RESUMEN

Treatment of post-transplant patients with immunosuppressive drugs targeting the calcineurin-nuclear factor of activated T cells (NFAT) pathway, including cyclosporine A or tacrolimus, is commonly associated with a higher incidence of opportunistic infections, such as Aspergillus fumigatus, which can lead to severe life-threatening conditions. A component of the A. fumigatus cell wall, ß-glucan, is recognized by dendritic cells (DCs) via the Dectin-1 receptor, triggering downstream signaling that leads to calcineurin-NFAT binding, NFAT translocation, and transcription of NFAT-regulated genes. Here, we address the question of whether calcineurin signaling in CD11c-expressing cells, such as DCs, has a specific role in the innate control of A. fumigatus. Impairment of calcineurin in CD11c-expressing cells (CD11ccrecnb1loxP) significantly increased susceptibility to systemic A. fumigatus infection and to intranasal infection in irradiated mice undergoing bone marrow transplant. Global expression profiling of bone marrow-derived DCs identified calcineurin-regulated processes in the immune response to infection, including expression of pentraxin-3, an important antifungal defense protein. These results suggest that calcineurin inhibition directly impairs important immunoprotective functions of myeloid cells, as shown by the higher susceptibility of CD11ccrecnbloxP mice in models of systemic and invasive pulmonary aspergillosis, including after allogeneic bone marrow transplantation. These findings are relevant to the clinical management of transplant patients with severe Aspergillus infections.

Asunto(s)

Aspergilosis/inmunología , Aspergillus fumigatus/inmunología , Trasplante de Médula Ósea , Proteína C-Reactiva/metabolismo , Calcineurina/metabolismo , Células Dendríticas/inmunología , Inmunosupresores/efectos adversos , Componente Amiloide P Sérico/metabolismo , Animales , Proteína C-Reactiva/genética , Antígeno CD11c/metabolismo , Calcineurina/genética , Inhibidores de la Calcineurina/efectos adversos , Inhibidores de la Calcineurina/uso terapéutico , Células Cultivadas , Susceptibilidad a Enfermedades , Regulación hacia Abajo , Rechazo de Injerto/prevención & control , Humanos , Inmunosupresores/uso terapéutico , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Componente Amiloide P Sérico/genética , Transducción de Señal

RESUMEN

A crosstalk between commensals, gut immune cells, and colonic epithelia is required for a proper function of intestinal mucosal barrier. Here we investigated the importance of two distinct intestinal dendritic cell (DC) subsets in controlling intestinal inflammation. We show that Clec9A-diphtheria toxin receptor (DTR) mice after depletion of CD103(+)CD11b(-) DCs developed severe, low-dose dextran sodium sulfate (DSS)-induced colitis, whereas the lack of CD103(+)CD11b(+) DCs in Clec4a4-DTR mice did not exacerbate intestinal inflammation. The CD103(+)CD11b(-) DC subset has gained a functional specialization that able them to repress inflammation via several epithelial interferon-γ (IFN-γ)-induced proteins. Among others, we identified that epithelial IDO1 and interleukin-18-binding protein (IL-18bp) were strongly modulated by CD103(+)CD11b(-) DCs. Through its preferential property to express IL-12 and IL-15, this particular DC subset can induce lymphocytes in colonic lamina propria and in epithelia to secrete IFN-γ that then can trigger a reversible early anti-inflammatory response in intestinal epithelial cells.

Asunto(s)

Antígenos CD/inmunología , Antígeno CD11b/inmunología , Colitis/inmunología , Células Dendríticas/inmunología , Resistencia a la Enfermedad/inmunología , Cadenas alfa de Integrinas/inmunología , Interferón gamma/inmunología , Animales , Antígenos CD/genética , Antígeno CD11b/genética , Colitis/inducido químicamente , Colitis/genética , Colitis/patología , Colon/inmunología , Colon/patología , Células Dendríticas/patología , Sulfato de Dextran , Células Epiteliales/inmunología , Células Epiteliales/patología , Femenino , Regulación de la Expresión Génica , Factor de Crecimiento Similar a EGF de Unión a Heparina/genética , Factor de Crecimiento Similar a EGF de Unión a Heparina/inmunología , Indolamina-Pirrol 2,3,-Dioxigenasa/genética , Indolamina-Pirrol 2,3,-Dioxigenasa/inmunología , Cadenas alfa de Integrinas/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/inmunología , Interferón gamma/genética , Interleucina-12/genética , Interleucina-12/inmunología , Interleucina-15/genética , Interleucina-15/inmunología , Mucosa Intestinal/inmunología , Mucosa Intestinal/patología , Lectinas Tipo C/genética , Lectinas Tipo C/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Receptores Inmunológicos/genética , Receptores Inmunológicos/inmunología , Transducción de Señal

