Vaccination with yeast-expressed cottontail rabbit papillomavirus (CRPV) virus-like particles protects rabbits from CRPV-induced papilloma formation

doi:10.1016/0264-410X(95)00103-8

Vaccine

Volume 13, Issue 16, 1995, Pages 1509-1514

https://doi.org/10.1016/0264-410X(95)00103-8 Get rights and content

Abstract

Papillomaviruses infect epithelia of the skin and mucous membranes and cause benign or malignant tumours in animals and in humans. The viruses are highly species-specific, and cell culture systems for propagating human papillomaviruses (HPVs) do not exist. However, there are several animal papillomavirus models. In the cottontail rabbit papillomavirus (CRPV) system, we demonstrated that recombinant CRPV virus-like particles (VLPs) consisting of the capsid proteins L1 or L1+L2 can be produced in the yeast Saccharomyces cerevisiae. Three immunizations with L1 VLPs formulated on aluminum adjuvant at 1–100 μg dose⁻¹ efficiently protected rabbits from challenge with CRPV. Sera of immunized rabbits were shown to contain high-titered serum antibodies to CRPV L1 VLPs and to neutralize CRPV in vitro. Our results suggest that recombinant yeast-derived VLPs could be the basis for a candidate HPV vaccine.

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Cited by (167)

