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. 2009 May 26;27(25-26):3385-90.
doi: 10.1016/j.vaccine.2009.01.061. Epub 2009 Feb 5.

Recombinant measles virus-HPV vaccine candidates for prevention of cervical carcinoma

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Free PMC article

Recombinant measles virus-HPV vaccine candidates for prevention of cervical carcinoma

Giuseppina Cantarella et al. Vaccine. .
Free PMC article

Abstract

Cervical cancer is mainly associated with HPV genotype 16 infection. Recombinant measles virus (rMV) expressing HPV genotype 16 L1 capsid protein was generated by construction of an antigenomic plasmid, followed by rescue using the human "helper" cell line 293-3-46. In cell cultures the recombinant MV-L1 virus replicated practically as efficiently as the standard attenuated MV established as commercial vaccine, devoid of the transgene. The high genetic stability of MVb2-L1 was confirmed by 10 serial viral transfers in cell culture. In transgenic mice expressing the MV receptor CD46 the recombinant induced strong humoral immune responses against both MV and HPV; the antibodies against L1 exhibited mainly neutralizing capacity. Our data suggest that MV is a promising vehicle for development of inexpensive and efficient vaccines protecting from HPV infection.

Figures

Fig. 1
Fig. 1
Construction of recombinant p(+)RMVb2-HPV-L1 plasmid. The HPV16-L1 gene was amplified by PCR and the resulting product was cloned into the MV genome context via BssHII and AatII.
Fig. 2
Fig. 2
Immunofluorescence analysis for expression of HPV-L1 protein by recombinant MVs. Monolayers of Vero cells grown on tissue culture chamber slides were infected either with MVb (A) either with rMVb2-HPV-L1 (B) at MOI 0.05 for 48 h and processed for immunofluorescence. Cells were stained with a monoclonal anti-HPV-L1 antibody subsequently with a goat anti-mouse coupled to FITC (green). The same slides were stained with DAPI and visualized by an inverted Leica microscope under normal and fluorescent light, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of the article.)
Fig. 3
Fig. 3
Western blot analysis for expression of HPV-L1 protein by recombinant MV. Vero cells were infected with rMVs at MOI 0.05 for 48 h. Expression of L1 from lysed Vero cells and medium was determined by Western blot analysis. Lane 1: medium from non-infected cells; lane 2: medium from cells infected with MVb; lane 3: medium from cells infected with rMVb2-HPV-L1; lane 4: lysed non-infected cells; lane 5: lysed cells infected with MVb; lane 6: lysed cells infected with rMVb2-HPV-L1.
Fig. 4
Fig. 4
Growth curve comparison of MVb and rMVb2-HPV-L1. MRC5 cells on six-well plates were infected with MVb or rMVb2-HBV-L1 at MOI 0.05 for 6 days. At each time point indicated, media (cell-free (CF) virus) and cells (cell-associated (CA) virus) were collected separately. Virus titers were determined by plaque assay.

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