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.
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References
Leitner WW, Bergmann-Leitner ES, Angov E (2010) Comparison of Plasmodium berghei challenge models for the evaluation of pre-erythrocytic malaria vaccines and their effect on perceived vaccine efficacy. Malar J 9:145
Sauerwein RW, Roestenberg M, Moorthy VS (2011) Experimental human challenge infections can accelerate clinical malaria vaccine development. Nat Rev Immunol 11(1):57–64
Macchiarini F et al (2005) Humanized mice: are we there yet? J Exp Med 202(10):1307–1311
Meuleman P, Leroux-Roels G (2009) HCV animal models: a journey of more than 30 years. Viruses 1(2):222–240
Kaushansky A et al (2014) Of men in mice: the success and promise of humanized mouse models for human malaria parasite infections. Cell Microbiol 16(5):602–611
Meuleman P et al (2005) Morphological and biochemical characterization of a human liver in a uPA-SCID mouse chimera. Hepatology 41(4):847–856
Azuma H et al (2007) Robust expansion of human hepatocytes in Fah-/-/Rag2-/-/Il2rg-/-mice. Nat Biotechnol 25(8):903–910
Bissig KD et al (2007) Repopulation of adult and neonatal mice with human hepatocytes: a chimeric animal model. Proc Natl Acad Sci U S A 104(51):20507–20511
Vanwolleghem T et al (2010) Factors determining successful engraftment of hepatocytes and susceptibility to hepatitis B and C virus infection in uPA-SCID mice. J Hepatol 53(3):468–476
Tateno C et al (2013) Morphological and microarray analyses of human hepatocytes from xenogeneic host livers. Lab Investig 93(1):54–71
Braun KM, Sandgren EP (1998) Liver disease and compensatory growth: unexpected lessons from genetically altered mice. Int J Dev Biol 42(7):935–942
Grompe M et al (1993) Loss of fumarylacetoacetate hydrolase is responsible for the neonatal hepatic dysfunction phenotype of lethal albino mice. Genes Dev 7(12A):2298–2307
Wilson EM et al (2014) Extensive double humanization of both liver and hematopoiesis in FRGN mice. Stem Cell Res 13(3PA):404–412
Vaughan AM et al (2012) Development of humanized mouse models to study human malaria parasite infection. Future Microbiol 7(5):657–665
Sacci JB Jr et al (2006) Plasmodium falciparum infection and exoerythrocytic development in mice with chimeric human livers. Int J Parasitol 36(3):353–360
Vaughan AM et al (2012) Complete Plasmodium falciparum liver-stage development in liver-chimeric mice. J Clin Invest 122(10):3618–3628
Foquet L et al (2014) Vaccine-induced monoclonal antibodies targeting circumsporozoite protein prevent Plasmodium falciparum infection. J Clin Invest 124(1):140–144
Sack BK et al. (2013) A model for in vivo assessment of humoral protection against malaria sporozoite challenge by passive transfer of monoclonal antibodies and immune serum. Infect Immun. p. IAI. 01249-13
Foquet L et al (2013) Molecular detection and quantification of Plasmodium falciparum-infected human hepatocytes in chimeric immune-deficient mice. Malar J 12(1):430
Vaughan A et al (2012) A transgenic Plasmodium falciparum NF54 strain that expresses GFP-luciferase throughout the parasite life cycle. Mol Biochem Parasitol 186(2):143–147
Teirlinck AC et al (2013) NF135.C10: a new Plasmodium falciparum clone for controlled human malaria infections. J Infect Dis 207(4):656–660
Tateno C et al (2004) Near completely humanized liver in mice shows human-type metabolic responses to drugs. Am J Pathol 165(1fb9c63d-0fa7-8bd3-eba4-7a81d8a986dc):901–913
Mota MM et al (2001) Migration of Plasmodium sporozoites through cells before infection. Science 291(5501):141–144
Rosenberg R et al (1990) An estimation of the number of malaria sporozoites ejected by a feeding mosquito. Trans R Soc Trop Med Hyg 84(2):209–212
Jin Y, Kebaier C, Vanderberg J (2007) Direct microscopic quantification of dynamics of Plasmodium berghei sporozoite transmission from mosquitoes to mice. Infect Immun 75(11):5532–5539
Hermsen CC et al (2001) Detection of Plasmodium falciparum malaria parasites in vivo by real-time quantitative PCR. Mol Biochem Parasitol 118(2):247–251
Bijker EM et al (2014) Sporozoite immunization of human volunteers under mefloquine prophylaxis is safe, immunogenic and protective: a double-blind randomized controlled clinical trial. PLoS One 9(11), e112910
Alcoser SY et al (2011) Real-time PCR-based assay to quantify the relative amount of human and mouse tissue present in tumor xenografts. BMC Biotechnol 11:124
Marcos R, Monteiro RA, Rocha E (2006) Design-based stereological estimation of hepatocyte number, by combining the smooth optical fractionator and immunocytochemistry with anti-carcinoembryonic antigen polyclonal antibodies. Liver Int 26(1):116–124
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Foquet, L., Meuleman, P., Hermsen, C.C., Sauerwein, R., Leroux-Roels, G. (2015). Assessment of Parasite Liver-Stage Burden in Human-Liver Chimeric Mice. In: Vaughan, A. (eds) Malaria Vaccines. Methods in Molecular Biology, vol 1325. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2815-6_5
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DOI: https://doi.org/10.1007/978-1-4939-2815-6_5
Publisher Name: Humana Press, New York, NY
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