RESUMO
CIGB-230, a mixture of a DNA plasmid expressing hepatitis C virus (HCV) structural antigens and a HCV recombinant capsid protein, has demonstrated to elicit strong immune responses in animals. The present study evaluated the plasmid biodistribution after the administration of CIGB-230 in mice, as well as toxicity of this vaccine candidate in rats. In the biodistribution study, mice received single or repeated intramuscular injections of CIGB-230, 50 microg of plasmid DNA mixed with 5 microg of Co.120 protein. Plasmid presence was assessed in ovaries, kidney, liver, pancreas, mesenteric ganglion, blood, and muscle of the injection site by a qualitative polymerase chain reaction. The toxicology evaluation included treatment groups receiving doses 5, 15, or 50 times higher, according to the body weight, than the expected therapeutic clinical dose. During the first hour after repeated inoculation, a promiscuous distribution was observed. However, 3 months later, plasmid could not be detected in any tissue. There was an absence of detectable adverse effects on key toxicology parameters and no damage evidenced in inspected organs and tissues. These results indicate that CIGB-230 is nontoxic at local and systemic levels and no concerns about persistence are observed, which support clinical testing of this vaccine candidate against HCV.
Assuntos
Hepacivirus/imunologia , Hepatite C/prevenção & controle , Vacinas de DNA/farmacocinética , Vacinas de DNA/toxicidade , Vacinas contra Hepatite Viral/farmacocinética , Vacinas contra Hepatite Viral/toxicidade , Animais , Feminino , Hepacivirus/genética , Antígenos de Hepatite/genética , Antígenos de Hepatite/imunologia , Hepatite C/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ratos Sprague-Dawley , Distribuição Tecidual , Testes de Toxicidade , Proteínas do Core Viral/genética , Proteínas do Core Viral/imunologiaRESUMO
In the present study, we evaluated the capability of the plasmid pIDKE2, encoding the HCV (hepatitis C virus) structural proteins Core, E1 and E2, to induce immune response against HCV antigens after injection into rabbits and Macaca irus (crab-eating macaque). Animals were immunized intramuscularly with different amounts of plasmid on weeks 0, 3 and 8. Monkeys received a booster dose on week 46. All rabbits immunized with pIDKE2 generated a positive antibody response and, particularly in rabbits immunized with 2 mg, antibody titres reached values above 1:1500 and 1:400 against the core and the envelope proteins, respectively, 28 weeks after primary immunization. The antibody response in monkeys developed slowly, but antibody titres greater than 1:3500 against HCV structural antigens were detected at week 52. Moreover, anti-E2 antibodies recognized synthetic peptides covering the HVR-1 (hypervariable region-1) from different isolates corresponding to different genotypes. Additionally, a specific lymphoproliferative response against Core and E2 was detected in two out of three monkeys immunized with pIDKE2. The other monkey had a specific proliferative response to E1. Taking all these data together, immunization with pIDKE2 is able to elicit both humoral and cellular immunity against HCV structural antigens in animal models other than mice.
Assuntos
Antígenos Virais/imunologia , Proteínas do Capsídeo/imunologia , Anticorpos Anti-Hepatite C/imunologia , Vacinação/métodos , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia , Proteínas do Envelope Viral/metabolismo , Animais , Antígenos Virais/administração & dosagem , Feminino , Técnicas de Transferência de Genes , Terapia Genética/métodos , Hepatite C/imunologia , Hepatite C/prevenção & controle , Imunização/métodos , Imunoterapia/métodos , Macaca , Masculino , Coelhos , Especificidade da EspécieRESUMO
The in vitro self-assembly properties of the entire hepatitis C virus core protein (HCcAg) obtained from Pichia pastoris cells and the induction of specific antibody immune response were studied. HCcAg was purified as a low-molecular-weight species by electroelution under denaturing conditions for confirmation of its self-assembly properties. After renaturalization, electron microscopy showed that HCcAg assembled into spherical particles of 30 nm. HCcAg also showed homogeneity and was specifically recognized by serum from a chronic HCV carrier patient. The data indicated that in vitro assembly of HCcAg, into virus-like particles resembling HCV nucleocapsid particles at a mature stage, is an intrinsic quality of this protein. Finally, HCcAg generated a strong antibody immune response in sheep, suggesting its usefulness for stimulating the host immune response against HCV.