Carrot cells expressing the VP1 and VP2 poliovirus proteins effectively elicited mucosal immunity

Bolaños-Martínez, Omayra C.; Govea‐Alonso, Dania O.; Fragoso, Gladis; Sciutto, Edda; Rosales‐Mendoza, Sergio

doi:10.1007/s11240-021-02205-0

Cited by 4 publications

(6 citation statements)

References 49 publications

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“…The estimated levels of VP2 protein in transgenic SMC-1 line callus were similar to those reached for poliovirus VP1 and VP2 proteins expressed in tobacco, obtaining expression levels of 0.3 µg VP1/g, 6.16 – 16.85 µg VP2/g, 2.72 – 4.33 µg VP3/g, and 0.96 – 6 µg VP4/g fresh leaf tissue 29 . On other hand, Daucus carota is another of the systems implemented to produce poliovirus heterologous proteins; recently, the expression of VP1 (1.17 – 3.57 µg/g FW) and VP2 (4.24 – 13.86 µg/g FW) proteins was reported using a medium enriched with amino acids, urea, and other supplements 30 .…”

Section: Discussionmentioning

confidence: 99%

“…In this work, the VP2 transgenic carrot line named the SMC-1 line was used because of its higher accumulation levels of the VP2 protein (13.86 µg/g FW), a medium enriched with amino acids, urea, and other supplements. A non-transformed line (WT) was used as negative control [30]…”

Section: Transgenic Carrot Calluses Expressing Vp2mentioning

confidence: 99%

“…The VP2 protein level expression of the SMC-1 carrot line was assessed by ELISA according to a previously reported protocol [30]. To extract proteins from the lyophilized biomass from either wildtype (WT) or transgenic cell lines, 100 mg of the tissue was resuspended in 300 µL of extraction buffer (25 mM sodium phosphate at pH 6.6, 100 mM NaCl, 0.5 % Triton X-100 (v/v), and 1 mM PMSF) and subjected to sonication using 5 pulses of 5 s each at 25 % amplitude.…”

Section: Elisa and Dot Blotmentioning

confidence: 99%

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Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Morales-Aguilar,

Bolaños-Martínez,

Maldonado

et al. 2023

Planta Med

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The development of virus-free, oral vaccines against poliovirus capable of inducing mucosal protective immunity is needed to safely combat this pathogen. In the present study, a carrot cell line expressing the poliovirus VP2 antigen was established at the level of callus and cell suspensions, exploring the effects of culture media (MS and B5), supplementation with urea, phytoregulators (2,4-D : KIN), and light conditions (continuous light, photoperiod, and total darkness). The best callus growth was obtained on B5 medium supplemented with 2 mg/L of 2,4-D + 2 mg/L kinetin and 0.0136 g/L of urea and in continuous light conditions. Suspension cultures of the SMC-1 line in 250 mL Erlenmeyer flasks had a maximum growth of 16.07 ± 0.03 g/L DW on day 12 with a growth rate of µ=0.3/d and a doubling time of 2.3 days. In a 2 L airlift bioreactor, the biomass yield achieved was 25.6 ± 0.05 g/L DW at day 10 with a growth rate of µ= 0.58/d and doubling time of 1.38 d. Cell growth was 1.5 times higher in bioreactors than in shake flasks, highlighting that both systems resulted in the accumulation of VP2 throughout the time in culture. The maximum VP2 yield in flasks was 387.8 µg/g DW at day 21, while in the reactor it was 550.2 µg/g DW at day 18. In conclusion, bioreactor-based production of the VP2 protein by the SMC-1 suspension cell line offers a higher productivity when compared to flask cultures, offering a key perspective to produce low-cost vaccines against poliomyelitis.

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Section: Discussionmentioning

confidence: 99%

Section: Transgenic Carrot Calluses Expressing Vp2mentioning

confidence: 99%

Section: Elisa and Dot Blotmentioning

confidence: 99%

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Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Morales-Aguilar,

Bolaños-Martínez,

Maldonado

et al. 2023

Planta Med

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“…Additionally, generally, these vaccines are expensive and inaccessible to poor communities and are highmaintenance as their stability must be preserved. In the continued effort to address such shortcomings, recently, polio viral proteins (VPs) 1 and 2 were expressed in carrot cell lines [79]. Oral immunization of mice resulted in elevated levels of S-IgA and IgG as detected in the fecal excrement of the mice.…”

Section: Cancer Vaccinesmentioning

confidence: 99%

Recent Progress on Vaccines Produced in Transgenic Plants

et al. 2022

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The development of vaccines from plants has been going on for over two decades now. Vaccine production in plants requires time and a lot of effort. Despite global efforts in plant-made vaccine development, there are still challenges that hinder the realization of the final objective of manufacturing approved and safe products. Despite delays in the commercialization of plant-made vaccines, there are some human vaccines that are in clinical trials. The novel coronavirus (SARS-CoV-2) and its resultant disease, coronavirus disease 2019 (COVID-19), have reminded the global scientific community of the importance of vaccines. Plant-made vaccines could not be more important in tackling such unexpected pandemics as COVID-19. In this review, we explore current progress in the development of vaccines manufactured in transgenic plants for different human diseases over the past 5 years. However, we first explore the different host species and plant expression systems during recombinant protein production, including their shortcomings and benefits. Lastly, we address the optimization of existing plant-dependent vaccine production protocols that are aimed at improving the recovery and purification of these recombinant proteins.

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“…In order to investigate the capacity of a different edible plant to yield poliovirus VPs and advance to the development of an oral vaccine candidate, the VP1 and VP2 Sabin 1 proteins were expressed in carrot cells ( Bolaños-Martínez et al., 2022 ). The obtained yields ranged from 1.17 to 3.57 µg/g fresh weight for the VP1 protein and 4.24 to 13.86 µg/g for the VP2 protein.…”

Section: Plant-made Poliovirus Vaccine Candidatesmentioning

confidence: 99%

Plant-made poliovirus vaccines – Safe alternatives for global vaccination

Bolaños-Martínez¹,

Strasser²

2022

Front. Plant Sci.

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Human polioviruses are highly infectious viruses that are spread mainly through the fecal-oral route. Infection of the central nervous system frequently results in irreversible paralysis, a disease called poliomyelitis. Children under five years are mainly affected if they have not acquired immunity through natural infection or via vaccination. Current polio vaccines comprise the injectable inactivated polio vaccine (IPV, also called the Salk vaccine) and the live-attenuated oral polio vaccine (OPV, also called the Sabin vaccine). The main limitations of the IPV are the reduced protection at the intestinal mucosa, the site of virus replication, and the high costs for manufacturing due to use of live viruses. While the OPV is more effective and stimulates mucosal immunity, it is manufactured using live-attenuated strains that can revert into pathogenic viruses resulting in major safety concerns and vaccine-derived outbreaks. During the last fifteen years, plant-based poliovirus vaccines have been explored by several groups as a safe and low-cost alternative, and promising results in protection against challenges with viruses and induction of neutralizing antibodies have been obtained. However, low yields and a high frequency in dose administration highlight the need for improvements in polioviral antigen production. In this review, we provide insights into recent efforts to develop plant-made poliovirus candidates, with an emphasis on strategies to optimize the production of viral antigens.

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Carrot cells expressing the VP1 and VP2 poliovirus proteins effectively elicited mucosal immunity

Cited by 4 publications

References 49 publications

Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Recent Progress on Vaccines Produced in Transgenic Plants

Plant-made poliovirus vaccines – Safe alternatives for global vaccination

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