RESUMEN
The class Mollicutes (trivial name "mycoplasma") is composed of wall-less bacteria with reduced genomes whose evolution was long thought to be only driven by gene losses. Recent evidences of massive horizontal gene transfer (HGT) within and across species provided a new frame to understand the successful adaptation of these minimal bacteria to a broad range of hosts. Mobile genetic elements are being identified in a growing number of mycoplasma species, but integrative and conjugative elements (ICEs) are emerging as pivotal in HGT. While sharing common traits with other bacterial ICEs, such as their chromosomal integration and the use of a type IV secretion system to mediate horizontal dissemination, mycoplasma ICEs (MICEs) revealed unique features: their chromosomal integration is totally random and driven by a DDE recombinase related to the Mutator-like superfamily. Mycoplasma conjugation is not restricted to ICE transmission, but also involves the transfer of large chromosomal fragments that generates progenies with mosaic genomes, nearly every position of chromosome being mobile. Mycoplasmas have thus developed efficient ways to gain access to a considerable reservoir of genetic resources distributed among a vast number of species expanding the concept of minimal cell to the broader context of flowing information.
Asunto(s)
Transferencia de Gen Horizontal , Mycoplasma/fisiología , Tenericutes/fisiología , Cromosomas Bacterianos , Conjugación Genética , Evolución Molecular , Mycoplasma/clasificación , Elementos de Respuesta , Tenericutes/clasificaciónRESUMEN
UNLABELLED: Mycoplasmas are minimal, wall-less bacteria but have retained the ability to secrete complex carbohydrate polymers that constitute a glycocalyx. In members of the Mycoplasma mycoides cluster, which are important ruminant pathogens, the glycocalyx includes both cell-attached and cell-free polysaccharides. This report explores the potential secretion of polysaccharides by M. agalactiae, another ruminant pathogen that belongs to a distant phylogenetic group. Comparative genomic analyses showed that M. agalactiae possesses all the genes required for polysaccharide secretion. Notably, a putative synthase gene (gsmA) was identified, by in silico reconstruction of the biosynthetic pathway, that could be involved in both polymerization and export of the carbohydrate polymers. M. agalactiae polysaccharides were then purified in vitro and found to be mainly cell attached, with a linear ß-(1â6)-glucopyranose structure [ß-(1â6)-glucan]. Secretion of ß-(1â6)-glucan was further shown to rely on the presence of a functional gsmA gene, whose expression is subjected to high-frequency phase variation. This event is governed by the spontaneous intraclonal variation in length of a poly(G) tract located in the gsmA coding sequence and was shown to occur in most of the M. agalactiae clinical isolates tested in this study. M. agalactiae susceptibility to serum-killing activity appeared to be dictated by ON/OFF switching of ß-(1â6)-glucan secretion, suggesting a role of this phenomenon in survival of the pathogen when it invades the host bloodstream. Finally, ß-(1â6)-glucan secretion was not restricted to M. agalactiae but was detected also in M. mycoides subsp. capri PG3(T), another pathogen of small ruminants. IMPORTANCE: Many if not all bacteria are able to secrete polysaccharides, either attached to the cell surface or exported unbound into the extracellular environment. Both types of polysaccharides can play a role in bacterium-host interactions. Mycoplasmas are no exception despite their poor overall metabolic capacity. We showed here that M. agalactiae secretes a capsular ß-(1â6)-glucopyranose thanks to a specific glycosyltransferase with synthase activity. This secretion is governed by high-frequency ON/OFF phase variation that might be crucial in mycoplasma host dissemination, as cell-attached ß-(1â6)-glucopyranose increases serum-killing susceptibility. Our results provide functional genetic data about mycoplasmal glycosyltransferases with dual functions, i.e., assembly and export of the sugar polymers across the cell membrane. Furthermore, we demonstrated that nonprotein epitopes can be subjected to surface antigenic variation in mycoplasmas. Finally, the present report contributes to unravel the role of secreted polysaccharides in the virulence and pathogenicity of these peculiar bacteria.
