos/uso terapêutico , Sepse/tratamento farmacológico , Idoso , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Neutrófilos/imunologia , Neutrófilos/metabolismo , Fagócitos/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Estudos Prospectivos , Sepse/metabolismo , Streptococcus pneumoniae/imunologiaRESUMO
Bordetella pertussis (Bp) is the etiologic agent of pertussis (whooping cough), a highly communicable infection. Although pertussis is vaccine preventable, in recent years there has been increased incidence, despite high vaccine coverage. Possible reasons for the rise in cases include the following: Bp strain adaptation, waning vaccine immunity, increased surveillance, and improved clinical diagnostics. A pertussis outbreak impacted California (USA) in 2010; children and preadolescents were the most affected but the burden of disease fell mainly on infants. To identify protein biomarkers associated with this pertussis outbreak, we report a whole cellular protein characterization of six Bp isolates plus the pertussis acellular vaccine strain Bp Tohama I (T), utilizing gel-free proteomics-based mass spectrometry (MS). MS/MS tryptic peptide detection and protein database searching combined with western blot analysis revealed three Bp isolates in this study had markedly reduced detection of pertactin (Prn), a subunit of pertussis acellular vaccines. Additionally, antibody affinity capture technologies were implemented using anti-Bp T rabbit polyclonal antisera and whole cellular proteins to identify putative immunogens. Proteome profiling could shed light on pathogenesis and potentially lay the foundation for reduced infection transmission strategies and improved clinical diagnostics.
RESUMO
Pneumococcal surface adhesin A (PsaA) is a multifunctional lipoprotein known to bind nasopharyngeal epithelial cells, and is significantly involved in bacterial adherence and virulence. Identification of PsaA peptides that optimally bind human leucocyte antigen (HLA) and elicit a potent immune response would be of great importance to vaccine development. However, this is hindered by the multitude of HLA polymorphisms in humans. To identify the conserved immunodominant epitopes, we used an experimental dataset of 28 PsaA synthetic peptides and in silico methods to predict specific peptide-binding to HLA and murine MHC class II molecules. We also characterized spleen and cervical lymph node (CLN) -derived T helper (Th) lymphocyte cytokine responses to these peptides after Streptococcus pneumoniae strain EF3030 challenge in mice. Individual, yet overlapping, peptides 15 amino acids in length revealed residues of PsaA that consistently caused the highest interferon-γ, interleukin-2 (IL-2), IL-5 and IL-17 responses and proliferation as well as moderate IL-10 and IL-4 responses by ex vivo re-stimulated splenic and CLN CD4⺠T cells isolated from S. pneumoniae strain EF3030-challenged F1 (B6 × BALB/c) mice. In silico analysis revealed that peptides from PsaA may interact with a broad range of HLA-DP, -DQ and -DR alleles, due in part to regions lacking ß-turns and asparagine endopeptidase sites. These data suggest that Th cell peptides (7, 19, 20, 22, 23 and 24) screened for secondary structures and MHC class II peptide-binding affinities can elicit T helper cytokine and proliferative responses to PsaA peptides.
Assuntos
Proteínas de Bactérias/imunologia , Mapeamento de Epitopos , Epitopos de Linfócito T , Epitopos Imunodominantes , Infecções Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Proliferação de Células , Técnicas de Cocultura , Células Alimentadoras , Feminino , Antígenos HLA/imunologia , Antígenos HLA/metabolismo , Antígenos de Histocompatibilidade Classe II/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Humanos , Interferon gama/metabolismo , Interleucinas/metabolismo , Linfonodos/imunologia , Linfonodos/microbiologia , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos BALB C , Modelos Moleculares , Dados de Sequência Molecular , Infecções Pneumocócicas/metabolismo , Infecções Pneumocócicas/microbiologia , Ligação Proteica , Estrutura Secundária de Proteína , Baço/imunologia , Baço/microbiologia , Streptococcus pneumoniae/metabolismo , Linfócitos T Auxiliares-Indutores/microbiologiaRESUMO
New treatment strategies are urgently needed to overcome early mortality in acute bacterial infections. Previous studies have shown that administration of a novel immunoactivating peptide (P4) alongside passive immunotherapy prevents the onset of septicemia and rescues mice from lethal invasive disease models of pneumococcal pneumonia and sepsis. In this study, using two diverse populations of adult volunteers, we determined whether P4 treatment of human alveolar macrophages would upregulate phagocytic killing of Streptococcus pneumoniae ex vivo. We also measured macrophage intracellular oxidation, cytokine secretion, and surface marker expression following stimulation. Peptide treatment showed enhanced bacterial killing in the absence of nonspecific inflammation, consistent with therapeutic potential. This is the first demonstration of P4 efficacy on ex vivo-derived human lung cells.
