RESUMO
Amoxicillin is a broad-spectrum antibiotic used to treat a variety of gram-positive and gram-negative infections, such as infections of the ear, nose, and throat, genitourinary tract, skin, and lower respiratory tract; gonorrhea; and Helicobacter pylori. The prophylactic benefit of both amoxicillin and Augmentin (amoxicillin-clavulanate for use against ß-lactamase-expressing bacteria) was evaluated for inhalation anthrax in cynomolgus macaques in 2 studies. A pilot study on amoxicillin-clavulanate that used a portion of the study animals demonstrated empirically that dosing twice a day was efficacious. In a subsequent study on both amoxicillin and amoxicillin-clavulanate that used the remaining study animals, the animals were treated orally every 12 hours on days 1-28 postchallenge and followed for an additional 60 days (total of 88 days from day of aerosol challenge to when the animals were culled). The animals from each treatment arm of the 2 studies were completely protected. All untreated animals succumbed to the infection. The degree of protection observed in this study suggests that both amoxicillin and amoxicillin-clavulanate, administered prophylactically over a period of 28 days after a lethal exposure to Bacillus anthracis spores, is sufficient for full protection.
Assuntos
Bacillus anthracis , Amoxicilina/farmacologia , Amoxicilina/uso terapêutico , Combinação Amoxicilina e Clavulanato de Potássio/uso terapêutico , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Macaca , Projetos Piloto , beta-LactamasesRESUMO
Left untreated, inhalation anthrax is usually fatal. Vegetative forms of Bacillus anthracis survive in blood and tissues during infection due to elaboration of a protective poly-γ-D-glutamic acid (PDGA) capsule that permits uncontrolled bacterial growth in vivo, eventually leading to overwhelming bacillosis and death. As a measure to counter threats from multidrug-resistant strains, we are evaluating the prophylactic and therapeutic potential of the PDGA depolymerase EnvD, a stable and potent enzyme which rapidly and selectively removes the capsule from the surface of vegetative cells. Repeated intravenous administration of 10 mg/kg recombinant EnvD (rEnvD) to mice infected with lethal doses of B. anthracis Ames spores by inhalation prevented the emergence of symptoms of anthrax and death; all animals survived the 5-day treatment period, and 70% survived to the end of the 14-day observation period. In contrast to results in sham-treated animals, the lungs and spleen of rEnvD-dosed animals were free of gross pathological changes. We conclude that rEnvD has potential as an agent to prevent the emergence of inhalation anthrax in infected animals and is likely to be effective against drug-resistant forms of the pathogen.
Assuntos
Antraz/prevenção & controle , Antibacterianos/uso terapêutico , Cápsulas Bacterianas/efeitos dos fármacos , Peptídeo Hidrolases/uso terapêutico , Infecções Respiratórias/prevenção & controle , Administração Intravenosa , Aerossóis , Animais , Antibacterianos/administração & dosagem , Bacillus anthracis/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla , Feminino , Meia-Vida , Camundongos Endogâmicos BALB C , Peptídeo Hidrolases/administração & dosagem , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/uso terapêuticoRESUMO
To support the licensure of a new and safer vaccine to protect people against smallpox, a monkeypox model of infection in cynomolgus macaques, which simulates smallpox in humans, was used to evaluate two vaccines, Acam2000 and Imvamune, for protection against disease. Animals vaccinated with a single immunization of Imvamune were not protected completely from severe and/or lethal infection, whereas those receiving either a prime and boost of Imvamune or a single immunization with Acam2000 were protected completely. Additional parameters, including clinical observations, radiographs, viral load in blood, throat swabs, and selected tissues, vaccinia virus-specific antibody responses, immunophenotyping, extracellular cytokine levels, and histopathology were assessed. There was no significant difference (P > 0.05) between the levels of neutralizing antibody in animals vaccinated with a single immunization of Acam2000 (132 U/ml) and the prime-boost Imvamune regime (69 U/ml) prior to challenge with monkeypox virus. After challenge, there was evidence of viral excretion from the throats of 2 of 6 animals in the prime-boost Imvamune group, whereas there was no confirmation of excreted live virus in the Acam2000 group. This evaluation of different human smallpox vaccines in cynomolgus macaques helps to provide information about optimal vaccine strategies in the absence of human challenge studies.
