RESUMO
Vaccines against yellow fever currently recommended by the World Health Organization contain either virus sub-strains 17D or 17DD. In adults, the 17DD vaccine demonstrated high seroconversion and similar performance to vaccines manufactured with the WHO 17D-213/77 seed-lot. In another study, 17DD vaccine showed lower seroconversion rates in children younger than 2 years. Data also suggested lower seroconversion with simultaneous application of measles vaccine. This finding in very young children is not consistent with data from studies with 17D vaccines. A multicenter, randomized, double-blind clinical trial was designed (1) to compare the immunogenicity and reactogenicity of two yellow fever vaccines: 17DD (licensed product) and 17D-213/77 (investigational product) in children aged 9-23 months; (2) to assess the effect of simultaneous administration of yellow fever and the measles-mumps-rubella vaccines; and (3) to investigate the interference of maternal antibodies in the response to yellow fever vaccination. The anticipated implications of the results are changes in vaccine sub-strains used in manufacturing YF vaccine used in several countries and changes in the yellow fever vaccination schedule recommendations in national immunization programs.
Assuntos
Programas de Imunização , Vacina contra Febre Amarela/imunologia , Febre Amarela/imunologia , Vírus da Febre Amarela/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Brasil , Método Duplo-Cego , Contaminação de Medicamentos , Humanos , Lactente , Oligonucleotídeos/análise , Oligorribonucleotídeos/análise , Estudos Prospectivos , Vacinas Virais/administração & dosagem , Vacinas Virais/normas , Febre Amarela/prevenção & controle , Vacina contra Febre Amarela/administração & dosagem , Vacina contra Febre Amarela/efeitos adversosRESUMO
A rapid nucleic acid hybridization procedure was developed for examining the genotypic variation of dengue type 2 viruses (DEN 2) having distinct RNase T1 fingerprints and isolated from different geographical areas. Synthetic DNA hybridization probes were constructed complementary in nucleotide sequence to common and unique RNase T1 oligonucleotides of topotype viruses from Puerto Rico/South Pacific, Jamaica, the Seychelles, Thailand/Burma and Africa. Hybridization probes with both type- and topotype-specific reactivities were observed, as were probes specific for two or more of the DEN 2 topotypes. These results confirm geographical movement of topotype virus strains and suggest possible origins. Detection of DEN 2 RNA by hybridization is a rapid and reproducible method that can be modified and applied as a viable alternative to the laborious T1 oligonucleotide fingerprinting.
Assuntos
Vírus da Dengue/genética , Hibridização de Ácido Nucleico , RNA Viral/análise , África , Ásia , Sequência de Bases , DNA , Vírus da Dengue/classificação , Vírus da Dengue/isolamento & purificação , Exorribonucleases , Genes Virais , Genótipo , Mapeamento de Nucleotídeos , Oligodesoxirribonucleotídeos , Oligorribonucleotídeos/análise , RNA Viral/genética , Índias OcidentaisRESUMO
A rapid nucleic acid hybridization procedure was developed for examining the genotypic variation of dengue type 2 viruses (DEN 2) having distinct RNase T1 fingerprints and isolated from different geographical areas. Synthetic DNA hybridization probes were constructed complementary in nucleotide sequence to common and unique RNase T1 oligonecleotides of topotype viruses from Puerto Rico/South Pacific, Jamaica, the Seychelles, Thailand/Burma and Africa. Hybridization probes with both type- and topotype-specific reactivities were observed, as were probes specific for two or more of the DEN 2 topotypes. These results confirm geographical movement of topotype virus strains and suggest possible origins. Detection of DEN 2 RNA by hybridization is a rapid and reproducible method that can be modified and applied as a viable alternative to the labourious T1 oligonucleotide fingerprinting.(AU)
Assuntos
Vírus da Dengue/genética , Hibridização de Ácido Nucleico , RNA Viral , África , Ásia , Sequência de Bases , Vírus da Dengue/classificação , Vírus da Dengue/isolamento & purificação , DNA , Oligodesoxirribonucleotídeos , Exorribonucleases , Genes Virais , Genótipo , Mapeamento de Nucleotídeos , Oligorribonucleotídeos/análise , RNA Viral/genética , Índias OcidentaisRESUMO
Dengue-2 virus strains from different locations were compared by T1-RNAse-resistant oligonucleotide fingerprinting and antigen signature analysis. The latter technique involved construction of radioimmunoassays using monoclonal antibodies that recognize nine distinct dengue-2 type-specific and flavivirus cross-reactive epitopes over a range of antigen concentrations. A statistical method was used to align unknown dengue antigen concentrations in different strain preparations, allowing comparison of binding profiles. Twenty-six dengue-2 virus strains were separated into five distinct groups (topotypes) on the basis of unique RNA fingerprints. Two of these were represented by New Guinea C, the prototype virus isolated in 1944, and a Philippine strain; others were segregated on the basis of greater than or equal to 80% shared oligonucleotides into similarity groups representing Burma/Thailand (8 strains), Puerto Rico (12 strains), and Jamaica (4 strains). Signature analysis of the prototype and four geographic topotype strains revealed striking antigenic differences. In contrast, a high degree of antigenic similarity was found among strains from the same geographic region. Variation between antigenically distinct strains occurred at both type-specific and group-reactive epitopes, but the widest differences appeared at group-reactive determinants. Signature analysis provides a more rapid and simpler means than RNA fingerprinting of monitoring changes or new introductions of dengue virus populations in a geographic region.
