A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection

Béatrice M.F. Winkel, Clarize M. de Korne, Matthias N. van Oosterom, Diego Staphorst, Anton Bunschoten, Marijke C.C. Langenberg, Séverine C. Chevalley-Maurel, Chris J. Janse, Blandine Franke-Fayard, Fijs W.B. van Leeuwen, Meta Roestenberg*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

Introduction: The skin stage of malaria is a vital and vulnerable migratory life stage of the parasite. It has been characterised in rodent models, but remains wholly uninvestigated for human malaria parasites. To enable in depth analysis of not genetically modified (non-GMO) Plasmodium falciparum (Pf) sporozoite behaviour in human skin, we devised a labelling technology (Cy5M2, targeting the sporozoite mitochondrion) that supports tracking of individual non-GMO sporozoites in human skin. Methods: Sporozoite labelling with Cy5M2 was performed in vitro as well as via the feed of infected Anopheles mosquitos. Labelling was validated using confocal microscopy and flow cytometry and the fitness of labelled sporozoites was determined by analysis of infectivity to human hepatocytes in vitro, and in vivo in a rodent infection model. Using confocal video microscopy and custom software, single-sporozoite tracking studies in human skin-explants were performed. Results: Both in vitro and in mosquito labelling strategies yielded brightly fluorescent sporozoites of three different Plasmodium species. Cy5M2 uptake colocalized with MitoTracker® green and could be blocked using the known Translocator protein (TSPO)-inhibitor PK11195. This method supported the visualization and subsequent quantitative analysis of the migration patterns of individual non-GMO Pf sporozoites in human skin and did not affect the fitness of sporozoites. Conclusions: The ability to label and image non-GMO Plasmodium sporozoites provides the basis for detailed studies on the human skin stage of malaria with potential for in vivo translation. As such, it is an important tool for development of vaccines based on attenuated sporozoites and their route of administration.

Original languageEnglish
Pages (from-to)2768-2778
Number of pages11
JournalTheranostics
Volume9
Issue number10
DOIs
Publication statusPublished - 2019

Fingerprint

Sporozoites
Falciparum Malaria
Parasites
Skin
Infection
Malaria
Plasmodium
Plasmodium falciparum
Culicidae
Confocal Microscopy
Rodentia
Video Microscopy
Aptitude
Anopheles
Hepatocytes
Flow Cytometry
Mitochondria
Software
Vaccines

Keywords

  • Cell tracking
  • Malaria
  • Molecular Imaging
  • Skin
  • Sporozoites

Cite this

Winkel, B. M. F., de Korne, C. M., van Oosterom, M. N., Staphorst, D., Bunschoten, A., Langenberg, M. C. C., ... Roestenberg, M. (2019). A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection. Theranostics, 9(10), 2768-2778. https://doi.org/10.7150/thno.33467
Winkel, Béatrice M.F. ; de Korne, Clarize M. ; van Oosterom, Matthias N. ; Staphorst, Diego ; Bunschoten, Anton ; Langenberg, Marijke C.C. ; Chevalley-Maurel, Séverine C. ; Janse, Chris J. ; Franke-Fayard, Blandine ; van Leeuwen, Fijs W.B. ; Roestenberg, Meta. / A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection. In: Theranostics. 2019 ; Vol. 9, No. 10. pp. 2768-2778.
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title = "A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection",
abstract = "Introduction: The skin stage of malaria is a vital and vulnerable migratory life stage of the parasite. It has been characterised in rodent models, but remains wholly uninvestigated for human malaria parasites. To enable in depth analysis of not genetically modified (non-GMO) Plasmodium falciparum (Pf) sporozoite behaviour in human skin, we devised a labelling technology (Cy5M2, targeting the sporozoite mitochondrion) that supports tracking of individual non-GMO sporozoites in human skin. Methods: Sporozoite labelling with Cy5M2 was performed in vitro as well as via the feed of infected Anopheles mosquitos. Labelling was validated using confocal microscopy and flow cytometry and the fitness of labelled sporozoites was determined by analysis of infectivity to human hepatocytes in vitro, and in vivo in a rodent infection model. Using confocal video microscopy and custom software, single-sporozoite tracking studies in human skin-explants were performed. Results: Both in vitro and in mosquito labelling strategies yielded brightly fluorescent sporozoites of three different Plasmodium species. Cy5M2 uptake colocalized with MitoTracker{\circledR} green and could be blocked using the known Translocator protein (TSPO)-inhibitor PK11195. This method supported the visualization and subsequent quantitative analysis of the migration patterns of individual non-GMO Pf sporozoites in human skin and did not affect the fitness of sporozoites. Conclusions: The ability to label and image non-GMO Plasmodium sporozoites provides the basis for detailed studies on the human skin stage of malaria with potential for in vivo translation. As such, it is an important tool for development of vaccines based on attenuated sporozoites and their route of administration.",
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author = "Winkel, {B{\'e}atrice M.F.} and {de Korne}, {Clarize M.} and {van Oosterom}, {Matthias N.} and Diego Staphorst and Anton Bunschoten and Langenberg, {Marijke C.C.} and Chevalley-Maurel, {S{\'e}verine C.} and Janse, {Chris J.} and Blandine Franke-Fayard and {van Leeuwen}, {Fijs W.B.} and Meta Roestenberg",
year = "2019",
doi = "10.7150/thno.33467",
language = "English",
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Winkel, BMF, de Korne, CM, van Oosterom, MN, Staphorst, D, Bunschoten, A, Langenberg, MCC, Chevalley-Maurel, SC, Janse, CJ, Franke-Fayard, B, van Leeuwen, FWB & Roestenberg, M 2019, 'A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection', Theranostics, vol. 9, no. 10, pp. 2768-2778. https://doi.org/10.7150/thno.33467