Optimized production strategy of the major capsid protein HPV 16L1 non-assembly variant in E. coli
2020, Protein Expression and Purification
Citation Excerpt :
They can efficiently prevent HPV infections of the respective HPV types, they consist of [6]. Each L1 protein of the respective HPV-types is expressed in either yeast, insect cells or E. coli [7–10]. Available vaccines include Cervarix® produced in insect cells (GSK, HPV 16 and 18), Gardasil® produced in yeast (Merck&Co., HPV 6, 11, 16, 18), Gardasil-9® produced in yeast (Merck&Co., HPV 6, 11, 16, 18, 31, 33, 45, 52, 58) and Cecolin® produced in E. coli (Innovax, HPV 16 and 18) [10–14].
The capsid of human papillomavirus (HPV) consists of two capsid proteins - the major capsid protein L1 and the minor capsid protein L2. Assembled virus-like particles, which only consist of L1 proteins, are successfully applied as prophylactic vaccines against HPV infections. The capsid subunits are L1-pentamers, which are also reported to protect efficiently against HPV infections in animals.
The recombinant production of L1 has been previously shown in E. coli, yeast, insect cells, plants and mammalian cell culture. Principally, in E. coli-based expression system L1 shows high expression yields but the protein is largely insoluble. In order to overcome this problem reported strategies address fusion proteins and overexpression of bacterial chaperones. However, an insufficient cleavage of the fusion proteins and removal of co-purified chaperones can hamper subsequent down streaming.
We report a significant improvement in the production of soluble L1-pentamers by combining (I) a fusion of a N-terminal SUMO-tag to L1, (II) the heterologous co-expression of the chaperon system GroEL/ES and (III) low expression temperature. The fusion construct was purified in a 2-step protein purification including efficient removal of GroEL/ES and complete removal of the N-terminal SUMO-tag. The expression strategy was transferred to process-controlled high-cell-density fermentation with defined media according to the guidelines of good manufacturing practice. The produced L1 protein is highly pure (>95%), free of DNA (260:280 = 0.5) and pentameric. The production strategy yielded 5.73 mg of purified L1-pentamers per gram dry biomass. The optimized strategy is a suitable alternative for high yield L1-pentamer production and purification as a cheaper process for vaccine production.
Prevention of cervical cancer: Journey to develop the first human papillomavirus virus-like particle vaccine and the next generation vaccine
2016, Current Opinion in Chemical Biology
Citation Excerpt :
Several labs have adopted and licensed the Kreider method which became important in the testing of HPV vaccine candidates. In 1995, three animal papillomavirus challenge studies were published [23,24,25]. These studies provided critical information which supported the continued advancement of the HPV vaccine program.
In 2006, the first human papillomavirus (HPV) virus-like particle (VLP) vaccine was licensed. Gardasil^®, the quadrivalent HPV 6, 11, 16 and 18 recombinant VLP vaccine (4vHPV), developed by Merck demonstrated remarkable efficacy in prevention of important clinical pre-cursors to cervical cancer and genital warts. The vaccine was designed to protect against HPV 16 and 18 that cause ∼70% of cervical cancers and HPV 6 and 11 that cause ∼90% of genital warts. Initially, Gardasil^® was indicated in the United States for women 9–26 years of age for the prevention of HPV 16 and 18-related cervical, vulvar and vaginal cancer, HPV 6, 11, 16 and 18-related genital intraepithelial neoplasia and the prevention of HPV 6 and 11-related genital warts. Subsequently, a bivalent HPV 16 and 18 VLP vaccine, Cervarix (2vHPV) developed by GlaxoSmithKline was licensed. Since the original licensures, the indications for Gardasil^® have been expanded to include males and a vaccine with extended HPV coverage, Gardasil 9 (9vHPV), licensed in 2014.
Development of a recombinant toxin fragment vaccine for Clostridium difficile infection
2014, Vaccine
Clostridium difficile infection (CDI) is the major cause of antibiotic-associated diarrhea and pseudomembranous colitis, a disease associated with significant morbidity and mortality. The disease is mostly of nosocomial origin, with elderly patients undergoing anti-microbial therapy being particularly at risk. C. difficile produces two large toxins: Toxin A (TcdA) and Toxin B (TcdB). The two toxins act synergistically to damage and impair the colonic epithelium, and are primarily responsible for the pathogenesis associated with CDI. The feasibility of toxin-based vaccination against C. difficile is being vigorously investigated. A vaccine based on formaldehyde-inactivated Toxin A and Toxin B (toxoids) was reported to be safe and immunogenic in healthy volunteers and is now undergoing evaluation in clinical efficacy trials. In order to eliminate cytotoxic effects, a chemical inactivation step must be included in the manufacturing process of this toxin-based vaccine. In addition, the large-scale production of highly toxic antigens could be a challenging and costly process.
Vaccines based on non-toxic fragments of genetically engineered versions of the toxins alleviate most of these limitations. We have evaluated a vaccine assembled from two recombinant fragments of TcdB and explored their potential as components of a novel experimental vaccine against CDI. Golden Syrian hamsters vaccinated with recombinant fragments of TcdB combined with full length TcdA (Toxoid A) developed high titer IgG responses and potent neutralizing antibody titers. We also show here that the recombinant vaccine protected animals against lethal challenge with C. difficile spores, with efficacy equivalent to the toxoid vaccine. The development of a two-segment recombinant vaccine could provide several advantages over toxoid TcdA/TcdB such as improvements in manufacturability.
Prophylactic papillomavirus vaccines
2014, Clinics in Dermatology
Human papillomaviruses (HPV) are the causative agents of cervical cancer, the third most common cancer in women. The development of prophylactic HPV vaccines Gardasil® and Cervarix® targeting the major oncogenic HPV types is now the frontline of cervical cancer prevention. Both vaccines have been proven to be highly effective and safe although there are still open questions about their target population, cross-protection, and long-term efficacy. The main limitation for a worldwide implementation of Gardasil® and Cervarix® is their high cost. To develop more affordable vaccines research groups are concentrated in new formulations with different antigens including capsomeres, the minor capsid protein L2 and DNA. In this article we describe the vaccines' impact on HPV-associated disease, the main open questions about the marketed vaccines, and current efforts for the development of second-generation vaccines.
Virus-like particles as a highly efficient vaccine platform: Diversity of targets and production systems and advances in clinical development
2012, Vaccine
Citation Excerpt :
HPV L1 VLPs have been shown to be immunogenic in mice [135] and also to induce both systemic and mucosal neutralizing antibodies as well as Th1 and Th2 cytokine responses in vaccinated non-human primates [136,141]. Furthermore, in a proof-of-principle study, cottontail rabbit papillomavirus L1 VLPs produced in S. cerevisiae successfully protected animals from virus-induced papilloma formation [142]. The first licensed HPV vaccine approved by the U.S. FDA, Gardasil® (Merck, USA), is a quadrivalent vaccine containing L1 VLPs from HPV types 6, 11, 16 and 18 produced in S. cerevisiae and administered intramuscularly (IM) in three doses with aluminum hydroxyphosphate sulphate as an adjuvant (Table 2) [13].
Virus-like particles (VLPs) are a class of subunit vaccines that differentiate themselves from soluble recombinant antigens by stronger protective immunogenicity associated with the VLP structure. Like parental viruses, VLPs can be either non-enveloped or enveloped, and they can form following expression of one or several viral structural proteins in a recombinant heterologous system. Depending on the complexity of the VLP, it can be produced in either a prokaryotic or eukaryotic expression system using target-encoding recombinant vectors, or in some cases can be assembled in cell-free conditions. To date, a wide variety of VLP-based candidate vaccines targeting various viral, bacterial, parasitic and fungal pathogens, as well as non-infectious diseases, have been produced in different expression systems. Some VLPs have entered clinical development and a few have been licensed and commercialized. This article reviews VLP-based vaccines produced in different systems, their immunogenicity in animal models and their status in clinical development.
Prophylactic human papillomavirus vaccines: Past, present and future
2012, Pathology
Human papillomavirus (HPV) is a highly transmissible infection responsible for a range of diseases in women including cervical carcinomas, vulval carcinomas, anogenital carcinomas and genital warts. In men it is associated with penile carcinomas, anogenital carcinomas and oropharyngeal carcinomas. The history of the development of HPV vaccines includes a significant Australian input and represents a tremendous advancement in our understanding of HPV virology as well as further elucidating the overall contribution of viruses to carcinogenesis. Prophylactic HPV vaccines were licensed for use in Australia in 2007 in order to protect against development of future cases of cervical carcinoma and early results are promising. The benefit of the vaccine will not be restricted to cervical lesions and cross protection amongst a variety of HPV subtypes is described. The development of the HPV vaccine and its ultimate incorporation into our National Immunisation Schedule is reviewed.

