Plant-based Vaccines Research Articles

The heat-labile toxin B subunit of E. coli fused with VP6 from GCRV (Grass carp reovirus) was expressed and folded into an active protein in rice calli

Grass carp reovirus (GCRV) is one of the most serious pathogens threatening grass carp (Ctenopharyngon idellus) production in China. VP6 could be suitable for developing vaccine for the control of GCRV. Transgenic plants are an attractive bioreactor for their safety and ability to make economical vaccines. The B subunit of Escherichia coli heat-labile enterotoxin (LTB) fused to VP6 (LTB-VP6) was transformed into rice calli by Agrobacterium tumefaciens-mediated gene transformation. Transgenic rice calli was confirmed by PCR analysis separately. The copy numbers of LTB-VP6 inserted into the rice genome are between 1 and 2. The expression level of LTB-VP6 in rice calli was 0.0005–0.0019%, an average of 0.0011% of the TSP(total soluble proteins). LTB-VP6 was folded and assembled into a pentameric form of approximately 305 kDa capable of binding monosialoganglioside (GM1). The suitable concentration of LTB-VP6 in TSP was 0.4 μg/μl. LTB-VP6 is stable and highly active at room temperature. LTB-VP6 binding to GM1 is affected with different affinities under different temperatures. LTB-VP6 had a strong binding affinity at 25 °C and pH 8.4. Our results showed that LTB-VP6 is capable of forming an active pentameric form protein. It provides an ideal alternative to plant-based vaccines against GCRV in aquaculture.

Efficacy and Safety of a Recombinant Plant-Based Adjuvanted Covid-19 Vaccine

BackgroundCoronavirus-like particles (CoVLP) that are produced in plants and display the prefusion spike glycoprotein of the original strain of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are combined with an adjuvant (Adjuvant System 03 [AS03]) to form the candidate vaccine.MethodsIn this phase 3, multinational, randomized, placebo-controlled trial conducted at 85 centers, we assigned adults (≥18 years of age) in a 1:1 ratio to receive two intramuscular injections of the CoVLP+AS03 vaccine or placebo 21 days apart. The primary objective of the trial was to determine the efficacy of the CoVLP+AS03 vaccine in preventing symptomatic coronavirus disease 2019 (Covid-19) beginning at least 7 days after the second injection, with the analysis performed after the detection of at least 160 cases.ResultsA total of 24,141 volunteers participated in the trial; the median age of the participants was 29 years. Covid-19 was confirmed by polymerase-chain-reaction assay in 165 participants in the intention-to-treat population; all viral samples that could be sequenced contained variants of the original strain. Vaccine efficacy was 69.5% (95% confidence interval [CI], 56.7 to 78.8) against any symptomatic Covid-19 caused by five variants that were identified by sequencing. In a post hoc analysis, vaccine efficacy was 78.8% (95% CI, 55.8 to 90.8) against moderate-to-severe disease and 74.0% (95% CI, 62.1 to 82.5) among the participants who were seronegative at baseline. No severe cases of Covid-19 occurred in the vaccine group, in which the median viral load for breakthrough cases was lower than that in the placebo group by a factor of more than 100. Solicited adverse events were mostly mild or moderate and transient and were more frequent in the vaccine group than in the placebo group; local adverse events occurred in 92.3% and 45.5% of participants, respectively, and systemic adverse events in 87.3% and 65.0%. The incidence of unsolicited adverse events was similar in the two groups up to 21 days after each dose (22.7% and 20.4%) and from day 43 through day 201 (4.2% and 4.0%).ConclusionsThe CoVLP+AS03 vaccine was effective in preventing Covid-19 caused by a spectrum of variants, with efficacy ranging from 69.5% against symptomatic infection to 78.8% against moderate-to-severe disease. (Funded by Medicago; ClinicalTrials.gov number, NCT04636697.)

A review on the recent advances and application of vaccines against fish pathogens in aquaculture.