Assuntos
Imunização/métodos , Orthopoxvirus/imunologia , Infecções por Poxviridae/prevenção & controle , Vacina Antivariólica/farmacologia , Animais , Anticorpos Neutralizantes/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Macaca fascicularis , Masculino , Reação em Cadeia da Polimerase em Tempo Real , Vacinas Atenuadas/farmacologia , Eliminação de Partículas Virais/imunologiaRESUMO
A selection of previously identified protective Mycobacterium tuberculosis DNA vaccines were re-formulated as proteins and administered with a Th1-inducing adjuvant to help stimulate the relevant immune responses necessary for protection. All three candidate-vaccines conferred high levels of antigen-specific cellular and humoral responses, as indicated by lymphocyte proliferation and serum IgG levels. Protective efficacy was also assessed in comparison with the current vaccine, BCG (the 'gold-standard' against which new vaccines are tested), and a saline (negative) control. One candidate (Rv1806-1807) induced protection in the guinea pig aerosol infection model 30 days post-challenge on the basis of reducing the bacterial burden of M. tuberculosis in the lungs.
Assuntos
Vacinas contra a Tuberculose/imunologia , Tuberculose Pulmonar/prevenção & controle , Vacinas de DNA/imunologia , Adjuvantes Imunológicos , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Epitopos , Escherichia coli/genética , Feminino , Genes Bacterianos , Vetores Genéticos , Cobaias , Imunidade Celular , Imunoglobulina G/sangue , Pulmão/microbiologia , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/imunologia , Mycobacterium tuberculosis/isolamento & purificação , Células Th1/imunologia , Vacinas contra a Tuberculose/administração & dosagem , Tuberculose Pulmonar/imunologia , Vacinas de DNA/administração & dosagem , Vacinas de Subunidades Antigênicas/administração & dosagem , Vacinas de Subunidades Antigênicas/imunologiaRESUMO
The aim of this study was to establish an assay to compare Mycobacterium tuberculosis strains, and cells grown under different growth conditions, in terms of their ability to cause a lung infection and disseminate to the spleen. M. tuberculosis strains H37Rv, Erdman, South Indian (TMC120, SI) and H37Rv cells grown aerobically or under low oxygen/iron limitation in a chemostat were assayed for infectivity. Groups of 8 animals were challenged with 3 different doses of each strain. Lung and spleen bacteriology was assessed at 16 days post-infection for all strains. Bacteriology and lung pathology at day 56 was studied for H37Rv, Erdman and SI. Strains H37Rv and Erdman had a statistically significantly higher pathogenic potential than SI and this was confirmed by analysis of lung pathology performed at 8 weeks post-infection, although the Erdman strain caused more extensive caseation without calcification and little encapsulation. The model could discriminate between cells grown under different growth conditions; low-oxygen/iron-limited cells had a significantly higher infectivity than those grown aerobically. This study presents a quick and reliable method for comparing with statistical confidence, the pathogenic potential of M. tuberculosis strains and the impact of in vitro growth conditions on the infectivity of M. tuberculosis in vivo.
Assuntos
Mycobacterium tuberculosis/patogenicidade , Tuberculose Pulmonar/microbiologia , Aerobiose , Aerossóis , Anaerobiose , Animais , Contagem de Colônia Microbiana , Meios de Cultura , Compostos Férricos , Cobaias , Mycobacterium tuberculosis/classificação , Mycobacterium tuberculosis/crescimento & desenvolvimento , Baço/microbiologia , Tuberculose Pulmonar/patologia , VirulênciaRESUMO
The TB Vaccine Cluster project funded by the EU Fifth Framework programme aims to provide novel vaccines against tuberculosis that are suitable for evaluation in humans. This paper describes the studies of the protective efficacy of vaccines in a guinea pig aerosol-infection model of primary tuberculosis. The objective was to conduct comparative evaluations of vaccines that had previously demonstrated efficacy in other animal models. Groups of 6 guinea pigs were immunized with vaccines provided by the relevant EU Vaccine Cluster partners. Survival over 17 or 26 weeks was used as the principal measure of vaccine efficacy following aerosol challenge with H37Rv. Counts of mycobacteria in lungs and spleens, and histopathological changes in the lungs, were also used to provide evidence of protection. A total of 24 vaccines were evaluated in 4 experiments each of a different design. A heterologous prime-boost strategy of DNA and MVA, each expressing Ag85A and a fusion protein of ESAT-6 and Ag85B in adjuvant, protected the guinea pigs to the same extent as BCG. Genetically modified BCG vaccines and boosted BCG strategies also protected guinea pigs to the same extent as BCG but not statistically significantly better. A relatively high aerosol-challenge dose and evaluation over a protracted time post-challenge allowed superior protection over BCG to be demonstrated by BCG boosted with MVA and fowl pox vectors expressing Ag85A.
