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Anti-tumor effects of genetic vaccines against HPV major oncogenes.

Cordeiro MN, Paolini F, Massa S, Curzio G, Illiano E, Duarte Silva AJ, Franconi R, Bissa M, Morghen Cde G, de Freitas AC, Venuti A - Hum Vaccin Immunother (2014)

Bottom Line: The last one is a long multi epitope gene designed as a harmless E5 version.Both E5 genes were codon optimized for mammalian expression.These E7 and E6 vaccines were made with mutated oncogenes, the E7GGG mutant that does not bind pRb and the E6F47R mutant that is less effective in inhibiting p53, respectively.

View Article: PubMed Central - PubMed

Affiliation: a Federal University of Pernambuco; Department of Genetics; Laboratory of Molecular Studies and Experimental Therapy (LEMTE); Pernambuco, Brazil.

ABSTRACT
Expression of HPV E5, E6 and E7 oncogenes are likely to overcome the regulation of cell proliferation and to escape immunological control, allowing uncontrolled growth and providing the potential for malignant transformation. Thus, their three oncogenic products may represent ideal target antigens for immunotherapeutic strategies. In previous attempts, we demonstrated that genetic vaccines against recombinant HPV16 E7 antigen were able to affect the tumor growth in a pre-clinical mouse model. To improve this anti-HPV strategy we developed a novel approach in which we explored the effects of E5-based genetic immunization. We designed novel HPV16 E5 genetic vaccines based on two different gene versions: whole E5 gene and E5Multi. The last one is a long multi epitope gene designed as a harmless E5 version. Both E5 genes were codon optimized for mammalian expression. In addition, we demonstrated that HPV 16 E5 oncogene is expressed in C3 mouse cell line making it an elective model for the study of E5 based vaccine. In this mouse model the immunological and biological activity of the E5 vaccines were assessed in parallel with the activity of anti-E7 and anti-E6 vaccines already reported to be effective in an immunotherapeutic setting. These E7 and E6 vaccines were made with mutated oncogenes, the E7GGG mutant that does not bind pRb and the E6F47R mutant that is less effective in inhibiting p53, respectively. Results confirmed the immunological activity of genetic formulations based on attenuated HPV16 oncogenes and showed that E5-based genetic immunization provided notable anti-tumor effects.

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Related in: MedlinePlus

Figure 3. HPV16 E5 and E5Multi genes. The schematic representations of the E5 HPV16 gene (252 bp) with the position of CTL epitope sequences (cassettes A and B) and of the synthetic E5Multi gene (224 bp) with the position of the duplicated CTL epitope sequences (cassettes A and B) are shown together with the restriction sites for directional cloning. The aminoacid sequences of both epitopes are also indicated.
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Figure 3: Figure 3. HPV16 E5 and E5Multi genes. The schematic representations of the E5 HPV16 gene (252 bp) with the position of CTL epitope sequences (cassettes A and B) and of the synthetic E5Multi gene (224 bp) with the position of the duplicated CTL epitope sequences (cassettes A and B) are shown together with the restriction sites for directional cloning. The aminoacid sequences of both epitopes are also indicated.

Mentions: In premalignant lesions, when E5 is still expressed, a vaccine targeted to E5-expressing cells may be a good strategy to prevent premalignant lesions from progressing to invasive cervical cancers. We designed two different constructs to be utilized as genetic vaccine: the whole E5 gene and a synthetic harmless version. Indeed, the E5 protein can be harmful in humans due its oncogenic activity, and therefore we designed an E5-based gene, the E5Multi, which contains two previous described coding sequences for immune epitopes, in duplicate.29,30 By this way, this E5Multi gene should increase the immunogenicity of the antigen and in the meantime could eliminate any possible oncogenic activity. An illustrated scheme of E5 and E5Multi genes is described in Figure 3.


Anti-tumor effects of genetic vaccines against HPV major oncogenes.

Cordeiro MN, Paolini F, Massa S, Curzio G, Illiano E, Duarte Silva AJ, Franconi R, Bissa M, Morghen Cde G, de Freitas AC, Venuti A - Hum Vaccin Immunother (2014)

Figure 3. HPV16 E5 and E5Multi genes. The schematic representations of the E5 HPV16 gene (252 bp) with the position of CTL epitope sequences (cassettes A and B) and of the synthetic E5Multi gene (224 bp) with the position of the duplicated CTL epitope sequences (cassettes A and B) are shown together with the restriction sites for directional cloning. The aminoacid sequences of both epitopes are also indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4514265&req=5

Figure 3: Figure 3. HPV16 E5 and E5Multi genes. The schematic representations of the E5 HPV16 gene (252 bp) with the position of CTL epitope sequences (cassettes A and B) and of the synthetic E5Multi gene (224 bp) with the position of the duplicated CTL epitope sequences (cassettes A and B) are shown together with the restriction sites for directional cloning. The aminoacid sequences of both epitopes are also indicated.
Mentions: In premalignant lesions, when E5 is still expressed, a vaccine targeted to E5-expressing cells may be a good strategy to prevent premalignant lesions from progressing to invasive cervical cancers. We designed two different constructs to be utilized as genetic vaccine: the whole E5 gene and a synthetic harmless version. Indeed, the E5 protein can be harmful in humans due its oncogenic activity, and therefore we designed an E5-based gene, the E5Multi, which contains two previous described coding sequences for immune epitopes, in duplicate.29,30 By this way, this E5Multi gene should increase the immunogenicity of the antigen and in the meantime could eliminate any possible oncogenic activity. An illustrated scheme of E5 and E5Multi genes is described in Figure 3.

Bottom Line: The last one is a long multi epitope gene designed as a harmless E5 version.Both E5 genes were codon optimized for mammalian expression.These E7 and E6 vaccines were made with mutated oncogenes, the E7GGG mutant that does not bind pRb and the E6F47R mutant that is less effective in inhibiting p53, respectively.

View Article: PubMed Central - PubMed

Affiliation: a Federal University of Pernambuco; Department of Genetics; Laboratory of Molecular Studies and Experimental Therapy (LEMTE); Pernambuco, Brazil.

ABSTRACT
Expression of HPV E5, E6 and E7 oncogenes are likely to overcome the regulation of cell proliferation and to escape immunological control, allowing uncontrolled growth and providing the potential for malignant transformation. Thus, their three oncogenic products may represent ideal target antigens for immunotherapeutic strategies. In previous attempts, we demonstrated that genetic vaccines against recombinant HPV16 E7 antigen were able to affect the tumor growth in a pre-clinical mouse model. To improve this anti-HPV strategy we developed a novel approach in which we explored the effects of E5-based genetic immunization. We designed novel HPV16 E5 genetic vaccines based on two different gene versions: whole E5 gene and E5Multi. The last one is a long multi epitope gene designed as a harmless E5 version. Both E5 genes were codon optimized for mammalian expression. In addition, we demonstrated that HPV 16 E5 oncogene is expressed in C3 mouse cell line making it an elective model for the study of E5 based vaccine. In this mouse model the immunological and biological activity of the E5 vaccines were assessed in parallel with the activity of anti-E7 and anti-E6 vaccines already reported to be effective in an immunotherapeutic setting. These E7 and E6 vaccines were made with mutated oncogenes, the E7GGG mutant that does not bind pRb and the E6F47R mutant that is less effective in inhibiting p53, respectively. Results confirmed the immunological activity of genetic formulations based on attenuated HPV16 oncogenes and showed that E5-based genetic immunization provided notable anti-tumor effects.

Show MeSH
Related in: MedlinePlus