RESUMEN
Burkholderia pseudomallei is the causative agent of melioidosis, which is endemic primarily in Southeast Asia and northern Australia but is increasingly being seen in other tropical and subtropical regions of the world. Melioidosis is associated with high morbidity and mortality rates, which is mediated by the wide range of virulence factors encoded by B. pseudomallei. These virulence determinants include surface polysaccharides such as lipopolysaccharide (LPS) and capsular polysaccharides (CPS). Here, we investigated a predicted arabinose-5-phosphate isomerase (API) similar to KdsD in B. pseudomallei strain K96243. KdsD is required for the production of the highly conserved 3-deoxy-d-manno-octulosonic acid (Kdo), a key sugar in the core region of LPS. Recombinant KdsD was expressed and purified, and API activity was determined. Although a putative API paralogue (KpsF) is also predicted to be encoded, the deletion of kdsD resulted in growth defects, loss of motility, reduced survival in RAW 264.7 murine macrophages, and attenuation in a BALB/c mouse model of melioidosis. Suppressor mutations were observed during a phenotypic screen for motility, revealing single nucleotide polymorphisms or indels located in the poorly understood CPS type IV cluster. Crucially, suppressor mutations did not result in reversion of attenuation in vivo. This study demonstrates the importance of KdsD for B. pseudomallei virulence and highlights further the complex nature of the polysaccharides it produces. IMPORTANCE The intrinsic resistance of B. pseudomallei to many antibiotics complicates treatment. This opportunistic pathogen possesses a wide range of virulence factors, resulting in severe and potentially fatal disease. Virulence factors as targets for drug development offer an alternative approach to combat pathogenic bacteria. Prior to initiating early drug discovery approaches, it is important to demonstrate that disruption of the target gene will prevent the development of disease. This study highlights the fact that KdsD is crucial for virulence of B. pseudomallei in an animal model of infection and provides supportive phenotypic characterization that builds a foundation for future therapeutic development.
Asunto(s)
Isomerasas Aldosa-Cetosa , Burkholderia pseudomallei , Melioidosis , Animales , Ratones , Burkholderia pseudomallei/genética , Melioidosis/tratamiento farmacológico , Melioidosis/microbiología , Melioidosis/patología , Virulencia/genética , Lipopolisacáridos , Isomerasas Aldosa-Cetosa/genética , Factores de Virulencia/genética , PolisacáridosRESUMEN
Burkholderia pseudomallei is a gram-negative, facultative intracellular bacterium, which causes a disease known as melioidosis. Professional phagocytes represent a crucial first line of innate defense against invading pathogens. Uptake of pathogens by these cells involves the formation of a phagosome that matures by fusing with early and late endocytic vesicles, resulting in killing of ingested microbes. Host Rab GTPases are central regulators of vesicular trafficking following pathogen phagocytosis. However, it is unclear how Rab GTPases interact with B. pseudomallei to regulate the transport and maturation of bacterial-containing phagosomes. Here, we showed that the host Rab32 plays an important role in mediating antimicrobial activity by promoting phagosome maturation at an early phase of infection with B. pseudomallei. And we demonstrated that the expression level of Rab32 is increased through the downregulation of the synthesis of miR-30b/30c in B. pseudomallei infected macrophages. Subsequently, we showed that B. pseudomallei resides temporarily in Rab32-positive compartments with late endocytic features. And Rab32 enhances phagosome acidification and promotes the fusion of B. pseudomallei-containing phagosomes with lysosomes to activate cathepsin D, resulting in restricted intracellular growth of B. pseudomallei. Additionally, Rab32 mediates phagosome maturation depending on its guanosine triphosphate/guanosine diphosphate (GTP/GDP) binding state. Finally, we report the previously unrecognized role of miR-30b/30c in regulating B. pseudomallei-containing phagosome maturation by targeting Rab32 in macrophages. Altogether, we provide a novel insight into the host immune-regulated cellular pathway against B. pseudomallei infection is partially dependent on Rab32 trafficking pathway, which regulates phagosome maturation and enhances the killing of this bacterium in macrophages.
Asunto(s)
Burkholderia pseudomallei/inmunología , Melioidosis/inmunología , MicroARNs/inmunología , Fagosomas/inmunología , Proteínas de Unión al GTP rab/inmunología , Animales , Burkholderia pseudomallei/patogenicidad , Melioidosis/patología , Ratones , Viabilidad Microbiana/inmunología , Fagosomas/microbiología , Fagosomas/patología , Células RAW 264.7RESUMEN
BACKGROUND: Melioidosis is an infectious disease caused by Burkholderia pseudomallei. In Mexico, the disease is rarely diagnosed in humans and there is no evidence of simultaneous environmental isolation of the pathogen. Here, we describe clinical profiles of fatal cases of melioidosis in two children, in a region without history of that disease. CASE PRESENTATION: About 48 h before onset of symptoms, patients swam in a natural body of water, and thereafter they rapidly developed fatal septicemic illness. Upon necropsy, samples from liver, spleen, lung, cerebrospinal fluid, and bronchial aspirate tissues contained Burkholderia pseudomallei. Environmental samples collected from the locations where the children swam also contained B. pseudomallei. All the clinical and environmental strains showed the same BOX-PCR pattern, suggesting that infection originated from the area where the patients were swimming. CONCLUSIONS: The identification of B. pseudomallei confirmed that melioidosis disease exists in Sonora, Mexico. The presence of B. pseudomallei in the environment may suggest endemicity of the pathogen in the region. This study highlights the importance of strengthening laboratory capacity to prevent and control future melioidosis cases.
