RESUMEN
A vaccine consisting of DNA priming followed by recombinant modified vaccinia Ankara (rMVA) boosting has achieved long-term control of a pathogenic challenge with a chimera of simian and human immunodeficiency viruses (SHIV-89.6P) in rhesus macaques. Based on these results, clade B HIV-1 DNA and rMVA immunogens have been developed for trials in humans. We conducted a first-time in humans phase I safety trial using the pGA2/JS2 (JS2) HIV-1 DNA priming vector expressing Gag, Pol, Env, Tat, Rev, and Vpu. Thirty HIV-uninfected adults were vaccinated with 0.3 or 3 mg of JS2 DNA, or a saline placebo, by intramuscular injection at months 0 and 2. Both doses of DNA were safe and well-tolerated with no differences between the control, 0.3 mg, or 3 mg groups (n = 6, 12, and 12, respectively) through 12 months of postvaccination follow- up. A chromium-release assay using fresh peripheral blood mononuclear cells (PBMCs) and a validated IFN-gamma ELISpot assay with frozen PBMCs failed to detect CD4(+) or CD8(+) HIV-1-specific T cell responses. HIV-specific neutralizing antibodies were also not detected. The vaccine is being further developed as a priming vector for a combined DNA plus rMVA prime/boost HIV vaccination regimen.
Asunto(s)
Vacunas contra el SIDA/efectos adversos , VIH-1/inmunología , Plásmidos/efectos adversos , Vacunas de ADN/efectos adversos , Virus Vaccinia/inmunología , Vacunas contra el SIDA/inmunología , Adulto , Método Doble Ciego , Ensayo de Inmunoadsorción Enzimática , Femenino , Vectores Genéticos/efectos adversos , Vectores Genéticos/inmunología , Anticuerpos Anti-VIH/metabolismo , Humanos , Interferón gamma/metabolismo , Masculino , Plásmidos/inmunología , Estadísticas no Paramétricas , Linfocitos T Citotóxicos/inmunología , Vacunas de ADN/inmunología , Virus Vaccinia/genéticaRESUMEN
The microsatellites HEL5, HEL9, INRA063, and BM2113 were used to analyze genetic similarities and differences of geographically isolated Criollo cattle herds in Mexico. Criollo cattle from five counties within the state of Chihuahua and one county from the state of Tamaulipas (n = 60) were sampled. The five counties in Chihuahua included Cerocahui (n = 14), Chinipas (n = 10), Guachochi (n = 15), Morelos (n = 30), and Temoris (n = 9). Samples of DNA were amplified by PCR and separated on a 7% polyacrylamide gel. Microsatellite size was established by comparison to M13mp18 DNA ladder and a documented set of four bovine controls. Allele frequencies and genotypic deviations from Hardy-Weinberg equilibrium were tested using the GENEPOP program. Eleven alleles were generated at HEL5 for the populations sampled (149 to 169 bp). Allele frequencies were greatest for the 163-bp allele in Criollo cattle from Cerocahui, Chinipas, Moralos, and Tamaulipas (0.23 to 0.5). Cattle from Guachochi had an allele frequency of 0.38 for the 151-bp allele, and cattle from Temoris had an allele frequency of 0.25 for the 149- and 167-bp alleles, with no 163-bp allele. Amplification with HEL9 produced 12 alleles (145, 149 to 169 bp) and showed common high-frequency alleles at 149, 157, and 159 bp for animals from all regions. The Chinipas population showed a moderate allele frequency at 145 bp; no other regions contained this allele. For INRA063 there were five alleles with 182 and 184 bp in low frequency. For BM2113 there were 10 alleles in the Criollo cattle (125 to 143 bp), with an equal distribution of frequencies for all alleles. In two regions, Guachochi and Morelos, genotypic frequencies deviated from Hardy-Weinberg equilibrium. Cattle from the Temoris region were genetically most distant from Criollo cattle of the other five regions.
Asunto(s)
Bovinos/genética , Variación Genética , Animales , Evolución Biológica , Frecuencia de los Genes , México , Repeticiones de MicrosatéliteRESUMEN
Infections with Streptococcus pneumoniae remain a significant cause of morbidity and mortality. To gain insight into structure-function relationships for human antibodies to pneumococcal capsular polysaccharide (PPS), we studied the response of transgenic mice reconstituted with human immunoglobulin loci, XenoMouse, to PPS antigens in a pneumococcal vaccine. Enzyme-linked immunosorbent assays of sera from mice vaccinated with a 23-valent pneumococcal vaccine revealed that they produced serotype-specific human antibodies, with the greatest response being to the PPS of serotype 3 (PPS 3). Molecular sequence analysis of three monoclonal antibodies (MAbs) to PPS 3 generated from lymphoid cells from mice vaccinated with a 23-valent pneumococcal vaccine or a PPS 3-bovine serum albumin conjugate revealed that they all used heavy-chain immunoglobulin genes from the V(H)3 family, two expressed light chain genes from the human Vkappa1 family, and one expressed a mouse lambda light chain. The protective efficacy of the two MAbs was examined in mice. A 10-microgram dose of both, and a 1-microgram dose of one, significantly prolonged survival from a lethal serotype 3 infection in CBA/N mice. Our data show that XenoMouse mice produced protective, serotype-specific human antibodies to PPS 3, and they lend support to the proposal that these animals represent a useful model to study the human antibody response to PPS antigens.