Leishmania modulates phagosome fonctions by altering certain SNAREs in a GP63-dependent pathway

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Université du Québec INRS-Institut Armand Frappier

Leishmania

modulates phagosome fonctions by altering certain SNAREs in a

GP63-dependent pathway

By Neda Moradin

A thesis subrnitted in partial fulfillment ofthe requirements of the degree of Philosophiae Doctor (PhD) in lmmunology and Virology

Internai examiner Externat examiners

Thesis director

Evaluation Committee

Maritza Jaramillo, INRS-Institut Annand-Frappier Armando Jardim, McGill University

Sachiko Sato, Laval University

Albert Descoteaux, INRS-Institut Annand-Frappier

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The most beautiful thing we cao experience is the mysterious. It is the source of ali true art and science.

(Albert Einstein)

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Abstract

Leishmania is the parasite responsible for the leishmaniases. Like severa! other

intracellular pathogens, they evolved multiple strategies to escape the immune system. In macrophages, phagosomes mature through sequential fusion and fission events with various compartments of the endocytic pathway. This process is interrupted by Leishmania promastigotes when they block aquisition of microbicidal molecules from late endosomes and lysosomes white amastigotes-harboring vacuoles are hybrid compartments composed of both endoplasmic reticulum and endocytic pathway components. The N-ethylmaleimide sensitive factor (NSF) attachment receptor proteins (SNAREs) act in intracellular trafficking pathways such as phagolysosome biogenesis. Inhibition of phagosome remodeling through different virulenece factors including LPG and GP63 is one of the most important strategies by Leishmania to survive and replicate in host cells.

In the first project, we showed that Leishmania cleaves V AMP8 protein which is one of the vesicular SNAREs on phagosome membranes. We also demonstrated that V AMP8 plays a key role in cross-presentation, which is inhibited by Leishmania through GP63-dependent cleavage of V AMP8. It had been already shown that gp91pl•ox is one of the crucial subunits in the NADPH oxidase complex that regulates cross-presentation in dendritic cells. We found that V AMP8 is necessary for the recruitment of gp91pho.T on the phagosome membrane whereby the NADPH oxidase regulates cross-presentation in dendritic cells.

In the second project, we investigated the effect of Leishmania on other SNAREs. lt is already known that Sec22b is a regulator for cross-presentation and in this study we showed that Sec22b (ER-SNARE) is excluded from phagosomes harbouring Leishmania in a

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