Assuntos
Macrófagos Alveolares/efeitos dos fármacos , Oligopeptídeos/farmacologia , Fagocitose/efeitos dos fármacos , Adulto , Biomarcadores/metabolismo , Células Cultivadas , Citocinas/biossíntese , Feminino , Expressão Gênica , Voluntários Saudáveis , Humanos , Macrófagos Alveolares/citologia , Macrófagos Alveolares/imunologia , Malaui , Masculino , Oxirredução , Fagocitose/imunologia , Streptococcus pneumoniae/crescimento & desenvolvimento , Reino UnidoRESUMO
New treatments against severe bacterial infections are needed because the response to antibiotic treatment is slow in acute settings and is becoming less effective owing to the emergence of antibiotic-resistant pathogens. P4-mediated antibody therapy offers a unique treatment strategy that combines exogenous immunoglobulin with the immunoactivating peptide P4. In an acute model of pneumococcal disease, mice were infected with Streptococcus pneumoniae and treated intravenously or intranasally with P4 and intravenous immunoglobulin (IVIG). Survival of P4-IVIG-treated mice increased from 0% to 60% among those that received intravenous treatment and from 0% to 100% among those that received intranasal treatment. Importantly, intranasal administration of P4 at an early stage of infection prevented the onset of bacteremia and sepsis. Increased survival was associated with reduced bacterial burden in affected tissues and with recruitment and activation of professional phagocytes, as manifested by increased expression of Fc-γ receptors. In vitro studies involving P4-stimulated alveolar, peritoneal, and J774.2 murine macrophages showed an increased ability of these immune cells to phagocytose pneumococci independent of capsule. The use of adjunct antibody therapies to treat infectious diseases shows promise.
Assuntos
Imunização Passiva/métodos , Imunoglobulinas Intravenosas/uso terapêutico , Fatores Imunológicos/uso terapêutico , Infecções Pneumocócicas/tratamento farmacológico , Doença Aguda , Animais , Antibacterianos/uso terapêutico , Anticorpos Antibacterianos/imunologia , Bacteriemia/imunologia , Bacteriemia/prevenção & controle , Linhagem Celular , Modelos Animais de Doenças , Feminino , Citometria de Fluxo , Imunoglobulinas Intravenosas/imunologia , Fatores Imunológicos/imunologia , Camundongos , Fagócitos/imunologia , Infecções Pneumocócicas/imunologia , Sepse/imunologia , Sepse/prevenção & controleRESUMO
During the 2009 influenza pandemic, the Centers for Disease Control and Prevention provided antiviral susceptibility testing for patients infected with suspected drug-resistant viruses. Specimens from 72 patients admitted to an intensive care unit or with a severe immunocompromising condition, who failed to clinically improve after oseltamivir treatment, were accepted for testing. Respiratory specimens were tested for the presence of the oseltamivir resistance-conferring H275Y substitution in the neuraminidase (NA) by pyrosequencing. Virus isolates propagated in MDCK cells were tested in phenotypic NA inhibition (NI) assays using licensed NA inhibitors (NAIs), zanamivir and oseltamivir, and investigational NAIs, peramivir and laninamivir. Conventional sequencing and plaque purification were conducted on a subset of viruses. Pyrosequencing data were obtained for 87 specimens collected from 58 of the 72 (81%) patients. Of all patients, 27 (38%) had at least one specimen in which H275Y was detected. Analysis of sequential samples from nine patients revealed intra-treatment emergence of H275Y variant and a shift from wildtype-to-H275Y in quasispecies during oseltamivir therapy. A shift in the H275Y proportion was observed as a result of virus propagation in MDCK cells. Overall, the NI method was less sensitive than pyrosequencing in detecting the presence of H275Y variants in virus isolates. Using the NI method, isolates containing H275Y variant at⩾50% exhibited resistance to oseltamivir and peramivir, but retained full susceptibility to zanamivir. H275Y viruses recovered from two patients had an additional substitution I223K or I223R that conferred a 38-52- and 33-97-fold enhancement in oseltamivir- and peramivir-resistance, respectively. These viruses also showed decreased susceptibility to zanamivir and laninamivir. These data suggest that pyrosequencing is a powerful tool for timely detection of NAI resistant viruses and that NI assays are needed for comprehensive testing to detect novel resistance substitutions.