RESUMEN

BACKGROUND: Southeast Asian populations are increasingly affected by allergic airway diseases. Etiology and specific causes, however, are still unknown. The aim of this study is therefore to identify allergens and risk factors for the high prevalence of allergic airway disease in the tropical urban environment. METHODS: Symptoms of allergic rhinitis (AR), asthma, and allergic dermatitis were recorded in two independent cohorts of 576 and 7373 ethnic Chinese individuals living in Singapore. Reactivity against common allergens was determined by skin prick tests (SPT); specific immunoglobulin E (sIgE) titers against 12 common allergens, as well as total serum IgE (tIgE), were measured in the smaller cohort. RESULTS: Immunoglobulin E sensitization was almost exclusively directed against house dust mite (HDM) allergens. More than 80% of individuals were HDM-sIgE positive. Of these, less than 30% also had sIgE for other allergens, and similarly, few of the HDM-sIgE-negative individuals reacted to other allergens. Titers for HDM-sIgE were 8-30 times higher than other non-HDM allergen titers and correlated directly with total serum tIgE levels. Migrants from nontropical countries typically arrived with low or undetectable HDM-sIgE but developed substantial titers in a time-dependent fashion. Importantly, prolonged stay in Singapore also resulted in the manifestation of AR and asthma symptoms, contributing to some of the highest national prevalence rates worldwide. CONCLUSION: In a tropical urban environment, the allergic response is dominated by a single allergen class. The mono-specific IgE sensitization against HDM translates into increased prevalence of allergic airway diseases, which now impact a large proportion of the population in Singapore.

Asunto(s)

Alérgenos/inmunología , Pyroglyphidae/inmunología , Hipersensibilidad Respiratoria/epidemiología , Hipersensibilidad Respiratoria/etiología , Clima Tropical , Población Urbana , Adolescente , Adulto , Animales , Especificidad de Anticuerpos , Niño , Estudios de Cohortes , Femenino , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Masculino , Persona de Mediana Edad , Prevalencia , Hipersensibilidad Respiratoria/diagnóstico , Factores de Riesgo , Pruebas Cutáneas , Adulto Joven

RESUMEN

Until recently, West Nile (WN) and Kunjin (KUN) viruses were classified as distinct types in the Flavivirus genus. However, genetic and antigenic studies on isolates of these two viruses indicate that the relationship between them is more complex. To better define this relationship, we performed sequence analyses on 32 isolates of KUN virus and 28 isolates of WN virus from different geographic areas, including a WN isolate from the recent outbreak in New York. Sequence comparisons showed that the KUN virus isolates from Australia were tightly grouped but that the WN virus isolates exhibited substantial divergence and could be differentiated into four distinct groups. KUN virus isolates from Australia were antigenically homologous and distinct from the WN isolates and a Malaysian KUN virus. Our results suggest that KUN and WN viruses comprise a group of closely related viruses that can be differentiated into subgroups on the basis of genetic and antigenic analyses.

