Intranasal Vaccination with a Recombinant Vesicular Stomatitis Virus Expressing Cottontail Rabbit Papillomavirus L1 Protein Provides Complete Protection against Papillomavirus-Induced Disease

Abstract

Immunizations with live recombinant vesicular stomatitis viruses (rVSV) expressing foreign viral proteins have successfully protected animals from challenges with several heterologous viruses. We developed an rVSV expressing the major capsid protein (L1) of cottontail rabbit papillomavirus (CRPV) and tested the efficacy of protection following CRPV challenge. An rVSV expressing L1 of CRPV (VSV-L1) was characterized for the protective ability afforded by intranasal, intradermal, or intramuscular vaccination in rabbits subsequently challenged with CRPV. Protein expression of L1 in VSV-L1 was confirmed by radioimmunoprecipitation assays. Nuclear localization of L1 was demonstrated by indirect immunofluorescence assays. Immunized rabbits elicited significant VSV neutralization and VLP-L1 enzyme-linked immunosorbent assay titers. VSV-L1 vaccination was not associated with weight loss or any other adverse clinical signs in the rabbit model. VSV shedding in nasal secretions occurred in some rabbits, peaking at 4 to 6 days after intranasal vaccination, with no further shedding after day 6. Specific humoral immunity to the L1 protein was consistently seen after a single VSV-L1 vaccination when administered through an intradermal or intramuscular route or after a boost via the intranasal route. Rabbits were completely protected from CRPV-induced papillomas after VSV-L1 vaccination and boost given intranasally or intramuscularly. Vaccination with VSV-L1 is a novel approach to prevent papillomavirus-induced disease and demonstrates a potential strategy for developing a human papillomavirus vaccine that can be given without injection.High-risk human papillomavirus (HPV) is the etiologic agent associated with over 90% of cervical cancer cases (41). Human papillomaviruses comprise more than 80 distinct types, some of which are sexually transmitted and predominantly infect genital skin and mucosa. Most genital warts are caused by sexually transmitted HPV types 6 and 11, whereas types 16 and 18 are most frequently associated with cervical cancer (2). HPV-associated malignancies are leading causes of cancerrelated deaths in the world, and among women, cervical cancer is second only to breast cancer in incidence and mortality (23). In developing nations, cervical cancer is, in fact, the leading cause of death among women between the ages of 35 and 45 (40). More than 500,000 new cases of cervical cancer are reported worldwide each year. HPVs are also associated with penile, vulvar, anal, respiratory, and cutaneous neoplasms (24, 40). Unfortunately, current therapies for premalignant neoplasms associated with HPV infection are inadequate (2, 7). The cost burden for screening for HPV-induced cervical neoplasia exceeds 5 billion dollars per year in the United States alone (D. R. Lowy, unpublished data).Prophylactic vaccination has been the most effective public health measure to reduce morbidity and mortality associated with viral infections. Since HPV infections cause cervical neoplasia, vaccines targeting HPV antigens should be effe...

Can genital-tract human papillomavirus infection and cervical cancer be prevented with a vaccine?

“…99). An rVSV expressing L1 of CRPV (VSV-L1) was characterised for the protective ability afforded by intranasal, intradermal or intramuscular vaccination in rabbits subsequently challenged with CRPV.…”

CD4+T-Cell Reconstitution Reduces Cytomegalovirus in the Immunocompromised Brain

“…Cells were harvested, semipurified, titered using plaque assays, and stored in aliquots at Ϫ70°C. Recombinant vesicular stomatitis virus (VSV) was grown and titered as previously described for recombinant VSV-HA (55,57). Plaque-purified recombinant VSV was grown and titered on baby hamster kidney (BHK-21, American Type Culture Collection) cells.…”

Cervarix™: a vaccine for the prevention of HPV 16, 18-associated cervical cancer

“…Clinical trials are currently being planned to evaluate the safety and immunogenicity of L1 capsomere vaccines formulated in alum. Alternatively, the inclusion of L1 in other vaccines, such as typhoid, tuberculosis or measles vaccines might represent a cost-effective and practical alternative that could also provide immunity to HPV (Reuter et al 2002;Baud et al 2004a,b;Govan et al 2006). In addition, the use of live vectors for the delivery of L1 VLPs is attractive in remote and low-resource areas as immunity could be spread, but safety remains an issue with such vectors.…”