Fowlpox virus recombinants expressing HPV-16 E6 and E7 oncogenes for the therapy of cervical carcinoma elicit humoral and cell-mediated responses in rabbits

Radaelli, Antonia; Pozzi, Eleana; Pacchioni, Sole; Zanotto, Carlo; Morghen, Carlo De Giuli

doi:10.1186/1479-5876-8-40

Cited by 9 publications

(3 citation statements)

References 57 publications

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“…Recombinant FWPVs (rFWPVs) are being developed clinically as vaccine vectors because of their impeccable safety profile and large cloning capacity (enabling the expression of multiple heterologous genes) as well as their ability to induce cell-mediated immunity (15; reviewed in reference 4). Recombinant FWPV-vectored vaccines have delivered promising preclinical and clinical results, particularly when used as part of homologous or heterologous prime-boost immunization regimens (18,21,25,27,28,40,55,58,62). Most recently, in humans, and following antiretroviral drug therapy withdrawal, homologous primeboost vaccination with rFWPV encoding human immunodeficiency virus (HIV) Gag-Pol and human gamma interferon (hIFN-␥) was associated with a partial control of HIV replication in a subset of individuals (22).…”

mentioning

confidence: 99%

Antigen-Specific T-Cell Responses to a Recombinant Fowlpox Virus Are Dependent on MyD88 and Interleukin-18 and Independent of Toll-Like Receptor 7 (TLR7)- and TLR9-Mediated Innate Immune Recognition

Lousberg

Diener

Fraser

et al. 2011

J Virol

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Fowlpox virus (FWPV) is a double-stranded DNA virus long used as a live-attenuated vaccine against poultry diseases, but more recent interest has focused on its use as a mammalian vaccine vector. Here, in a mouse model system using FWPV encoding the nominal target antigen chicken ovalbumin (OVA) (FWPV OVA ), we describe for the first time some of the fundamental processes by which FWPV engages both the innate and adaptive immune systems. We show that Toll-like receptor 7 (TLR7) and TLR9 are important for type I interferon secretion by dendritic cells, while TLR9 is solely required for proinflammatory cytokine secretion. Despite this functional role for TLR7 and TLR9 in vitro, only the adapter protein myeloid differentiation primary response gene 88 (MyD88) was shown to be essential for the formation of adaptive immunity to FWPV OVA in vivo. The dependence on MyD88 was confined only to the T-cell compartment and was not related to its contribution to TLR signaling, dendritic cell maturation, or the capture and presentation of FWPVderived OVA antigen. We demonstrate that this is not by means of mediating T-cell-dependent interleukin-1 (IL-1) signaling, but rather, we suggest that MyD88 functions to support T-cell-specific IL-18 receptor signaling, which in turn is essential for the formation of adaptive immunity to FWPV-encoded OVA.

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mentioning

confidence: 99%

Antigen-Specific T-Cell Responses to a Recombinant Fowlpox Virus Are Dependent on MyD88 and Interleukin-18 and Independent of Toll-Like Receptor 7 (TLR7)- and TLR9-Mediated Innate Immune Recognition

Lousberg

Diener

Fraser

et al. 2011

J Virol

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“…The expression of the E6 F47R gene was also verified after transfection with the pcDNA3E6 F47R plasmid by the PolyJet™ transfection reagent (SignaGen Lab., Rockville, MD, USA). Briefly, cells were overlaid with 1 μg reagent in 1.1 ml medium per 4-cm-diameter Petri dish without serum and antibiotics for 16–24 h, and replaced with the complete medium for an additional 24 h. The samples were then treated for 1 h with the 1:100-diluted mouse AbE6/Gi or with the 1:300-diluted rabbit Rpool polyclonal antibodies, obtained in our laboratory [ 50 ] followed by the 1:100-diluted FITC-labeled goat anti-mouse or sheep anti-rabbit sera (DakoCytomation) for 1 h. FPwt and pcDNA3 empty plasmid were used as the negative controls to infect or transfect the different cell lines. The samples were viewed under a Zeiss Axioskop fluorescence microscope.…”