Asunto(s)
Melioidosis/complicaciones , Neumonía Bacteriana/etiología , Adolescente , Burkholderia pseudomallei/aislamiento & purificación , Niño , Resultado Fatal , Femenino , Humanos , Masculino , Melioidosis/diagnóstico , Melioidosis/patología , Melioidosis/fisiopatología , México , Neumonía Bacteriana/diagnóstico , Neumonía Bacteriana/patología , Neumonía Bacteriana/fisiopatología , Sepsis/microbiología , NataciónRESUMEN
Melioidosis is a tropical infectious disease with diverse clinical presentations. We aimed to investigate the characteristics and mortality risk factors of patients diagnosed with melioidosis in the past 10 years. This was a retrospective cohort study conducted at a quaternary care centre in South India. Clinical, demographic, and biochemical data in patients diagnosed with melioidosis with cultures were collected between January 2011 and December 2020 from medical records. Logistic regression analysis was performed to screen mortality risk factors of melioidosis in addition to descriptive statistics and chi-square analysis. Seventy-three melioidosis patients' records were analysed, and the most common comorbidity was type 2 diabetes mellitus (n = 53, 72.6%). The patients showed diverse presentations: pulmonary involvement, 30 (41.1%); splenomegaly, 29 (39.7%); abscesses and cutaneous involvement, 18 (24.7%); lymph node, 10 (13.7%); arthritis and osteomyelitis, 9 (12.3%); and genitourinary infection, 4 (5.4%). The mortality was noted to be 15 (20.5%). Logistic regression analysis indicated that chronic kidney disease (OR = 14.0), CRP >100 IU/L (OR = 6.964), and S. albumin <3 gm/dl (OR = 8.0) were risk factors associated with mortality and can guide in risk stratification. Hypoalbuminemia is a novel mortality risk factor, detected in this study, and requires further investigation to validate its utility as a prognostic marker and reveal possible therapeutic benefits in clinical correction.
Asunto(s)
Melioidosis/mortalidad , Burkholderia pseudomallei/efectos de los fármacos , Distribución de Chi-Cuadrado , Comorbilidad , Diabetes Mellitus Tipo 2/epidemiología , Femenino , Humanos , India/epidemiología , Modelos Logísticos , Masculino , Melioidosis/diagnóstico , Melioidosis/microbiología , Melioidosis/patología , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Pronóstico , Insuficiencia Renal Crónica/epidemiología , Estudios Retrospectivos , Factores de Riesgo , Albúmina Sérica/análisisRESUMEN
Endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) is routinely performed for diagnostic evaluation of mediastinal lymphadenopathy due to various etiologies with excellent sensitivity and specificity. Melioidosis can have atypical features like isolated mediastinal lymphadenopathy mimicking as tuberculosis or lymphoma. Differentiation of such atypical melioidosis presentation become difficult due to similar clinical, radiological and even similar EBUS lymph node characteristics. Role of EBUS TBNA in diagnosing melioidosis is under investigated and sparsely reported. We describe two cases of melioidosis diagnosed by point of care rapid lateral flow immunoassay antigen testing and culture of EBUS-TBNA samples from necrotic mediastinal lymph nodes.