Assuntos
Antivirais/farmacologia , Farmacorresistência Viral , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/virologia , Oseltamivir/farmacologia , Animais , Linhagem Celular , Humanos , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A Subtipo H1N1/enzimologia , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/epidemiologia , Mutação de Sentido Incorreto , Neuraminidase/genética , Neuraminidase/metabolismo , Pandemias , Estados Unidos/epidemiologia , Proteínas Virais/genética , Proteínas Virais/metabolismoRESUMO
Mass spectrometry (MS) coupled with 1-D and 2-D electrophoresis can be utilized to detect and identify immunogenic proteins, but these methods are laborious and time-consuming. We describe an alternative, simple, rapid gel-free strategy to identify multiple immunogenic proteins from Bordetella pertussis (Bp). It couples immunoprecipitation to nano liquid chromatography- tandem mass spectrometry (IP-nLC-MS/MS) and is significantly both time- and labor-saving. We developed a gel-free magnetic bead-based immunoprecipitation (IP) method using different NP-40/PBS concentrations in which solubilized proteins of Bp Tohama I membrane fractions were precipitated with polyclonal rabbit anti-Bp whole cell immune sera. Immune complexes were analyzed by MS and Scaffold analysis (>95% protein identification probability). Total immunoproteins identified were 50, 63 and 49 for 0.90%, 0.45% and 0.22% NP-40/PBS buffer concentrations respectively. Known Bp proteins identified included pertactin, serotype 2 fimbrial subunit and filamentous hemagglutinin. As proof of concept that this gel-free protein immunoprecipitation method enabled the capture of multiple immunogenic proteins, IP samples were also analyzed by SDS-PAGE and immunoblotting. Bypassing gels and subjecting immunoprecipitated proteins directly to MS is a simple and rapid antigen identification method with relatively high throughput. IP-nLC-MS/MS provides a novel alternative approach for current methods used for the identification of immunogenic proteins.
Assuntos
Antígenos de Bactérias/isolamento & purificação , Proteínas de Bactérias/isolamento & purificação , Bordetella pertussis/imunologia , Animais , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Imunoprecipitação/métodos , Nanotecnologia , Proteômica/métodos , Coelhos , Espectrometria de Massas em TandemRESUMO
Staphylococcus aureus (SA) is a major community-acquired pathogen. The emergence of drug-resistant strains like, methicillin-resistant SA (MRSA), poses stiff challenges to therapeutic intervention. Passive immune-therapy with specific antibodies is being actively examined to treat fulminant infections with limited success. In this study, we demonstrate that P4, a 28-amino acid peptide, derived from pneumococcal surface adhesin A along with pathogen-specific antibody (IVIG; P4 therapy) is successful in enhancing the opsonophagocytic killing (OPK) of S. aureus in vitro. We questioned if it is possible to expand P4 therapy to treat staphylococcal infections in vivo. P4 therapy in combination with IVIG rescued 7/10 morbidly ill S. aureus-infected mice while only 2/10 survived in the control group.
RESUMO
Polymerase chain reaction (PCR) offers promise in pneumococcal diagnostics, but whether nasopharyngeal carriage causes false-positive results in people colonized with Streptococcus pneumoniae is unknown. We found in serum no positive samples for pneumococcal DNA in 100 carriers and noncarriers using 2 different real-time PCR assays targeting lytA and psaA genes.
Assuntos
Adesinas Bacterianas/genética , Técnicas Bacteriológicas/métodos , Lipoproteínas/genética , Técnicas de Diagnóstico Molecular/métodos , N-Acetil-Muramil-L-Alanina Amidase/genética , Infecções Pneumocócicas/diagnóstico , Reação em Cadeia da Polimerase/métodos , Streptococcus pneumoniae/isolamento & purificação , Portador Sadio/diagnóstico , Portador Sadio/microbiologia , Criança , Pré-Escolar , DNA Bacteriano/sangue , DNA Bacteriano/genética , Feminino , Humanos , Masculino , Infecções Pneumocócicas/microbiologia , Sensibilidade e Especificidade , Streptococcus pneumoniae/genéticaRESUMO
Alternate therapies are needed for treatment of secondary bacterial pneumonia following influenza. The immunomodulatory peptide P4 has shown promise in mouse models of primary pneumococcal infection. Mice infected with influenza virus and then challenged with Streptococcus pneumoniae were treated with a combination of P4 peptide and intravenous immune globulin. Survival was improved from 20% to 80% in treated mice relative to controls. Clinical cure correlated with increased clearance of bacteria and decreased lung consolidation. Greater trafficking of professional phagocytic cells to the site of pneumococcal infection coupled with enhanced opsonophagocytosis as manifest by decreased surface display of Fcγ receptors (FcγR) on neutrophils and macrophages were associated with P4 peptide treatment. This suggests that the mechanism of action for improved clearance of bacteria engendered by P4 is through improved uptake by phagocytes mediated by IgG Fc-Fcγ receptor interactions following antibody-mediated opsonophagocytosis of bacteria. Antibody-based therapies, when coupled with immune modulators, such as P4 peptide, may be an effective tool together with antibiotics in our armamentarium against severe pneumonia.