A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection. / Winkel, Béatrice M.F.; de Korne, Clarize M.; van Oosterom, Matthias N.; Staphorst, Diego; Bunschoten, Anton; Langenberg, Marijke C.C.; Chevalley-Maurel, Séverine C.; Janse, Chris J.; Franke-Fayard, Blandine; van Leeuwen, Fijs W.B.; Roestenberg, Meta.

In: Theranostics, Vol. 9, No. 10, 2019, p. 2768-2778.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - A tracer-based method enables tracking of plasmodium falciparum malaria parasites during human skin infection

AU - Winkel, Béatrice M.F.

AU - de Korne, Clarize M.

AU - van Oosterom, Matthias N.

AU - Staphorst, Diego

AU - Bunschoten, Anton

AU - Langenberg, Marijke C.C.

AU - Chevalley-Maurel, Séverine C.

AU - Janse, Chris J.

AU - Franke-Fayard, Blandine

AU - van Leeuwen, Fijs W.B.

AU - Roestenberg, Meta

PY - 2019

Y1 - 2019

N2 - Introduction: The skin stage of malaria is a vital and vulnerable migratory life stage of the parasite. It has been characterised in rodent models, but remains wholly uninvestigated for human malaria parasites. To enable in depth analysis of not genetically modified (non-GMO) Plasmodium falciparum (Pf) sporozoite behaviour in human skin, we devised a labelling technology (Cy5M2, targeting the sporozoite mitochondrion) that supports tracking of individual non-GMO sporozoites in human skin. Methods: Sporozoite labelling with Cy5M2 was performed in vitro as well as via the feed of infected Anopheles mosquitos. Labelling was validated using confocal microscopy and flow cytometry and the fitness of labelled sporozoites was determined by analysis of infectivity to human hepatocytes in vitro, and in vivo in a rodent infection model. Using confocal video microscopy and custom software, single-sporozoite tracking studies in human skin-explants were performed. Results: Both in vitro and in mosquito labelling strategies yielded brightly fluorescent sporozoites of three different Plasmodium species. Cy5M2 uptake colocalized with MitoTracker® green and could be blocked using the known Translocator protein (TSPO)-inhibitor PK11195. This method supported the visualization and subsequent quantitative analysis of the migration patterns of individual non-GMO Pf sporozoites in human skin and did not affect the fitness of sporozoites. Conclusions: The ability to label and image non-GMO Plasmodium sporozoites provides the basis for detailed studies on the human skin stage of malaria with potential for in vivo translation. As such, it is an important tool for development of vaccines based on attenuated sporozoites and their route of administration.

AB - Introduction: The skin stage of malaria is a vital and vulnerable migratory life stage of the parasite. It has been characterised in rodent models, but remains wholly uninvestigated for human malaria parasites. To enable in depth analysis of not genetically modified (non-GMO) Plasmodium falciparum (Pf) sporozoite behaviour in human skin, we devised a labelling technology (Cy5M2, targeting the sporozoite mitochondrion) that supports tracking of individual non-GMO sporozoites in human skin. Methods: Sporozoite labelling with Cy5M2 was performed in vitro as well as via the feed of infected Anopheles mosquitos. Labelling was validated using confocal microscopy and flow cytometry and the fitness of labelled sporozoites was determined by analysis of infectivity to human hepatocytes in vitro, and in vivo in a rodent infection model. Using confocal video microscopy and custom software, single-sporozoite tracking studies in human skin-explants were performed. Results: Both in vitro and in mosquito labelling strategies yielded brightly fluorescent sporozoites of three different Plasmodium species. Cy5M2 uptake colocalized with MitoTracker® green and could be blocked using the known Translocator protein (TSPO)-inhibitor PK11195. This method supported the visualization and subsequent quantitative analysis of the migration patterns of individual non-GMO Pf sporozoites in human skin and did not affect the fitness of sporozoites. Conclusions: The ability to label and image non-GMO Plasmodium sporozoites provides the basis for detailed studies on the human skin stage of malaria with potential for in vivo translation. As such, it is an important tool for development of vaccines based on attenuated sporozoites and their route of administration.

KW - Cell tracking

KW - Malaria

KW - Molecular Imaging

KW - Skin

KW - Sporozoites

U2 - 10.7150/thno.33467

DO - 10.7150/thno.33467

M3 - Article

VL - 9

SP - 2768

EP - 2778

JO - Theranostics

JF - Theranostics

SN - 1838-7640

IS - 10

ER -