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Co-culture models illustrate the digestion of Gemmata spp. by phagocytes

Gemmata spp. bacteria thrive in the same aquatic environments as free-living amoebae. DNA-based detection of Gemmata spp. sequences in the microbiota of the human digestive tract and blood further questioned the susceptibility of Gemmata spp. to phagocytes. Here, Gemmata obscuriglobus and Gemmata ma...

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Autores principales: Kaboré, Odilon D., Loukil, Ahmed, Godreuil, Sylvain, Drancourt, Michel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6127157/
https://www.ncbi.nlm.nih.gov/pubmed/30190504
http://dx.doi.org/10.1038/s41598-018-31667-0
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author Kaboré, Odilon D.
Loukil, Ahmed
Godreuil, Sylvain
Drancourt, Michel
author_facet Kaboré, Odilon D.
Loukil, Ahmed
Godreuil, Sylvain
Drancourt, Michel
author_sort Kaboré, Odilon D.
collection PubMed
description Gemmata spp. bacteria thrive in the same aquatic environments as free-living amoebae. DNA-based detection of Gemmata spp. sequences in the microbiota of the human digestive tract and blood further questioned the susceptibility of Gemmata spp. to phagocytes. Here, Gemmata obscuriglobus and Gemmata massiliana were co-cultured with the amoebae Acanthamoeba polyphaga, Acanthamoeba castellanii, Acanthamoeba griffini and THP-1 macrophage-like phagocytes. All experiments were performed in five independant replicates. The ratio amoeba/bacteria was 1:20 and the ratio THP-1/bacteria was 1:10. After a 2-hour co-culture, extracellular bacteria were killed by kanamycin or amikacin and eliminated. The intracellular location of Gemmata bacteria was specified by confocal microscopy. Microscopic enumerations and culture-based enumerations of colony-forming units were performed at T = 0, 1, 2, 3, 4, 8, 16, 24, 48 and 72 hours post-infection. Then, Gemmata bacteria were engulfed into the phagocytes’ cytoplasmic vacuoles, more than (98 ± 2)% of Gemmata bacteria, compared to controls, were destroyed by phagocytic cells after a 48-h co-culture according to microscopy and culture results, and no positive culture was observed at T = 72-hours. Under our co-culture conditions, Gemmata bacteria were therefore susceptible to the environmental and host phagocytes here investigated. These data suggest that these Acanthamoeba species and THP-1 cells cannot be used to isolate G. massiliana and G. obscuriglobus under the co-culture conditions applied in this study. Although the THP-1 response can point towards potential responses that might occur in vivo, these responses should first bevalidated by in vivo studies to draw definite conclusions.
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spelling pubmed-61271572018-09-10 Co-culture models illustrate the digestion of Gemmata spp. by phagocytes Kaboré, Odilon D. Loukil, Ahmed Godreuil, Sylvain Drancourt, Michel Sci Rep Article Gemmata spp. bacteria thrive in the same aquatic environments as free-living amoebae. DNA-based detection of Gemmata spp. sequences in the microbiota of the human digestive tract and blood further questioned the susceptibility of Gemmata spp. to phagocytes. Here, Gemmata obscuriglobus and Gemmata massiliana were co-cultured with the amoebae Acanthamoeba polyphaga, Acanthamoeba castellanii, Acanthamoeba griffini and THP-1 macrophage-like phagocytes. All experiments were performed in five independant replicates. The ratio amoeba/bacteria was 1:20 and the ratio THP-1/bacteria was 1:10. After a 2-hour co-culture, extracellular bacteria were killed by kanamycin or amikacin and eliminated. The intracellular location of Gemmata bacteria was specified by confocal microscopy. Microscopic enumerations and culture-based enumerations of colony-forming units were performed at T = 0, 1, 2, 3, 4, 8, 16, 24, 48 and 72 hours post-infection. Then, Gemmata bacteria were engulfed into the phagocytes’ cytoplasmic vacuoles, more than (98 ± 2)% of Gemmata bacteria, compared to controls, were destroyed by phagocytic cells after a 48-h co-culture according to microscopy and culture results, and no positive culture was observed at T = 72-hours. Under our co-culture conditions, Gemmata bacteria were therefore susceptible to the environmental and host phagocytes here investigated. These data suggest that these Acanthamoeba species and THP-1 cells cannot be used to isolate G. massiliana and G. obscuriglobus under the co-culture conditions applied in this study. Although the THP-1 response can point towards potential responses that might occur in vivo, these responses should first bevalidated by in vivo studies to draw definite conclusions. Nature Publishing Group UK 2018-09-06 /pmc/articles/PMC6127157/ /pubmed/30190504 http://dx.doi.org/10.1038/s41598-018-31667-0 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Kaboré, Odilon D.
Loukil, Ahmed
Godreuil, Sylvain
Drancourt, Michel
Co-culture models illustrate the digestion of Gemmata spp. by phagocytes
title Co-culture models illustrate the digestion of Gemmata spp. by phagocytes
title_full Co-culture models illustrate the digestion of Gemmata spp. by phagocytes
title_fullStr Co-culture models illustrate the digestion of Gemmata spp. by phagocytes
title_full_unstemmed Co-culture models illustrate the digestion of Gemmata spp. by phagocytes
title_short Co-culture models illustrate the digestion of Gemmata spp. by phagocytes
title_sort co-culture models illustrate the digestion of gemmata spp. by phagocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6127157/
https://www.ncbi.nlm.nih.gov/pubmed/30190504
http://dx.doi.org/10.1038/s41598-018-31667-0
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