Limits...
Novel entries in a fungal biofilm matrix encyclopedia.

Zarnowski R, Westler WM, Lacmbouh GA, Marita JM, Bothe JR, Bernhardt J, Lounes-Hadj Sahraoui A, Fontaine J, Sanchez H, Hatfield RD, Ntambi JM, Nett JE, Mitchell AP, Andes DR - MBio (2014)

Bottom Line: Newly described, more abundant polysaccharides included α-1,2 branched α-1,6-mannans (87%) associated with unbranched β-1,6-glucans (13%) in an apparent mannan-glucan complex (MGCx).Importance: This report is the first to decipher the complex and unique macromolecular composition of the Candida biofilm matrix, demonstrate the clinical relevance of matrix components, and show that multiple matrix components are needed for protection from antifungal drugs.The availability of these biochemical analyses provides a unique resource for further functional investigation of the biofilm matrix, a defining trait of this lifestyle.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Infectious Diseases, University of Wisconsin, Madison, Wisconsin, USA.

Show MeSH

Related in: MedlinePlus

Detection of α-1,2 branched α-1,6-mannan, β-1,6-glucan, and β-1,3-glucan in the C. albicans biofilm matrix using an in vitro biofilm model (A to D) and the in vivo rat catheter biofilm model (E to H). The scale bars in panels A, E, F, and G are 50 µm. (A) In vitro biofilm community of yeast and hyphae visualized using a calcofluor white stain. (B) Detection of α-1,2 branched α-1,6-mannan in the in vitro biofilm matrix using anti-α-1,2 branched α-1,6-mannan monoclonal antibody fluorescently labeled with FITC. (C) Detection of β-1,6-glucan in the in vitro biofilm matrix using anti-β-1,6-glucan monoclonal antibody fluorescently labeled with rhodamine. (D) An overlay of both stains showed extracellular staining throughout the in vitro biofilm thickness, and the yellow color is consistent with colocalization. The scale bar is 200 µm. (E) Detection of α-1,2 branched α-1,6-mannan in the in vivo rat catheter biofilm matrix. CL indicates catheter lumen. (F) Detection of β-1,6-glucan in the in vivo rat catheter biofilm matrix. (G) An overlay of both stains showed extracellular staining throughout the biofilm thickness. (H) Immuno-TEM of biofilm scraped from the catheter model identified β-1,3-glucan using gold particle-labeled antibodies. As indicated by arrows, β-1,3-glucan was sparsely scattered throughout the extracellular matrix, consistent with the modest concentration of this polysaccharide relative to other components. The scale bar is 0.5 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4128356&req=5

fig4: Detection of α-1,2 branched α-1,6-mannan, β-1,6-glucan, and β-1,3-glucan in the C. albicans biofilm matrix using an in vitro biofilm model (A to D) and the in vivo rat catheter biofilm model (E to H). The scale bars in panels A, E, F, and G are 50 µm. (A) In vitro biofilm community of yeast and hyphae visualized using a calcofluor white stain. (B) Detection of α-1,2 branched α-1,6-mannan in the in vitro biofilm matrix using anti-α-1,2 branched α-1,6-mannan monoclonal antibody fluorescently labeled with FITC. (C) Detection of β-1,6-glucan in the in vitro biofilm matrix using anti-β-1,6-glucan monoclonal antibody fluorescently labeled with rhodamine. (D) An overlay of both stains showed extracellular staining throughout the in vitro biofilm thickness, and the yellow color is consistent with colocalization. The scale bar is 200 µm. (E) Detection of α-1,2 branched α-1,6-mannan in the in vivo rat catheter biofilm matrix. CL indicates catheter lumen. (F) Detection of β-1,6-glucan in the in vivo rat catheter biofilm matrix. (G) An overlay of both stains showed extracellular staining throughout the biofilm thickness. (H) Immuno-TEM of biofilm scraped from the catheter model identified β-1,3-glucan using gold particle-labeled antibodies. As indicated by arrows, β-1,3-glucan was sparsely scattered throughout the extracellular matrix, consistent with the modest concentration of this polysaccharide relative to other components. The scale bar is 0.5 µm.

Mentions: Confocal imaging of in vitro biofilms was undertaken to examine the distribution of mannan and β-1,6-glucan matrix components by labeling with monoclonal antibodies developed against and specific for purified matrix components (data not shown). The images demonstrated an extracellular location for each polysaccharide and a distribution throughout the biofilm depth in Fig. 4A to C. The two-channel image color is suggestive of polysaccharide colocalization and consistent with an MGCx (Fig. 4D).