Asunto(s)
Mycoplasma agalactiae/metabolismo , Polisacáridos Bacterianos/metabolismo , beta-Glucanos/metabolismo , Biología Computacional , Redes y Vías Metabólicas/genética , Mycoplasma agalactiae/genéticaRESUMEN
BACKGROUND: The Global Rating Scale, defined by the Joint Advisory Group for Gastrointestinal Endoscopy, requires monitoring of endoscopic performance indicators. There are known variations in colonoscopic performance, and investigation of factors causing this is needed. This study aimed to analyse the impact of endoscopist specialty and procedural volume on the quality of colonoscopy. METHODS: Data collected prospectively from a UK hospital endoscopy service between June 2007 and January 2010 were analysed. The main endpoint was the adenoma detection rate (ADR). Secondary endpoints were polyp detection rate (PDR), reported caecal intubation rate (CIR) and reported complications. Multivariable binary regression models were built to adjust for confounding patient-level and endoscopist-level variation. RESULTS: A total of 10,026 colonoscopies were included, with an overall ADR of 19.2 per cent, a CIR of 90.2 per cent and a perforation rate of 0.06 per cent. In univariable analyses, surgeons had a higher ADR and higher PDR, but lower CIR, compared with physicians. Surgeons had a significantly different case mix in terms of age, sex and indication for colonoscopy. After adjusting for this case mix in multivariable analysis, specialty was no longer a significant predictor of ADR; however, surgeons retained their higher PDR and physicians their higher CIR. Endoscopists accredited for screening and those performing more than 100 colonoscopies per year had a higher ADR. CONCLUSION: Adjusting for case mix, physicians and surgeons performed equally well in terms of ADR. Accreditation and a higher annual number of colonoscopies were more important factors in achieving quality standards.
Asunto(s)
Adenoma/diagnóstico , Competencia Clínica/normas , Neoplasias del Colon/diagnóstico , Colonoscopía/normas , Cirugía Colorrectal/normas , Acreditación , Anciano , Pólipos del Colon/diagnóstico , Colonoscopía/efectos adversos , Colonoscopía/estadística & datos numéricos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Calidad de la Atención de Salud , Análisis de Regresión , Especialización , Carga de TrabajoRESUMEN
The association in young females of long-standing primary hypothyroidism, isosexual precocious pseudopuberty and multicystic enlarged ovaries was first described in 1960 by Van Wyk and Grumbach. Since then, sporadic case reports have contributed to clarifying the key features of this syndrome. The unique elements that lead to this diagnosis are FSH-dominated sexual precocity combined with a delayed bone age in the presence of hypothyroidism. It is important to recognise this syndrome because initiating simple thyroid hormone replacement completely resolves symptoms and hormone abnormalities, avoiding unnecessary investigations for malignancies or surgical intervention. We describe an 8-year-old girl with autoimmune thyroiditis and severe long-standing hypothyroidism presenting with the clinical features of Van Wyk-Grumbach syndrome, a secondary TSH-secreting adenoma and hyperprolactinaemia. In addition, this girl presented with microcytic anaemia, elevated erythrocyte sedimentation rate (ESR) and two unusual features - a newly developed streaky hyperpigmented skin lesion and parathyroid hormone suppression despite vitamin D deficiency. Thyroxine replacement normalised all hormone abnormalities and shrunk the pituitary adenoma within 9 months, but the new skin lesion persisted. We review the literature and explore the pathophysiology of known and new features that give rise to speculation indicating stimulation of the FSH G protein-coupled receptor by excessive TSH, but LH suppression by hyperprolactinaemia.
Asunto(s)
Hipotiroidismo/diagnóstico por imagen , Quistes Ováricos/diagnóstico por imagen , Pubertad Precoz/diagnóstico por imagen , Niño , Diagnóstico Diferencial , Femenino , Humanos , Hipotiroidismo/complicaciones , Quistes Ováricos/complicaciones , Pubertad Precoz/complicaciones , UltrasonografíaRESUMEN
Despite reducing disease, vaccination rarely protects against infection and many pathogens persist within vaccinated animal populations. Circulation of viral pathogens within vaccinated populations may favour the development of vaccine resistance with implications for the evolution of virus pathogenicity and the emergence of variant viruses. The high rate of mutations during replication of ribonucleic acid (RNA) viruses is conducive to the development of escape mutants. In vaccinated cattle, unusual mutations have been found in the major antigenic site of foot and mouth disease virus, which is also involved in receptor recognition. Likewise, atypical changes have been detected in the immunodominant region of bovine respiratory syncytial virus. Large deoxyribonucleic acid (DNA) viruses are able to recombine, generating new genotypes, as shown by the potential of glycoprotein E-negative vaccine strains of bovine herpesvirus-1 to recombine with wild-type strains. Marek's disease virus is often quoted as an example of vaccine-induced change in pathogenicity. The reasons for this increase in virulence have not been elucidated and possible explanations are discussed.
