Expression of full-length Plasmodium falciparum P48/45 in P. berghei blood stages: A method to express and evaluate vaccine antigens

Mol Biochem Parasitol. 2018 Sep:224:44-49. doi: 10.1016/j.molbiopara.2018.07.009. Epub 2018 Jul 24.

Abstract

The transmission-blocking vaccine candidate Pfs48/45 from the human malaria parasite Plasmodium falciparum is known to be difficult to express in heterologous systems, either as full-length protein or as correctly folded protein fragments that retain conformational epitopes. In this study we express full-length Pfs48/45 in the rodent parasite P. berghei. Pfs48/45 is expressed as a transgene under control of the strong P. berghei schizont-specific msp1 gene promoter (Pfs48/45@PbMSP1). Pfs48/45@PbMSP1 schizont-infected red blood cells produced full-length Pfs48/45 and the structural integrity of Pfs48/45 was confirmed using a panel of conformation-specific monoclonal antibodies that bind to different Pfs48/45 epitopes. Sera from mice immunized with transgenic Pfs48/45@PbMSP1 schizonts showed strong transmission-reducing activity in mosquitoes infected with P. falciparum using standard membrane feeding. These results demonstrate that transgenic rodent malaria parasites expressing human malaria antigens may be used as means to evaluate immunogenicity and functionality of difficult to express malaria vaccine candidate antigens.

Keywords: Immunization; Pfs48/45; Plasmodium; Standard membrane feeding; Transgene-expression; Transmission blocking.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Protozoan / biosynthesis
  • Antigens, Protozoan / genetics
  • Antigens, Protozoan / immunology*
  • Disease Models, Animal
  • Disease Transmission, Infectious / prevention & control
  • Gene Expression
  • Malaria / prevention & control
  • Malaria Vaccines / immunology*
  • Membrane Proteins / biosynthesis
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology*
  • Mice
  • Plasmodium berghei / genetics*
  • Plasmodium falciparum / genetics*
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology*
  • Transgenes

Substances

  • Antigens, Protozoan
  • Malaria Vaccines
  • Membrane Proteins
  • Recombinant Proteins