Asunto(s)

Fiebre del Nilo Occidental/virología , Virus del Nilo Occidental/clasificación , Aedes/citología , Secuencia de Aminoácidos , Animales , Antígenos Virales/análisis , Australia , Secuencia de Bases , Línea Celular , Chlorocebus aethiops , ADN Viral , Humanos , Malasia , Datos de Secuencia Molecular , New York/epidemiología , Filogenia , ARN Viral/análisis , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Células Vero , Fiebre del Nilo Occidental/epidemiología , Virus del Nilo Occidental/genética , Virus del Nilo Occidental/inmunología , Virus del Nilo Occidental/aislamiento & purificación

RESUMEN

MOTIVATION: Sequence databases represent an enormous resource of phylogenetic information, but there is a lack of tools for accessing that information in order to assess the amount of evolutionary information in these databases that may be suitable for phylogenetic reconstruction and for identifying areas of the taxonomy that are under-represented for specific gene sequences. RESULTS: We have developed TreeGeneBrowser which allows inspection and evaluation of gene sequence data for phylogenetic reconstruction. This program improves the efficiency of identification of genes that may be useful for particular phylogenetic studies and identifies taxa and taxonomic branches that are under-represented in sequence databases.

Asunto(s)

Mapeo Cromosómico , Bases de Datos Factuales , Biblioteca de Genes , Almacenamiento y Recuperación de la Información/métodos , Programas Informáticos , Algoritmos , Clasificación , Internet , National Library of Medicine (U.S.) , Filogenia , Estados Unidos

RESUMEN

BACKGROUND: Hepatitis G Virus (HGV)/GB Virus-C (GBV-C) is a newly discovered RNA virus. Nucleotide sequence comparison and phylogenetic studies of the 5' untranslated region (5'UTR) within the viral genome have identified at least three different types which have provisionally been classified as type 1 (West African origin), type 2 (North American origin) and type 3 (Asian origin). METHODS AND RESULTS: The products of RT-PCR were sequenced by using blood donors and patients infected with HGV/GBV-C in Australia, Papua New Guinea and the Solomon Islands to investigate the genotype distribution in this area of the world. All the Australian isolates showed strong sequence homology with type 2, while the Papua New Guinea and Solomon Islands sequences were more closely related, but differ from type 3, which has previously been reported from isolates studied within Asia. CONCLUSIONS: Phylogenetic analysis suggests that these latter sequences are either a new HGV/GBV-C Pacific type or a subtype of the Asian type RNA virus. Isolates homologous with type 1 were not identified in these population groups.

Asunto(s)

Flaviviridae/genética , Hepatitis Viral Humana/genética , Australia/epidemiología , Variación Genética , Genoma Viral , Genotipo , Humanos , Melanesia/epidemiología , Papúa Nueva Guinea/epidemiología , Filogenia , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Análisis de Secuencia de ARN

RESUMEN

We describe herein the molecular epidemiology and phylogeny of Kokobera (KOK) virus, a flavivirus found in Australia and Papua New Guinea. We sequenced a region encompassing the 200 nucleotides of the 3' terminus of the NS5 gene, and the first 300 nucleotides of the 3' untranslated region (UTR). The study included 25 isolates of the virus, including an isolate from PNG, and several recent isolates from the south-west of Western Australia (WA), where the virus had not previously been detected. We found that the KOK isolates clustered according to geographic location and time of isolation into three distinct topotypes: one covering Queensland and New South Wales; another represented by the single isolate from PNG; and a third covering the Northern Territory and WA. This latter group was further subdivided into northern and south-west isolates. This molecular epidemiology is significantly different from other Australian flaviviruses, such as Murray Valley encephalitis (MVE) and Kunjin (KUN) viruses, which exist as single genetic types across the entire Australian continent. However, it is similar to the molecular epidemiology of the alphavirus Ross River (RR) virus. This may be explained by the fact that MVE and KUN viruses are known to have birds as their main vertebrate hosts, whereas RR virus utilises macropods, which have also been implicated as the vertebrate host for KOK virus. In addition, the south-west isolates exhibited a degree of sequence heterogeneity, including one isolate that has a nine nucleotide deletion in the 3'UTR. This suggests that KOK virus has been in the south-west of WA for some time, and was not recently introduced.