Assuntos
Modelos Animais de Doenças , Vacinas contra a Tuberculose/uso terapêutico , Tuberculose/prevenção & controle , Aerossóis , Animais , Vacina BCG/uso terapêutico , Contagem de Colônia Microbiana/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , União Europeia , Cobaias , Humanos , Pulmão/microbiologia , Mycobacterium tuberculosis/imunologia , Mycobacterium tuberculosis/isolamento & purificação , Baço/microbiologia , Análise de Sobrevida , Tuberculose/imunologia , Vacinas contra a Tuberculose/imunologia , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/prevenção & controle , Vacinação/métodosRESUMO
We investigated how Mycobacterium tuberculosis responded to a reduced oxygen tension in terms of its pathogenicity and gene expression by growing cells under either aerobic or low-oxygen conditions in chemostat culture. The chemostat enabled us to control and vary the oxygen tension independently of other environmental parameters, so that true cause-and-effect relationships of reduced oxygen availability could be established. Cells grown under low oxygen were more pathogenic for guinea pigs than those grown aerobically. The effect of reduced oxygen on global gene expression was determined using DNA microarray. Spearman rank correlation confirmed that microarray expression profiles were highly reproducible between repeat cultures. Using microarray analysis we have identified genes that respond to a low-oxygen environment without the influence of other parameters such as nutrient depletion. Some of these genes appear to be involved in the biosynthesis of cell wall precursors and their induction may have contributed to increased infectivity in the guinea pig. This study has shown that a combination of chemostat culture and microarray presents a biologically robust and statistically reliable experimental approach for studying the effect of relevant and specific environmental stimuli on mycobacterial virulence and gene expression.
Assuntos
Regulação Bacteriana da Expressão Gênica/fisiologia , Mycobacterium tuberculosis/patogenicidade , Tuberculose/microbiologia , Anaerobiose , Animais , DNA Bacteriano/genética , Perfilação da Expressão Gênica , Genes Bacterianos/genética , Cobaias , Mycobacterium tuberculosis/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodosRESUMO
The Mycobacterium tuberculosis phoP mutant strain SO2 has previously been shown to have reduced multiplication in mouse macrophages and in vivo using the mouse intravenous-infection model. In this study we demonstrate that the M. tuberculosis SO2 is highly attenuated when compared with the parental M. tuberculosis MT103 strain and also more attenuated than BCG in severe combined immunodeficiency disease (SCID) mice. Complementation of the M. tuberculosis SO2 with the wild-type phoP gene restored the virulence of the strain in the SCID mice, confirming that the attenuated phenotype is due to the phoP mutation. In Balb/c mice subcutaneously vaccinated with either M. tuberculosis SO2 or BCG, the proportions of CD4+ and CD8+ populations measured in the spleen were significantly higher in the M. tuberculosis SO2 vaccinated group. In addition, the proportion of antigen-stimulated CD4+/CD8+ cells expressing IFN-gamma was significantly higher in the M. tuberculosis SO2 vaccinated group when compared with the BCG group. Balb/c mice subcutaneously vaccinated with the M. tuberculosis SO2 strain were also protected against intra-venous challenge with M. tuberculosis H37Rv at levels comparable to mice vaccinated with BCG, as measured by reduced bacterial counts in lung and spleens. Guinea pigs subcutaneously vaccinated with the M. tuberculosis SO2 strain were protected against aerosol challenge with M. tuberculosis H37Rv delivered at different doses. A high dose aerosol challenge of M. tuberculosis SO2 vaccinated guinea pigs resulted in superior levels of protection when compared with BCG vaccination, as measured by guinea pig survival and reduction in disease severity in the lung.
Assuntos
Vacina BCG/imunologia , Proteínas de Bactérias/genética , Vacinas contra a Tuberculose/imunologia , Tuberculose/prevenção & controle , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Cobaias , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Mutação , Vacinação , Vacinas Atenuadas/imunologiaRESUMO
Putative TB vaccine candidates were selected from lists of genes induced in response to in vivo-like stimuli, such as low oxygen and carbon starvation or growth in macrophages, and tested as plasmid DNA vaccines for their ability to protect against Mycobacterium tuberculosis challenge in a guinea pig aerosol infection model. This vaccination method was chosen as it induces the Th1 cell-mediated immune response required against intracellular pathogens such as M. tuberculosis. Protection was assessed in the guinea pig model in terms of mycobacteria present in the lungs at 30 days post-challenge. Protection achieved by the novel candidates was compared to BCG (positive control) and saline (negative control). Four vaccines encoding for proteins such as PE and PPE proteins, a zinc metalloprotease and an acyltransferase, gave a level of protection that was statistically better than saline in the lungs. These findings have enabled us to focus on a sub-set of vaccine candidates for further evaluation using additional vaccination strategies.