RESUMEN
OBJECTIVE: In order to gain further insight into the function of the enteric adenovirus short fiber (SF), we have constructed a recombinant dodecahedron containing the SF protein of HAdV-41 and the HAdV-3 penton base. METHODS: Recombinant baculoviruses expressing the HAdV-41 SF protein and HAdV-3 penton base were cloned and amplified in Sf9 insect cells. Recombinant dodecahedra were expressed by coinfection of High Five™ cells with both baculoviruses, 72 h post-infection. Cell lysate was centrifuged on sucrose density gradient and the purified recombinant dodecahedra were recovered. RESULTS: Analysis by negative staining electron microscopy demonstrated that chimeric dodecahedra made of the HAdV-3 penton base and decorated with the HAdV-41 SF were successfully generated. Next, recombinant dodecahedra were digested with pepsin and analyzed by Western blot. A 'site-specific' proteolysis of the HAdV-41 SF was observed, while the HAdV-3 penton base core was completely digested. CONCLUSION: These results show that, in vitro, the HAdV-41 SF likely undergoes proteolysis in the gastrointestinal tract, its natural environment, which may facilitate the recognition of receptors in intestinal cells. The results obtained in the present study may be the basis for the development of gene therapy vectors towards the intestinal epithelium, as well as orally administered vaccine vectors, but also for the HAdV-41 SF partner identification.
Asunto(s)
Adenovirus Humanos/genética , Proteínas de la Cápside/genética , Proteínas de la Cápside/ultraestructura , Sustancias Macromoleculares/ultraestructura , Virosomas/genética , Virosomas/ultraestructura , Animales , Baculoviridae/genética , Proteínas de la Cápside/metabolismo , Línea Celular , Clonación Molecular , Vectores Genéticos , Insectos , Sustancias Macromoleculares/metabolismo , Microscopía Electrónica , Pepsina A , Multimerización de Proteína , Proteolisis , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/ultraestructura , Virosomas/metabolismoRESUMEN
Human adenovirus type 7 (HAdV-7) is an important cause of acute respiratory disease (ARD). Different genomic variants of HAdV-7 have been described, designated 7a-7l. In a previous study to investigate risk factors for ARD and wheezing, nasopharyngeal samples were collected from 90 ill children seeking medical attention in Ribeirão Preto, São Paulo, Brazil. HAdVs were identified in 31 samples and were characterized by serum neutralization and genome restriction analysis. Eleven HAdVs were identified as being HAdV-7, five of which were classified as being of genome type 7p (Gomen). Six other HAdV-7 isolates gave new restriction profiles with all enzymes used and were classified as being a new genomic variant, 7m. These isolates were further characterized by sequencing. The hexon and fiber genes of the 7m variant were nearly identical to the prototype, 7p. However, nucleotide sequences from the E3 cassette revealed a 1743 bp deletion affecting the 16.1K, 19K, 20.1K and 20.5K ORFs.
Asunto(s)
Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/genética , Adenovirus Humanos/aislamiento & purificación , Eliminación de Secuencia , Proteínas E3 de Adenovirus , Adenovirus Humanos/clasificación , Brasil , Niño , Preescolar , ADN Viral/química , ADN Viral/genética , Femenino , Humanos , Lactante , Masculino , Datos de Secuencia Molecular , Nasofaringe/virología , Pruebas de Neutralización , Infecciones del Sistema Respiratorio/virología , Análisis de Secuencia de ADN , SerotipificaciónRESUMEN
Hantavirus pulmonary syndrome (HPS) is an increasing health problem in Brazil because of encroachment of sprawling urban, agricultural, and cattle-raising areas into habitats of subfamily Sigmodontinae rodents, which serve as hantavirus reservoirs. From 1993 through June 2007, a total of 884 cases of HPS were reported in Brazil (case-fatality rate 39%). To better understand this emerging disease, we collected 89 human serum samples and 68 rodent lung samples containing antibodies to hantavirus from a 2,500-km-wide area in Brazil. RNA was isolated from human samples and rodent tissues and subjected to reverse transcription-PCR. Partial sequences of nucleocapsid protein and glycoprotein genes from 22 human and 16 rodent sources indicated only Araraquara virus and Juquitiba virus lineages. The case-fatality rate of HPS was higher in the area with Araraquara virus. This virus, which may be the most virulent hantavirus in Brazil, was associated with areas that have had greater anthropogenic changes.