Assuntos
Antígenos Virais/análise , Vírus da Dengue/classificação , Anticorpos Monoclonais , Anticorpos Antivirais , Antígenos Virais/imunologia , Reações Cruzadas , Vírus da Dengue/genética , Vírus da Dengue/imunologia , Epitopos , Jamaica , Mianmar , Nova Guiné , Oligorribonucleotídeos/análise , Filipinas , Porto Rico , RNA Viral/análise , TailândiaRESUMO
Genetic variation in dengue 2 isolates from various geographic areas was examined by oligonucleotide fingerprinting of the 40 S genome RNA. Oligonucleotide maps of geographically isolated and epidemiologically unrelated viruses were very distinct. Direct comparison of the oligonucleotide map of the dengue 2 prototype New Guinea 2 virus, isolated in 1944, with the fingerprints of more recent isolates from the South Pacific indicated that the genome of dengue 2 virus had undergone extensive change although the viruses are serologically indistinguishable. The oligonucleotide map of an isolate from a recent case in Jamaica and a mosquito isolate from Upper Volta, Africa, were recognized to be almost identical, suggesting that virus may have been introduced into the Caribbean from West Africa. Likewise, the fingerprints of isolates from Puerto Rico and the South Pacific shared 80 to 95% of their large oligonucleotides, suggesting that the virus involved in these epidemics may have spread throughout Tahiti, American Samoa, Fiji, and to Puerto Rico in the Caribbean or vice versa. On the basis of these studies, five genetic variants or topotypes of dengue 2 virus have been established: (1) Puerto Rico-South Pacific, (2) Burma-Thailand, (3) the Seychelles, (4) the Philippines, and (5) Jamaica-West Africa. Oligonucleotide fingerprinting offers a highly sensitive and reproducible technical approach to the investigation of dengue 2 virus intratypic variation and possibly to the understanding of the biological variation associated with dengue fever and hemorrhagic disease.
Assuntos
Vírus da Dengue/genética , Genes Virais , Variação Genética , RNA Viral/análise , África Ocidental , Ásia , Vírus da Dengue/classificação , Vírus da Dengue/isolamento & purificação , Ilhas do Oceano Índico , Jamaica , Oligorribonucleotídeos/análise , Ilhas do Pacífico , Porto RicoRESUMO
Unclassified Venezuelan equine encephalitis (VEE) viruses Tonate (TON), Bijou Bridge (BB), Paramana (PARA), 71D-1252 and Cabassou (CAB) were characterized serologically and biochemically. The envelope glycoproteins of these and nine other VEE viruses representing VEE subtype variants I-AB, I-C, I-D, I-E, II, III and IV were separated by column isoelectric focusing. The E1 and E2 glycoproteins of all the Zwittergent-dissociated VEE viruses focused at pI 6.3 to 6.9 and pI 8.6 to 9.3 respectively. Haemagglutination-inhibition and neutralization tests using rabbit sera to the E2 glycoprotein of TON, BB and PARA viruses showed them to be indistinguishable from each other and closely related to prototype subtype III virus Mucambo (MUC). VEE strain 71D-1252 was also serologically closely related to prototype MUC virus. We proposed that MUC, TON and 71D-1252 VEE viruses be classified subtype III viruses, designated variants III-A, III-B and III-C respectively. CAB virus, which is not closely related to other VEE isolates, may represent a new VEE subtype (V). SDS-PAGE resolved the capsid protein (35 to 36 kdal) and two major envelope glycoproteins of 50 to 51 kdal (E1) and 51 to 58 kdal (E2) for all VEE viruses except CAB; the two glycoproteins of CAB virus co-migrated by PAGE with apparent identical mol. wt. of 51 kdal. Limited digestion of SDS-dissociated virus proteins with Staphylococcus aureus V8 protease produced identical peptide maps for serologically indistinguishable viruses. Oligonucleotide fingerprinting of virus RNA supported the close serological relationships observed at the genome level.