RESUMEN
Runx2 is an essential factor for skeletogenesis and heterozygous loss causes cleidocranial dysplasia in humans and a corresponding phenotype in the mouse. Homozygous Runx2-deficient mice lack hypertrophic cartilage and bone. We compared the expression profiles of E14.5 wildtype and Runx2(-/-) murine embryonal humeri to identify new transcripts potentially involved in cartilage and bone development. Seventy-one differentially expressed genes were identified by two independent oligonucleotide-microarray hybridizations and quantitative RT-PCR experiments. Gene Ontology analysis demonstrated an enrichment of the differentially regulated genes in annotations to terms such as extracellular, skeletal development, and ossification. In situ hybridization on E15.5 limb sections was performed for all 71 differentially regulated genes. For 54 genes conclusive in situ hybridization results were obtained and all of them showed skeletal expression. Co-expression with Runx2 was demonstrated for 44 genes. While 41 of the 71 differentially expressed genes have a known role in bone and cartilage, we identified 21 known genes that have not yet been implicated in skeletal development and 9 entirely new transcripts. Expression in the developing skeleton was demonstrated for 21 of these genes.
Asunto(s)
Desarrollo Óseo/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/deficiencia , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Animales , Desarrollo Óseo/fisiología , Displasia Cleidocraneal/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/fisiología , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Humanos , Hibridación in Situ , Ratones , Ratones Noqueados , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenotipo , Reacción en Cadena de la PolimerasaRESUMEN
Molecular analysis of 103 serogroup A Neisseria meningitidis strains isolated in Moscow from 1969 to 1997 showed that four independent clonal groupings were responsible for successive waves of meningococcal disease. An epidemic from 1969 to the mid-1970s was caused by genocloud 2 of subgroup III, possibly imported from China. Subsequent endemic disease through the early 1990s was caused by subgroup X and then by subgroup VI, which has also caused endemic disease elsewhere in eastern Europe. A 1996 epidemic was part of the pandemic spread from Asia of genocloud 8 of subgroup III. Recent genocloud 8 epidemic disease in Moscow may represent an early warning for spread of these bacteria to other countries in Europe.
Asunto(s)
Meningitis Meningocócica/epidemiología , Neisseria meningitidis/genética , Técnicas de Tipificación Bacteriana , Humanos , Meningitis Meningocócica/microbiología , Neisseria meningitidis/clasificación , Filogenia , Técnica del ADN Polimorfo Amplificado Aleatorio , Federación de Rusia/epidemiología , Factores de TiempoRESUMEN
The genetic variability at six polymorphic loci was examined within a global collection of 502 isolates of subgroup III, serogroup A Neisseria meningitidis. Nine "genoclouds" were identified, consisting of genotypes that were isolated repeatedly plus 48 descendent genotypes that were isolated rarely. These genoclouds have caused three pandemic waves of disease since the mid-1960s, the most recent of which was imported from East Asia to Europe and Africa in the mid-1990s. Many of the genotypes are escape variants, resulting from positive selection that we attribute to herd immunity. Despite positive selection, most escape variants are less fit than their parents and are lost because of competition and bottlenecks during spread from country to country. Competition between fit genotypes results in dramatic changes in population composition over short time periods.