Asunto(s)
Enfermedades Transmisibles Emergentes/epidemiología , Síndrome Pulmonar por Hantavirus/epidemiología , Animales , Anticuerpos Antivirales/sangre , Secuencia de Bases , Brasil/epidemiología , Enfermedades Transmisibles Emergentes/inmunología , Enfermedades Transmisibles Emergentes/mortalidad , Enfermedades Transmisibles Emergentes/virología , Cartilla de ADN/genética , Genes Virales , Orthohantavirus/clasificación , Orthohantavirus/genética , Orthohantavirus/aislamiento & purificación , Orthohantavirus/patogenicidad , Síndrome Pulmonar por Hantavirus/inmunología , Síndrome Pulmonar por Hantavirus/mortalidad , Síndrome Pulmonar por Hantavirus/virología , Humanos , Proteínas de la Nucleocápside/genética , Filogenia , ARN Viral/genética , ARN Viral/aislamiento & purificación , Roedores/virología , Proteínas Virales/genética , Virulencia/genéticaRESUMEN
In a study of acute respiratory disease, two collections of nasopharyngeal aspirates (NPA) were obtained from children hospitalized at the Pediatric Clinic of the University Hospital, São Paulo, in 1995 and 2000. Adenovirus was detected in 33 (8.2%) of 401 children followed. These viruses were isolated in HEp-2, HEK-293, or NCI-H292 cells and serotyped by neutralization. The genome types were determined after restriction analyses of the genomic DNA extracted from infected cells. Nineteen isolates were characterized as Human adenovirus B, genome types HAdV-3a, HAdV-7h, and HAdV-7h1; 11 as Human adenovirus C, genome types HAdV-1D10, HAdV-2D25, HAdV-5D2, and HAdV-6D3. Our findings show that species C adenoviruses present an endemic infection pattern, with co-circulation of different serotypes and genome types; no new genomic variant was observed. Species B adenoviruses showed epidemic infection patterns, with shifts in the predominant genome type. The isolates from 1995 belong to genome type 7h, or the variant 7h1, with a clear substitution of the type 7b, prevalent in São Paulo for more then 10 years. In 2000, the variant 7h1 predominated and the emergence of the type 3a was observed. Almost 10 years passed between the identification of HAdV-7h in Argentina and its detection in São Paulo. The geographic isolation of these two countries was reduced by the increase in population mobility due to growing commercial relationships.
Asunto(s)
Infecciones por Adenovirus Humanos/epidemiología , Adenovirus Humanos/clasificación , Adenovirus Humanos/genética , Infecciones del Sistema Respiratorio/epidemiología , Enfermedad Aguda , Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/aislamiento & purificación , Adolescente , Brasil/epidemiología , Línea Celular , Niño , Preescolar , Genotipo , Hospitalización , Humanos , Epidemiología Molecular , Nasofaringe/virología , Pruebas de Neutralización , Infecciones del Sistema Respiratorio/virología , Serotipificación , Cultivo de VirusRESUMEN
BACKGROUND: Risk factors for acute wheezing among children in subtropical areas are largely unknown. OBJECTIVE: To investigate the role of viral infections, allergen sensitization, and exposure to indoor allergens as risk factors for acute wheezing in children 0 to 12 years old. METHODS: One hundred thirty-two children 0 to 12 years of age who sought emergency department care for wheezing and 65 children with no history of wheezing were enrolled in this case-control study. Detection of respiratory syncytial virus antigen, rhinovirus and coronavirus RNA, adenovirus, influenza, and parainfluenza antigens was performed in nasal washes. Total IgE and specific IgE to mites, cockroach, cat, and dog were measured with the CAP system. Major allergens from mites, cockroach, cat, and dog were quantified in dust samples by ELISA. Univariate and multivariate analyses were performed by logistic regression. RESULTS: In children under 2 years of age, infection with respiratory viruses and family history of allergy were independently associated with wheezing (odds ratio, 15.5 and 4.2; P = .0001 and P = .008, respectively). Among children 2 to 12 years old, sensitization to inhalant allergens was the major risk factor for wheezing (odds ratio, 2.7; P = .03). High-level allergen exposure, exposure to tobacco smoke, and lack of breast-feeding showed no association with wheezing. CONCLUSIONS: Some risk factors for wheezing previously identified in temperate climates were present in a subtropical area, including respiratory syncytial virus infection in infants and allergy in children older than 2 years. Rhinovirus was not associated with wheezing and did not appear to be a trigger for asthma exacerbations.