Imaging malaria

(In vivo) imaging of parasite-host interactions

The Leiden Malaria Research Group (LMRG) is studying interactions between malaria parasites and their host in live animals using the P. berghei rodent model of malaria (see also Research Themes). The main thrust of the work is specifically geared to gain insights on key molecular interactions, between host and pathogen, which govern malaria associated pathology. For this research we make use of the latest real-time (in vivo) bio-imaging technologies and an array of transgenic (fluorescent and/or bio-luminescent) malaria parasites.

MILL
The Leiden Malaria Research Group participates in MILL, a technology platform dedicated to molecular imaging technology development and application.

Advances in genetic modification of malaria parasites and imaging technologies for visualizing cells expressing reporter genes have significantly broadened the possibilities for in vitro and in vivo studies of host parasite interactions. Transgenic malaria parasites expressing fluorescent or bioluminescent reporter proteins (i.e. luciferase) are used in real-time imaging experiments with either live animals or in isolated organs.

Equipment/facilities
(Optical) Imaging of is performed using the following equipment/facilities:

• IVIS® Imaging System 100 series (Xenogen/Caliper). Bioluminescence-based in-vivo (whole body) molecular imaging. This system is available in the DMIII facilities
• IVIS® Imaging System 200 series; Spectrum (Xenogen/Caliper) Fluorescence/Bioluminescence-based in-vivo (whole body) molecular imaging with option for 3D and spectral unmixing.
• Intravital multiphoton laser microscope (see here for more information)

References

-  Plasmepsin 4-deficient Plasmodium berghei are virulence attenuated and induce protective immunity against experimental malaria. Spaccapelo R, Janse CJ, Caterbi S, Franke-Fayard B, Bonilla JA, Syphard LM, Di Cristina M, Dottorini T, Savarino A, Cassone A, Bistoni F, Waters AP, Dame JB, Crisanti A. Am J Pathol. 2010;176(1):205-17.

-  Visualisation and Quantitative Analysis of the Rodent Malaria Liver Stage by Real Time Imaging. Ploemen IHJ, Prudêncio M, Douradinha BG, Ramesar J, Fonager J, van Gemert GJ, Luty AJF, Hermsen CC, Sauerwein RW, Baptista FG, Mota MM, Waters AP, Que I, Lowik CWGM, Khan SM, Janse CJ, Franke-Fayard BMD. PLoS ONE 2009 4(11): e7881.

-  Going live: A comparative analysis of the suitability of the RFP derivatives RedStar, mCherry and tdTomato for intravital and in vitro live imaging of Plasmodium parasites. Graewe S, Retzlaff S, Struck N, Janse CJ, Heussler VT. Biotechnol J. 2009;4(6):895-902.

-  Simple and sensitive antimalarial drug screening in vitro and in vivo using transgenic luciferase expressing Plasmodium berghei parasites. Franke-Fayard B, Djokovic D, Dooren MW, Ramesar J, Waters AP, Falade MO, Kranendonk M, Martinelli A, Cravo P, Janse CJ. Int J Parasitol. 2008, 38:1651-62.

-  A role for natural regulatory T cells in the pathogenesis of experimental cerebral malaria. Amante FH, Stanley AC, Randall LM, Zhou Y, Haque A, McSweeney K, Waters AP, Janse CJ, Good MF, Hill GR, Engwerda CR. Am J Pathol. 2007, 171:548-59.

-  Franke-Fayard B, Waters AP, Janse CJ. Real-time in vivo imaging of transgenic bioluminescent blood stages of rodent malaria parasites in mice. Nat Protoc. 2006;1(1):476-85.

-  Franke-Fayard B, Janse CJ, Cunha-Rodrigues M, Ramesar J, Buscher P, Que I, Lowik C, Voshol PJ, den Boer MA, van Duinen SG, Febbraio M, Mota MM, Waters AP.(2005). Murine malaria parasite sequestration: CD36 is the major receptor, but cerebral pathology is unlinked to sequestration.Proc Natl Acad Sci USA. 102. 11468-73