Globally, aquaculture has faced serious economic problems due to bacterial, viral, and various other infectious diseases of different origins. Even though such diseases are being detected and simultaneously treated with several therapeutic and prophylactic methods, the broad-spectrum activity of vaccines plays a vital role as a preventive measure in aquaculture. However, treatments like use of antibiotics and probiotics seem to be less effective when new mutant strains develop and disease causing pathogens become resistant to commonly used antibiotics. Therefore, vaccines developed by using recent advanced molecular techniques can be considered as an effective way of treating disease causing pathogens in aquatic organisms. The present review emphasizes on the current advances in technology and future outlook with reference to different types of vaccines used in the aquaculture industries. Beginning with traditional killed/inactivated and live attenuated vaccines, this work culminates in the review of modern new generation ones including recombinant, synthetic peptides, mucosal and DNA, subunit, nanoparticle-based and plant-based edible vaccines, reverse vaccinology, and monovalent and polyvalent vaccines.

Design and assembly of plant-based COVID-19 candidate vaccines: reсent development and future prospects.

An outbreak of a new variant of the coronavirus infection, known as COVID-19, occurred at the end of 2019 in China, in the city of Wuhan. It was caused by the SARS-CoV-2 virus. This variant of the virus is characterized by a high degree of variability and, as the current situation with its spread across different regions of the globe shows, it can lead to a progressive spread of infection among the human population and become the cause of a pandemic. The world scientific community is making tremendous efforts to develop means of protection,prevention and treatment of this disease based on modern advances in molecular biology, immunology and vaccinology. This review provides information on the current state of research in the field of vaccine development against COVID-19 with an emphasis on the role of plants in solving this complex problem. Although plants have long been used by mankind as sources of various medicinal substances, in a pandemic, plant expression systems become attractive as biofactories or bioreactors for the production of artificially created protein molecules that include protective antigens against viral infection. The design and creation of such artificial molecules underlies the development of recombinant subunit vaccines aimed at a rapid response against the spread of infections with a high degree of variability. The review presents the state of research covering a period of just over two years, i. e. since the emergence of the new outbreak of coronavirus infection. The authors tried to emphasize the importance of rapid response of research groups from various scientific fields towards the use of existing developments to create means of protection against various pathogens. With two plant expression systems – stable and transient – as examples, the development of work on the creation of recombinant subunit vaccines against COVID-19 in various laboratories and commercial companies is shown. The authors emphasize that plant expression systems have promise for the development of not only protective means under conditions of rapid response (subunit vaccines), but also therapeutic agents in the form of monoclonal antibodies against COVID-19 synthesized in plant cells.

Development of Plant-Based Vaccines for Prevention of Avian Influenza and Newcastle Disease in Poultry.

Viral diseases, including avian influenza (AI) and Newcastle disease (ND), are an important cause of morbidity and mortality in poultry, resulting in significant economic losses. Despite the availability of commercial vaccines for the major viral diseases of poultry, these diseases continue to pose a significant risk to global food security. There are multiple factors for this: vaccine costs may be prohibitive, cold chain storage for attenuated live-virus vaccines may not be achievable, and commercial vaccines may protect poorly against local emerging strains. The development of transient gene expression systems in plants provides a versatile and robust tool to generate a high yield of recombinant proteins with superior speed while managing to achieve cost-efficient production. Plant-derived vaccines offer good stability and safety these include both subunit and virus-like particle (VLP) vaccines. VLPs offer potential benefits compared to currently available traditional vaccines, including significant reductions in virus shedding and the ability to differentiate between infected and vaccinated birds (DIVA). This review discusses the current state of plant-based vaccines for prevention of the AI and ND in poultry, challenges in their development, and potential for expanding their use in low- and middle-income countries.

Novel Strategies of Immunization against COVID-19

COVID-19 manifested itself as a global pandemic in 2019 but even in 2021, it is still not successfully contained. This virus has claimed millions of lives worldwide and rendered many more jobless. Apart from causing mild to severe pneumonia, the virus has also caused a loss of livelihood for thousands globally, along with widespread trauma and depression. Since the transmission rate of the virus is so high, temporary prophylaxis relied on sanitization, wearing masks and physical distancing. However, a long-term solution for stopping viral spread is vaccination. Apart from being the fastest way to induce immunity against the virus, vaccination is also the cheapest and most practical way. However, a vaccine can only be commercially available after it has passed through various clinical trial phases. So far, more than two hundred potential vaccine candidates underwent different phases of the clinical trial, and some of the front-runners have shown more than 90% efficacy. This review has compiled all such vaccine candidates, their types, their modes of action, and the associated pros and cons. The current advances in clinical trials of vaccines have also been discussed, such as plant-based and cocktail vaccines that have recently emerged. Nowadays, novel strains like Delta plus are also emerging and posing a threat. Thus, it is mandatory to get vaccinated and choose a vaccine that provides long-term protection against multiple strains.