Assuntos
Imunoglobulinas Intravenosas/uso terapêutico , Pneumonia Pneumocócica/tratamento farmacológico , Animais , Feminino , Imunoterapia , Camundongos , Camundongos Endogâmicos BALB C , Pneumonia Pneumocócica/microbiologia , Streptococcus pneumoniae/efeitos dos fármacosAssuntos
Adesinas Bacterianas/administração & dosagem , Anticorpos Antibacterianos/administração & dosagem , Lipoproteínas/administração & dosagem , Infecções Pneumocócicas/tratamento farmacológico , Infecções Pneumocócicas/imunologia , Sepse/tratamento farmacológico , Sepse/imunologia , Adesinas Bacterianas/imunologia , Fatores Etários , Animais , Anticorpos Antibacterianos/imunologia , Imunoglobulina G/administração & dosagem , Imunoglobulina G/imunologia , Lipoproteínas/imunologia , Camundongos , Infecções Pneumocócicas/mortalidade , Sepse/mortalidade , Análise de SobrevidaRESUMO
In the early 1900s, passive immunization/antibody therapy was used to treat a variety of human ailments such as hypoimmunoglobulinemia, cancer and infectious disease. The advent of antibiotic therapy had relegated this type of therapy obsolete for treatment of infectious diseases. Emergence of multi-drug resistant pathogens along with novel monoclonal antibody production techniques has rekindled the interest in passive immunization (PI). An increase in the number of monoclonal antibody patent applications in the recent past suggests a renewed commercial interest in PI. Despite these developments, antibody therapy for infectious diseases has limitations including the need for large or frequent dosages. P4, a 28-amino acid peptide is a multi-lineage cellular activator. P4, along with infectious disease (i.e. Pathogen) specific immunoglobulin, has been shown in vitro and in vivo in mice to potentiate innate immunity. This review will discuss the progress made in passive antibody therapy, the challenges still to be surmounted, and the potential expanded role of an immune-potentiating peptide (bio-molecule) in the quest to utilize and revitalize passive immunization.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Imunização Passiva/métodos , Animais , Humanos , Modelos Imunológicos , Patentes como Assunto , Fagocitose/efeitos dos fármacosRESUMO
A murine colonization model was used to determine the effect of co-administering 7-valent polysaccharide-protein conjugate vaccine and pneumococcal surface adhesin A. Mice were challenged intranasally with either PCV7 serotypes, 4 or 14, or a non-PCV7 serotype, 19A. Post-challenge samples were evaluated for IgG antibody levels, opsonophagocytic activity, and nasopharyngeal colonization. No interference was observed between immune responses from the concomitant and individual immunizations. Concomitant immunizations reduced carriage for tested serotypes; largest reduction was observed for 19A. From these mouse studies, co-administering pneumococcal antigens appear to expand coverage and reduce colonization against a non-PCV7 serotype without inhibiting immunogenicity to other serotypes.
