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Scientific paper - Original scientific paper
Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages
Frontiers in Cellular and Infection Microbiology, 7 (2017); 1-11. https://doi.org/10.3389/fcimb.2017.00303
Marečić, Valentina; Shevchuk, Olga; Ožanič, Mateja; Mihelčić, Mirna; Steinert, Michael; Jurak Begonja, Antonija; Abu Kwaik, Yousef; Šantić, Marina

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Marečić, V., Shevchuk, O., Ožanič, M., Mihelčić, M., Steinert, M., Jurak Begonja, A. ... Šantić, M. (2017). Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages. Frontiers in Cellular and Infection Microbiology, 7., 1-11. doi: 10.3389/fcimb.2017.00303

Marečić, Valentina, et al. "Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages." Frontiers in Cellular and Infection Microbiology, vol. 7, 2017, pp. 1-11. https://doi.org/10.3389/fcimb.2017.00303

Marečić, Valentina, Olga Shevchuk, Mateja Ožanič, Mirna Mihelčić, Michael Steinert, Antonija Jurak Begonja, Yousef Abu Kwaik and Marina Šantić. "Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages." Frontiers in Cellular and Infection Microbiology 7 (2017): 1-11. https://doi.org/10.3389/fcimb.2017.00303

Marečić, V., et al. (2017) 'Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages', Frontiers in Cellular and Infection Microbiology, 7, pp. 1-11. doi: 10.3389/fcimb.2017.00303

Marečić V, Shevchuk O, Ožanič M, Mihelčić M, Steinert M, Jurak Begonja A, and sur.. Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages. Frontiers in Cellular and Infection Microbiology [Internet]. 2017 [cited 2024 April 23];7:1-11. doi: 10.3389/fcimb.2017.00303

V. Marečić, et al., "Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages", Frontiers in Cellular and Infection Microbiology, vol. 7, pp. 1-11, 2017. [Online]. Available at: https://urn.nsk.hr/urn:nbn:hr:184:589065. [Accessed: 23 April 2024]

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Metadata
Title (english)Isolation of F. novicida-Containing Phagosome from Infected Human Monocyte Derived Macrophages
AuthorValentina Marečić
AuthorOlga Shevchuk
AuthorMateja Ožanič
AuthorMirna Mihelčić
AuthorMichael Steinert
AuthorAntonija Jurak Begonja
AuthorYousef Abu Kwaik
AuthorMarina Šantić
Author's institutionUniversity of Rijeka
Faculty of Medicine
(Department of Microbiology and Parasitology)
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Clinical Medical Sciences
Medical Microbiology
Abstract (english)
Francisella is a gram-negative bacterial pathogen, which causes tularemia in humans and animals. A crucial step of Francisella infection is its invasion of macrophage cells. Biogenesis of the Francisella-containing phagosome (FCP) is arrested for ∼15min at the endosomal stage, followed by gradual bacterial escape into the cytosol, where the microbe proliferates. The crucial step in pathogenesis of tularemia is short and transient presence of the bacterium within phagosome. Isolation of FCPs for further studies has been challenging due to the short period of time of bacterial residence in it and the characteristics of the FCP. Here, we will for the first time present the method for isolation of the FCPs from infected human monocytes-derived macrophages (hMDMs). For elimination of lysosomal compartment these organelles were pre-loaded with dextran coated colloidal iron particles prior infection and eliminated by magnetic separation of the post-nuclear supernatant (PNS). We encountered the challenge that mitochondria has similar density to the FCP. To separate the FCP in the PNS from mitochondria, we utilized iodophenylnitrophenyltetrazolium, which is converted by the mitochondrial succinate dehydrogenase into formazan, leading to increased density of the mitochondria and allowing separation by the discontinuous sucrose density gradient ultracentrifugation. The purity of the FCP preparation and its acquisition of early endosomal markers was confirmed by Western blots, confocal and transmission electron microscopy. Our strategy to isolate highly pure FCPs from macrophages should facilitate studies on the FCP and its biogenesis.13
Keywords (english)
Languageenglish
Publication typeScientific paper - Original scientific paper
Publication statusPublished
Peer reviewPeer review - international
Publication versionPublished version
Journal titleFrontiers in Cellular and Infection Microbiology
Numberingvol. 7, pp. 1-11
p-ISSN2235-2988
DOIhttps://doi.org/10.3389/fcimb.2017.00303
URN:NBNurn:nbn:hr:184:589065
Publication2017
ProjectNumber: IP-2016-06-9003
Title (croatian): Uloga unutarstaničnog života Francisella tularensis u patogenezi eksperimentalne tularemije
Acronym: FRANCYCLE
Leader: Marina Šantić
Jurisdiction: Croatia
Funding stream: IP
Document URLhttp://bib.irb.hr/884138
Type of resourceText
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2017-09-26 19:42:53