Surface Expression of Major Allergens on Plant-Made eBioparticle Combines Hypo-Allergenicity with Potent Immune Activation

Allergen immunotherapy is the only disease-modifying treatment for allergic diseases with proven sustained efficacy, but long treatment duration, chronic exposure to aluminum salts as well as allergic side-effects are major drawbacks. We hereby evaluate an innovative plant-based vaccine platform of in vivo assembled micro-sized bioparticles (BPs) expressing recombinant allergens for improved safety and immunogenicity.

Plant-Derived Recombinant Vaccines against Zoonotic Viruses.

Emerging and re-emerging zoonotic diseases cause serious illness with billions of cases, and millions of deaths. The most effective way to restrict the spread of zoonotic viruses among humans and animals and prevent disease is vaccination. Recombinant proteins produced in plants offer an alternative approach for the development of safe, effective, inexpensive candidate vaccines. Current strategies are focused on the production of highly immunogenic structural proteins, which mimic the organizations of the native virion but lack the viral genetic material. These include chimeric viral peptides, subunit virus proteins, and virus-like particles (VLPs). The latter, with their ability to self-assemble and thus resemble the form of virus particles, are gaining traction among plant-based candidate vaccines against many infectious diseases. In this review, we summarized the main zoonotic diseases and followed the progress in using plant expression systems for the production of recombinant proteins and VLPs used in the development of plant-based vaccines against zoonotic viruses.

Plant-Based Vaccines in Combat against Coronavirus Diseases.

Coronavirus (CoV) diseases, including Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS) have gained in importance worldwide, especially with the current COVID-19 pandemic caused by SARS-CoV-2. Due to the huge global demand, various types of vaccines have been developed, such as more traditional attenuated or inactivated viruses, subunit and VLP-based vaccines, as well as novel DNA and RNA vaccines. Nonetheless, emerging new COVID-19 variants are necessitating continuous research on vaccines, including these produced in plants, either via stable expression in transgenic or transplastomic plants or transient expression using viral vectors or agroinfection. Plant systems provide low cost, high scalability, safety and capacity to produce multimeric or glycosylated proteins. To date, from among CoVs antigens, spike and capsid proteins have been produced in plants, mostly using transient expression systems, at the additional advantage of rapid production. Immunogenicity of plant-produced CoVs proteins was positively evaluated after injection of purified antigens. However, this review indicates that plant-produced CoVs proteins or their carrier-fused immunodominant epitopes can be potentially applied also as mucosal vaccines, either after purification to be administered to particular membranes (nasal, bronchus mucosa) associated with the respiratory system, or as oral vaccines obtained from partly processed plant tissue.

Immunological characteristics of CoVID-19 and its implications

CoVID-19 is the most formidable unequaled global challenge invading 220 countries and territories in this millennium to uncountable saga of mortality, disability as humanity is witnessing devastation of socio-economy with more than 4 million deaths till date. The natural history of CoVID-19 from transmission through varied clinical features to overt complications is still under global research and research groups are on the run to trace its ramifications. This ranges from primary involvement of the pulmonary system to multisystem involvement through web of immunological pathways associated with susceptibility, clinical presentations, and severity of COVID-19. It has been hypothesized that the safe and effective mass vaccination program across the globe can ensure flattening of the pandemic curve to prepandemic normal life. This research group explored the basic and applied researches on molecular and immune mechanisms of SARS COV-2 virus. A sincere attempt has been made in futuristic research vision to find potential strengths, shortfalls, and efficacy of the plant-based immunotherapy, antibodies, and vaccine. Different research groups have hypothesized for the best possible use of these indigenous, stable, secure, efficacious natural products by searching their potential to accomplish emergency demands in this trying time. There is an urgent need to understand the inherent immunological predictors of the natural history of the disease spread over the spectrum from mild to severe forms of the disease and harp on these issues. In the wake of multiple waves with worse situations of evolving clinical features with the “variants of concern” and “variants of interest” and innovative interventions, this research group believes in optimum mix of microbial-derived biologicals with immune modifying drugs will broaden the preventive and curative spectrum.