Asunto(s)
Biopsia por Aspiración con Aguja Fina Guiada por Ultrasonido Endoscópico/métodos , Endosonografía/instrumentación , Melioidosis/patología , Administración Intravenosa , Administración Oral , Anciano , Antibacterianos/administración & dosificación , Antibacterianos/uso terapéutico , Burkholderia pseudomallei/inmunología , Doxiciclina/administración & dosificación , Doxiciclina/uso terapéutico , Quimioterapia Combinada , Humanos , Inmunoensayo/métodos , Ganglios Linfáticos/patología , Linfadenopatía/diagnóstico , Masculino , Enfermedades del Mediastino/patología , Melioidosis/diagnóstico , Melioidosis/inmunología , Melioidosis/microbiología , Meropenem/administración & dosificación , Meropenem/uso terapéutico , Sensibilidad y Especificidad , Resultado del Tratamiento , Combinación Trimetoprim y Sulfametoxazol/administración & dosificación , Combinación Trimetoprim y Sulfametoxazol/uso terapéuticoRESUMEN
Background: Infection with the gram-negative bacillus Burkholderia pseudomallei (melioidosis) is an important cause of pneumosepsis in Southeast Asia and has a mortality of up to 40%. We aimed to assess the role of platelets in the host response against B. pseudomallei infection. Methods: Association between platelet counts and mortality was determined in 1160 patients with culture-proven melioidosis. Mice treated with (low- or high-dose) platelet-depleting antibody were inoculated intranasally with B. pseudomallei and killed. Additional studies using functional glycoprotein Ibα-deficient mice were conducted. Results: Thrombocytopenia was present in 31% of patients at admission and predicted mortality in melioidosis patients even after adjustment for confounders. In our murine-melioidosis model, platelet counts decreased, and mice treated with a platelet-depleting antibody showed enhanced mortality and higher bacterial loads compared to mice with normal platelet counts. Low platelet counts had a modest impact on early-pulmonary neutrophil influx. Reminiscent of their role in hemostasis, platelet depletion impaired vascular integrity, resulting in early lung bleeding. Glycoprotein Ibα-deficient mice had reduced platelet counts during B. pseudomallei infection together with an impaired local host defense in the lung. Conclusions: Thrombocytopenia predicts mortality in melioidosis patients and, during experimental melioidosis, platelets play a protective role in both innate immunity and vascular integrity.
Asunto(s)
Burkholderia pseudomallei/inmunología , Melioidosis/complicaciones , Melioidosis/patología , Trombocitopenia/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Asia Sudoriental , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Melioidosis/inmunología , Melioidosis/mortalidad , Ratones , Persona de Mediana Edad , Estudios Prospectivos , Análisis de Supervivencia , Trombocitopenia/inmunología , Adulto JovenRESUMEN
Burkholderia pseudomallei is the causative agent of melioidosis, a disease endemic to Southeast Asia and northern Australia. Mortality rates in these areas are high even with antimicrobial treatment, and there are few options for effective therapy. Therefore, there is a need to identify antibacterial targets for the development of novel treatments. Cyclophilins are a family of highly conserved enzymes important in multiple cellular processes. Cyclophilins catalyze the cis-trans isomerization of xaa-proline bonds, a rate-limiting step in protein folding which has been shown to be important for bacterial virulence. B. pseudomallei carries a putative cyclophilin B gene, ppiB, the role of which was investigated. A B. pseudomalleiΔppiB (BpsΔppiB) mutant strain demonstrates impaired biofilm formation and reduced motility. Macrophage invasion and survival assays showed that although the BpsΔppiB strain retained the ability to infect macrophages, it had reduced survival and lacked the ability to spread cell to cell, indicating ppiB is essential for B. pseudomallei virulence. This is reflected in the BALB/c mouse infection model, demonstrating the requirement of ppiB for in vivo disease dissemination and progression. Proteomic analysis demonstrates that the loss of PpiB leads to pleiotropic effects, supporting the role of PpiB in maintaining proteome homeostasis. The loss of PpiB leads to decreased abundance of multiple virulence determinants, including flagellar machinery and alterations in type VI secretion system proteins. In addition, the loss of ppiB leads to increased sensitivity toward multiple antibiotics, including meropenem and doxycycline, highlighting ppiB inhibition as a promising antivirulence target to both treat B. pseudomallei infections and increase antibiotic efficacy.
Asunto(s)
Proteínas Bacterianas/genética , Burkholderia pseudomallei/genética , Burkholderia pseudomallei/patogenicidad , Ciclofilinas/genética , Melioidosis/microbiología , Proteoma/genética , Animales , Antibacterianos/farmacología , Proteínas Bacterianas/clasificación , Proteínas Bacterianas/metabolismo , Burkholderia pseudomallei/efectos de los fármacos , Burkholderia pseudomallei/metabolismo , Línea Celular , Ciclofilinas/deficiencia , Femenino , Eliminación de Gen , Expresión Génica , Homeostasis/genética , Macrófagos/microbiología , Melioidosis/tratamiento farmacológico , Melioidosis/mortalidad , Melioidosis/patología , Ratones , Ratones Endogámicos BALB C , Viabilidad Microbiana/efectos de los fármacos , Proteoma/clasificación , Proteoma/metabolismo , Análisis de Supervivencia , VirulenciaRESUMEN
BACKGROUND: Every year, 90,000 people may die from melioidosis. Vaccine candidates have not proceeded past animal studies, partly due to uncertainty around the potential market size. This study aims to estimate the potential impact, cost-effectiveness and market size for melioidosis vaccines. METHODS: Age-structured decision tree models with country-specific inputs were used to estimate net costs and health benefits of vaccination, with health measured in quality-adjusted life years (QALYs). Four target groups of people living in endemic regions were considered: (i) people aged over 45 years with chronic renal disease, (ii) people aged over 45 years with diabetes, (iii) people aged over 45 years with diabetes and/or chronic renal disease, (iv) everyone aged over 45 years. Melioidosis risk was estimated using Bayesian evidence synthesis of 12 observational studies. In the base case, vaccines were assumed to have 80% efficacy, to have 5-year mean protective duration and to cost USD10.20-338.20 per vaccine. RESULTS: Vaccination could be cost-effective (with incremental cost-effectiveness ratio below GDP per capita) in 61/83 countries/territories with local melioidosis transmission. In these 61 countries/territories, vaccination could avert 68,000 lost QALYs, 8300 cases and 4400 deaths per vaccinated age cohort, at an incremental cost of USD59.6 million. Strategy (ii) was optimal in most regions. The vaccine market may be worth USD268 million per year at its threshold cost-effective price in each country/territory. CONCLUSIONS: There is a viable melioidosis vaccine market, with cost-effective vaccine strategies in most countries/territories with local transmission.