Asunto(s)

Infecciones por Flavivirus/epidemiología , Infecciones por Flavivirus/virología , Flavivirus/genética , Regiones no Traducidas 3'/genética , Aedes/virología , Secuencia de Aminoácidos , Animales , Culex/virología , Flavivirus/aislamiento & purificación , Epidemiología Molecular , Datos de Secuencia Molecular , Papúa Nueva Guinea/epidemiología , Filogenia , Análisis de Secuencia de ADN , Proteínas no Estructurales Virales/genética , Australia Occidental/epidemiología

RESUMEN

Viral cultures were identified recently that contained both Kunjin virus and the closely related flavivirus West Nile. The observation that the KUN virus population grew more efficiently in a mosquito cell line (C6/36) while the WN population replicated more effectively in mammalian cells (Vero) allowed enrichment for either virus by culturing the mixture in the appropriate cell line. Limit dilution of the enriched virus preparations was then performed by infecting microtitre cultures with serial ten fold dilutions. Culture wells that contained a pure population of virus were then identified by immunostaining fixed cell monolayers with virus-specific monoclonal antibodies. Subsequent passage of the 'cloned' viruses in either C6/36 or Vero cells and analysis of the infected cultures by specific monoclonal antibody staining, PCR and nucleotide sequencing confirmed the identity of the virus and that in each case an homogeneous virus population had been obtained. This procedure is particularly useful for isolating virus populations from heterogeneous mixtures that fail to develop discrete plaques in infected cell monolayers.

Asunto(s)

Arbovirus/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática/métodos , Virología/métodos , Animales , Anticuerpos Monoclonales , Arbovirus/genética , Arbovirus/inmunología , Secuencia de Bases , Línea Celular , Chlorocebus aethiops , Culicidae , ADN Viral/genética , Virus de la Encefalitis Japonesa (Subgrupo)/genética , Virus de la Encefalitis Japonesa (Subgrupo)/inmunología , Virus de la Encefalitis Japonesa (Subgrupo)/aislamiento & purificación , Estudios de Evaluación como Asunto , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Ácido Nucleico , Especificidad de la Especie , Células Vero , Cultivo de Virus , Virus del Nilo Occidental/genética , Virus del Nilo Occidental/inmunología , Virus del Nilo Occidental/aislamiento & purificación

RESUMEN

The molecular epidemiology and evolution of Sindbis (SIN) virus in Australia was examined. Several SIN virus strains isolated from other countries were also included in the analysis. Two regions of the virus genome were sequenced including a 418 bp region of the E2 gene and a 484 bp region containing part of the junction region and the 5' end of the C gene. Analysis of the nucleotide and deduced amino acid sequence data from 40 SIN virus isolates clearly separated the Paleoarctic/Ethiopian and Oriental/Australian genetic types of SIN virus. Examination of the Australian strains showed a temporal rather than geographic relationship. This is consistent with the virus having migratory birds as the major vertebrate host, as it allows for movement of virus over vast areas of the continent over a relatively short period of time. The results suggest that the virus is being periodically redistributed over the continent from an enzootic focus of evolving SIN virus. However, SIN virus strains isolated from mosquitoes collected in the south-west of Australia appear to represent a new SIN virus lineage, which is distinct from the Paleoarctic/Ethiopian and Oriental/Australian lineages. Given the widespread geographic dispersal of the Paleoarctic/Ethiopian and Oriental/Australian lineages, it is surprising that the South-west genetic type is so restricted in its area of circulation. Nucleotide sequence data from the C gene of the prototype strain of the alphavirus Whataroa were also determined. This virus was found to be genetically distinct from the SIN virus isolates included in the present study; however, it is clearly SIN-like and appears to have evolved from a SIN-like ancestral virus.

Asunto(s)

Infecciones por Alphavirus/epidemiología , Evolución Molecular , Virus Sindbis/genética , África/epidemiología , Alphavirus/clasificación , Alphavirus/genética , Infecciones por Alphavirus/virología , Secuencia de Aminoácidos , Animales , Australia/epidemiología , Aves/virología , Cápside/química , Cápside/genética , Culicidae/virología , Análisis Mutacional de ADN , Genotipo , Humanos , Epidemiología Molecular , Datos de Secuencia Molecular , Islas del Pacífico/epidemiología , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Virus Sindbis/clasificación , Factores de Tiempo , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/genética

