Evaluation of immunity against malaria using luciferase-expressing Plasmodium berghei parasites

Malar J. 2011 Dec 9:10:350. doi: 10.1186/1475-2875-10-350.

Abstract

Background: Measurement of liver stage development is of key interest in malaria biology and vaccine studies. Parasite development in liver cells can be visualized in real-time, both in culture and in live mice, using a transgenic Plasmodium berghei parasite, PbGFP-Luccon, expressing the bioluminescent reporter luciferase. This study explores the benefit of using these parasites for the evaluation of immunity against malaria, compared to qRT-PCR techniques in vivo and in vitro.

Methods: Mice were immunized with either radiation attenuated sporozoites (RAS) or wildtype sporozoites under chloroquine prophylaxis (CPS) and challenged with PbGFP-Luccon. The in vitro transgenic sporozoites neutralization assay (TSNA) was adapted by replacing PbCS(Pf) parasites for PbGFP-Luccon parasites.

Results: Application of PbGFP-Luccon transgenic parasites provides live quantitative visual information about the relation between parasite liver load and protection. Moreover, fast and reproducible results are obtained by using these parasites in the transgenic sporozoites neutralization assay, measuring functional antibody-mediated immune responses.

Conclusions: PbGFP-Luccon parasites are a straightforward and valuable tool for comprehension of the biological and immunological principles underlying protection against malaria.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Anopheles / parasitology
  • Chloroquine / administration & dosage
  • Diagnostic Imaging / methods*
  • Female
  • Genes, Reporter
  • Green Fluorescent Proteins / metabolism
  • Hepatocytes / parasitology
  • Immunity, Humoral*
  • Liver / immunology
  • Liver / parasitology
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Malaria / immunology*
  • Malaria / parasitology
  • Mice
  • Mice, Inbred C57BL
  • Neutralization Tests
  • Organisms, Genetically Modified / genetics
  • Organisms, Genetically Modified / immunology
  • Organisms, Genetically Modified / metabolism
  • Parasite Load
  • Plasmodium berghei / immunology*
  • Plasmodium berghei / pathogenicity
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sporozoites / immunology
  • Sporozoites / metabolism
  • Sporozoites / radiation effects

Substances

  • Green Fluorescent Proteins
  • Chloroquine
  • Luciferases