Asunto(s)
Melioidosis/tratamiento farmacológico , Vacunación/economía , Análisis Costo-Beneficio , Femenino , Humanos , Masculino , Melioidosis/patología , Persona de Mediana EdadRESUMEN
BACKGROUND: Melioidosis is a tropical infectious disease which is being increasingly recognised throughout the globe. Infection occurs in humans and animals, typically through direct exposure to soil or water containing the environmental bacterium Burkholderia pseudomallei. Case clusters of melioidosis have been described in humans following severe weather events and in exotic animals imported into melioidosis endemic zones. Direct transmission of B. pseudomallei between animals and/or humans has been documented but is considered extremely rare. Between March 2015 and October 2016 eight fatal cases of melioidosis were reported in slender-tailed meerkats (Suricata suricatta) on display at a Wildlife Park in Northern Australia. To further investigate the melioidosis case cluster we sampled the meerkat enclosure and adjacent park areas and performed whole-genome sequencing (WGS) on all culture-positive B. pseudomallei environmental and clinical isolates. RESULTS: WGS confirmed that the fatalities were caused by two different B. pseudomallei sequence types (STs) but that seven of the meerkat isolates were highly similar on the whole-genome level. Used concurrently with detailed pathology data, our results demonstrate that the seven cases originated from a single original source, but routes of infection varied amongst meerkats belonging to the clonal outbreak cluster. Moreover, in some instances direct transmission may have transpired through wounds inflicted while fighting. CONCLUSIONS: Collectively, this study supports the use of high-resolution WGS to enhance epidemiological investigations into transmission modalities and pathogenesis of melioidosis, especially in the instance of a possible clonal outbreak scenario in exotic zoological collections. Such findings from an animal outbreak have important One Health implications.
Asunto(s)
Burkholderia pseudomallei/genética , Herpestidae/microbiología , Melioidosis/veterinaria , Animales , Animales de Zoológico , Australia , Brotes de Enfermedades/veterinaria , Microbiología Ambiental , Femenino , Masculino , Melioidosis/mortalidad , Melioidosis/patología , Melioidosis/transmisión , Secuenciación Completa del GenomaRESUMEN
The naturally antibiotic-resistant bacterium Burkholderia pseudomallei is the causative agent of melioidosis, a disease with stubbornly high mortality and a complex, protracted treatment regimen. The worldwide incidence of melioidosis is likely grossly underreported, though it is known to be highly endemic in northern Australia and Southeast Asia. Bacterial disulfide bond (DSB) proteins catalyze the oxidative folding and isomerization of disulfide bonds in substrate proteins. In the present study, we demonstrate that B. pseudomallei membrane protein disulfide bond protein B (BpsDsbB) forms a functional redox relay with the previously characterized virulence mediator B. pseudomallei disulfide bond protein A (BpsDsbA). Genomic analysis of diverse B. pseudomallei clinical isolates demonstrated that dsbB is a highly conserved core gene. Critically, we show that DsbB is required for virulence in B. pseudomallei A panel of B. pseudomalleidsbB deletion strains (K96243, 576, MSHR2511, MSHR0305b, and MSHR5858) were phenotypically diverse according to the results of in vitro assays that assess hallmarks of virulence. Irrespective of their in vitro virulence phenotypes, two deletion strains were attenuated in a BALB/c mouse model of infection. A crystal structure of a DsbB-derived peptide complexed with BpsDsbA provides the first molecular characterization of their interaction. This work contributes to our broader understanding of DSB redox biology and will support the design of antimicrobial drugs active against this important family of bacterial virulence targets.