RESUMEN

OBJECTIVES: To investigate two outbreaks of dengue type 2 in north Queensland, one in the Torres Strait beginning in late 1996, the other in a Cairns suburb in early 1997. DESIGN: Epidemiological investigation of all laboratory-confirmed cases of dengue, entomological investigation of the local environment, and laboratory analysis of the isolated dengue viruses. MAIN OUTCOME MEASURES: Numbers of confirmed and of locally acquired cases; virus serotype; comparison of nucleotide sequences between viruses isolated from the two outbreaks; and Breteau Index (BI = number of containers with larvae of the mosquito vector Aedes aegypti found per 100 houses investigated) on the affected islands and in the Cairns suburb. RESULTS: There were 201 confirmed cases of dengue in the Torres Strait outbreak, which lasted nearly seven months, and seven confirmed cases in the Cairns outbreak, which lasted about nearly 11 weeks. Most (190) were confirmed as dengue type 2. Nucleotide sequencing of viruses isolated from the two outbreaks showed they were identical. Ae. aegypti breeding sites were very common on the five Torres Strait islands surveyed (BIs, 73-219--high risk), but less so in the Cairns suburb (BI, 23). The most common breeding sites were water storage reservoirs, particularly rainwater tanks, on the outer Torres Strait islands, discarded containers (such as plastic containers, buckets and tyres) on Thursday Island, and garden items (such as flowerpot bases and jars) in Cairns. CONCLUSIONS: The virus responsible for the Cairns outbreak was most probably introduced from the Torres Strait, whereas the virus responsible for the Torres Strait outbreak was imported from Papua New Guinea. Preventive strategies tailored to specific locations are needed to reduce breeding of Ae. aegypti in north Queensland, and the consequent risk of future outbreaks of dengue.

Asunto(s)

Virus del Dengue/clasificación , Dengue/epidemiología , Brotes de Enfermedades/estadística & datos numéricos , Dengue Grave/epidemiología , Adolescente , Adulto , Aedes/virología , Anciano , Animales , Secuencia de Bases , Niño , Preescolar , Estudios Transversales , Dengue/transmisión , Dengue/virología , Virus del Dengue/genética , Virus del Dengue/inmunología , Femenino , Humanos , Inmunoglobulina M/sangre , Incidencia , Lactante , Masculino , Persona de Mediana Edad , Queensland/epidemiología , ARN Viral/genética , Dengue Grave/transmisión , Dengue Grave/virología

RESUMEN

An increase in the incidence of polyarthritis caused by Barmah Forest (BF) virus, and its recent emergence into Western Australia, prompted a study of the molecular epidemiology of this Australian mosquito-borne alphavirus. The nucleotide sequence of a 500-basepair region of the 3' end of the envelope (E2) gene of the prototype BF virus strain (BH2193) was compared with other members of the alphavirus genus, and to a panel of isolates of BF virus collected more for than 20 years from different geographic regions of Australia. The BF virus was shown to be genetically distinct from other members of the alphavirus genus. A high degree of sequence homology (98-100%) was found between the BF isolates, with no evidence of geographic or temporal divergence. This nucleotide homogeneity was similar to that observed with other Australian mosquito-borne viruses with avian vertebrate hosts, such as Sindbis, Murray Valley, and Kunjin viruses, but it contrasts to the heterogeneity reported for Ross River virus, an alphavirus with mammalian vertebrate hosts.

Asunto(s)

Infecciones por Alphavirus/epidemiología , Alphavirus/genética , Artritis Infecciosa/virología , Epidemiología Molecular , Proteínas del Envoltorio Viral/genética , Secuencia de Aminoácidos , Animales , Artritis Infecciosa/epidemiología , Australia/epidemiología , Células Cultivadas , Chlorocebus aethiops , Biblioteca de Genes , Humanos , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Células Vero

RESUMEN

Mosquitoes were collected using CO2 and octenol-baited light traps during an outbreak of Japanese encephalitis (JE) on Badu Island in the Torres Strait, Australia in April 1995. A total of 13,300 mosquitoes comprising 12 species were processed for virus isolation. Eight isolates of JE virus were obtained from Culex annulirostris, with a carriage rate of 2.97:1,000; this mosquito also yielded one Sindbis virus isolate. A reverse transcriptase-polymerase chain reaction was used to sequence the JE viruses, which were compared with published sequences. The eight isolates were 90% homologous with known JE strains but only 68% homologous with other flaviviruses. Among the isolates, 99% homology was obtained, suggesting a common point of origin.

