Broadly neutralizing antiviral responses induced by a single-molecule HPV vaccine based on thermostable thioredoxin-L2 multiepitope nanoparticles

Spagnoli, Gloria; Pouyanfard, Somayeh; Cavazzini, D.; Canali, Elena; Maggi, Stefano; Tommasino, Massimo; Bolchi, Angelo; Müller, Martin; Ottonello, Simone

doi:10.1038/s41598-017-18177-1

Cited by 28 publications

(46 citation statements)

References 52 publications

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“…Staining of formalin-fixed, paraffin-embedded tumors was performed as previously described ( Hasche et al, 2017 ). Briefly, deparaffinized sections were heated in citrate buffer pH6.0 prior to blocking with 5% goat serum/5% FCS/1% BSA in PBS and incubation with primary antibodies (serum of a VLP-vaccinated Mastomys ( Vinzón et al, 2014 ), serum of a mouse immunized with the N-terminal 31 aa of MnPV-L1 LONG in the OVX313 platform ( Spagnoli et al, 2017 ) or the respective pre-immune sera, cross-reactive anti-L2 (K18L2) ( Rubio et al, 2011 ) or serum of a guinea pig immunized with Gardasil9 (unpublished) overnight at 4°C. Detection of L1 isoforms was achieved with the Dako REAL Detection System, Peroxidase/AEC, Rabbit/Mouse.…”

Section: Methodsmentioning

confidence: 99%

Expression of different L1 isoforms of Mastomys natalensis papillomavirus as mechanism to circumvent adaptive immunity

Cao

Schäfer

et al. 2020

eLife

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Although many high-risk mucosal and cutaneous human papillomaviruses (HPVs) theoretically have the potential to synthesize L1 isoforms differing in length, previous seroepidemiological studies only focused on the short L1 variants, co-assembling with L2 to infectious virions. Using the multimammate mouse Mastomys coucha as preclinical model, this is the first study demonstrating seroconversion against different L1 isoforms during the natural course of papillomavirus infection. Intriguingly, positivity with the cutaneous MnPV was accompanied by a strong seroresponse against a longer L1 isoform, but to our surprise, the raised antibodies were non-neutralizing. Only after a delay of around 4 months, protecting antibodies against the short L1 appeared, enabling the virus to successfully establish an infection. This argues for a novel humoral immune escape mechanism that may also have important implications on the interpretation of epidemiological data in terms of seropositivity and protection of PV infections in general.

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Section: Methodsmentioning

confidence: 99%

Expression of different L1 isoforms of Mastomys natalensis papillomavirus as mechanism to circumvent adaptive immunity

Cao

Schäfer

et al. 2020

eLife

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“…endotoxin levels below 8 IU per ml. Details of the OVX313 protein purification and characterization will be published elsewhere (53). Mouse and guinea pig immunization.…”

Section: Figmentioning

confidence: 99%

Minor Capsid Protein L2 Polytope Induces Broad Protection against Oncogenic and Mucosal Human Papillomaviruses

Pouyanfard

Spagnoli

Bulli

et al. 2018

J Virol

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The amino terminus of the human papillomavirus (HPV) minor capsid protein L2 contains a major cross-neutralization epitope which provides the basis for the development of a broadly protecting HPV vaccine. A wide range of protection against different HPV types would eliminate one of the major drawbacks of the commercial, L1-based prophylactic vaccines. Previously, we have reported that insertion of the L2 epitope into a scaffold composed of bacterial thioredoxin protein generates a potent antigen inducing comprehensive protection against different animal and human papillomaviruses. We also reported, however, that although protection is broad, some oncogenic HPV types escape the neutralizing antibody response, if L2 epitopes from single HPV types are used as immunogen. We were able to compensate for this by applying a mix of thioredoxin proteins carrying L2 epitopes from HPV16, -31, and -51. As the development of a cost-efficient HPV prophylactic vaccines is one of our objectives, this approach is not feasible as it requires the development of multiple good manufacturing production processes in combination with a complex vaccine formulation. Here, we report the development of a thermostable thioredoxin-based single-peptide vaccine carrying an L2 polytope of up to 11 different HPV types. The L2 polytope antigens have excellent abilities in respect to broadness of protection and robustness of induced immune responses. To further increase immunogenicity, we fused the thioredoxin L2 polytope antigen with a heptamerization domain. In the final vaccine design, we achieve protective responses against all 14 oncogenic HPV types that we have analyzed plus the low-risk HPVs 6 and 11 and a number of cutaneous HPVs. Infections by a large number of human papillomaviruses lead to malignant and nonmalignant disease. Current commercial vaccines based on virus-like particles (VLPs) effectively protect against some HPV types but fail to do so for most others. Further, only about a third of all countries have access to the VLP vaccines. The minor capsid protein L2 has been shown to contain so-called neutralization epitopes within its N terminus. We designed polytopes comprising the L2 epitope amino acids 20 to 38 of up to 11 different mucosal HPV types and inserted them into the scaffold of thioredoxin derived from a thermophile archaebacterium. The antigen induced neutralizing antibody responses in mice and guinea pigs against 26 mucosal and cutaneous HPV types. Further, addition of a heptamerization domain significantly increased the immunogenicity. The final vaccine design comprising a heptamerized L2 8-mer thioredoxin single-peptide antigen with excellent thermal stability might overcome some of the limitations of the current VLP vaccines.