Asunto(s)
Burkholderia pseudomallei/genética , Burkholderia pseudomallei/patogenicidad , Melioidosis/patología , Proteínas de la Membrana/inmunología , Ratones Endogámicos BALB C/inmunología , Oxidorreductasas/inmunología , Virulencia/genética , Animales , Australia , Burkholderia pseudomallei/inmunología , Modelos Animales de Enfermedad , Melioidosis/genética , Melioidosis/microbiología , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Virulencia/inmunologíaRESUMEN
BACKGROUND: Melioidosis is a tropical disease caused by Burkholderia pseudomallei (B. pseudomallei). It can infect any organ system and lead to multiple abscesses. A few studies reported that central nervous system (CNS) is also involved. We present a diabetic patient with multi-systemic melioidosis that affected the CNS, thorax, and spleen. The aim was to study the clinical and radiological features of melioidosis and enhance understanding of the disease. CASE PRESENTATION: A 38-year-old male presented with cough and expectoration mixed with blood for several days. Chest computed tomography (CT) showed a patchy opacity in his left lung, and multiple low-density lesions in his spleen. After 10 days of antibiotics treatment, his clinical symptoms improved and he was discharged from the hospital. But 8 months later, the patient experienced sudden onset of left limb weakness and seizure and was re-admitted to the hospital. Brain CT indicated a low-density lesion over the right frontal lobe, and magnetic resonance imaging (MRI) indicated a well-enhanced lobulated lesion with multiple diffusion restriction areas in the lesion. He had a neuronavigation-guided open surgery but no malignancy was found. B. pseudomallei was cultured from the operative samples. After 4 months of systemic and intraventricular antibiotic administration treatment, he recovered complete consciousness with left hemiparesis. CONCLUSIONS: Multi-systemic melioidosis may present atypical clinical, neurological, and radiological manifestations. It is extremely important to accurately diagnose before treatment is selected. CNS melioidosis in early stage manifests similar symptoms to malignancy or stroke. It might mislead to a false diagnose. Diffusion weighted imaging (DWI) can help in differentiate abscesses from cystic tumours.
Asunto(s)
Infecciones Bacterianas del Sistema Nervioso Central/diagnóstico , Complicaciones de la Diabetes/diagnóstico , Melioidosis/diagnóstico , Absceso/diagnóstico , Absceso/tratamiento farmacológico , Absceso/microbiología , Adulto , Antibacterianos/uso terapéutico , Encéfalo/diagnóstico por imagen , Encéfalo/microbiología , Encéfalo/patología , Burkholderia pseudomallei/aislamiento & purificación , Infecciones Bacterianas del Sistema Nervioso Central/complicaciones , Infecciones Bacterianas del Sistema Nervioso Central/tratamiento farmacológico , Infecciones Bacterianas del Sistema Nervioso Central/patología , China , Complicaciones de la Diabetes/tratamiento farmacológico , Complicaciones de la Diabetes/patología , Humanos , Imagen por Resonancia Magnética , Masculino , Melioidosis/tratamiento farmacológico , Melioidosis/patología , Radiografía , Radiografía Torácica , Bazo/diagnóstico por imagen , Bazo/microbiología , Bazo/patología , Tomografía Computarizada por Rayos XRESUMEN
The environmental bacterium and potential biothreat agent Burkholderia pseudomallei causes melioidosis, an often fatal infectious disease. Increased serum bilirubin has been shown to be a negative predictive factor in melioidosis patients. We therefore investigated the role of heme oxygenase-1 (HO-1), which catalyzes the degradation of heme into the bilirubin precursor biliverdin, ferrous iron, and CO during B. pseudomallei infection. We found that infection of murine macrophages induces HO-1 expression, involving activation of several protein kinases and the transcription factor nuclear erythroid-related factor 2 (Nrf2). Deficiency of Nrf2 improved B. pseudomallei clearance by macrophages, whereas Nrf2 activation by sulforaphane and tert-butylhydroquinone with subsequent HO-1 induction enhanced intracellular bacterial growth. The HO-1 inducer cobalt protoporphyrin IX diminished proinflammatory cytokine levels, leading to an increased bacterial burden in macrophages. In contrast, HO-1 gene knockdown reduced the survival of intramacrophage B. pseudomallei Pharmacological administration of cobalt protoporphyrin IX to mice resulted in an enhanced bacterial load in various organs and was associated with higher mortality of intranasally infected mice. The unfavorable outcome of B. pseudomallei infection after HO-1 induction was associated with higher serum IL-6, TNF-α, and MCP-1 levels but decreased secretion of IFN-γ. Finally, we demonstrate that the CO-releasing molecule CORM-2 increases the B. pseudomallei load in macrophages and mice. Thus, our data suggest that the B. pseudomallei-mediated induction of HO-1 and the release of its metabolite CO impair bacterial clearance in macrophages and during murine melioidosis.