Asunto(s)

Culex/virología , Virus de la Encefalitis Japonesa (Especie)/aislamiento & purificación , Insectos Vectores/virología , Aedes/virología , Animales , Secuencia de Bases , Cartilla de ADN/química , ADN Viral/química , Brotes de Enfermedades , Virus de la Encefalitis Japonesa (Especie)/clasificación , Virus de la Encefalitis Japonesa (Especie)/genética , Encefalitis Japonesa/epidemiología , Encefalitis Japonesa/transmisión , Técnica del Anticuerpo Fluorescente Indirecta , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Queensland/epidemiología , ARN Viral/química , Homología de Secuencia de Ácido Nucleico

RESUMEN

OBJECTIVES: To determine the distribution of virus infection during an outbreak of Japanese encephalitis (JE) in the Torres Strait, and to describe the environmental factors facilitating the outbreak. DESIGN: Human and porcine serological surveys for JE virus activity throughout the Torres Strait, and mosquito and household surveys on the island of Badu. SETTING: The island of Badu (where the clinical cases occurred) and the other islands of the Torres Strait, Australia, during April-May 1995. RESULTS: The serological surveys identified recent JE virus infection among residents or domestic pigs on at least nine outer Torres Strait islands. A JE virus, confirmed by nucleotide sequencing, was isolated from two asymptomatic Badu residents. Virus isolations and mosquito surveys implicated Culex annulirostris as the major vector involved in the outbreak. There was prolific Cx. annulirostris breeding in a variety of water bodies close to and within the Badu community. Over half (53%) of the households kept pigs in pens, and many (63%) of the pigpens were situated near standing water; in 56% of these "wet" pigpens Cx. annulirostris was breeding. CONCLUSIONS: There was evidence of widespread JE virus activity throughout the outer islands of the Torres Strait. We suggest that migratory birds and/or wind-blown mosquitoes could have imported the virus into the Torres Strait from a focus of viral activity, possibly in Papua New Guinea, thereby initiating the outbreak. A combination of environmental factors, with large numbers of domestic pigs in close proximity to human dwellings and mosquito breeding sites, undoubtedly facilitated the outbreak on Badu.

Asunto(s)

Brotes de Enfermedades , Encefalitis Japonesa/epidemiología , Adulto , Animales , Australia/epidemiología , Secuencia de Bases , Culicidae/virología , Cartilla de ADN , Enfermedades de los Perros/virología , Perros , Virus de la Encefalitis Japonesa (Especie)/genética , Virus de la Encefalitis Japonesa (Especie)/aislamiento & purificación , Encefalitis Japonesa/transmisión , Encefalitis Japonesa/virología , Microbiología Ambiental , Genes Virales/genética , Enfermedades de los Caballos/virología , Caballos , Vivienda para Animales , Humanos , Insectos Vectores , Datos de Secuencia Molecular , Homología de Secuencia de Ácido Nucleico , Porcinos , Enfermedades de los Porcinos/virología

RESUMEN

The Japanese encephalitis (JE) serocomplex of flaviviruses comprises 10 members, 9 of which: Alfuy (ALF); Koutango (KOU); Kokobera (KOK); Kunjin (KUN); Murray Valley encephalitis (MVE); JE; Stratford (STR); Usutu (USU); and West Nile (WN) have been isolated from Africa, southern Europe, Middle East, Asia, and Australia. The tenth member, St. Louis encephalitis (SLE) virus, is confined to North, Central, and South America. For ALF, KOK, KOU, STR, and USU, no sequence data have as yet been reported, and little molecular phylogeny has been determined for this complex as a whole. Using a rapid, one-step RT-PCR and universal primers, we have amplified and sequenced a 450-600 base pair region of the virus genome encompassing the N terminus of the nonstructural protein NS5 and the 5' end of the 3' noncoding region, for several strains of all of these viruses, except USU and SLE viruses. These data, as well as published sequence data for other flaviviruses, were analyzed with the ClustalW and Phylip computer packages. The resultant phylogenetic data were consistent with some of the current flavivirus serological classification, showing a close relationship between ALF and MVE viruses and between KOK and STR viruses, but suggested that KOK and STR are distantly related to the other viruses and should perhaps be reclassified in their own serocomplex. The data also confirmed the close relationship between KUN and WN viruses and showed that an isolate of KUN virus from Sarawak may represent a "link" between these two virus species. In addition, the primary sequence data revealed a polymorphic region just downstream of the stop codon in the 3' end of the viral genomes.

