Design of a multiepitopic Zaire ebolavirus protein and its expression in plant cells

Nieto-Gómez, Ricardo; Angulo, Carlos; Monreal‐Escalante, Elizabeth; Govea‐Alonso, Dania O.; Groot, Anne S. De; Rosales‐Mendoza, Sergio

doi:10.1016/j.jbiotec.2019.02.003

Cited by 12 publications

(6 citation statements)

References 41 publications

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“…The research on development of plant‐based vaccine needs to be expanded for the diseases common all over the world, and especially in developing countries, such as TB. Plant‐based expression of foreign proteins of pharmaceutical importance has several advantages including ease to cultivate in large areas, high yield of recombinant proteins, easy storage at room temperature, and low cost .…”

Section: Discussionmentioning

confidence: 99%

“…Due to safety issues of heat killed or whole cell antigen vaccines, production of plant‐based vaccines has become a striking strategy for the development of cost‐effective vaccines due to some important features such as: high scalability, low health risks, minimum need of cooling chain, and low cost of downstream processing . Currently, delivery of vaccines via plant systems has gained a high consideration platform for oral administration and its renowned potential to stimulate immunity in human and animals .…”

Section: Introductionmentioning

confidence: 99%

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Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Saba

Sameeullah

Asghar

et al. 2020

Biotech and App Biochem

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Tuberculosis (TB) is one of the major infectious diseases caused by Mycobacterium tuberculosis. The development of an effective and economical vaccine for controlling TB is essential especially for developing countries. Edible plants can serve as biofactories to produce vaccine antigens. In this study, 6 kDa early secretory antigenic target (ESAT‐6) of M. tuberculosis was expressed in Brassica oleracea var. italica via Agrobacterium‐mediated transformation to facilitate oral delivery of antigen. ESAT‐6 gene was cloned using Gateway® cloning strategy. Transformation and presence of transgene was confirmed through PCR. Expression level of transgene was calculated via quantitative real‐time PCR (qRT‐PCR) and the maximum integrated transgene number was two. Maximum amount of total soluble fraction of ESAT‐6 was evaluated by immunoblotting, estimated to accumulate up to 0.5% of total soluble protein. The recombinant ESAT‐6 protein was further purified and detected using silver staining and Western blotting. ESAT‐6 protein induced humoral immune response in mice immunized orally and subcutaneously. The expression of M. tuberculosis antigen in edible plants could aid in the development of cost‐effective and oral delivery of an antigen‐based subunit vaccine against TB. To the best our knowledge, it is the first report of expression of a vaccine antigen in broccoli.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Saba

Sameeullah

Asghar

et al. 2020

Biotech and App Biochem

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“…no. WP_012846869), (c) a proline‐glycine containing linker (GPGP, 29,30 ), (d) the full‐length sequence of CdeM (CD1581; GenBank acc. no.…”

Section: Methodsmentioning

confidence: 99%

Transgenic plants expressing a Clostridium difficile spore antigen as an approach to develop low‐cost oral vaccines

Rosales‐Mendoza

Cervantes-Rincón

Romero-Maldonado

et al. 2021

Biotechnology Progress

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Gastrointestinal infections caused by Clostridium difficile lead to significant impact in terms of morbidity and mortality, causing from mild symptoms, such as a lowgrade fever, watery stools, and minor abdominal cramping as well as more severe symptoms such as bloody diarrhea, pseudomembrane colitis, and toxic megacolon.Vaccination is a viable approach to fight against C. difficile and several efforts in this direction are ongoing. Plants are promising vaccine biofactories offering low cost, enhanced safety, and allow for the formulation of oral vaccines. Herein, the CdeM protein, which is a spore antigen associated with immunoprotection against C. difficile, was selected to begin the development of plant-based vaccine candidates. The vaccine antigen is based in a fusion protein (LTB-CdeM), carrying the CdeM antigen, fused to the carboxi-terminus of the B subunit of the Escherichia coli heat-labile enterotoxin (LTB) as a mucosal immunogenic carrier. LTB-CdeM was produced in plants using a synthetic optimized gene according codon usage and mRNA stability criteria. The obtained transformed tobacco lines produced the LTB-CdeM antigen in the range of 52-90 μg/g dry weight leaf tissues. The antigenicity of the plant-made LTB-CdeM antigen was evidenced by GM1-ELISA and immunogenicity assessment performed in test mice revealed that the LTB-CdeM antigen is orally immunogenic inducing humoral responses against CdeM epitopes. This report constitutes the first step in the development of plant-based vaccines against C. difficile infection. K E Y W O R D S C. difficile infection, chimeric antigen, heat labile E. coli enterotoxin B subunit, molecular farming, oral immunization 1 | INTRODUCTION Clostridium difficile is a spore-forming, Gram-positive bacteria that is the leading cause of antibiotic-associated infectious diarrhea in the United States and is one the most common hospital bacterial acquired infection. 1 C. difficile infection (CDI) is a gastrointestinal disease believed to be causally related to perturbations in the intestinal microbiota and is the largest single cause of morbidity and mortality among hospitalacquired infections. 2,3 CDI is generally acquired when patients with predisposing factors, such as advanced age and antibiotic use, are exposed to C. difficile spores emanating from other hospitalized infected patients. 4 CDI cause mild symptoms, such as a low-grade fever, watery Sergio Rosales-Mendoza and Tomás Cervantes-Rincón contributed equally to this study.

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“…There are, however, examples from other families of enveloped viruses which deserve mention. For example, a synthetic multiepitope peptide from ebolavirus ( Filoviridae ) surface glycoprotein (GP) was expressed in transgenic tobacco [ 173 ], and the GP1 protein was expressed in N. benthamiana as part of an immune complex strategy (similar to what has been described above for flaviviruses) and was found to be protective in a mouse lethal challenge model [ 174 , 175 ]. There have also been attempts at making vaccine candidates against human respiratory syncytial virus ( Pneumoviridae ) by fusing small epitopes of the G protein to alfalfa mosaic virus coat protein [ 176 , 177 ].…”

Section: Other Virus Familiesmentioning

confidence: 99%

Producing Vaccines against Enveloped Viruses in Plants: Making the Impossible, Difficult

et al. 2021

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The past 30 years have seen the growth of plant molecular farming as an approach to the production of recombinant proteins for pharmaceutical and biotechnological uses. Much of this effort has focused on producing vaccine candidates against viral diseases, including those caused by enveloped viruses. These represent a particular challenge given the difficulties associated with expressing and purifying membrane-bound proteins and achieving correct assembly. Despite this, there have been notable successes both from a biochemical and a clinical perspective, with a number of clinical trials showing great promise. This review will explore the history and current status of plant-produced vaccine candidates against enveloped viruses to date, with a particular focus on virus-like particles (VLPs), which mimic authentic virus structures but do not contain infectious genetic material.

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Design of a multiepitopic Zaire ebolavirus protein and its expression in plant cells

Cited by 12 publications

References 41 publications

Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Transgenic plants expressing a Clostridium difficile spore antigen as an approach to develop low‐cost oral vaccines

Producing Vaccines against Enveloped Viruses in Plants: Making the Impossible, Difficult

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