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“…For example, immunizations with an L2 peptide conjugated to thioredoxin [44,45], immunizations with concatemers of L2 protein fused to a self-adjuvanting protein (flagellin) [46,47] as well as immunizations with a concatemer of L2 proteins derived from different HPV types [48,49] have been used. More recently, an L2 polypeptide and a heptamerizing coiled-coil polypeptide OVX313 fused to a nanoparticle derived from a thermostable thioredoxin has been used [50][51][52]. These approaches have enhanced the immunogenicity of L2 peptides, especially immunization with the nanoparticle thioredoxin-L2-OVX313 candidate vaccine.…”

Section: L2 Proteinmentioning

confidence: 99%

“…Sequence alignment of L2 (aa 1-135) from different HPV types. Sequence alignment was done using sequences from both high-risk (HPVs 16,18,26,31,33,34,35,39,45,[51][52][53]56,58,59,66,68,73,and 82) and low-risk HPV types (HPVs 6, 11, 43, and 44). Alignment was done using Jalview software.…”

mentioning

confidence: 99%

Virus-like Particle-Based L2 Vaccines against HPVs: Where Are We Today?

Yadav

Zhai

Tumban

2019

Viruses

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Human papillomaviruses (HPVs) are the most common sexually transmitted infections worldwide. Ninety percent of infected individuals clear the infection within two years; however, in the remaining 10% of infected individuals, the infection(s) persists and ultimately leads to cancers (anogenital cancers and head and neck cancers) and genital warts. Fortunately, three prophylactic vaccines have been approved to protect against HPV infections. The most recent HPV vaccine, Gardasil-9 (a nonavalent vaccine), protects against seven HPV types associated with ~90% of cervical cancer and against two HPV types associated with ~90% genital warts with little cross-protection against non-vaccine HPV types. The current vaccines are based on virus-like particles (VLPs) derived from the major capsid protein, L1. The L1 protein is not conserved among HPV types. The minor capsid protein, L2, on the other hand, is highly conserved among HPV types and has been an alternative target antigen, for over two decades, to develop a broadly protective HPV vaccine. The L2 protein, unlike the L1, cannot form VLPs and as such, it is less immunogenic. This review summarizes current studies aimed at developing HPV L2 vaccines by multivalently displaying L2 peptides on VLPs derived from bacteriophages and eukaryotic viruses. Recent data show that a monovalent HPV L1 VLP as well as bivalent MS2 VLPs displaying HPV L2 peptides (representing amino acids 17–36 and/or consensus amino acids 69–86) elicit robust broadly protective antibodies against diverse HPV types (6/11/16/18/26/31/33/34/35/39/43/44/45/51/52/53/56/58/59/66/68/73) associated with cancers and genital warts. Thus, VLP-based L2 vaccines look promising and may be favorable, in the near future, over current L1-based HPV vaccines and should be explored further.

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Broadly neutralizing antiviral responses induced by a single-molecule HPV vaccine based on thermostable thioredoxin-L2 multiepitope nanoparticles

Cited by 28 publications

References 52 publications

Expression of different L1 isoforms of Mastomys natalensis papillomavirus as mechanism to circumvent adaptive immunity

Expression of different L1 isoforms of Mastomys natalensis papillomavirus as mechanism to circumvent adaptive immunity

Minor Capsid Protein L2 Polytope Induces Broad Protection against Oncogenic and Mucosal Human Papillomaviruses

Virus-like Particle-Based L2 Vaccines against HPVs: Where Are We Today?

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