Asunto(s)
Vacunas Virales/inmunología , Virosis/veterinaria , Virus/patogenicidad , Animales , Mutación , Resultado del Tratamiento , Vacunas Virales/administración & dosificación , Vacunas Virales/efectos adversos , Virulencia/fisiología , Virosis/inmunología , Virosis/prevención & control , Virosis/virologíaRESUMEN
The Arg-Gly-Asp (RGD) triplet found in the G-H loop of capsid protein VP1 of foot-and-mouth disease virus (FMDV) is critically involved in the interaction of FMDV with integrin receptors and with neutralizing antibodies. Multiplication of FMDV C-S8c1 in baby hamster kidney 21 (BHK-21) cells selected variant viruses exploiting alternative mechanisms of cell recognition that rendered the RGD integrin-binding triplet dispensable for infectivity. By constructing chimeric viruses, we show that dispensability of the RGD in these variant FMDVs can be extended to surrounding amino acid residues. Replacement of eight amino acid residues within the G-H loop of VP1 by an unrelated FLAG marker yielded infectious virus. Evolution of FLAG-containing viruses in BHK-21 cells generated complex quasispecies in which individual mutants included amino acid replacements at other antigenic sites of FMDV. Inclusion of such replacements in the parental FLAG clone resulted in an increase of relative fitness of the viruses. These results suggest structural or functional connections between antigenic sites of FMDV and underscore the value of mutant spectrum analysis for the identification of fitness-promoting genetic modifications in viral populations. The possibility of producing viable viruses lacking antigenic site A may find application in the design of new anti-FMD vaccines.
Asunto(s)
Adaptación Fisiológica/inmunología , Antígenos Virales/inmunología , Cápside/inmunología , Virus de la Fiebre Aftosa/inmunología , Adaptación Fisiológica/genética , Secuencia de Aminoácidos , Animales , Antígenos Virales/genética , Sitios de Unión , Cápside/genética , Proteínas de la Cápside , Línea Celular , Cricetinae , Virus de la Fiebre Aftosa/genética , Virus de la Fiebre Aftosa/fisiología , Datos de Secuencia Molecular , Mutagénesis , Oligopéptidos , Péptidos , Replicación ViralRESUMEN
The effects of the vaccination of neonatal calves with a glycoprotein E (gE)-negative bovine herpesvirus type 1 (BHV-1) were investigated in naïve and passively immunised calves either with the recommended dose or a 5-fold concentrated one. After inoculation (PI), all calves excreted the virus vaccine except three passively immunised calves inoculated with the lower titre. No antibody response could be detected in passively immunised calves, whatever the dose used, and they all became BHV-1 seronegative and remained so after dexamethasone treatment (PDT). Nevertheless, as shown by a gamma-interferon assay, all calves that excreted the vaccine PI developed a cell-mediated immune response and a booster response was observed PDT, suggesting viral reactivation. The vaccine virus was recovered PDT from nasal secretions in two calves and BHV-1 DNA were detected in trigeminal ganglia from five calves belonging to all inoculated groups. The results show that the BHV-1 gE-negative vaccine can establish latency not only in naïve but also in passively immunised neonatal calves after a single intranasal inoculation. Moreover, this study shows for the first time that the gE-negative vaccine, when used in passively immunised calves, can lead to seronegative vaccine virus carriers.
