Assessment of Parasite Liver-Stage Burden in Human-Liver Chimeric Mice

Methods Mol Biol. 2015:1325:59-68. doi: 10.1007/978-1-4939-2815-6_5.

Abstract

Humanized mice with a chimeric liver are a promising tool to evaluate the "in vivo" efficacy of novel compounds or vaccine-induced antibodies directed against the pre-erythrocytic stages of Plasmodium falciparum. The absence of human red blood cells in these humanized mice precludes the transition from liver to blood stage. The qPCR-based method described below allows for a sensitive and reliable quantification of parasite DNA in the chimeric liver following a challenge via infected mosquito bite or intravenous injection of sporozoites. With this method approximately 25 % of the total chimeric liver is examined and a single infected hepatocyte can be detected in the analyzed tissue. The use of appropriate species-specific probes can also allow for the detection of other Plasmodium species in vivo.

Keywords: Humanized mouse model; In vivo; Liver stage; Malaria; Plasmodium falciparum; Sporozoite; qPCR.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Anopheles / parasitology
  • Erythrocytes / immunology
  • Hepatocytes / immunology
  • Hepatocytes / pathology
  • Humans
  • Liver / immunology*
  • Liver / parasitology
  • Liver / pathology
  • Malaria / immunology*
  • Malaria / parasitology
  • Malaria / prevention & control
  • Mice
  • Molecular Biology / methods*
  • Plasmodium falciparum / immunology*
  • Plasmodium falciparum / pathogenicity
  • Plasmodium yoelii / immunology
  • Plasmodium yoelii / pathogenicity
  • Sporozoites / immunology