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Vaccine

Abstract

References (27)

The open reading frame L2 of cottontail rabbit papillomavirus contains antibody-inducing neutralizing epitopes

Virology

Studies on vaccination against papillomaviruses: prophylactic and therapeutic vaccination with recombinant structural proteins

Virology

Effective vaccination against papilloma development by immunization with L1 or L2 structural protein of cottontail rabbit papillomavirus

Virology

Papillomavirus L1 major capsid protein self-assembles into virus-like particles that are highly immunogenic

The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase

J. Biol. Chem.

Transformation of yeast by a replicating hybrid plasmid

Nature

Summary of clinical findings on Engenerix-B, a genetically engineered yeast derived hepatitis B vaccine

J. Postgrad. Med.

Heterogeneity of the human papillomavirus group

J. Virol.

Papillomavirus in anogenital cancer: the dilemma of epidemiologic approaches

J. Natl Cancer Inst.

Human papillomavirus types 6 and 11 DNA sequences in genital and laryngeal papillomas and in some cervical cancers

Epidemiology of genital human papillomavirus infection

Epidemiol. Rev.

Papillomaviruses

Laboratory production in vivo of infectious human papillomavirus type 11

J. Virol.

Cited by (167)

Optimized production strategy of the major capsid protein HPV 16L1 non-assembly variant in E. coli

Prevention of cervical cancer: Journey to develop the first human papillomavirus virus-like particle vaccine and the next generation vaccine

Development of a recombinant toxin fragment vaccine for Clostridium difficile infection

Prophylactic papillomavirus vaccines

Virus-like particles as a highly efficient vaccine platform: Diversity of targets and production systems and advances in clinical development

Prophylactic human papillomavirus vaccines: Past, present and future

PaperVaccination with yeast-expressed cottontail rabbit papillomavirus (CRPV) virus-like particles protects rabbits from CRPV-induced papilloma formation

Abstract

Virology

Virology

Virology

J. Biol. Chem.

Nature

J. Postgrad. Med.

Heterogeneity of the human papillomavirus group

J. Virol.

Papillomavirus in anogenital cancer: the dilemma of epidemiologic approaches

J. Natl Cancer Inst.

Human papillomavirus types 6 and 11 DNA sequences in genital and laryngeal papillomas and in some cervical cancers

Epidemiology of genital human papillomavirus infection

Epidemiol. Rev.

Papillomaviruses

Laboratory production in vivo of infectious human papillomavirus type 11

J. Virol.

Paper
Vaccination with yeast-expressed cottontail rabbit papillomavirus (CRPV) virus-like particles protects rabbits from CRPV-induced papilloma formation