Asunto(s)
Anticuerpos Antivirales/inmunología , Infecciones por Herpesviridae/veterinaria , Herpesvirus Bovino 1/inmunología , Inmunidad Materno-Adquirida , Rinotraqueítis Infecciosa Bovina/prevención & control , Vacunación/veterinaria , Vacunas Marcadoras/inmunología , Vacunas Virales/inmunología , Animales , Bovinos , Línea Celular , Calostro/inmunología , Dexametasona/farmacología , Relación Dosis-Respuesta Inmunológica , Ensayo de Inmunoadsorción Enzimática , Femenino , Infecciones por Herpesviridae/inmunología , Infecciones por Herpesviridae/prevención & control , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/crecimiento & desarrollo , Herpesvirus Bovino 1/aislamiento & purificación , Herpesvirus Bovino 1/fisiología , Inmunidad Celular , Inmunización Pasiva , Inmunización Secundaria , Rinotraqueítis Infecciosa Bovina/inmunología , Interferón gamma/sangre , Pruebas de Neutralización , Embarazo , Factores de Tiempo , Ganglio del Trigémino/virología , Vacunas Atenuadas/inmunología , Proteínas del Envoltorio Viral/deficiencia , Proteínas del Envoltorio Viral/genética , Proteínas Virales , Activación Viral/efectos de los fármacos , Latencia del VirusRESUMEN
Evolution of receptor specificity by viruses has several implications for viral pathogenesis, host range, virus-mediated gene targeting, and viral adaptation after organ transplantation and xenotransplantation, as well as for the emergence of viral diseases. Recent evidence suggests that minimal changes in viral genomes may trigger a shift in receptor usage for virus entry, even into the same cell type. A capacity to exploit alternative entry pathways may reflect the ancient evolutionary origins of viruses and a possible role as agents of horizontal gene transfers among cells.
Asunto(s)
Evolución Biológica , Receptores Virales/metabolismo , Fenómenos Fisiológicos de los Virus , Animales , Terapia Genética , Humanos , Especificidad de Órganos , Especificidad por Sustrato , Virosis/epidemiología , Virosis/virología , Virus/genética , Virus/inmunología , Virus/metabolismo , Virus/patogenicidadRESUMEN
Foot-and-mouth disease virus (FMDV) was the first animal virus identified. Since then, FMDV has become a model system in animal virology and a considerable amount of information on its structure, biology and vaccinology has been obtained. However, the disease that this virus produces (FMD) still constitutes one of the main animal health concerns. In this review, we have attempted to summarise the state of the knowledge in different basic and applied areas of FMDV research, with emphasis on those aspects relevant to the control of the disease.
Asunto(s)
Aphthovirus , Brotes de Enfermedades/veterinaria , Fiebre Aftosa/prevención & control , Fiebre Aftosa/virología , Vacunación/veterinaria , Animales , Variación Antigénica , Antígenos Virales/genética , Antígenos Virales/inmunología , Aphthovirus/química , Aphthovirus/genética , Aphthovirus/inmunología , Brotes de Enfermedades/prevención & control , Fiebre Aftosa/inmunología , Genotipo , Fenotipo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinariaRESUMEN
The genetic changes selected during the adaptation of a clonal population of foot-and-mouth disease virus (FMDV) to the guinea pig have been analyzed. FMDV clone C-S8c1 was adapted to the guinea pig by serial passage in the animals until secondary lesions were observed. Analysis of the virus directly recovered from the lesions developed by the animals revealed the selection of variants with two amino acid substitutions in nonstructural proteins, I(248)-->T in 2C and Q(44)-->R in 3A. On further passages, an additional mutation, L(147)-->P, was selected in an important antigenic site located in the G-H loop of capsid protein VP1. The amino acid substitution Q(44)-->R in 3A, either alone or in combination with the replacement I(248)-->T in 2C, was sufficient to give FMDV the ability to produce lesions. This was shown by using infectious transcripts which generated chimeric viruses with the relevant amino acid substitutions. Clinical symptoms produced by the artificial chimeras were similar to those produced by the naturally adapted virus. These results obtained with FMDV imply that one or very few replacements in nonstructural viral proteins, which should be within reach of the mutant spectra of replicating viral quasispecies, may result in adaptation of a virus to a new animal host.
