Cooperation between apoptotic and viable metacyclics enhances the pathogenesis of Leishmaniasis

• Published in: PloS one Vol. 4; no. 5; p. e5733
• Main Authors: Wanderley, João Luiz Mendes, Pinto da Silva, Lucia Helena, Deolindo, Poliana, Soong, Lynn, Borges, Valéria Matos, Prates, Deboraci Brito, de Souza, Ana Paula Almeida, Barral, Aldina, Balanco, José Mario de Freitas, do Nascimento, Michelle Tanny Cunha, Saraiva, Elvira Maria, Barcinski, Marcello André
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 29-05-2009; Public Library of Science (PLoS)
• Subjects: Altruism; Amastigotes; Animals; Apoptosis; Autophagy; Biodegradation; Cancer; Caspase inhibitors; Cell Biology/Cell Signaling; Cell Biology/Microbial Growth and Development; Cell culture; Cell cycle; Cooperation; Cytoplasm; Death; Deoxyribonucleic acid; Development and progression; Digestive System - cytology; Digestive System - parasitology; Digestive System - ultrastructure; Disease transmission; DNA; DNA fragmentation; Electron microscopy; Exposure; Foregut; Health aspects; Immunology; Immunology/Immunity to Infections; In Situ Nick-End Labeling; Infections; Infectious Diseases/Neglected Tropical Diseases; Infectious Diseases/Protozoal Infections; Inflammation; Inflammatory response; Leishmania; Leishmania donovani; Leishmania major; Leishmania mexicana; Leishmania mexicana - cytology; Leishmania mexicana - growth & development; Leishmania mexicana - pathogenicity; Leishmania mexicana - ultrastructure; Leishmaniasis; Leishmaniasis - parasitology; Leishmaniasis - pathology; Lesions; Life Cycle Stages; Macrophages; Mammals; Medicine; Mice; Microbiology/Cellular Microbiology and Pathogenesis; Microbiology/Immunity to Infections; Microbiology/Innate Immunity; Microbiology/Parasitology; Midgut; Mimicry; Mortality; Organisms; Parasites; Parasitic diseases; Parasitology; Pathogenesis; Phosphatidylserine; Phosphatidylserines - metabolism; Population; Promastigotes; Protozoa; Psychodidae - cytology; Psychodidae - parasitology; Psychodidae - ultrastructure; Transmission electron microscopy; Trends; Tropical diseases; Vector-borne diseases; Vectors; Viruses; Windows (intervals); Brazil; Rio de Janeiro Brazil
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0005733

Mimicking mammalian apoptotic cells by exposing phosphatidylserine (PS) is a strategy used by virus and parasitic protozoa to escape host protective inflammatory responses. With Leishmania amazonensis (La), apoptotic mimicry is a prerogative of the intramacrophagic amastigote form of the parasite and is modulated by the host. Now we show that differently from what happens with amastigotes, promastigotes exposing PS are non-viable, non-infective cells, undergoing apoptotic death. As part of the normal metacyclogenic process occurring in axenic cultures and in the gut of sand fly vectors, a sub-population of metacyclic promastigotes exposes PS. Apoptotic death of the purified PS-positive (PS(POS)) sub-population was confirmed by TUNEL staining and DNA laddering. Transmission electron microscopy revealed morphological alterations in PS(POS) metacyclics such as DNA condensation, cytoplasm degradation and mitochondrion and kinetoplast destruction, both in in vitro cultures and in sand fly guts. TUNEL(POS) promastigotes were detected only in the anterior midgut to foregut boundary of infected sand flies. Interestingly, caspase inhibitors modulated parasite death and PS exposure, when added to parasite cultures in a specific time window. Efficient in vitro macrophage infections and in vivo lesions only occur when PS(POS) and PS-negative (PS(NEG)) parasites were simultaneously added to the cell culture or inoculated in the mammalian host. The viable PS(NEG) promastigote was the infective form, as shown by following the fate of fluorescently labeled parasites, while the PS(POS) apoptotic sub-population inhibited host macrophage inflammatory response. PS exposure and macrophage inhibition by a subpopulation of promastigotes is a different mechanism than the one previously described with amastigotes, where the entire population exposes PS. Both mechanisms co-exist and play a role in the transmission and development of the disease in case of infection by La. Since both processes confer selective advantages to the infective microorganism they justify the occurrence of apoptotic features in a unicellular pathogen.