PMID- 38033163 OWN - NLM STAT- MEDLINE DCOM- 20231216 LR - 20231216 IS - 1553-7374 (Electronic) IS - 1553-7366 (Print) IS - 1553-7366 (Linking) VI - 19 IP - 11 DP - 2023 Nov TI - Kicking sleepers out of bed: Macrophages promote reactivation of dormant Cryptococcus neoformans by extracellular vesicle release and non-lytic exocytosis. PG - e1011841 LID - 10.1371/journal.ppat.1011841 [doi] LID - e1011841 AB - Macrophages play a key role in disseminated cryptococcosis, a deadly fungal disease caused by Cryptococcus neoformans. This opportunistic infection can arise following the reactivation of a poorly characterized latent infection attributed to dormant C. neoformans. Here, we investigated the mechanisms underlying reactivation of dormant C. neoformans using an in vitro co-culture model of viable but non-culturable (VBNC; equivalent of dormant) yeast cells with bone marrow-derived murine macrophages (BMDMs). Comparative transcriptome analysis of BMDMs incubated with log, stationary phase or VBNC cells of C. neoformans showed that VBNC cells elicited a reduced transcriptional modification of the macrophage but retaining the ability to regulate genes important for immune response, such as NLRP3 inflammasome-related genes. We further confirmed the maintenance of the low immunostimulatory capacity of VBNC cells using multiplex cytokine profiling, and analysis of cell wall composition and dectin-1 ligands exposure. In addition, we evaluated the effects of classic (M1) or alternative (M2) macrophage polarization on VBNC cells. We observed that intracellular residence sustained dormancy, regardless of the polarization state of macrophages and despite indirect detection of pantothenic acid (or its derivatives), a known reactivator for VBNC cells, in the C. neoformans-containing phagolysosome. Notably, M0 and M2, but not M1 macrophages, induced extracellular reactivation of VBNC cells by the secretion of extracellular vesicles and non-lytic exocytosis. Our results indicate that VBNC cells retain the low immunostimulatory profile required for persistence of C. neoformans in the host. We also describe a pro-pathogen role of macrophage-derived extracellular vesicles in C. neoformans infection and reinforce the impact of non-lytic exocytosis and the macrophage profile on the pathophysiology of cryptococcosis. CI - Copyright: (c) 2023 de Castro et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. FAU - de Castro, Raffael Junio Araujo AU - de Castro RJA AUID- ORCID: 0000-0002-5207-8892 AD - Translational Mycology Research Group, National Reference Center for Invasive Mycoses and Antifungals, Mycology Department, Institut Pasteur, Universite Paris Cite, Paris, France. AD - Laboratory of Applied Immunology, Department of Cell Biology, Institute of Biological Sciences, University of Brasilia, Brasilia, Distrito Federal, Brazil. FAU - Marina, Clara Luna AU - Marina CL AD - Laboratory of Applied Immunology, Department of Cell Biology, Institute of Biological Sciences, University of Brasilia, Brasilia, Distrito Federal, Brazil. FAU - Sturny-Leclere, Aude AU - Sturny-Leclere A AD - Translational Mycology Research Group, National Reference Center for Invasive Mycoses and Antifungals, Mycology Department, Institut Pasteur, Universite Paris Cite, Paris, France. FAU - Hoffmann, Christian AU - Hoffmann C AD - Food Research Center, Department of Food Sciences and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo, Sao Paulo, Brazil. FAU - Burgel, Pedro Henrique AU - Burgel PH AD - Laboratory of Applied Immunology, Department of Cell Biology, Institute of Biological Sciences, University of Brasilia, Brasilia, Distrito Federal, Brazil. FAU - Wong, Sarah Sze Wah AU - Wong SSW AD - Immunobiology of Aspergillus, Institut Pasteur, Universite Paris Cite, Paris, France. FAU - Aimanianda, Vishukumar AU - Aimanianda V AD - Immunobiology of Aspergillus, Institut Pasteur, Universite Paris Cite, Paris, France. FAU - Varet, Hugo AU - Varet H AD - Plate-forme Technologique Biomics, Institut Pasteur, Universite Paris Cite, Paris, France. FAU - Agrawal, Ruchi AU - Agrawal R AD - Translational Mycology Research Group, National Reference Center for Invasive Mycoses and Antifungals, Mycology Department, Institut Pasteur, Universite Paris Cite, Paris, France. FAU - Bocca, Anamelia Lorenzetti AU - Bocca AL AD - Laboratory of Applied Immunology, Department of Cell Biology, Institute of Biological Sciences, University of Brasilia, Brasilia, Distrito Federal, Brazil. FAU - Alanio, Alexandre AU - Alanio A AUID- ORCID: 0000-0001-9726-3082 AD - Translational Mycology Research Group, National Reference Center for Invasive Mycoses and Antifungals, Mycology Department, Institut Pasteur, Universite Paris Cite, Paris, France. AD - Laboratoire de parasitologie-mycologie, AP-HP, Hopital Saint-Louis, Paris, France. LA - eng PT - Journal Article DEP - 20231130 PL - United States TA - PLoS Pathog JT - PLoS pathogens JID - 101238921 SB - IM MH - Animals MH - Mice MH - *Cryptococcus neoformans/genetics MH - *Cryptococcosis/microbiology MH - Macrophages MH - Exocytosis MH - *Extracellular Vesicles PMC - PMC10715671 COIS- The authors have declared that no competing interests exist. EDAT- 2023/11/30 18:45 MHDA- 2023/12/17 13:19 PMCR- 2023/11/30 CRDT- 2023/11/30 14:23 PHST- 2023/06/16 00:00 [received] PHST- 2023/11/18 00:00 [accepted] PHST- 2023/12/12 00:00 [revised] PHST- 2023/12/17 13:19 [medline] PHST- 2023/11/30 18:45 [pubmed] PHST- 2023/11/30 14:23 [entrez] PHST- 2023/11/30 00:00 [pmc-release] AID - PPATHOGENS-D-23-00969 [pii] AID - 10.1371/journal.ppat.1011841 [doi] PST - epublish SO - PLoS Pathog. 2023 Nov 30;19(11):e1011841. doi: 10.1371/journal.ppat.1011841. eCollection 2023 Nov.