Plant-derived VLP: a worthy platform to produce vaccine against SARS-CoV-2.

After its emergence in late 2019 SARS-CoV-2 was declared a pandemic by the World Health Organization on 11 March 2020 and has claimed more than 2.8 million lives. There has been a massive global effort to develop vaccines against SARS-CoV-2 and the rapid and low cost production of large quantities of vaccine is urgently needed to ensure adequate supply to both developed and developing countries. Virus-like particles (VLPs) are composed of viral antigens that self-assemble into structures that mimic the structure of native viruses but lack the viral genome. Thus they are not only a safer alternative to attenuated or inactivated vaccines but are also able to induce potent cellular and humoral immune responses and can be manufactured recombinantly in expression systems that do not require viral replication. VLPs have successfully been produced in bacteria, yeast, insect and mammalian cell cultures, each production platform with its own advantages and limitations. Plants offer a number of advantages in one production platform, including proper eukaryotic protein modification and assembly, increased safety, low cost, high scalability as well as rapid production speed, a critical factor needed to control outbreaks of potential pandemics. Plant-based VLP-based viral vaccines currently in clinical trials include, amongst others, Hepatitis B virus, Influenza virus and SARS-CoV-2 vaccines. Here we discuss the importance of plants as a next generation expression system for the fast, scalable and low cost production of VLP-based vaccines.

Global Innovation Trends for Plant-Based Vaccines Production: A Patent Analysis.

The use of plants as biofactories for the production of medical products and vaccines has a long history, but the recent COVID-19 pandemic has caused this set of technologies, for their potential to contribute to the development of innovative solutions for tackling pandemic spread worldwide, to rise in prominence. The purpose of this paper is to analyze the global innovation scenario of plant-based vaccine production. Methods: Patent search using a specific set of technical classification codes and keywords was performed using the Questel-Orbit database, with a final output of 180 patent families, corresponding to 1397 single patents. Results: Plant-based vaccines production is an innovation sector with positive development especially in the last five-year period (30% growth). Fifty percent of the patents were registered in the United States, standing out as the most attractive patent system worldwide. The inventive activity was led by private firms owning the 49% of the patent families, and the key-players group includes the companies that successfully developed plant-based COVID-19 vaccine candidates, indicating a strong connection between the expertise in innovation production and the capacity to adapt inventions to the current pandemic vaccine demand. Virus-like particles technology has increased in importance over the past few years. Conclusion: Patent data confirm their relevant role as indicators of innovation and technological evolution. Plant-based vaccines are expected to acquire an increasing role over the next few years as the current pandemic acts as an innovation catalyst.

Oral Immunization With Plant-Based Vaccine Induces a Protective Response Against Infectious Bursal Disease.

Infectious bursal disease virus (IBDV) is the etiological agent of an immunosuppressive and highly contagious disease that affects young birds causing important economic losses in the poultry industry worldwide. We have previously developed a plant-based vaccine candidate for infectious bursal disease (IBD) that is able to protect against infection with IBDV when administered through intramuscular (im) route. Given that oral vaccination is non-invasive and stimulates the immunity of the mucosal gastrointestinal surface, the initial site of contact and entry of IBDV, the aim of this work was to study if our immunogen was also able to elicit a protective immune response when orally administered. We demonstrated that 85% of the animals that received two oral doses of the vaccine formulation and all animals that were orally boosted after an im prime scheme developed virus neutralizing antibodies and were protected against IBDV infection, evidenced by the bursa/body weight (BB) ratio, absence of T-cell infiltration, and low viral load in bursa. Although mild to moderate bursal damage was observed in some of these animals, these lesions were not as severe as the ones observed in challenged control groups, which also presented signs of acute inflammation, bursal atrophy, T-cell infiltration, and absence of viral clearance. These results show that two immunizations with our recombinant immunogen are able to induce a specific and protective immune response in chicken against IBDV when orally administered in a prime/boost scheme or when the oral boost follows an im prime scheme. In conclusion, our oral plant-based vaccine candidate could represent a viable alternative to conventional vaccines and is of great interest to the poultry industry.