Asunto(s)
Adaptación Fisiológica/genética , Sustitución de Aminoácidos/genética , Aphthovirus/genética , Aphthovirus/fisiología , Cobayas/virología , Mutación/genética , Proteínas no Estructurales Virales/metabolismo , Animales , Aphthovirus/clasificación , Aphthovirus/patogenicidad , Clonación Molecular , Fiebre Aftosa/patología , Fiebre Aftosa/virología , Genoma Viral , Masculino , Fenotipo , ARN Viral/genética , Selección Genética , Proteínas no Estructurales Virales/genética , Replicación ViralRESUMEN
We present sequence data from two genomic regions of foot-and-mouth disease virus (FMDV) subjected to several experimental passage regimens. Maximum-likelihood estimates of the nonsynonymous-to-synonymous rate ratio parameter (d(N)/d(S)) suggested the action of positive selection on some antigenic sites of the FMDV capsid during some experimental passages. These antigenic sites showed an accumulation of convergent amino acid replacements during massive serial cytolytic passages and also in persistent infections of FMDV in cell culture. This accumulation was most significant at the antigenic site A (the G-H loop of capsid VP1), which includes an Arg-Gly-Asp (RGD) cellular recognition motif. Our analyses also identified a subregion of VP3, part of the fivefold axis of FMDV particles, that also appeared to be subjected to positive selection of amino acid replacements. From these results, we can conclude that under the restrictive conditions imposed either by the presence of the monoclonal antibodies, by the persistent infections, or by the competition processes established between different variants of the viral population, amino acid replacement in some capsid-coding regions can be positively selected toward an increase of those mutants with a higher capability to infect the cell.
Asunto(s)
Aphthovirus/genética , Cápside/genética , Evolución Molecular , Selección Genética , Modelos Genéticos , Datos de Secuencia Molecular , Filogenia , Pase SeriadoRESUMEN
The consequences of the vaccination of neonatal calves with the widely used live-attenuated temperature-sensitive (ts) bovine herpesvirus type 1 (BHV-1) were investigated. The ts strain established acute and latent infections in all vaccinated calves either with or without passive immunity. Four of seven calves vaccinated under passive immunity became clearly BHV-1 seronegative by different serological tests, as did uninfected control calves after the disappearance of maternal antibodies, and they remained so for long periods. A cell-mediated immune response was detected by a BHV-1 gamma interferon assay, but this test failed to detect the seronegative latent carriers (SNLCs). While they are not detected, SNLCs represent a threat for BHV-1-free herds or countries. This study demonstrates that SNLCs can be easily obtained by inoculation with a live-attenuated BHV-1 under passive immunity and that latent carrier animals without any antibody do exist. Consequently, this situation could represent a good model to experimentally produce SNLCs.
Asunto(s)
Anticuerpos Antivirales/sangre , Portador Sano/veterinaria , Enfermedades de los Bovinos/virología , Infecciones por Herpesviridae/veterinaria , Herpesvirus Bovino 1/inmunología , Vacunas Virales/inmunología , Animales , Portador Sano/virología , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/prevención & control , Infecciones por Herpesviridae/inmunología , Infecciones por Herpesviridae/prevención & control , Infecciones por Herpesviridae/virología , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/aislamiento & purificación , Inmunidad Materno-Adquirida , Inmunización Pasiva , Interferón gamma/biosíntesis , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/inmunología , Vacunas Virales/administración & dosificación , Latencia del VirusRESUMEN
Biological adaptive systems share some common features: variation among their constituent elements and continuity of core information. Some of them, such as the immune system, are endowed with memory of past events. In this study we provide direct evidence that evolving viral quasispecies possess a molecular memory in the form of minority components that populate their mutant spectra. The experiments have involved foot-and-mouth disease virus populations with known evolutionary histories. The composition and behavior of the viral population in response to a selective constraint were influenced by past evolutionary history in a way that could not be predicted from examination of consensus nucleotide sequences of the viral populations. The molecular memory of the viral quasispecies influenced both the nature and the intensity of the response of the virus to a selective constraint.