Asunto(s)
Monóxido de Carbono/metabolismo , Hemo-Oxigenasa 1/biosíntesis , Macrófagos/metabolismo , Macrófagos/microbiología , Melioidosis/metabolismo , Animales , Western Blotting , Burkholderia pseudomallei , Citocinas/biosíntesis , Modelos Animales de Enfermedad , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Técnicas de Silenciamiento del Gen , Melioidosis/patología , Ratones , Ratones Endogámicos C57BL , Compuestos Organometálicos/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Neurologic melioidosis is a serious, potentially fatal form of Burkholderia pseudomallei infection. Recently, we reported that a subset of clinical isolates of B. pseudomallei from Australia have heightened virulence and potential for dissemination to the central nervous system. In this study, we demonstrate that this subset has a B. mallei-like sequence variation of the actin-based motility gene, bimA. Compared with B. pseudomallei isolates having typical bimA alleles, isolates that contain the B. mallei-like variation demonstrate increased persistence in phagocytic cells and increased virulence with rapid systemic dissemination and replication within multiple tissues, including the brain and spinal cord, in an experimental model. These findings highlight the implications of bimA variation on disease progression of B. pseudomallei infection and have considerable clinical and public health implications with respect to the degree of neurotropic threat posed to human health.
Asunto(s)
Burkholderia pseudomallei/genética , Infecciones Bacterianas del Sistema Nervioso Central/microbiología , Variación Genética , Melioidosis/microbiología , Proteínas de Microfilamentos/genética , Animales , Australia , Burkholderia mallei/genética , Burkholderia pseudomallei/aislamiento & purificación , Infecciones Bacterianas del Sistema Nervioso Central/mortalidad , Infecciones Bacterianas del Sistema Nervioso Central/patología , Enfermedades Transmisibles Emergentes/microbiología , Enfermedades Transmisibles Emergentes/mortalidad , Enfermedades Transmisibles Emergentes/patología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Muermo/microbiología , Humanos , Melioidosis/mortalidad , Melioidosis/patología , Ratones , Mucosa Nasal/microbiología , Fagocitos/inmunología , Fagocitos/microbiología , Virulencia/genéticaRESUMEN
Burkholderia pseudomallei is a Gram-negative intracellular bacterium that causes the disease melioidosis. The disease can be fatal if left untreated or when antibiotic therapy is delayed and total clearance of the pathogen from the host is often not accomplished with current therapies. Thus, new therapeutic approaches for the treatment of infections caused by B. pseudomallei are required. To better understand host responses to B. pseudomallei infection, the activation of key proteins involved in the TLR inflammatory cascade was measured by western blotting. Activation of the mitogen-activated protein kinases (MAPKs) p38 and ERK were both significantly altered during both in vitro and in vivo infection. In considering an approach for therapy of B. pseudomallei infection the inhibition of ERK was achieved in vitro using the inhibitor PD0325901, along with decreased TNF-α production. However, the reduction in phosphorylated ERK and TNF-α release did not correspond with decreased bacterial replication or enhance clearance from infected macrophages. Despite this apparent lack of effect on the intracellular growth of B. pseudomallei in vitro, it is not clear what effect inhibition of ERK activation might have on outcome of disease in vivo. It may be that decreasing the levels of TNF-α in vivo could aid in reducing the overactive immune response that is known to ensue following B. pseudomallei infection, thereby increasing host survival.
Asunto(s)
Burkholderia pseudomallei/crecimiento & desarrollo , Quimiocina CCL2/biosíntesis , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Melioidosis/patología , Factor de Necrosis Tumoral alfa/biosíntesis , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Benzamidas/farmacología , Burkholderia pseudomallei/inmunología , Burkholderia pseudomallei/metabolismo , Línea Celular , Difenilamina/análogos & derivados , Difenilamina/farmacología , Activación Enzimática , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Femenino , Macrófagos/microbiología , Melioidosis/inmunología , Melioidosis/microbiología , Ratones , Ratones Endogámicos BALB CRESUMEN
Melioidosis, a severe human disease caused by the bacterium Burkholderia pseudomallei, has a wide spectrum of clinical manifestations ranging from acute septicemia to chronic localized illness or latent infection. Murine models have been widely used to study the pathogenesis of infection and to evaluate novel therapies or vaccines, but how faithfully they recapitulate the biology of human melioidosis at a molecular level is not known. In this study, mice were intranasally infected with either high or low doses of B. pseudomallei to generate either acute, chronic, or latent infection and host blood and tissue transcriptional profiles were generated. Acute infection was accompanied by a homogeneous signature associated with induction of multiple innate immune response pathways, such as IL-10, TREM1, and IFN signaling, largely found in both blood and tissue. The transcriptional profile in blood reflected the heterogeneity of chronic infection and quantitatively reflected the severity of disease. Genes associated with fibrosis and tissue remodeling, including matrix metalloproteases and collagen, were upregulated in chronically infected mice with severe disease. Transcriptional signatures of both acute and chronic melioidosis revealed upregulation of iNOS in tissue, consistent with the expression of IFN-γ, but also Arginase-1, a functional antagonist of the iNOS pathway, and was confirmed by immunohistochemistry. Comparison of these mouse blood datasets by pathway and modular analysis with the blood transcriptional signature of patients with melioidosis showed that many genes were similarly perturbed, including Arginase-1, IL-10, TREM1, and IFN signaling, revealing the common immune response occurring in both mice and humans.
