Expressing the immunodominant projection domain of infectious bursal disease virus fused to the fragment crystallizable of chicken IgY in yellow maize for a prospective edible vaccine

Mol Immunol. 2020 Feb:118:132-141. doi: 10.1016/j.molimm.2019.12.015. Epub 2019 Dec 24.

Abstract

Control of Infectious bursal disease virus (IBDV) in endemic countries has been based on early immunization of chicks using conventional live or inactivated vaccines that became not fully effectual and have biosafety concerns. This endeavor seeks generating a recombinant chimeric protein merging the projection domain (PD) of IBDV VP2 capsid with the fragment crystallizable (Fc) of avian IgY (FcIgY), in maize as a prospective poultry edible vaccine. The PD sequence was built on the basis of very virulent IBDV isolates circulating in Egypt. After optimization of codon-usage in maize, sequences of PD and FcIgY were effectively expressed in two elites of yellow maize via bombardment transformation in immature embryos. Chimeric protein amount in stable transgenic samples ranged from1.36% to 3.03% of the total soluble protein based on tissue age and maize cultivar. IBDV VP2 coding sequence was amplified from viral RNA, cloned, and expressed in E. coli. A group of Balb/C mice were hyper-immunized with purified recombinant VP2 protein for raising anti- recombinant VP2 antibodies (anti-rVP2 Ab). Proper expression in maize and immunoreactivity of the chimeric protein (PD-FcIgY) to chicken anti- IBDV and anti-rVP2 Ab were confirmed by both direct and indirect double antibody sandwich (DAS)-ELISAs as well as western blotting. Seeds of regenerated transgenic maize will be validated for chickens as edible vaccination in further studies.

Keywords: Biolistic bombardment; Chimeric protein; IgY fragment crystallizable (Fc); Immature embryos; Infectious bursal disease virus (IBDV); Transgenic maize; VP2 projection domain (PD).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Viral / immunology
  • Birnaviridae Infections / immunology
  • Capsid Proteins / immunology
  • Chickens / immunology*
  • Egypt
  • Escherichia coli / immunology
  • Female
  • Immunoglobulins / immunology*
  • Infectious bursal disease virus / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Poultry Diseases / immunology
  • Prospective Studies
  • Recombinant Proteins / immunology
  • Sequence Alignment
  • Vaccination / methods
  • Vaccines, Edible / immunology*
  • Vaccines, Inactivated / immunology
  • Viral Structural Proteins / immunology
  • Viral Vaccines / immunology
  • Zea mays

Substances

  • Antibodies, Viral
  • Capsid Proteins
  • IgY
  • Immunoglobulins
  • Recombinant Proteins
  • Vaccines, Edible
  • Vaccines, Inactivated
  • Viral Structural Proteins
  • Viral Vaccines