Asunto(s)
Aphthovirus/genética , Aphthovirus/fisiología , Genoma Viral , Animales , Aphthovirus/clasificación , Secuencia de Bases , Línea Celular , Clonación Molecular , Cricetinae , Evolución Molecular , Mutación , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas Virales/química , Proteínas Virales/genéticaRESUMEN
Cell surface molecules that can act as virus receptors may exert an important selective pressure on RNA viral quasispecies. Large population passages of foot-and-mouth disease virus (FMDV) in cell culture select for mutant viruses that render dispensable a highly conserved Arg-Gly-Asp (RGD) motif responsible for integrin receptor recognition. Here, we provide evidence that viability of recombinant FMDVs including a Asp-143-->Gly change at the RGD motif was conditioned by a number of capsid substitutions selected upon FMDV evolution in cell culture. Multiply passaged FMDVs acquired the ability to infect human K-562 cells, which do not express integrin alpha(v)beta(3). In contrast to previously described cell culture-adapted FMDVs, the RGD-independent infection did not require binding to the surface glycosaminoglycan heparan sulfate (HS). Viruses which do not bind HS and lack the RGD integrin-binding motif replicate efficiently in BHK-21 cells. Interestingly, FMDV mutants selected from the quasispecies for the inability to bind heparin regained sensitivity to inhibition by a synthetic peptide that represents the G-H loop of VP1. Thus, a single amino acid replacement leading to loss of HS recognition can shift preferential receptor usage of FMDV from HS to integrin. These results indicate at least three different mechanisms for cell recognition by FMDV and suggest a potential for this virus to use multiple, alternative receptors for entry even into the same cell type.
Asunto(s)
Aphthovirus/metabolismo , Cápside/metabolismo , Oligopéptidos/metabolismo , Receptores Virales/metabolismo , Receptores de Vitronectina/metabolismo , Secuencia de Aminoácidos , Animales , Aphthovirus/genética , Aphthovirus/fisiología , Sitios de Unión , Células CHO , Cápside/genética , Proteínas de la Cápside , Línea Celular , Cricetinae , Heparitina Sulfato/metabolismo , Humanos , Células K562 , Datos de Secuencia Molecular , Oligopéptidos/genética , Recombinación Genética , Replicación ViralRESUMEN
Ribonucleic acid (RNA) viruses evolve as complex distributions of genetically different but closely related variants termed viral quasispecies. The precise genome of a quasispecies cannot be defined, since the consensus genome is an average of many variants. The dynamics of quasispecies has considerable implications for the understanding of the adaptability and pathogenic potential of viruses, and in addition, for the design of preventive and therapeutic measures for the diseases caused by these viruses. The authors summarise current knowledge on the structure of quasispecies, and the biological implications of this structure.
Asunto(s)
Evolución Molecular , Infecciones por Virus ARN/virología , Virus ARN/clasificación , Adaptación Biológica/genética , Animales , Ecología , Variación Genética , Genoma Viral , Humanos , Mutación , Fenotipo , Infecciones por Virus ARN/terapia , Virus ARN/genética , Virus ARN/patogenicidad , ARN Viral/químicaRESUMEN
The function of a loop exposed on the aphthovirus capsid (the G-H loop of protein VP1) has been explored by combining genetic and structural studies with viral mutants. The loop displays a dual function of receptor recognition and interaction with neutralizing antibodies. Remarkably, some amino acid residues play a critical role in both such disparate functions. Therefore residues subjected to antibody pressure for variation may nevertheless maintain a role in receptor recognition for which invariance is a requirement. Evolution of FMDV in cell culture may relax the requirements at this site and allow further increase of antigenic diversification. Essential residues at one stage of virus evolution may become dispensable at another not very distant point in the evolutionary landscape. Implications for FMDV evolution and vaccine design are discussed.