P033 SURFACE EXPRESSION OF MAJOR ALLERGENS ON PLANT-MADE BIOPARTICLES COMBINES HYPO-ALLERGENICITY WITH POTENT IMMUNE ACTIVATION

Allergen immunotherapy is the only disease-modifying treatment for allergic diseases with proven sustained efficacy, but long treatment duration, chronic exposure to aluminum salts as well as allergic side-effects are major drawbacks. We hereby evaluate an innovative plant-based vaccine platform of in vivo assembled nano-sized bioparticles expressing recombinant allergens for improved safety and immunogenicity.

Plant-based Vaccines

The pandemic of novel coronavirus disease-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has stimulated scientists from different backgrounds to gear up on developing vaccines against the virus. Several antigenic epitopes of the virus have the potential to induce an immunogenic response, among which viral spike protein (“S” protein) is considered to be the most suitable vaccine candidate. In this review, the latest progress in the field of plant molecular pharming (PMF), its application, limitations, and commercial initiatives toward the production of the SARS-CoV-2 vaccine have been discussed. Vaccine production by PMF has gained considerable attention these days and can be used for large-scale commercial production of antigenic proteins, antibodies, and other biopharmaceuticals. New age plant breeding techniques facilitated by CRISPR-Cas-based genome editing technology and next-generation sequencing methods also help to achieve greater precision and rapidity. Several unique advantages are offered by plant-based vaccine production techniques over that of the microbial or mammalian cell culture system. Virus-like particles and Agrobacterium-mediated transient somatic expression systems have a high potential for the large scale, cost-effective, and robust production of plant-derived vaccines against SARS-CoV-2.

The Recent Developmental Platforms and Potential Targets of SARS CoV-2 Vaccines: A Comprehensive Review

The novel coronavirus disease is a rapidly spreading infection caused by recently discovered different variants of SARS CoV-2 viruses, causing mild to severe respiratory symptoms in the majority of people. The Coronavirus disease 2019 pandemic has spread almost all the nooks and corners of the world. There are significant possible approaches pharmaceutically to fight against COVID-19. Original full-text research articles were searched online in PubMed, ScienceDirect, ResearchGate, Google Scholar, Core and Wiley Online Library. Scientists throughout the world are working on different platforms and targeting certain proteins moieties against SARS CoV-2 for the development of methods of safety, efficacy and potential vaccine candidates. Many candidates showed efficacy in In-vitro studies but relatively few clinical studies proceeded through different vaccine development platforms, such as entire virus vaccines, plant-based and nucleic acid vaccines, recombinant protein subunit vaccines. This review study provides a short description of SARS-CoV-2 characteristics and deals with recent developments in the design of attempts to produce vaccines to combat COVID-19.

Challenges and Prospects of Plant-Derived Oral Vaccines against Hepatitis B and C Viruses.

Hepatitis B and C viruses chronically affect approximately 3.5% of the global population, causing more than 800,000 deaths yearly due to severe liver pathogenesis. Current HBV vaccines have significantly contributed to the reduction of chronic HBV infections, supporting the notion that virus eradication is a feasible public health objective in the near future. In contrast to HBV, a prophylactic vaccine against HCV infection is not available yet; however, intense research efforts within the last decade have significantly advanced the field and several vaccine candidates are shortlisted for clinical trials. A successful vaccine against an infectious disease of global importance must not only be efficient and safe, but also easy to produce, distribute, administer, and economically affordable to ensure appropriate coverage. Some of these requirements could be fulfilled by oral vaccines that could complement traditional immunization strategies. In this review, we discuss the potential of edible plant-based oral vaccines in assisting the worldwide fight against hepatitis B and C infections. We highlight the latest research efforts to reveal the potential of oral vaccines, discuss novel antigen designs and delivery strategies, as well as the limitations and controversies of oral administration that remain to be addressed to make this approach successful.

Production of a Bacteria-like Particle Vaccine Targeting Rock Bream (Oplegnathus fasciatus) Iridovirus Using Nicotiana benthamiana.