Section: Methodsmentioning

confidence: 99%

A prime/boost strategy using DNA/fowlpox recombinants expressing the genetically attenuated E6 protein as a putative vaccine against HPV-16-associated cancers

et al. 2015

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BackgroundConsidering the high number of new cases of cervical cancer each year that are caused by human papilloma viruses (HPVs), the development of an effective vaccine for prevention and therapy of HPV-associated cancers, and in particular against the high-risk HPV-16 genotype, remains a priority. Vaccines expressing the E6 and E7 proteins that are detectable in all HPV-positive pre-cancerous and cancer cells might support the treatment of HPV-related lesions and clear already established tumors.MethodsIn this study, DNA and fowlpox virus recombinants expressing the E6F47R mutant of the HPV-16 E6 oncoprotein were generated, and their correct expression verified by RT-PCR, Western blotting and immunofluorescence. Immunization protocols were tested in a preventive or therapeutic pre-clinical mouse model of HPV-16 tumorigenicity using heterologous (DNA/FP) or homologous (DNA/DNA and FP/FP) prime/boost regimens. The immune responses and therapeutic efficacy were evaluated by ELISA, ELISPOT assays, and challenge with TC-1* cells.ResultsIn the preventive protocol, while an anti-E6-specific humoral response was just detectable, a specific CD8+ cytotoxic T-cell response was elicited in immunized mice. After the challenge, there was a delay in cancer appearance and a significant reduction of tumor volume in the two groups of E6-immunized mice, thus confirming the pivotal role of the CD8+ T-cell response in the control of tumor growth in the absence of E6-specific antibodies. In the therapeutic protocol, in-vivo experiments resulted in a higher number of tumor-free mice after the homologous DNA/DNA or heterologous DNA/FP immunization.ConclusionsThese data establish a preliminary indication for the prevention and treatment of HPV-related tumors by the use of DNA and avipox constructs as safe and effective immunogens following a prime/boost strategy. The combined use of recombinants expressing both E6 and E7 proteins might improve the antitumor efficacy, and should represent an important approach to control HPV-associated cancers.

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“…The antigen was incubated overnight at 4 °C. ELISAs were performed in duplicate, essentially as described previously [55], using serum from each animal of both groups of mice (G1, control group; G2, experimental group) from T0 to T11. The sera dilutions were 1:1,000.…”

Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

Construction of a recombinant avipoxvirus expressing the env gene of Zika virus as a novel putative preventive vaccine

Zanotto

Paolini²,

Radaelli

et al. 2021

Virol J

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Background Zika virus (ZIKV) has been declared a public health emergency that requires development of an effective vaccine, as it might represent an international threat. Methods Here, two novel DNA-based (pVAXzenv) and fowlpox-based (FPzenv) recombinant putative vaccine candidates were constructed that contained the cPrME genes of ZIKV. The env gene inserted into the fowlpox vector was verified for correct transgene expression by Western blotting and by immunofluorescence in different cell lines. The production of virus-like particles as a result of env gene expression was also demonstrated by electron microscopy. BALB/c mice were immunosuppressed with dexamethasone and immunized following a prime–boost strategy in a heterologous protocol where pVAXzenv was followed by FPzenv, to evaluate the immunogenicity of the Env protein. The mice underwent a challenge with an epidemic ZIKV after the last boost. Results These data show that the ZIKV Env protein was correctly expressed in both normal human lung fibroblasts (MRC-5 cells) and green monkey kidney (Vero) cells infected with FPzenv, and that the transgene expression lasted for more than 2 weeks. After mucosal administration of FPzenv, the immunized mice showed specific and significantly higher humoral responses compared to the control mice. However, virus neutralizing antibodies were not detected using plaque reduction assays. Conclusions Although BALB/c mice appear to be an adequate model for ZIKV infection, as it mimics the natural mild infection in human beings, inadequate immune suppression seemed to occur by dexamethasone and different immune suppression strategies should be applied before challenge to reveal any protection of the mice.

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Fowlpox virus recombinants expressing HPV-16 E6 and E7 oncogenes for the therapy of cervical carcinoma elicit humoral and cell-mediated responses in rabbits

Cited by 9 publications

References 57 publications

Antigen-Specific T-Cell Responses to a Recombinant Fowlpox Virus Are Dependent on MyD88 and Interleukin-18 and Independent of Toll-Like Receptor 7 (TLR7)- and TLR9-Mediated Innate Immune Recognition

Antigen-Specific T-Cell Responses to a Recombinant Fowlpox Virus Are Dependent on MyD88 and Interleukin-18 and Independent of Toll-Like Receptor 7 (TLR7)- and TLR9-Mediated Innate Immune Recognition

A prime/boost strategy using DNA/fowlpox recombinants expressing the genetically attenuated E6 protein as a putative vaccine against HPV-16-associated cancers

Construction of a recombinant avipoxvirus expressing the env gene of Zika virus as a novel putative preventive vaccine

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