Asunto(s)
Anticuerpos Antivirales , Antígenos Virales/química , Aphthovirus/química , Aphthovirus/inmunología , Cápside/química , Animales , Anticuerpos Antivirales/biosíntesis , Antígenos Virales/inmunología , Cápside/inmunología , Proteínas de la Cápside , Células Cultivadas , Microscopía por Crioelectrón , Cristalografía por Rayos X , Epítopos , Humanos , Estructura Terciaria de Proteína , Receptores Virales/inmunologíaRESUMEN
In the context of infectious bovine rhinotracheitis (IBR) control programmes using glycoprotein E (gE) deleted marker vaccines, a PCR assay was developed to allow the genotypic differentiation between wildtype bovine herpesvirus type 1 (BoHV-1) and gE negative strains. This assay is based on the PCR amplification of a 281 bp DNA fragment within the gE gene. The specificity of the amplification was confirmed by restriction endonuclease analysis and nucleotide sequencing of the PCR product. Its ability to determine the gE genotype of BoHV-1 strains was demonstrated on isolates coming from 20 experimental calves infected with four different BoHV-1 strains. This PCR assay may be a useful tool for monitoring the spread of live marker vaccine and the gE genotype of viral field isolates.
Asunto(s)
Herpesvirus Bovino 1/aislamiento & purificación , Rinotraqueítis Infecciosa Bovina/virología , Reacción en Cadena de la Polimerasa/veterinaria , Proteínas del Envoltorio Viral/inmunología , Vacunas Virales/aislamiento & purificación , Animales , Bovinos , Células Cultivadas , Cartilla de ADN/química , ADN Viral/análisis , ADN Viral/química , Desoxirribonucleasas de Localización Especificada Tipo II/química , Electroforesis en Gel de Agar , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/inmunología , Rinotraqueítis Infecciosa Bovina/prevención & control , Riñón , Masculino , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Testículo , Proteínas del Envoltorio Viral/genética , Proteínas Virales , Vacunas Virales/genética , Vacunas Virales/inmunologíaRESUMEN
RNA virus populations consist of complex distributions of closely related but not identical genomes known as viral quasi-species. The quasi-species concept describes the dynamics of these genomes subjected to a continuous process of variation, competition, and selection. Quasi-species dynamics has broad implications not only in the understanding of the molecular mechanisms underlying adaptation of RNA viruses but also in the design of strategies for control and prevention of viral disease. Viral load and genetic heterogeneity have a determinant influence on the adaptation of RNA virus to their environment. Vaccines designed to control diseases caused by highly variable viruses must contain several B and T epitopes to provide an ample and diversified immune response. Similarly, antiviral drugs should be used in combination therapy to minimize selection of resistant viruses. The theoretical model of quasi-species has opened the way for new antiviral therapies based on augmentation of the mutation rate during replication of viral RNA. Finally the quasi-species concept provides the basis for defining the selective factors that could influence the evolution of RNA virus and promote the emergence or reemergence of viral diseases.
Asunto(s)
Virus ARN/fisiología , Antibiosis , Antivirales/uso terapéutico , Epítopos/inmunología , Variación Genética , Genoma Viral , Humanos , Biología Molecular , Mutación/genética , Infecciones por Virus ARN/prevención & control , Virus ARN/genética , Virus ARN/inmunología , Selección Genética , Carga Viral , Vacunas Virales/inmunología , Replicación Viral/genéticaRESUMEN
Viral quasispecies are closely related (but nonidentical) mutant and recombinant viral genomes subjected to continuous genetic variation, competition, and selection. Quasispecies structure and dynamics of replicating RNA enable virus populations to persist in their hosts and cause disease. We review mechanisms of viral persistence in cells, organisms, and populations of organisms and suggest that the critical interplay between host and viral influences (including in some cases the quasispecies organization) is the main driving force for long-term survival of viruses in nature.
Asunto(s)
Virus ARN/fisiología , Animales , Genoma Viral , Humanos , Virus ARN/genética , Latencia del VirusRESUMEN
RNA virus quasispecies are subjected to processes of positive Darwinian selection, to a very active and continuous negative selection and to random genetic drift. The course of RNA virus evolution is often unpredictable, and recent results suggest that even highly conserved motifs, once regarded as essential for infectivity, may be rendered dispensable by singular evolutionary events. An immediate consequence of the quasispecies genetic organization of RNA viruses is a surprising ability to gain fitness once a minimal replication ability is established in a biological environment. The unique features of RNA genetics should not be underestimated since they are at the basis of the emergence of new viral diseases and of the current difficulties to control many diseases associated variable viruses.