Viral diseases are extremely widespread infections that change constantly through mutations. To produce vaccines against viral diseases, transient expression systems are employed, and Nicotiana benthamiana (tobacco) plants are a rapidly expanding platform. In this study, we developed a recombinant protein vaccine targeting the major capsid protein (MCP) of iridovirus fused with the lysine motif (LysM) and coiled-coil domain of coronin 1 (ccCor1) for surface display using Lactococcus lactis. The protein was abundantly produced in N. benthamiana in its N-glycosylated form. Total soluble proteins isolated from infiltrated N. benthamiana leaves were treated sequentially with increasing ammonium sulfate solution, and recombinant MCP mainly precipitated at 40–60%. Additionally, affinity chromatography using Ni–NTA resin was applied for further purification. Native structure analysis using size exclusion chromatography showed that recombinant MCP existed in a large oligomeric form. A minimum OD600 value of 0.4 trichloroacetic acid (TCA)-treated L. lactis was required for efficient recombinant MCP display. Immunogenicity of recombinant MCP was assessed in a mouse model through enzyme-linked immunosorbent assay (ELISA) with serum-injected recombinant MCP-displaying L. lactis. In summary, we developed a plant-based recombinant vaccine production system combined with surface display on L. lactis.

Plant-Based COVID-19 Vaccines: Current Status, Design, and Development Strategies of Candidate Vaccines.

The prevalence of the coronavirus disease 2019 (COVID-19) pandemic in its second year has led to massive global human and economic losses. The high transmission rate and the emergence of diverse SARS-CoV-2 variants demand rapid and effective approaches to preventing the spread, diagnosing on time, and treating affected people. Several COVID-19 vaccines are being developed using different production systems, including plants, which promises the production of cheap, safe, stable, and effective vaccines. The potential of a plant-based system for rapid production at a commercial scale and for a quick response to an infectious disease outbreak has been demonstrated by the marketing of carrot-cell-produced taliglucerase alfa (Elelyso) for Gaucher disease and tobacco-produced monoclonal antibodies (ZMapp) for the 2014 Ebola outbreak. Currently, two plant-based COVID-19 vaccine candidates, coronavirus virus-like particle (CoVLP) and Kentucky Bioprocessing (KBP)-201, are in clinical trials, and many more are in the preclinical stage. Interim phase 2 clinical trial results have revealed the high safety and efficacy of the CoVLP vaccine, with 10 times more neutralizing antibody responses compared to those present in a convalescent patient’s plasma. The clinical trial of the CoVLP vaccine could be concluded by the end of 2021, and the vaccine could be available for public immunization thereafter. This review encapsulates the efforts made in plant-based COVID-19 vaccine development, the strategies and technologies implemented, and the progress accomplished in clinical trials and preclinical studies so far.

Combating Human Viral Diseases: Will Plant-Based Vaccines Be the Answer?

Molecular pharming or the technology of application of plants and plant cell culture to manufacture high-value recombinant proteins has progressed a long way over the last three decades. Whether generated in transgenic plants by stable expression or in plant virus-based transient expression systems, biopharmaceuticals have been produced to combat several human viral diseases that have impacted the world in pandemic proportions. Plants have been variously employed in expressing a host of viral antigens as well as monoclonal antibodies. Many of these biopharmaceuticals have shown great promise in animal models and several of them have performed successfully in clinical trials. The current review elaborates the strategies and successes achieved in generating plant-derived vaccines to target several virus-induced health concerns including highly communicable infectious viral diseases. Importantly, plant-made biopharmaceuticals against hepatitis B virus (HBV), hepatitis C virus (HCV), the cancer-causing virus human papillomavirus (HPV), human immunodeficiency virus (HIV), influenza virus, zika virus, and the emerging respiratory virus, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) have been discussed. The use of plant virus-derived nanoparticles (VNPs) and virus-like particles (VLPs) in generating plant-based vaccines are extensively addressed. The review closes with a critical look at the caveats of plant-based molecular pharming and future prospects towards further advancements in this technology. The use of biopharmed viral vaccines in human medicine and as part of emergency response vaccines and therapeutics in humans looks promising for the near future.