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Enhanced immunogenicity of human papillomavirus 16 L1 genetic vaccines fused to an ER-targeting secretory signal peptide and RANTES

Abstract

To increase the potency of human papillomavirus (HPV) DNA vaccines, we constructed a series of HPV16 L1 vaccines genetically fused with a secretion signal and/or immune cell-recruiting RANTES. The DNA vaccines encoding secretory HPV L1 were constructed by inserting HPV L1 gene into a vector with an ER-targeting secretory signal sequence. The expression plasmid encoding secretory HPV L1 (pER/L1) was fused with cDNA of RANTES, generating pER/L1/R. For comparison, HPV L1 genes were cloned into pVAX1 vector with no signal sequence (pL1), and further linked to the N-terminus (pL1/R) or C-terminus of RANTES (pR/L1). The secretion of L1 proteins was observed in the pER/L1, pER/L1/R, and pR/L1-transfected cells, except the pL1/R-transfected group. Cytoplasmic localization of L1 protein was observed in the cells transfected with pL1/R, but not with pER/L1/R at 48 h after transfection. In mice, RANTES-fused vaccines more effectively elicited the levels of HPV16 L1-specific IgG and IgG2a antibodies than pL1. Of RANTES-fused vaccines, pER/L1/R encoding the secreted fusion protein induced the highest humoral and CD8+ T-cell-stimulating responses. These results suggest that the immunogenicity of HPV L1 DNA vaccines could be enhanced by genetic fusion to a chemokine and secretory signal peptide sequences.

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References

  1. Tjiong MY et al. Epidemiologic and mucosal immunologic aspects of HPV infection and HPV-related cervical neoplasia in the lower female genital tract. Int J Gynecol Cancer 2001; 11: 9–17.

    Article  CAS  Google Scholar 

  2. Giannoudis CS, Herrington CS . Human papillomavirus variants and squamous neoplasia of the cervix. J Pathol 2001; 193: 295–302.

    Article  CAS  Google Scholar 

  3. Rocha-zavalta L, Alejandre JE, Garcia-carranca A . Parenteral and oral immunization with a plasmid DNA expressing the human papillomavirus 16 L1 gene induces systemic and mucosal antibodies and cytotoxic T lymphocyte responses. J Med Virol 2002; 66: 86–95.

    Article  Google Scholar 

  4. Mckenzie BS et al. Nucleic acid vaccines: tasks and tactics. Immunol Res 2001; 24: 225–244.

    Article  CAS  Google Scholar 

  5. Boyle JS, Koniaras C, Lew AM . Influence of cellular location of expressed antigen on the efficacy of DNA vaccination: cytotoxic T lymphocyte and antibody responses are suboptimal when antigen is cytoplasmic after intramuscular DNA immunization. Int Immunol 1997; 9: 1897–1906.

    Article  CAS  Google Scholar 

  6. Boyle JS, Brady JL, Lew AM . Enhanced responses to a DNA vaccine encoding a fusion antigen that is directed to sites of immune induction. Nature 1998; 392: 408–411.

    Article  CAS  Google Scholar 

  7. Biragyn A et al. Genetic fusion of chemokines to a self tumor antigen induces protective, T-cell dependent antitumor immunity. Nat Biotechnol 1999; 17: 253–258.

    Article  CAS  Google Scholar 

  8. Solari R et al. Receptor-mediated endocytosis of CC-chemokines. J Biol Chem 1997; 272: 9617–9620.

    Article  CAS  Google Scholar 

  9. Mueller A, Kelly E, Strange PG . Pathways for internalization and recycling of the chemokine receptor CCR5. Blood 2002; 99: 785–791.

    Article  CAS  Google Scholar 

  10. Oh YK, Swanson JA . Different fates of phagocytosed particles after delivery into macrophage lysosomes. J Cell Biol 1996; 132: 585–593.

    Article  CAS  Google Scholar 

  11. Sasagawa T et al. Synthesis and assembly of virus-like particles of human papillomaviruses type 6 and type 16 in fission yeast Schizosaccharomyces pombe. Virology 1995; 206: 126–135.

    Article  CAS  Google Scholar 

  12. Tobery TW et al. A simple and efficient method for the monitoring of antigen-specific T cell responses using peptide pool arrays in a modified ELISpot assay. J Immunol Methods 2001; 254: 59–66.

    Article  CAS  Google Scholar 

  13. Lillard JW Jr et al. RANTES potentiates antigen-specific mucosal immune responses. J Immunol 2001; 166: 162–169.

    Article  CAS  Google Scholar 

  14. Weiss R et al. Improvement of the immune response against plasmid DNA encoding OspC of Borrelia by an ER-targeting leader sequence. Vaccine 2000; 18: 815–824.

    Article  Google Scholar 

  15. Fischer FR et al. RANTES-induced chemokine cascade in dendritic cells. J Immunol 2001; 167: 1637–1643.

    Article  CAS  Google Scholar 

  16. Sadeghi H et al. Genetic fusion of human insulin B-chain to the B-subunit of cholera toxin enhances in vitro antigen presentation and induction of bystander suppression in vivo. Immunology 2002; 106: 237–245.

    Article  CAS  Google Scholar 

  17. Ward SG, Westwick J . Chemokines: understanding their role in T-lymphocyte biology. Biochem J 1998; 333: 457–470.

    Article  CAS  Google Scholar 

  18. Barash S, Wang W, Shi Y . Human secretory signal peptide description by hidden Markov model and generation of a strong artificial signal peptide for secreted protein expression. Biochem Biophys Res Commun 2002; 294: 835–842.

    Article  CAS  Google Scholar 

  19. Sin J et al. DNA vaccines encoding interleukin-8 and RANTES enhance antigen-specific Th1-type CD4(+) T-cell-mediated protective immunity against herpes simplex virus type 2 in vivo. J Virol 2000; 74: 11173–11180.

    Article  CAS  Google Scholar 

  20. Pete I et al. Detection of high-risk HPV (16, 18, 33) in situ cancer of the cervix by PCR technique. Eur J Gynaecol Oncol 2002; 23: 74–78.

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by Korea Research Foundation Grant (KRF 2001-015-FP0128).

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Kim, S., Lee, C., Lee, S. et al. Enhanced immunogenicity of human papillomavirus 16 L1 genetic vaccines fused to an ER-targeting secretory signal peptide and RANTES. Gene Ther 10, 1268–1273 (2003). https://doi.org/10.1038/sj.gt.3301997

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