Asunto(s)
Burkholderia pseudomallei/inmunología , Inmunidad Innata/inmunología , Melioidosis/inmunología , Animales , Arginasa/biosíntesis , Arginasa/sangre , Modelos Animales de Enfermedad , Femenino , Perfilación de la Expresión Génica , Humanos , Interferón gamma/biosíntesis , Interferón gamma/sangre , Interferón gamma/inmunología , Interleucina-10/sangre , Interleucina-10/genética , Interleucina-10/inmunología , Pulmón/inmunología , Pulmón/microbiología , Pulmón/patología , Metaloproteinasa 9 de la Matriz/sangre , Melioidosis/microbiología , Melioidosis/patología , Glicoproteínas de Membrana/sangre , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Óxido Nítrico Sintasa de Tipo II/genética , Receptores Inmunológicos/sangre , Receptores Inmunológicos/genética , Receptores Inmunológicos/inmunología , Transducción de Señal/inmunología , Transcriptoma/genética , Receptor Activador Expresado en Células Mieloides 1RESUMEN
The genus Burkholderia comprises metabolically diverse and adaptable Gram-negative bacteria, which thrive in often adversarial environments. A few members of the genus are prominent opportunistic pathogens. These include Burkholderia mallei and Burkholderia pseudomallei of the B. pseudomallei complex, which cause glanders and melioidosis, respectively. Burkholderia cenocepacia, Burkholderia multivorans, and Burkholderia vietnamiensis belong to the Burkholderia cepacia complex and affect mostly cystic fibrosis patients. Infections caused by these bacteria are difficult to treat because of significant antibiotic resistance. The first line of defense against antimicrobials in Burkholderia species is the outer membrane penetration barrier. Most Burkholderia contain a modified lipopolysaccharide that causes intrinsic polymyxin resistance. Contributing to reduced drug penetration are restrictive porin proteins. Efflux pumps of the resistance nodulation cell division family are major players in Burkholderia multidrug resistance. Third and fourth generation ß-lactam antibiotics are seminal for treatment of Burkholderia infections, but therapeutic efficacy is compromised by expression of several ß-lactamases and ceftazidime target mutations. Altered DNA gyrase and dihydrofolate reductase targets cause fluoroquinolone and trimethoprim resistance, respectively. Although antibiotic resistance hampers therapy of Burkholderia infections, the characterization of resistance mechanisms lags behind other non-enteric Gram-negative pathogens, especially ESKAPE bacteria such as Acinetobacter baumannii, Klebsiella pneumoniae and Pseudomonas aeruginosa.
Asunto(s)
Antibacterianos/farmacología , Burkholderia mallei/efectos de los fármacos , Burkholderia pseudomallei/efectos de los fármacos , Burkholderia/efectos de los fármacos , Farmacorresistencia Bacteriana Múltiple/genética , Regulación Bacteriana de la Expresión Génica , Genes MDR , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Burkholderia/genética , Burkholderia/crecimiento & desarrollo , Burkholderia/patogenicidad , Infecciones por Burkholderia/tratamiento farmacológico , Infecciones por Burkholderia/microbiología , Infecciones por Burkholderia/patología , Burkholderia mallei/genética , Burkholderia mallei/crecimiento & desarrollo , Burkholderia mallei/patogenicidad , Burkholderia pseudomallei/genética , Burkholderia pseudomallei/crecimiento & desarrollo , Burkholderia pseudomallei/patogenicidad , Girasa de ADN/genética , Girasa de ADN/metabolismo , Muermo/tratamiento farmacológico , Muermo/microbiología , Muermo/patología , Caballos , Humanos , Melioidosis/tratamiento farmacológico , Melioidosis/microbiología , Melioidosis/patología , Porinas/antagonistas & inhibidores , Porinas/genética , Porinas/metabolismo , Tetrahidrofolato Deshidrogenasa/genética , Tetrahidrofolato Deshidrogenasa/metabolismoRESUMEN
PURPOSE: Melioidosis is a potentially fatal infectious disease caused by Burkholderia pseudomallei. Neurologic involvement in pediatric age group is very rare, and only a handful of cases have been reported in literature. We sought to provide a systematic review of pediatric neurologic melioidosis. METHODS: Literature review was performed to analyze reported cases of pediatric neurologic melioidosis (≤16 years) by searching online database (PubMed/MEDLINE). RESULTS: Twenty-seven cases were analyzed. Mean age was 6.7 years (range 2 days-14 years) and around 50% were older children (>5 years). Cranial nerve palsies and fever were most common presenting features. Major manifestations were meningoencephalitis in 16 (59%) and cerebral abscesses in eight (29%) cases. Abscesses were mostly located in parietal lobe. Among older children, abscesses were common than meningeal disease, while being converse for neonates. Mean follow-up duration was 11.5 months. Ten cases showed good to excellent outcome, while eight cases had fair (incomplete recovery) outcomes. The overall mortality rate was 18.5% (5/27) and tends to decrease with age. Among the five deaths, 4 (80%) had septicemia or pneumonia. CONCLUSIONS: Pediatric neurologic melioidosis is very rare. Meningoencephalitis is the most common presentation. Mortality is the highest in neonates. Ceftazidime appears to be the drug of choice in intensive phase, although the best drug in maintenance phase cannot be commented upon, especially in very young children.
