Leishmania promastigotes lack phosphatidylserine but bind annexin V upon permeabilization or miltefosine treatment

Adrien Weingärtner; Gerdi Christine Kemmer; Frederic D. Müller; Ricardo Andrade Zampieri; Jürgen Schiller; Thomas Günther-Pomorski

doi:10.1371/journal.pone.0042070

Leishmania promastigotes lack phosphatidylserine but bind annexin V upon permeabilization or miltefosine treatment

Adrien Weingärtner, Gerdi Christine Kemmer, Frederic D. Müller, Ricardo Andrade Zampieri, Jürgen Schiller, Thomas Günther-Pomorski

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Abstract

The protozoan parasite Leishmania is an intracellular pathogen infecting and replicating inside vertebrate host macrophages. A recent model suggests that promastigote and amastigote forms of the parasite mimic mammalian apoptotic cells by exposing phosphatidylserine (PS) at the cell surface to trigger their phagocytic uptake into host macrophages. PS presentation at the cell surface is typically analyzed using fluorescence-labeled annexin V. Here we show that Leishmania promastigotes can be stained by fluorescence-labeled annexin V upon permeabilization or miltefosine treatment. However, combined lipid analysis by thin-layer chromatography, mass spectrometry and ³¹P nuclear magnetic resonance (NMR) spectroscopy revealed that Leishmania promastigotes lack any detectable amount of PS. Instead, we identified several other phospholipid classes such phosphatidic acid, phosphatidylethanolamine; phosphatidylglycerol and phosphatidylinositol as candidate lipids enabling annexin V staining.

Originalsprog	Engelsk
Tidsskrift	P L o S One
Vol/bind	7
Udgave nummer	8
Antal sider	11
ISSN	1932-6203
DOI	https://doi.org/10.1371/journal.pone.0042070
Status	Udgivet - 1 aug. 2012

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10.1371/journal.pone.0042070

Leishmania Promastigotes Lack Phosphatidylserine but Bind Annexin V upon Permeabilization or Miltefosine TreatmentForlagets udgivne version, 741 KB

Citationsformater

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abstract = "The protozoan parasite Leishmania is an intracellular pathogen infecting and replicating inside vertebrate host macrophages. A recent model suggests that promastigote and amastigote forms of the parasite mimic mammalian apoptotic cells by exposing phosphatidylserine (PS) at the cell surface to trigger their phagocytic uptake into host macrophages. PS presentation at the cell surface is typically analyzed using fluorescence-labeled annexin V. Here we show that Leishmania promastigotes can be stained by fluorescence-labeled annexin V upon permeabilization or miltefosine treatment. However, combined lipid analysis by thin-layer chromatography, mass spectrometry and 31P nuclear magnetic resonance (NMR) spectroscopy revealed that Leishmania promastigotes lack any detectable amount of PS. Instead, we identified several other phospholipid classes such phosphatidic acid, phosphatidylethanolamine; phosphatidylglycerol and phosphatidylinositol as candidate lipids enabling annexin V staining.",

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AU - Weingärtner, Adrien

AU - Kemmer, Gerdi Christine

AU - Müller, Frederic D.

AU - Zampieri, Ricardo Andrade

AU - Schiller, Jürgen

AU - Günther-Pomorski, Thomas

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AB - The protozoan parasite Leishmania is an intracellular pathogen infecting and replicating inside vertebrate host macrophages. A recent model suggests that promastigote and amastigote forms of the parasite mimic mammalian apoptotic cells by exposing phosphatidylserine (PS) at the cell surface to trigger their phagocytic uptake into host macrophages. PS presentation at the cell surface is typically analyzed using fluorescence-labeled annexin V. Here we show that Leishmania promastigotes can be stained by fluorescence-labeled annexin V upon permeabilization or miltefosine treatment. However, combined lipid analysis by thin-layer chromatography, mass spectrometry and 31P nuclear magnetic resonance (NMR) spectroscopy revealed that Leishmania promastigotes lack any detectable amount of PS. Instead, we identified several other phospholipid classes such phosphatidic acid, phosphatidylethanolamine; phosphatidylglycerol and phosphatidylinositol as candidate lipids enabling annexin V staining.

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Leishmania promastigotes lack phosphatidylserine but bind annexin V upon permeabilization or miltefosine treatment

Abstract

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