Novel entries in a fungal biofilm matrix encyclopedia.

Zarnowski R, Westler WM, Lacmbouh GA, Marita JM, Bothe JR, Bernhardt J, Lounes-Hadj Sahraoui A, Fontaine J, Sanchez H, Hatfield RD, Ntambi JM, Nett JE, Mitchell AP, Andes DR - MBio (2014)

Detection of α-1,2 branched α-1,6-mannan, β-1,6-glucan, and β-1,3-glucan in the C. albicans biofilm matrix using an in vitro biofilm model (A to D) and the in vivo rat catheter biofilm model (E to H). The scale bars in panels A, E, F, and G are 50 µm. (A) In vitro biofilm community of yeast and hyphae visualized using a calcofluor white stain. (B) Detection of α-1,2 branched α-1,6-mannan in the in vitro biofilm matrix using anti-α-1,2 branched α-1,6-mannan monoclonal antibody fluorescently labeled with FITC. (C) Detection of β-1,6-glucan in the in vitro biofilm matrix using anti-β-1,6-glucan monoclonal antibody fluorescently labeled with rhodamine. (D) An overlay of both stains showed extracellular staining throughout the in vitro biofilm thickness, and the yellow color is consistent with colocalization. The scale bar is 200 µm. (E) Detection of α-1,2 branched α-1,6-mannan in the in vivo rat catheter biofilm matrix. CL indicates catheter lumen. (F) Detection of β-1,6-glucan in the in vivo rat catheter biofilm matrix. (G) An overlay of both stains showed extracellular staining throughout the biofilm thickness. (H) Immuno-TEM of biofilm scraped from the catheter model identified β-1,3-glucan using gold particle-labeled antibodies. As indicated by arrows, β-1,3-glucan was sparsely scattered throughout the extracellular matrix, consistent with the modest concentration of this polysaccharide relative to other components. The scale bar is 0.5 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128356&req=5

fig4: Detection of α-1,2 branched α-1,6-mannan, β-1,6-glucan, and β-1,3-glucan in the C. albicans biofilm matrix using an in vitro biofilm model (A to D) and the in vivo rat catheter biofilm model (E to H). The scale bars in panels A, E, F, and G are 50 µm. (A) In vitro biofilm community of yeast and hyphae visualized using a calcofluor white stain. (B) Detection of α-1,2 branched α-1,6-mannan in the in vitro biofilm matrix using anti-α-1,2 branched α-1,6-mannan monoclonal antibody fluorescently labeled with FITC. (C) Detection of β-1,6-glucan in the in vitro biofilm matrix using anti-β-1,6-glucan monoclonal antibody fluorescently labeled with rhodamine. (D) An overlay of both stains showed extracellular staining throughout the in vitro biofilm thickness, and the yellow color is consistent with colocalization. The scale bar is 200 µm. (E) Detection of α-1,2 branched α-1,6-mannan in the in vivo rat catheter biofilm matrix. CL indicates catheter lumen. (F) Detection of β-1,6-glucan in the in vivo rat catheter biofilm matrix. (G) An overlay of both stains showed extracellular staining throughout the biofilm thickness. (H) Immuno-TEM of biofilm scraped from the catheter model identified β-1,3-glucan using gold particle-labeled antibodies. As indicated by arrows, β-1,3-glucan was sparsely scattered throughout the extracellular matrix, consistent with the modest concentration of this polysaccharide relative to other components. The scale bar is 0.5 µm.
Mentions: Confocal imaging of in vitro biofilms was undertaken to examine the distribution of mannan and β-1,6-glucan matrix components by labeling with monoclonal antibodies developed against and specific for purified matrix components (data not shown). The images demonstrated an extracellular location for each polysaccharide and a distribution throughout the biofilm depth in Fig. 4A to C. The two-channel image color is suggestive of polysaccharide colocalization and consistent with an MGCx (Fig. 4D).

Bottom Line: Newly described, more abundant polysaccharides included α-1,2 branched α-1,6-mannans (87%) associated with unbranched β-1,6-glucans (13%) in an apparent mannan-glucan complex (MGCx).Importance: This report is the first to decipher the complex and unique macromolecular composition of the Candida biofilm matrix, demonstrate the clinical relevance of matrix components, and show that multiple matrix components are needed for protection from antifungal drugs.The availability of these biochemical analyses provides a unique resource for further functional investigation of the biofilm matrix, a defining trait of this lifestyle.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Infectious Diseases, University of Wisconsin, Madison, Wisconsin, USA.

Show MeSH
Related in: MedlinePlus