RESUMO
Dendritic cells (DC) enhanced the immunogenicity of recombinant vaccinia viruses (rVV) expressing the E7 protein of HPV16, a tumor-associated antigen (TAA). Immunization with DC transduced by rVV generated from strain Praha or MVA induced better protection against the growth of transplanted TC-1 tumors in C57Bl/6 mice than did immunization with either of these rVVs administered alone by the same route. Interestingly, DC transduced with a double recombinant vaccinia virus expressing E7 protein together with the Th1-polarizing cytokine IL12, which has been shown to enhance the cellular response in several other systems, induced lower anti-tumor immunity than DC transduced with rVV expressing E7 protein alone. The inhibitory effect mediated by IL12 on immunization with rVV-infected DC was dose-dependent and was observed after immunization with DC transduced with IL12-expressing rVV even at low multiplicity.
Assuntos
Células Dendríticas/imunologia , Terapia Genética , Imunoterapia , Interleucina-12/uso terapêutico , Neoplasias Experimentais/terapia , Proteínas Oncogênicas Virais/uso terapêutico , Vaccinia virus/genética , Adjuvantes Imunológicos , Animais , Apresentação de Antígeno , Vacinas Anticâncer/imunologia , Vacinas Anticâncer/uso terapêutico , Feminino , Vetores Genéticos , Humanos , Imunização , Interleucina-12/genética , Interleucina-12/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/virologia , Proteínas Oncogênicas Virais/genética , Proteínas Oncogênicas Virais/imunologia , Papillomaviridae/fisiologia , Proteínas E7 de Papillomavirus , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/terapia , Taxa de Sobrevida , Células Th1/imunologia , Transdução GenéticaRESUMO
Viral CC chemokine inhibitor (vCCI) of the clone P13 vaccinia virus (VACV) strain PRAHA lacks eight amino acids in the signal peptide sequence. To study the influence of vCCI on virus biology, a virus with the vCCI gene coding for a prolonged signal sequence was prepared. We found that secreted vCCI attenuated the virus in vivo, and that it correlated with decreased levels of RANTES, eotaxin, TARC, and MDC in the blood in comparison with the parental virus. We determined the influence of vCCI on the CTL response against VACV E3((140-148)) (VGPSNSPTF) and HPV16 E7((49-57)) (RAHYNIVTF) H-2D(b)-restricted epitopes. The examination of the specific CTL response elicited by immunization with the recombinant VACV-expressing tumor-associated HPV16 E7 antigen by IFN-γ ELISPOT showed that the immunogenicity of the recombinant VACV-producing secretory vCCI was similar to that of the parent virus or deletion mutant in the C23L/B29R locus. Immunization with the secretory vCCI-producing recombinant virus has a lower therapeutic anti-tumor effect against TC-1 tumors. Viral CCI downregulated the E7-specific response induced by gene gun immunization with the DNA vaccines pBSC-SigE7 LAMP and pBSC-vCCI. We also observed that the immune response against vCCI elicited by the DNA vaccine did not affect the multiplication of VACV in vivo.
Assuntos
Quimiocinas CC/antagonistas & inibidores , Quimiocinas CC/sangue , Proteínas E7 de Papillomavirus/genética , Linfócitos T Citotóxicos/imunologia , Vaccinia virus/genética , Vaccinia virus/imunologia , Proteínas Virais/metabolismo , Vacinas Virais/imunologia , Proteínas ADAM/sangue , Animais , Vacinas Anticâncer/genética , Vacinas Anticâncer/imunologia , Linhagem Celular , Quimiocina CCL17/sangue , Quimiocina CCL5/sangue , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas E7 de Papillomavirus/imunologia , Deleção de Sequência , Proteínas Supressoras de Tumor/sangue , Vacinação , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vaccinia virus/patogenicidade , Proteínas Virais/genéticaRESUMO
Tattooing has been shown to be very efficient at inducing immunity by vaccination with DNA vaccines. In this study, we examined the usability of tattooing for delivery of peptide vaccines. We compared tattooing with subcutaneous (s.c.) needle injection using peptides derived from human papillomavirus type 16 (HPV16) proteins. We observed that higher peptide-specific immune responses were elicited after vaccination with the simple peptides (E7(44-62) and E7(49-57)) and keyhole limpet hemocyanin-(KLH)-conjugated peptides (E7(49-57), L2(18-38) and L2(108-120)) with a tattoo device compared to s.c. inoculation. The administration of the synthetic oligonucleotide containing immunostimulatory CpG motifs (ODN1826) enhanced the immune responses developed after s.c. injection of some peptides (E7(44-62), KLH-conjugated L2(18-38) and L2(108-120)) to levels close to or even comparable to those after tattoo delivery of identical peptides with ODN1826. The highest efficacy of tattooing was observed in combination with ODN1826 for the vaccination with the less immunogenic E6(48-57) peptide and KLH-conjugated and non-conjugated E7(49-57) peptides which form the visible aggregates that could negatively influence the development of immune responses after s.c. injection but probably not after tattooing. In summary, we first evidenced that tattoo administration of peptide vaccines that might be useful in some cases efficiently induced both humoral and cell-mediated immune responses.
Assuntos
Proteínas do Capsídeo/imunologia , Papillomavirus Humano 16/imunologia , Proteínas Oncogênicas Virais/imunologia , Vacinas contra Papillomavirus/imunologia , Fragmentos de Peptídeos/imunologia , Proteínas Repressoras/imunologia , Tatuagem/instrumentação , Vacinação , Sequência de Aminoácidos , Animais , Feminino , Hemocianinas/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Proteínas E7 de Papillomavirus , Linfócitos T Citotóxicos/imunologia , Vacinas de DNA/imunologiaRESUMO
The Bordetella adenylate cyclase toxoid (CyaA) targets cells expressing the alphaMbeta2 integrin receptor CD11b/CD18 (CR3 or Mac-1) and can penetrate into cytosol of professional antigen-presenting cells, such as dendritic cells. This allows us to use CyaA for delivery of passenger antigens into the cytosolic pathway of processing and MHC class I-restricted presentation, which can promote induction of antigen-specific CD8+ cytotoxic T-lymphocyte immune responses. We show here that vaccination with a genetically detoxified CyaA336/E7 protein, carrying the full-length oncoprotein E7 of the human papilloma virus 16 inserted at position 336 of the cell-invasive AC domain of CyaA, induces an E7-specific CD8+ T-cell immune response and confers on mice protective, as well as therapeutic immunity against challenge with TC-1 tumor cells expressing the E7 oncoprotein. The therapeutic efficacy of priming with the CyaA336/E7 vaccine could further be enhanced by a heterologous booster immunization with a highly attenuated modified vaccinia virus Ankara (MVA) expressing the E7 protein fused to the lysosome-associated membrane protein (LAMP1). These results establish the potential of CyaA as a new antigen delivery tool for prime/boost immunotherapy of tumors.