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Bat and pig IFN-induced transmembrane protein 3 restrict cell entry by influenza virus and lyssaviruses.

Benfield CT, Smith SE, Wright E, Wash RS, Ferrara F, Temperton NJ, Kellam P - J. Gen. Virol. (2015)

Bottom Line: Ectopically expressed pig and microbat IFITM3 co-localized with transferrin (early endosomes) and CD63 (late endosomes/multivesicular bodies).Pig and microbat IFITM3 restricted cell entry mediated by multiple influenza haemagglutinin subtypes and lyssavirus glycoproteins.In summary, we showed that IFITMs function as potent broad-spectrum antiviral effectors in two mammals - pigs and bats - identified as major reservoirs for emerging viruses.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Pathogen Biology, The Royal Veterinary College, Hatfield, UK cbenfield@rvc.ac.uk.

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Subcellular localization of IFITM3 proteins. A549 cells stably expressing HA-tagged IFITM3 proteins from human, pig or microbat (or untransduced cells) were fixed, permeabilized and stained for HA, CD63 (b) or LAMP1 (c). In (a) cells were incubated with Alexa Fluor 546-conjugated transferrin before fixation and staining with FITC-conjugated anti-HA. Nuclei were stained with DAPI and the coverslips examined by confocal microscopy. Images show representative staining patterns, and merged images are also shown. The right-hand column shows phase-contrast images. Bar, 10 µm. Arrows in (b) identify ‘ring-like’ staining of microbat IFITM3–HA and CD63.
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f2: Subcellular localization of IFITM3 proteins. A549 cells stably expressing HA-tagged IFITM3 proteins from human, pig or microbat (or untransduced cells) were fixed, permeabilized and stained for HA, CD63 (b) or LAMP1 (c). In (a) cells were incubated with Alexa Fluor 546-conjugated transferrin before fixation and staining with FITC-conjugated anti-HA. Nuclei were stained with DAPI and the coverslips examined by confocal microscopy. Images show representative staining patterns, and merged images are also shown. The right-hand column shows phase-contrast images. Bar, 10 µm. Arrows in (b) identify ‘ring-like’ staining of microbat IFITM3–HA and CD63.

Mentions: Similar to human IFITM3, both pig and microbat IFITM3 had a punctate intracellular distribution following cell fixation and permeabilization (Fig. 2) and co-localized with endocytosed transferrin (early endosomes) and with CD63, a marker for late endosomes/multivesicular bodies (MVBs), but not with the lysosomal marker LAMP1 (Fig. 2). There was an enlargement of CD63-positive structures in cells expressing microbat or human IFITM3 in comparison with the smaller CD63-positive vesicles seen in pig IFITM3-expressing or untransduced A549 cells (Fig. 2b). Enlargement of CD63-containing compartments was most marked in cells containing larger foci of IFITM3–HA staining. Microbat IFITM3–HA sometimes co-localized with CD63 in ‘hollow’ ring-like structures (arrows in Fig. 2b).


Bat and pig IFN-induced transmembrane protein 3 restrict cell entry by influenza virus and lyssaviruses.

Benfield CT, Smith SE, Wright E, Wash RS, Ferrara F, Temperton NJ, Kellam P - J. Gen. Virol. (2015)

Subcellular localization of IFITM3 proteins. A549 cells stably expressing HA-tagged IFITM3 proteins from human, pig or microbat (or untransduced cells) were fixed, permeabilized and stained for HA, CD63 (b) or LAMP1 (c). In (a) cells were incubated with Alexa Fluor 546-conjugated transferrin before fixation and staining with FITC-conjugated anti-HA. Nuclei were stained with DAPI and the coverslips examined by confocal microscopy. Images show representative staining patterns, and merged images are also shown. The right-hand column shows phase-contrast images. Bar, 10 µm. Arrows in (b) identify ‘ring-like’ staining of microbat IFITM3–HA and CD63.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4631062&req=5

f2: Subcellular localization of IFITM3 proteins. A549 cells stably expressing HA-tagged IFITM3 proteins from human, pig or microbat (or untransduced cells) were fixed, permeabilized and stained for HA, CD63 (b) or LAMP1 (c). In (a) cells were incubated with Alexa Fluor 546-conjugated transferrin before fixation and staining with FITC-conjugated anti-HA. Nuclei were stained with DAPI and the coverslips examined by confocal microscopy. Images show representative staining patterns, and merged images are also shown. The right-hand column shows phase-contrast images. Bar, 10 µm. Arrows in (b) identify ‘ring-like’ staining of microbat IFITM3–HA and CD63.
Mentions: Similar to human IFITM3, both pig and microbat IFITM3 had a punctate intracellular distribution following cell fixation and permeabilization (Fig. 2) and co-localized with endocytosed transferrin (early endosomes) and with CD63, a marker for late endosomes/multivesicular bodies (MVBs), but not with the lysosomal marker LAMP1 (Fig. 2). There was an enlargement of CD63-positive structures in cells expressing microbat or human IFITM3 in comparison with the smaller CD63-positive vesicles seen in pig IFITM3-expressing or untransduced A549 cells (Fig. 2b). Enlargement of CD63-containing compartments was most marked in cells containing larger foci of IFITM3–HA staining. Microbat IFITM3–HA sometimes co-localized with CD63 in ‘hollow’ ring-like structures (arrows in Fig. 2b).

Bottom Line: Ectopically expressed pig and microbat IFITM3 co-localized with transferrin (early endosomes) and CD63 (late endosomes/multivesicular bodies).Pig and microbat IFITM3 restricted cell entry mediated by multiple influenza haemagglutinin subtypes and lyssavirus glycoproteins.In summary, we showed that IFITMs function as potent broad-spectrum antiviral effectors in two mammals - pigs and bats - identified as major reservoirs for emerging viruses.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Pathogen Biology, The Royal Veterinary College, Hatfield, UK cbenfield@rvc.ac.uk.

Show MeSH
Related in: MedlinePlus