Assuntos
Adesinas Bacterianas/imunologia , Portador Sadio/prevenção & controle , Lipoproteínas/imunologia , Vacinas Pneumocócicas/imunologia , Infecções Estreptocócicas/prevenção & controle , Streptococcus pneumoniae/imunologia , Adesinas Bacterianas/administração & dosagem , Animais , Anticorpos Antibacterianos/sangue , Vacina Pneumocócica Conjugada Heptavalente , Lipoproteínas/administração & dosagem , Camundongos , Proteínas Opsonizantes/sangue , Fagocitose , Vacinas Pneumocócicas/administração & dosagem , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/imunologiaRESUMO
P4, a 28-amino-acid peptide, is a eukaryotic cellular activator that enhances specific in vitro opsonophagocytic killing of multiple bacterial pathogens. In a previous study, we successfully recreated this phenomenon in mice in vivo by using a two-dose regimen of P4 and pathogen-specific antibodies, which significantly reduced moribundity in mice. For the present study, we hypothesized that the inclusion of a low-dose antibiotic would make it possible to treat the infected mice with a single dose containing a mixture of P4 and a pathogen-specific antibody. A single dose consisting of P4, intravenous immunoglobulin (IVIG), and ceftriaxone effectively reduced moribundity compared to that of untreated controls (n = 10) by 75% (P < 0.05) and rescued all (10 of 10) infected animals (P < 0.05). If rescued animals were reinfected with Streptococcus pneumoniae and treated with a single dose containing P4, IVIG, and ceftriaxone, they could be rerescued. This observation of the repeated successful use of P4 combination therapy demonstrates a low risk of tolerance development. Additionally, we examined the polymorphonuclear leukocytes (PMN) derived from infected mice and observed that P4 enhanced in vitro opsonophagocytic killing (by >80% over the control level; P < 0.05). This finding supports our hypothesis that PMN are activated by P4 during opsonophagocytosis and the recovery of mice from pneumococcal infection. P4 peptide-based combination therapy may offer an alternative and rapid immunotherapy to treat fulminant pneumococcal infection.
Assuntos
Imunização Passiva/métodos , Fatores Imunológicos/uso terapêutico , Peptídeos/uso terapêutico , Infecções Pneumocócicas/terapia , Streptococcus pneumoniae/imunologia , Animais , Antibacterianos/uso terapêutico , Anticorpos Antibacterianos/uso terapêutico , Proteínas de Bactérias/síntese química , Proteínas de Bactérias/farmacologia , Proteínas de Bactérias/uso terapêutico , Ceftriaxona/uso terapêutico , Quimioterapia Combinada , Feminino , Imunoglobulinas Intravenosas/uso terapêutico , Fatores Imunológicos/síntese química , Fatores Imunológicos/farmacologia , Camundongos , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Peptídeos/síntese química , Peptídeos/farmacologia , Fagocitose/efeitos dos fármacos , Streptococcus pneumoniae/química , Análise de SobrevidaRESUMO
BACKGROUND: P4, a 28-aa peptide derived from pneumococcal surface adhesin A, is a multilineage cell activator in vitro. We hypothesized that P4-mediated activation of phagocytic cells could rapidly and substantially increase opsonophagocytosis of bacteria, which could be translated in vivo to reduced mouse morbidity from fatal pneumococcal infection. METHODS: Reference in vitro opsonophagocytic killing and uptake assays were used with suitable effector cells and pathogen-specific antibodies. P4 peptide solution was added at the preopsonization stage. ND4-SW mice were infected intranasally with Streptococcus pneumoniae serotype 3 (WU2). At 72 and 96 h, infected mice received intraperitoneal or intravenous injection of gamma globulin, followed by an injection of P4. RESULTS: P4 treatment enhanced in vitro opsonophagocytosis of bacterial pathogens by many fold, and this effect was dependent on complement, P4, and antibody concentrations. Treatment of highly virulent WU2-infected mice with the combination of P4 and serotype-specific antiserum resulted in 100% remission of bacteremia and rescued 80% of the animals (P < .05). CONCLUSION: P4 peptide in combination with pathogen-specific antibodies and complement enhances specific opsonophagocytosis and rescues mice from life-threatening pneumococcal infection. P4 peptide provides a fresh direction for therapeutic intervention through augmented passive immunotherapy.
Assuntos
Adesinas Bacterianas/química , Adesinas Bacterianas/imunologia , Proteínas de Bactérias/química , Imunização Passiva , Infecções Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Adesinas Bacterianas/metabolismo , Animais , Camundongos , Neutrófilos , Fagocitose/fisiologia , Infecções Pneumocócicas/terapia , gama-Globulinas/imunologiaRESUMO
Pneumococcal surface adhesin A (PsaA) is a surface-exposed common 37-kilodalton multi-functional lipoprotein detected on all known serotypes of Streptococcus pneumoniae. This lipoprotein belongs to the ABC-type transport protein complex that transports Mn(2+); it is also an adhesin that plays a major role in pneumococcal attachment to the host cell and virulence. PsaA is immunogenic and natural nasopharyngeal colonization of pneumococci elicits an increase in antibody towards PsaA. Hence, PsaA is being actively evaluated as a component of a vaccine in formulations composed of pneumococcal common proteins. PsaA has been expressed as an E. coli recombinant protein, purified, and evaluated in a phase one clinical trial. This article reviews PsaA, its structure and role in pneumococcal virulence, immunogenicity, and potential to reduce nasopharyngeal colonization (a major prerequisite for pneumococcal pathogenesis) as a component of a common pneumococcal protein vaccine.
