Using carrot cells as biofactories and oral delivery vehicles of LTB-Syn: A low-cost vaccine candidate against synucleinopathies

J Biotechnol. 2020 Feb 10:309:75-80. doi: 10.1016/j.jbiotec.2019.12.007. Epub 2019 Dec 13.

Abstract

Synucleinopathies are conditions that remain with no available effective treatments thus far. Immunotherapy is a possible path to fight against such pathologies by inducing antibodies against alpha-synuclein (α-Syn), which could induce the clearance of its pathologic form. Looking to develop a new low-cost, effective vaccine against synucleinopathies; we have designed a chimeric plant-made antigen comprising the subunit B of the enterotoxin from enterotoxigenic E. coli and three B cell epitopes from α-Syn, which is named LTB-Syn. In the present study, LTB-Syn was produced in carrot cell lines as appropriate platform for the formulation of oral vaccines not requiring purification. The development of transgenic carrot cell lines took 8 months and the LTB-Syn yield reached 2.3 μg/g dry biomass. The antigen encapsulated in lyophilized carrot cells was highly stable at room temperature over a six-month period and upon heating at 50 °C for 2 h. Moreover, LTB-Syn was able to prime immune responses that, in combination with parenteral boosting using an OVA-Syn conjugate, induced significant humoral resposes in mice. Thus the carrot-made oral LTB-Syn vaccine is a promising candidate that deserves further analyses to advance in its preclinical evaluation.

Keywords: Alpha-synuclein; Carrot cell lines; Humoral response; Oral priming; Plant-made vaccine; Synucleinopathies.

MeSH terms

  • Animals
  • Biomass
  • Cell Line
  • Daucus carota / chemistry*
  • Daucus carota / genetics
  • Disease Models, Animal
  • Enterotoxins / immunology
  • Epitopes, B-Lymphocyte
  • Escherichia coli / genetics
  • Escherichia coli Proteins / genetics
  • Female
  • Immunogenicity, Vaccine / immunology
  • Immunotherapy
  • Mice
  • Mice, Inbred BALB C
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Synucleinopathies / immunology
  • Synucleinopathies / prevention & control*
  • Vaccines / economics
  • Vaccines / immunology*
  • alpha-Synuclein / genetics
  • alpha-Synuclein / immunology*

Substances

  • Enterotoxins
  • Epitopes, B-Lymphocyte
  • Escherichia coli Proteins
  • Vaccines
  • alpha-Synuclein