Asunto(s)
Burkholderia pseudomallei/patogenicidad , Infecciones Bacterianas del Sistema Nervioso Central , Melioidosis , Adolescente , Antibacterianos/uso terapéutico , Infecciones Bacterianas del Sistema Nervioso Central/tratamiento farmacológico , Infecciones Bacterianas del Sistema Nervioso Central/patología , Niño , Preescolar , Femenino , Humanos , Recién Nacido , Masculino , Melioidosis/tratamiento farmacológico , Melioidosis/patologíaRESUMEN
Burkholderia pseudomallei is a causative agent of melioidosis. Clinical signs of melioidosis vary from acute septicemia to chronic inflammation or subclinical infection. This study investigated the role of B. pseudomallei biofilm in chronic inflammation in lungs of infected C57BL/6 mice. Low doses of B. pseudomallei H777 and its biofilm defective M10 mutant were fed intra-gastrically to C57BL/6 mice and inflammatory responses were investigated by histopathological techniques. Two hundred colony forming units (CFUs) of B. pseudomallei H777 induced chronic inflammatory responses in mice on day 20 post-infection, with discrete interstitial infiltration by mononuclear inflammatory cells. On day 40 postinfection, there were marked thickening of alveolar septa and congested capillaries, which increased in severity by day 60. On the other hand, mice infected with B. pseudomallei M10 showed less mononuclear infiltration. The results indicate that B. pseudomallei defective in biofilm production gave rise to less severe pathology, resulting a higher rate of survival in infected mice; and pulmonary melioidosis could be developed in C57BL/6 mice by intra-gastric feeding makes it a possible animal model of chronic human melioidosis.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Burkholderia pseudomallei/fisiología , Inflamación/microbiología , Melioidosis/patología , Animales , Enfermedad Crónica , Inflamación/patología , Melioidosis/microbiología , Ratones , Ratones Endogámicos C57BLRESUMEN
Melioidosis is caused by the saprophytic gram-negative bacillus Burkholderia pseudomallei, and has varied presentations, with cutaneous manifestations occurring in about 13% of cases. The usual histopathological features of melioidosis are suppurative to chronic granulomatous inflammation. Recommended treatment of melioidosis is sequential use of intravenous followed by oral antibiotics for a few months, although oral antibiotics alone can be used in primary cutaneous melioidosis. We report a case of cutaneous melioidosis in a healthy young man, with the unusual histopathological feature of lymphoplasmacytic inflammation, and failing an initial trial of oral antibiotics alone.
Asunto(s)
Melioidosis/patología , Enfermedades Cutáneas Bacterianas/patología , Administración Intravenosa , Adulto , Antibacterianos/administración & dosificación , Burkholderia pseudomallei/aislamiento & purificación , Humanos , Rodilla , Masculino , Resultado del TratamientoRESUMEN
Burkholderia pseudomallei (Bp), the causative agent of the often-deadly infectious disease melioidosis, contains one of the largest prokaryotic genomes sequenced to date, at 7.2 Mb with two large circular chromosomes (1 and 2). To comprehensively delineate the Bp transcriptome, we integrated whole-genome tiling array expression data of Bp exposed to >80 diverse physical, chemical, and biological conditions. Our results provide direct experimental support for the strand-specific expression of 5,467 Sanger protein-coding genes, 1,041 operons, and 766 non-coding RNAs. A large proportion of these transcripts displayed condition-dependent expression, consistent with them playing functional roles. The two Bp chromosomes exhibited dramatically different transcriptional landscapes--Chr 1 genes were highly and constitutively expressed, while Chr 2 genes exhibited mosaic expression where distinct subsets were expressed in a strongly condition-dependent manner. We identified dozens of cis-regulatory motifs associated with specific condition-dependent expression programs, and used the condition compendium to elucidate key biological processes associated with two complex pathogen phenotypes--quorum sensing and in vivo infection. Our results demonstrate the utility of a Bp condition-compendium as a community resource for biological discovery. Moreover, the observation that significant portions of the Bp virulence machinery can be activated by specific in vitro cues provides insights into Bp's capacity as an "accidental pathogen", where genetic pathways used by the bacterium to survive in environmental niches may have also facilitated its ability to colonize human hosts.