RESUMO
Pneumococcal surface adhesin A (PsaA) is a surface-exposed common 37-kilodalton multi-functional lipoprotein detected on all known serotypes of Streptococcus pneumoniae. This lipoprotein belongs to the ABC-type transport protein complex that transports Mn2+; it is also an adhesin that plays a major role in pneumococcal attachment to the host cell and virulence. PsaA is immunogenic and natural nasopharyngeal colonization of pneumococci elicits an increase in antibody towards PsaA. Hence, PsaA is being actively evaluated as a component of a vaccine in formulations composed of pneumococcal common proteins. PsaA has been expressed as an E. coli recombinant protein, purified, and evaluated in a phase one clinical trial. This article reviews PsaA, its structure and role in pneumococcal virulence, immunogenicity, and potential to reduce nasopharyngeal colonization (a major prerequisite for pneumococcal pathogenesis) as a component of a common pneumococcal protein vaccine.
Assuntos
Adesinas Bacterianas/imunologia , Adesinas Bacterianas/fisiologia , Lipoproteínas/imunologia , Lipoproteínas/fisiologia , Streptococcus pneumoniae/imunologia , Streptococcus pneumoniae/patogenicidade , Fatores de Virulência/imunologia , Fatores de Virulência/fisiologia , Adesinas Bacterianas/genética , Animais , Aderência Bacteriana , Humanos , Lipoproteínas/genética , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/imunologia , Proteínas de Membrana Transportadoras/fisiologia , Camundongos , Vacinas Estreptocócicas/imunologia , Streptococcus pneumoniae/genética , Fatores de Virulência/genéticaRESUMO
Better diagnostics for pneumococcal disease are urgently needed. In a murine model, real-time polymerase chain reaction was superior to conventional culture in detecting pneumococcus in blood, particularly in early disease and after antibiotic administration, and could distinguish between commensalism and infection.
Assuntos
Bacteriemia/diagnóstico , Modelos Animais de Doenças , Infecções Pneumocócicas/diagnóstico , Reação em Cadeia da Polimerase/métodos , Streptococcus pneumoniae/isolamento & purificação , Animais , Antibacterianos/uso terapêutico , Bacteriemia/tratamento farmacológico , Bacteriemia/microbiologia , Sangue/microbiologia , Meios de Cultura , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , Camundongos , Infecções Pneumocócicas/tratamento farmacológico , Infecções Pneumocócicas/microbiologia , Sensibilidade e Especificidade , Streptococcus pneumoniae/genética , Fatores de TempoRESUMO
Pneumococcal surface adhesin A (PsaA) is a putative pneumococcal (Pnc) adhesin known to bind to nasopharyngeal (NP) epithelial cells. This study evaluated the effect of peptides within a functional domain of PsaA on NP cells. Detroit 562 NP cells were treated with synthetic peptides derived from PsaA (P4, P6, and P7; 28, 12, and 16 amino acids, respectively). The P4 peptide also binds to NP cells. Analysis of P4-treated NP cells by transmission electron microscopy revealed major cytological changes. Of 9 cytokines analyzed, a 6-fold increase in FGFb secretion at 3 and 6h (11-fold at 12h) was found post-P4 treatment of NP cells. There was a simultaneous reduction in the secreted levels of IL-6, IL-8, and VEGF. We observed enhancement in the adherence of Pnc strains to P4-treated NP cells (2-38-fold increase). Enhancement in adherence (2-fold increase) to P4-treated NP cells was also recorded with other streptococcal species (Streptococcus mitis and Streptococcus pyogenes). Internalization experiments demonstrated that 45% of the adherent bacteria were actually internalized after pretreatment with P4 peptide as compared to controls. Peptide fragments of P4, P6 and P7 did not activate NP cells to the extent of P4 peptide. The P4-mediated enhancement of Pnc adherence was blocked (100%) by anti-P4 antibodies, confirming the specificity of the P4 sequence for NP cell activation. Our data suggests that this functional domain of PsaA contained within the P4 sequence binds and activates NP cells to facilitate Pnc invasion.