RESUMO
<p><b>OBJECTIVE</b>To evaluate the immune potency of recombinant adenovirus combined with rAAV1 vector expressing HPV16L1 protein in mice.</p><p><b>METHODS</b>The rAdV and rAAV1 vector containing codon-modified HPV16L1 gene was constructed using Admax and AAVmax packaging system respectively. C57 BL/6 mice were immunized with purified rAdV and rAAV1 vector through intramuscular and intranasal inoculation routes, and the titer of neutralizing antibody was determined by neutralization assay based HPV16 pseudovirus.</p><p><b>RESULTS</b>Intramuscular immunization by rAAV1-mod. HPV16L1 or combined with rAd-mod. HPV16L1 can induce higher titer of neutralizing antibody in serum than that of other groups. The titer of neutralizing antibody of intranasal groups is significantly lower than that of intramuscular group, although the prime-boost strategy using in intranasal group was effective to enhance the specific humoral immunity.</p><p><b>CONCLUSION</b>The rAAV1-mod. HPV16L1 combined with rAd-mod. HPV16L1 can induce higher titer of neutralizing antibody in serum through intramuscular route than that of other groups at the 16th week after the first immunization.</p>
Assuntos
Animais , Camundongos , Adenoviridae , Genética , Alergia e Imunologia , Infecções por Adenoviridae , Alergia e Imunologia , Anticorpos Antivirais , Alergia e Imunologia , Dependovirus , Genética , Alergia e Imunologia , Fenômenos do Sistema Imunitário , Imunização , Camundongos Endogâmicos C57BL , Proteínas de Fusão Oncogênica , Alergia e Imunologia , Proteínas Oncogênicas Virais , Alergia e Imunologia , Proteínas Recombinantes , Genética , Alergia e ImunologiaRESUMO
<p><b>OBJECTIVE</b>To construct recombinant adenovirus containing codon-modified HPV16L1 gene, and evaluate systemic and mucosal immunological responses induced after immunization with the recombinant virus.</p><p><b>METHODS</b>The recombinant adenovirus rAd-mod.HPV16L1 was constructed by Admax kit. The C57 BL/6 mice were immunized by purified rAd-mod.HPV16L1 through different inoculation routes. The immunological effect was evaluated by testing the specific neutralizing antibodies in sera and vaginal secretions of immunized mice through indirect ELISA and neutralization assay based HPV pseudovirus.</p><p><b>RESULTS</b>The result showed that intramuscular immunization could induce good systemic immunity, but the mucosal immunity was too weak, and immunization via intranasal route could induce satisfactory immunity both in sera and vaginal secretions, while intravaginal immunization failed to induce any specific immunological responses either in sera or vaginal secretions.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing codon- modified HPV16L1 gene was successfully constructed. Immunization through intranasal route could induce satisfactory immunity both in sera and vaginal secretions, while intramuscular immunization could only induce high titer of neutralizing antibodies in sera.</p>
Assuntos
Animais , Feminino , Camundongos , Adenoviridae , Genética , Anticorpos Antivirais , Alergia e Imunologia , Especificidade de Anticorpos , Proteínas do Capsídeo , Genética , Alergia e Imunologia , Códon , Genética , DNA Recombinante , Genética , Engenharia Genética , Papillomavirus Humano 16 , Genética , Alergia e Imunologia , Camundongos Endogâmicos C57BL , Proteínas Oncogênicas Virais , Genética , Alergia e Imunologia , VacinaçãoRESUMO
To investigate the feasibility of using recombinant adeno-associated virus type 1 vector as prophylactic vaccine against HPV16 infection, rAAV1-mod. HPV16L1, the recombinant AAV1 vector containing codon-modified HPV16 L1 gene, was constructed. C57BL/6 mice were immunized with purified rAAV1 vector through intramuscular and intranasal inoculation routes, and the titer of neutralizing antibody was determined by neutralization assay based on HPV16 pseudovirus. The result shows that the single dose of rAAV1-mod. HPV16L1 can induce specific neutralizing antibody in serum through both inoculation routes. Compared with intranasal group, intramuscular group can induce higher titer of neutralizing antibody. Eliciting strong and prolonged neutralizing antibody in serum, the rAAV1-mod. HPV16L1 is one of promising HPV16 prophylactic vaccine candidates.