Asunto(s)

Virus de la Encefalitis Japonesa (Subgrupo)/genética , Filogenia , Secuencia de Aminoácidos , Secuencia de Bases , Cartilla de ADN , Brotes de Enfermedades , Encefalitis por Arbovirus/virología , Humanos , Datos de Secuencia Molecular , ARN Viral/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico

RESUMEN

Three distinct patterns in the molecular epidemiology and evolution are evident among the alphaviruses and flaviviruses enzootic in Australia. One pattern, exemplified by MVE and KUN viruses, is of a single genetic type evolving slowly and uniformly in geographically widely separated regions of Australia with no evidence of independent divergence. The second pattern, exemplified by RR virus, is of separate genotypes evolving in different geographic regions with significant nucleotide divergence between genotypes. The third pattern, exemplified by SIN virus, is of a succession of temporally related genotypes that extend over most of the Australian continent, with relatively low levels of nucleotide divergence within a genotype, and which are each replaced by the subsequent genotype. These patterns are associated in part due to the nature and dispersal of their vertebrate hosts. Nucleotide divergence rates for Australian alphaviruses are similar to those reported elsewhere. Genomic relationships between Australian flavivirus members of the JE virus serological complex and between Australian alphaviruses are discussed, and evidence is presented for a possible new genomic lineage of SIN virus.

Asunto(s)

Alphavirus/genética , Culicidae/virología , Evolución Molecular , Flavivirus/genética , Animales , Australia , Humanos , Epidemiología Molecular

Asunto(s)

Priones/genética , Animales , Línea Celular , Clonación Molecular , Citomegalovirus/genética , Vectores Genéticos , Glicosilfosfatidilinositoles/metabolismo , Ratones , Mutación , Fosfatidilinositol Diacilglicerol-Liasa , Hidrolasas Diéster Fosfóricas/metabolismo , Regiones Promotoras Genéticas , Virus 40 de los Simios/genética

RESUMEN

The sequence of the coding regions of the PrP genes of the Arabian oryx and greater kudu have been determined and compared with the related sheep and bovine PrP gene sequences. The oryx gene sequence was found to be very closely related to that of the sheep, encoding just one amino acid difference. The greater kudu gene sequence was found to be more closely related to the bovine, encoding four amino acid differences. The possible influence that the gene sequences have on the transmission of spongiform encephalopathy to these antelope species is discussed.

Asunto(s)

Antílopes/genética , Enfermedades por Prión/veterinaria , Priones/genética , Animales , Secuencia de Bases , Bovinos , ADN , Predisposición Genética a la Enfermedad , Datos de Secuencia Molecular , Enfermedades por Prión/inmunología , Enfermedades por Prión/microbiología , Ovinos

RESUMEN

Murray Valley encephalitis (MVE) virus strain OR2 was serially passaged on Vero cells to establish a persistent infection which was maintained for over 300 days. Supernatants from infected cells protected Vero cells from c.p.e. and caused up to a 95% reduction of wild-type virus yield. These protective and interfering effects suggest that defective interfering (DI) particles are responsible for the establishment and maintenance of the MVE virus persistent infection. The persistently infected cell supernatant preparations shared several features with DI particle preparations from other viral systems, such as their amplification to detectable levels after two to four passages of virus. However, results from this study suggest that DI particles of MVE virus differ from other studied systems in that they are able to affect only moderately the yield of infectious wild-type virus. The genetic drift of the parental virus during the course of a long term persistent infection in vitro appears to be minimal.

Asunto(s)

Virus Defectuosos/fisiología , Flavivirus/fisiología , Animales , Efecto Citopatogénico Viral , Virus Defectuosos/genética , Flavivirus/genética , Técnica del Anticuerpo Fluorescente , Frecuencia de los Genes , ARN Viral/análisis , Pase Seriado , Células Vero