Asunto(s)
Variación Genética/genética , Meningitis Meningocócica/epidemiología , Meningitis Meningocócica/microbiología , Neisseria meningitidis/genética , Neisseria meningitidis/inmunología , Selección Genética , Alelos , Evolución Biológica , Frecuencia de los Genes/genética , Genes Dominantes/genética , Genotipo , Geografía , Humanos , Meningitis Meningocócica/inmunología , Meningitis Meningocócica/transmisión , Datos de Secuencia Molecular , Mutación/genética , Neisseria meningitidis/clasificación , Neisseria meningitidis/fisiología , Filogenia , Polimorfismo Genético/genéticaRESUMEN
The protein sequences of the IgA1 protease, TbpA and TbpB proteins differ between meningococci representative of serogroup A, subgroup IV-1 from epidemic disease in The Gambia and serogroup C, ET-37 complex from endemic disease in Mali. The uniformity of restriction endonuclease sites was determined for the iga, tbpA and thpB genes among strains of both clonal lineages. Rare isolates had acquired a variant thpAB operon by horizontal genetic exchange but all other strains were uniform within each clonal lineage. The quantitative levels of IgG to capsular polysaccharide, IgA1 protease and TBP complex were measured in paired acute phase and convalescent phase sera from The Gambia and from Mali using antigens from the homologous clonal lineages. IgG levels to these antigens were also measured in paired sera from healthy Gambians who permanently carried meningococci in the nasopharynx or did not. The results showed that disease stimulated IgG to each antigen in Mali and to all but TBP complex in The Gambia. Similarly, higher levels of IgG were found in sera from permanent carriers than in sera from permanent non-carriers. Acute phase sera from Mali contained low levels of IgG to C capsular polysaccharide (geometric mean value of 0.3 microg ml(-1)) while such sera from The Gambia contained higher and potentially protective levels of IgG to A polysaccharide (geometric mean of 5.5 microg ml(-1)). The concentrations of IgG to TBP complex in acute phase sera were higher and IgG to IgA1 protease was even higher, suggesting that intermediate levels of IgG to these proteins do not protect against disease.
Asunto(s)
Anticuerpos Antibacterianos/biosíntesis , Proteínas Portadoras/inmunología , Inmunoglobulina G/biosíntesis , Meningitis Meningocócica/inmunología , Neisseria meningitidis/inmunología , Polisacáridos Bacterianos/inmunología , Serina Endopeptidasas/inmunología , Secuencia de Aminoácidos , Antígenos Bacterianos/inmunología , Epítopos/inmunología , Gambia/epidemiología , Humanos , Inmunoglobulina G/inmunología , Proteínas de Unión a Hierro , Meningitis Meningocócica/sangre , Meningitis Meningocócica/epidemiología , Datos de Secuencia Molecular , Nasofaringe/microbiología , Neisseria meningitidis/clasificación , Neisseria meningitidis/aislamiento & purificación , Proteínas de Unión a TransferrinaRESUMEN
Sera were taken over a 5 year period from Gambian children vaccinated in 1983, when aged 1-4 years, with A + C meningococcal capsular polysaccharide, ELISA tests were devised to determine the concentrations of immunoglobulin A, G and M reacting with A polysaccharide and of IgG reacting with Opc protein, IgA1 protease and an internal 104 mer peptide derived from IgA1 protease. Vaccination resulted in a brief rise of antibodies to A polysaccharide followed by decline to pre-immunization levels. IgM levels were very high even before vaccination. Antibodies to Opc protein stimulated by natural exposure also declined over the 5 year period. In contrast, antibodies stimulated by natural exposure to IgA1 protease or to the internal peptide remained constant or increased (final geometric mean level of 47 micrograms IgG ml-1). We speculate that healthy carriage of Neisseria meningitidis or Haemophilus influenzae is responsible for this increase in IgG concentration.
Asunto(s)
Anticuerpos Antibacterianos/biosíntesis , Proteínas de la Membrana Bacteriana Externa/inmunología , Neisseria meningitidis/enzimología , Neisseria meningitidis/inmunología , Polisacáridos Bacterianos/inmunología , Serina Endopeptidasas/inmunología , Secuencia de Aminoácidos , Anticuerpos Antibacterianos/sangre , Antígenos Bacterianos/inmunología , Antígenos de Superficie/inmunología , Vacunas Bacterianas/inmunología , Preescolar , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/sangre , Lactante , Datos de Secuencia MolecularRESUMEN
Five murine epitopes were defined and mapped within IgA1 protease produced by Neisseria meningitidis. Epitopes 1 and 2 were present in IgA1 protease from all strains, and from Neisseria gonorrhoeae. Epitopes 3 through to 5 varied between subgroups of serogroup A meningococci, but have remained constant over decades within the subgroups, except for epitope 4, which changed between 1983 and 1987 during the spread of subgroup III meningococci from Asia to Africa. Binding of monoclonal antibodies to epitopes 1, 4 and 5 neutralized enzymatic function. Human sera containing antibodies to IgA1 protease as a result of natural infection inhibited binding of monoclonal antibodies to epitope 4 but not to the other epitopes.